Acetylcysteine contains not less than 99.
Acetylcysteine occurs as white, crystals or crys-talline powder. It is freely soluble in water and in ethanol (99.5). It dissolves in sodium hydroxide TS.
Determine the infrared absorption spectrum of Acetylcysteine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋21.0 – ＋27.09Weigh accu-rately an amount of Acetylcysteine, equivalent to about 2.5 g calculated on the dried basis, and dissolve with 2 mL of a so-lution of disodium dihydrogen ethylenediamine tetraacetate dihydrate (1 in 100) and 15 mL of a solution of sodium hy-droxide (1 in 25). To this solution add a solution, prepared by adjusting the pH to 7.0 of 500 mL of a solution of potas-sium dihydrogen phosphate (17 in 125) with sodium hydrox-ide TS and adding water to make 1000 mL, to make exactly 50 mL. Determine the optical rotation of this solution using ‌JP XVIII Official Monographs / Aciclovir 407 a 100-mm cell.
(1) Chloride <1.03>—Dissolve 0.40 g of Acetyl-cysteine in 25 mL of sodium hydroxide TS, add 4 mL of hydrogen peroxide (30), heat in a water bath for 45 minutes, and cool. Then add 5 mL of nitric acid, add water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.040z). Sulfate <1.14>—Perform the test with 0.8 g of Acetyl- cysteine. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.030z). Ammonium <1.02>—Perform the test with 0.10 g of Acetylcysteine, using the distillation under reduced pressure. Prepare the control solution with 2.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 1.0 g of Acetyl- cysteine in 40 mL of water, add 3 mL of sodium hydroxide TS, 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution as follows: To 1.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Acetylcysteine according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Acetyl-cysteine in 25 mL of the mobile phase, and use this solution as the sample solution. The sample solution is prepared before using. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the area of the peak other than acetylcysteine is not more than 0.3z, and the total area of the peak other than acetylcysteine is not more than 0.6z. Operating conditions— Detector: An ultraviolet absorption photometer (wave- the symmetry factor of the peak of acetylcysteine are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of acetylcysteine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.2 g of Acetylcysteine, place it in a stoppered flask, dissolve in 20 mL of water, add 4 g of potassium iodide and 5 mL of dilute hydrochloric acid, then add exactly 25 mL of 0.05 mol/L iodine VS, stop-per tightly, allow to stand for 20 minutes in an ice cold water in the dark, and titrate <2.50> the excess of iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 16.32 mg of C5H9NO3S
Cycloserine contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calcu-lated on the dried basis.
Cycloserine occurs as white to light yellow-white, crystals or crystalline powder. It is soluble in water, and sparingly soluble in ethanol (95). ‌824
Determine the infrared absorption spectrum of Cycloserine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Cycloserine RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋108 – ＋1149(2.5 g calcu-lated on the dried basis, 2 mol/L sodium hydroxide TS, 50 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cycloserine in 20 mL of water: the pH of the solution is between 5.0 and 7.4.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cycloserine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Condensation products—Dissolve 20 mg of Cycloser-ine in sodium hydroxide TS to make exactly 50 mL, and de-termine the absorbance of this solution at 285 nm as directed Cyproheptadine Hydrochloride Hydrate
Ethyl L-Cysteine Hydrochloride occurs as white, crystals or crystalline powder. It has a characteristic odor, and has a bitter taste at first with a burning aftertaste. It is very soluble in water, and freely soluble in ethanol (95). Melting point: about 1269C (with decomposition).
(1) Determine the infrared absorption spec-trum of Ethyl L-Cysteine Hydrochloride as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. (2) A solution of Ethyl L-Cysteine Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> (1) for chloride. D Optical rotation <2.49> [a]20: －10.0 – －13.09(after dry-ing, 2.0 g, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm).
(1) Sulfate <1.14>—Perform the test with 0.6 g of Ethyl L-Cysteine Hydrochloride. Prepare the the control so-lution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Heavy metals <1.07>—Proceed with 1.0 g of Ethyl L-Cysteine Hydrochloride according to Method 1, and per- form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure rapidly. Dissolve 0.05 g each of Ethyl L-Cysteine Hydrochloride and N-ethylmaleimide in 5 mL of mobile phase, allow to stand for 30 minutes, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of these solutions by the automatic integration method: a peak area obtained from the sample solution with the relative retention time to ethyl L-cysteine-N-ethylmaleimide complex from the standard so-lution being about 0.7 is not larger than the peak area of ethyl L-cysteine-N-ethylmaleimide complex from the stand-ard solution. Each area of all peaks other than ethyl L-cysteine-N-ethylmaleimide complex and N-ethylmaleimide from the sample solution is not larger than 1/3 times the peak area of ethyl L-cysteine N-ethylmaleimide complex from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column about 6 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogenphoshate TS and acetonitrile (2:1). Flow rate: Adjust so that the retention time of ethyl L-cysteine-N-ethylmaleimide complex is about 4 minutes. Selection of column: Dissolve 0.05 g of Ethyl L-Cysteine Hydrochloride, 0.01 g of L-cysteine hydrochloride and 0.05 g of N-ethylmaleimide in 25 mL of the mobile phase, and allow to stand for 30 minutes. Proceed with 2 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of L-cysteine-N-ethylmaleimide complex, ethyl L-cysteine-N-ethylmaleimide complex and N-ethylmaleimide in this order, complete reso-lution of each component, and the resolution of the peaks of L-cysteine-N-ethylmaleimide complex and ethyl L-cysteine- N-ethylmaleimide complex being not less than 3. Detection sensitivity: Adjust the detection sensitivity so that the peak height of ethyl L-cysteine-N-ethylmaleimide complex obtained from 2 mL of the standard solution is be-tween 10 mm and 20 mm. Time span of measurement: About 3 times as long as the retention time of ethyl L-cysteine-N-ethylmaleimide com-plex. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus oxide (V), 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Ethyl L-Cysteine Hydrochloride, previously dried, transfer into a glass-stoppered flask, and dissolve in 10 mL of water previously freshly boiled and cooled to a temperature not exceeding 59C in a stream of nitrogen. Add exactly 20 mL of 0.05 mol/L iodine VS, previously cooled to a temperature not exceeding 59C, and allow to stand for 30 seconds, then titrate <2.50> with 0.1 mol/L sodium thiosulfate VS, on cool-ing below 59C (indicator: 1 mL of starch TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.05 mol/L iodine VS ＝ 18.57 mg of C5H11NO2S.HCl
Glycine, when dried, contains not less than 98.
Glycine occurs as white, crystals or crystalline powder. It has a sweet taste. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (95). It shows crystal polymorphism.
Determine the infrared absorption spectrum of Glycine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry Gonadorelin Acetate
Weigh accurately about 80 mg of Glycine, previ-ously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 7.507 mg of C2H5NO2
L-Alanine, when dried, contains not less than 98.
L-Alanine occurs as white, crystals or crystal-line powder. It has a slightly sweet taste. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (99.5). It dissolves in 6 mol/L hydrochloric acid TS.
Determine the infrared absorption spectrum of L-Alanine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋13.5 – ＋15.59(after dry-ing, 2.5 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Alanine in 20 mL of water: the pH of the solution is between 5.7 and 6.7.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Alanine in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of L-Alanine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Alanine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Alanine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Alanine according to Method 1, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Alanine according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Weigh accurately about 0.5 g of L-Alanine, dissolve in 0.5 mL of hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the sample solution. Separately, accurately measure 2.5 mmol amounts of L-aspartic acid, L-threonine, L-serine, L-glutamic acid, glycine, L-alanine, L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammo-nium chloride, L-histidine and L-arginine, dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, add 0.02 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 4 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Based on the peak heights obtained from the sample solution and stand-ard solution, determine the mass of the amino acids other than alanine contained in 1 mL of the sample solution, and calculate the mass percent: the amount of each amino acid other than alanine is not more than 0.1z. Operating conditions— Detector: A visible spectrophotometer (wavelength: 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (Na type) com-posed with a sulfonated polystyrene copolymer (3 mm in particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Reaction time: About 1 minute. Mobile phase: Prepare mobile phases A, B, C, D and E according to the following table, and add 0.1 mL of capric acid to each mobile phase. ‌424‌ Mobile phase A Mobile phase B Mobile phase C Mobile phase D Mobile phase E Citric acid 19.80 g 22.00 g 12.80 g 6.10 g — Trisodium citrate 6.19 g dihydrate 7.74 g 13.31 g 26.67 g — Sodium chlo- ride 5.66 g 7.07 g 3.74 g 54.35 g — Sodium hydroxide — — — — 8.00 g Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL Thiodiglycol 5 mL 5 mL 5 mL — — Benzyl alcohol — — — 5 mL — Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL 4 mL monohydrate Water Appropriate Appropriate Appropriate Appropriate Appropriate Albumin Tannate
L-Arginine Hydrochloride, when dried, contains not less than 98.
L-Arginine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a slight, characteristic taste. It is freely soluble in water and in formic acid, and very slightly soluble in ethanol (95).
(1) Determine the infrared absorption spectrum of L-Arginine Hydrochloride, previously dried, as directed in the potassium bromide disk method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of L-Arginine Hydrochloride (1 in 50) responds to the Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: ＋21.5 – ＋23.59(after dry-ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Arginine Hydrochloride in 10 mL of water: the pH of this solution is between 4.7 and 6.2.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Arginine Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.6 g of L-Arginine Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of ‌L-Arginine Hydrochloride, using the distillation under reduced pressure. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z).‌ Heavy metals <1.07>—Proceed with 1.0 g of L-Arginine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of L-Arginine Hydrochloride according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of L-Arginine Hydrochloride in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 10 mL. Pipet 1 mL of this solu-tion, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5), water, 1-butanol and ammonia water (28) (2:1:1:1) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of L-Arginine Hydro-chloride, previously dried, dissolve in 2 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat on a water bath for 30 minutes. After cooling, add 45 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 10.53 mg of C6H14N4O2.HCl
L-Arginine, when dried, contains not less than 98.
L-Arginine occurs as white, crystals or crystal-line powder. It has a characteristic odor. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (99.5). It dissolves in dilute hydrochloric acid. It is hygroscopic.
Determine the infrared absorption spectrum of previously dried L-Arginine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋26.9 – ＋27.99(after dry-ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of L-Arginine in 10 mL of water is between 10.5 and 12.0.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of L-Arginine in 10 mL of water is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of L-Arginine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Arginine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Arginine, using the distillation under reduced pressure. Prepare the control solution with 5.0 mL of Standard Am-monium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 2.0 g of L-Arginine in 30 mL of water, add 1 drop of phenolphthalein TS, neutral- ize with dilute hydrochloric acid, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Arginine according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of L-Arginine in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-propanol and ammonia solution (28) (7:3) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly ninhydrin-butanol TS on the plate, and heat the plate at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 80 mg of L-Arginine, previ-ously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 8.710 mg of C6H14N4O2
L-Cysteine Hydrochloride Hydrate contains not less than 98.
L-Cysteine Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It has a characteristic odor and a strong acid taste. It is very soluble in water, and soluble in ethanol (99.5). It dissolves in 6 mol/L hydrochloric acid TS.
(1) Determine the infrared absorption spec-trum of L-Cysteine Hydrochloride Hydrate as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. (2) To 10 mL of a solution of L-Cysteine Hydrochloride Hydrate (1 in 50) add 1 mL of hydrogen peroxide (30), heat on a water bath for 20 minutes, and cool: the solution re-sponds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: ＋6.0 – ＋7.59(2 g, calculated on the dried basis, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of L-Cysteine Hydrochloride Hydrate in 100 mL of water is between 1.3 and 2.3.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of L-Cysteine Hydrochloride Hy-drate in 10 mL of water is clear and colorless. Sulfate <1.14>—Dissolve 0.8 g of L-Cysteine Hydro-chloride Hydrate in 30 mL of water and 3 mL of dilute hy- drochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution as follows: To 0.35 mL of 0.005 mol/L sulfuric acid VS add 3 mL of dilute hydrochloric acid and water to make 50 mL. To both of the test solution and the control solution add 4 mL of barium chloride TS (not more than 0.021z). Ammonium <1.02>—Perform the test with 0.25 g of L-Cysteine Hydrochloride Hydrate using the distillation under reduced pressure. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Cysteine Hydrochloride Hydrate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Cysteine Hydrochloride Hydrate according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of L-Cysteine Hydrochloride Hydrate in N-ethylmaleimide solution (1 in 50) to make 10 mL, allow to stand for 30 minutes, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add water to make exactly 10 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-Layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Then develop with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of methanol and acetic acid (100) (97:3) (1 in 100) on the plate, and then heat at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> 8.5 – 12.0z (1 g, in vacuum, phos-phorus (V) oxide, 20 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of L-Cysteine Hydro-chloride Hydrate, place in a glass-stoppered flask, and dis-solve in 20 mL of water. Dissolve 4 g of potassium iodide in this solution, soak immediately in ice cold water, add 5 mL of dilute hydrochloric acid and exactly 25 mL of 0.05 mol/L iodine VS, allow to stand for 20 minutes in a dark place, titrate <2.50> the excess of iodine with 0.1 mol/L sodium thiosulfate VS (indicator: starch TS). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.05 mol/L iodine VS ＝ 15.76 mg of C3H7NO2S.HCl
L-Cysteine occurs as white crystals or a white crystalline powder. It has a characteristic odor and a pun-gent taste. It is freely soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 1 mol/L hydrochloric acid TS.
Determine the infrared absorption spectrum of L-Cysteine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋8.0 – ＋10.09(2 g calcu-lated on the dried basis, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.25 g of L-Cysteine in 50 mL of water is 4.7 to 5.7.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Cysteine in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.30 g of L-Cysteine in 10 mL of diluted nitric acid (1 in 4), add 10 mL of hydrogen peroxide (30), heat for 20 minutes in a boiling water bath, cool, and then add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.35 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.041z). Sulfate <1.14>—Dissolve 0.6 g of L-Cysteine in 30 mL of water and 3 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.35 mL of 0.005 mol/L sulfuric acid VS add 3 mL of dilute hydrochloric acid and water to make 50 mL. Prepare the test mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of L-Cysteine in N-ethylmaleimide solution (1 in 50) to make exactly 10 mL, leave for 30 minutes, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 10 mL, pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution (1). Separately, dissolve 0.10 g of L-cystine in 0.5 mol/L hydrochloric acid TS to make exactly 20 mL. Pipet 1 mL of this solution, add water to make 100 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water, and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate for 30 minutes at 809C. Spray the plate evenly with a solution of ninhydrin in a mix-ture of methanol and acetic acid (100) (97:3) (1 in 100), and then heat at 809C for 10 minutes: the spot obtained from the sample solution corresponding to the spot from the standard solution (2) is not more intense than the spot from the stand-ard solution (2). Also, the spots other than the principal spot and the spots mentioned above from the sample solution are not more intense than the spot from the standard solution (1). Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of L-Cysteine, place it in a stoppered flask, and dissolve in 20 mL of water. Dis-solve 4 g of potassium iodide in this solution, immediately place in ice cold water, add 5 mL of dilute hydrochloric acid and exactly 25 mL of 0.05 mol/L iodine VS, leave in a dark place for 20 minutes, and then titrate <2.50> an excess amount of iodine with 0.1 mol/L sodium thiosulfate VS (in-dicator: starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 12.12 mg of C3H7NO2S
L-Histidine Hydrochloride Hydrate contains not less than 99.
L-Histidine Hydrochloride Hydrate occurs as white crystals or a white crystalline powder. It has an acid taste at first, and a slight bitter taste later. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (99.5). It dissolves in 6 mol/L hydrochloric acid TS.
(1) Determine the infrared absorption spec-trum of L-Histidine Hydrochloride Hydrate as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of L-Histidine Hydrochloride Hydrate (1 in 10) responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: ＋9.2 – ＋10.69(5.5 g calcu-lated on the anhydrous basis, 6 mol/L hydrochloric acid TS, 50 mL, 100 mm). pH <2.54> The pH of a solution of 1.0 g of L-Histidine Hy-drochloride Hydrate in 10 mL of water is between 3.5 and 4.5.
(1) Clarity and color of solution—A solution of 1.0 g of L-Histidine Hydrochloride Hydrate in 10 mL of water is clear and colorless. Sulfate <1.14>—Perform the test with 0.6 g of L-Histidine Hydrochloride Hydrate. Prepare the control so- <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy, develop the plate with a mixture of 1-propanol and am-monia solution (28) (67:33) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of methanol and acetic acid (100) (97:3) (1 in 100) to the plate, and heat the plate at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Water <2.48> 7.2 – 10.0z (0.12 g, volumetric titration, direct titration, using a mixture of methanol for water deter-mination and formamide for water determination (2:1) in-stead of methanol for water determination). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of L-Histidine Hydro-chloride Hydrate, dissolve in 3 mL of formic acid, add ex-actly 15 mL of 0.1 mol/L perchloric acid VS, and heat on a water bath for 30 minutes. After cooling, add 45 mL of acetic acid (100), and titrate <2.50> the excess of perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titra-tion). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 9.581 mg of C6H9N3O2.HCl
L-Histidine occurs as white, crystals or crystal-line powder, having a slight bitter taste. It is freely soluble in formic acid, and soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 6 mol/L hydrochloric acid TS. It shows crystal polymorphism.
Determine the infrared absorption spectrum of L-Histidine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve the sample with a little amount of water, evaporate the water at 609C under reduced pressure, dry the residue, and perform the test. D Optical rotation <2.49> [a]20: ＋11.8 – ＋12.89(5.5 g calcu-lated on the dried basis, 6 mol/L hydrochloric acid TS, 50 mL, 100 mm). pH <2.54> The pH of a solution of 1.0 g of L-Histidine in 50 mL of water is between 7.0 and 8.5.
(1) Clarity and color of solution—A solution of 0.40 g of L-Histidine in 20 mL of water is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of L-Histidine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Histidine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Histidine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 1.0 g of L-Histidine in 30 mL of water by warming. To this solution add 2.4 mL of dilute hydrochloric acid, 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 1.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Histidine according to Method 1, and perform the test ac-cording to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of L-Histidine in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 10 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of 1-propanol and ammonia solution (28) (67:33) to a dis-tance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of methanol and acetic acid (100) (97:3) (1 in 100) to the plate, and heat at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of L-Histidine, dis-solve in 2 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 15.52 mg of C6H9N3O2
L-Isoleucine, when dried, contains not less than 98.
L-Isoleucine occurs as white, crystals or crys-talline powder. It is odorless or has a faint characteristic odor, and has a slightly bitter taste. It is freely soluble in formic acid, sparingly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Isoleucine, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: ＋39.5 – ＋41.59(after dry-ing, 1 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Isoleucine in 100 mL of water: the pH of this solution is between 5.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.5 g of L-Isoleucine in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of L-Isoleucine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Isoleucine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Isoleucine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 1.0 g of L-Isoleucine in 40 mL of water and 2 mL of dilute acetic acid by warm- ing, cool, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of L-Isoleucine according to Method 2, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Isoleucine in 25 mL of water, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard ‌JP XVIII Official Monographs / L-Isoleucine, L-Leucine and L-Valine Granules 1197 solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly the plate with a solution of ninhydrin in acetone (1 in 50), and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.13 g of L-Isoleucine, pre-viously dried, and dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS Flow rate: Adjust so that the retention time of L-valine is about 2.5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, valine, isoleucine and leucine are eluted in this order, and the resolution between the peaks of isoleucine and leucine is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the reten-tion time of isoleucine, leucine and valine are not more than 1.0z, respectively. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose package meets the requirement of the Content uniformity test. To the total content of 1 package of L-Isoleucine, L-Leu-cine and L-Valine Granules add exactly V/25 mL of the in-ternal standard solution, and add 0.1 mol/L hydrochloric acid TS to make V mL so that each mL contains about 3.8 mg of L-isoleucine (C6H13NO2). To 2 mL of this solution add ＝ 13.12 mg of C6H 13NO2 0.02 mol/L hydrochloric acid TS to make 200 mL, and use this solution as the sample solution. Then, proceed as di-
L-Leucine, when dried, contains not less than 98.
L-Leucine occurs as white, crystals or crystal-line powder. It is odorless or has a faint characteristic odor, and has a slightly bitter taste. It is freely soluble in formic acid, sparingly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Leucine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋14.5 – ＋16.09(after dry-ing, 1 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Leucine in 100 mL of water: Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.13 g of L-Leucine, previ-ously dried, and dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 13.12 mg of C6H13NO2
L-Lysine Acetate, when dried, contains not less than 98.
(1) Determine the infrared absorption spec-trum of L-Lysine Acetate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of L-Lysine Acetate (1 in 20) responds to Qualitative Tests <1.09> (2) for acetate. D Optical rotation <2.49> [a]20: ＋8.5 – ＋10.09(after drying, 2.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Lysine Acetate in 10 mL of water: the pH of the solution is between 6.5 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Lysine Acetate in 10 mL of water: the solution is color-less and clear. Chloride <1.03>—Perform the test with 0.5 g of L-Lysine Acetate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Lysine Acetate. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Lysine Acetate. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Lysine Acetate according to Method 4, and perform the test. Pre- pare the control solution with 1.0 mL of Standard Lead So-lution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Lysine Acetate according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Weigh accurately about 0.5 g of L-Lysine Acetate, dissolve in 0.5 mL of hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solu-tion, add 0.02 mol/L hydrochloric acid to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately 2.5 mmol amounts of L-aspartic acid, L-threonine, L-serine, L-glutamic acid, glycine, L-alanine, L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammo-nium chloride, L-histidine and L-arginine, dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the standard stock solution. Pipet 5 mL of this solution, add 0.02 mol/L hydrochloric acid to make exactly 100 mL. Pipet 4 mL of this solution, add 0.02 mol/L hydrochloric acid to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Based on the peak heights of the amino acids obtained from the sample solution and standard solution, determine the mass of the amino acids other than lysine contained in 1 mL of the sample solution, and calcu-late the mass percent: the amount of each amino acids other than lysine is not more than 0.1z. Operating conditions— Detector: A visible spectrophotometer (wavelength: 570 nm). ‌1282 Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (Na type) com-posed with a sulfonated polystyrene copolymer (3 mm in particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B, C, D and E according to the following table, and to each phase add 0.1 mL of capric acid. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of L-Lysine Acetate, previously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 10.31 mg of C6H14N2O2.C2H4O2
L-Lysine Hydrochloride, when dried, contains not less than 98.
L-Lysine Hydrochloride occurs as a white pow-der. It has a slight, characteristic taste. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (95). It shows crystal polymorphism.
(1) Determine the infrared absorption spec-trum of L-Lysine Hydrochloride, previously dried, as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. If any differ-ence appears between the spectra, dissolve L-Lysine Hydro-chloride in water, evaporate the water to dryness at 609C, and repeat the test with the residue. (2) A solution of L-Lysine Hydrochloride (1 in 10) re-sponds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: ＋19.0 – ＋21.59(after dry-ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of L-Lysine Hydrochloride in 10 mL of water: the pH of this solution is between 5.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Lysine Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.6 g of L-Lysine Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Lysine Hydrochloride. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 2.0 g of L-Lysine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of L-Lysine Hydrochloride according to Method 1, and per- ‌JP XVIII Official Monographs / Lysozyme Hydrochloride 1283 form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Lysine Hy-drochloride in 25 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol and ammonia water (28) (67:33) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly the plate with a solution of ninhydrin in acetone (1 in 50) and heat the plate at 809C for 5 minutes: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of L-Lysine Hydro-chloride, previously dried, dissolve in 2 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat on a water bath for 30 minutes. After cooling, add 45 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 9.132 mg of C6H14N2O2.HCl
L-Methionine, when dried, contains not less than 98.
L-Methionine occurs as white, crystals or crys-talline powder. It has a characteristic odor. It is freely soluble in formic acid, soluble in water, and very slightly soluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Methionine, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: ＋21.0 – ＋25.09(after dry-ing, 0.5 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 0.5 g of L-Methionine in 20 mL of water: the pH of this solution is between 5.2 and 6.2.
(1) Clarity and color of solution—Dissolve 0.5 g of L-Methionine in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of L-Methionine in 20 mL of water, and add 6 mL of dilute nitric acid and water to make 40 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS, 6 mL of dilute nitric acid and water to make 40 mL. In this test, to the test solution and the control solution add 10 mL each of silver nitrate TS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Methionine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Methionine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 1.0 g of L-Methionine in 40 mL of water and 2 mL of dilute acetic acid, dissolve by warming, cool, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution as follows: to 2.0 mL of Standard Lead Solu-tion add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Transfer 1.0 g of L-Methionine to a 100-mL decomposition flask, add 5 mL of nitric acid and 2 mL of sulfuric acid, put a small funnel on the mouth of the flask, and heat carefully until white fumes are evolved. After cooling, add two 2-mL portions of nitric acid, heat, add 2-mL portions of hydrogen peroxide (30) several times, and heat until the solution becomes colorless or pale yellow. After cooling, add 2 mL of saturated ammonium oxalate monohydrate solution, and heat again until white fumes are evolved. After cooling, add water to make 5 mL, and per-form the test with this solution as the test solution (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Methionine in 10 mL of water, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. After air-drying, immedi-ately develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a so-lution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of L-Methionine, pre-viously dried, and dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 14.92 mg of C5H11NO2S
L-Phenylalanine, when dried, contains not less than 98.
L-Phenylalanine occurs as white, crystals or crystalline powder. It is odorless or has a faint characteristic odor, and has a slightly bitter taste. It is freely soluble in formic acid, sparingly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Phenylalanine, previously dried, as directed in the po-tassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －33.0 – －35.59(after dry-ing, 0.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.20 g of L-Phenylalanine in 20 mL of water: the pH of this solution is between 5.3 and 6.3.
(1) Clarity and color of solution—Dissolve 0.5 g of L-Phenylalanine in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of L-Phenylalanine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Phenylalanine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Phenylalanine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). ‌Heavy metals <1.07>—Dissolve 1.0 g of L-Phenylala-nine in 40 mL of water and 2 mL of dilute acetic acid by warming, cool, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution as follows: to 2.0 mL of Standard Lead Solu-tion add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of L-Phenylalanine in 5 mL of dilute hydrochloric acid and 15 mL of water, and perform the test with this solution as the test solution (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Phenylala-nine in 25 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.17 g of L-Phenylalanine, previously dried, and dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.52 mg of C9H11NO2
L-Proline occurs as white, crystals or crystal-line powder. It has a slightly sweet taste. It is very soluble in water and in formic acid, and slightly soluble in ethanol (99.5). It is deliquescent.
Determine the infrared absorption spectrum 1 cm Absorbance <2.24> E1z 4 mg, methanol, 250 mL). (225 nm): 384 – 414 (after drying, of L-Proline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra
(1) Heavy metals <1.07>—Proceed with 1.0 g of Proglumide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Proglumide add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) and 1.5 mL of hydrogen peroxide (30), burn the ethanol, and prepare the test solution according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Proglumide in 5 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －84.0 – －86.09(1 g calculated on the dried basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution of 1.0 g of L-Proline in 10 mL of water is 5.9 to 6.9. Purity (1) Clarity and color of solution—Dissolve 1.0 g of L-Proline in 10 mL of water: the solution is clear and col-orless. Chloride <1.03>—Perform the test with 0.5 g of L-Proline. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Pro-line. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Proline. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Pro-line according to Method 1, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Proline according to Method 1, and perform the test ac-cording to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Weigh accurately about 0.5 g of L-Proline, and dissolve in 0.5 mL of hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount, equivalent to 2.5 mmol, of L-aspartic acid, L-threonine, L-serine, L-glutamic acid, L-pro-line, glycine, L-alanine, L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammonium chloride, L-histidine and L-argi-nine, dissolve them in 0.1 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, and add 0.02 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 4 mL of this solution, add 0.02 mol/L hydro-chloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the mass percentage of each amino acid, using the mass of amino acid other than proline in 1 mL of the sample solution from the height of the peaks obtained from the sample and standard solution: the amount of each amino acid other than proline is not more than 0.1 z. Operating conditions— Detector: A visible absorption photometer (wavelength: 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with a sulfonated polystyrene (3 mm in particle diameter) (Na type). Column temperature: A constant temperature of about 579C. Chemical reaction vessel temperature: A constant temper-ature of about 1309C. Reaction time: About 1 minute. Mobile phase: Prepare the mobile phases A, B, C, D and E according to the following table, and add 0.1 mL each of caprylic acid. Mobile phase A B C D E Citric acid 19.80 g monohydrate 22.00 g 12.80 g 6.10 g — Trisodium citrate 6.19 g dihydrate 7.74 g 13.31 g 26.67 g — Sodium chlo- ride 5.66 g 7.07 g 3.74 g 54.35 g — Sodium hydroxide — — — — 8.00 g Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL Thiodiglycol 5 mL 5 mL 5 mL — — Benzyl alcohol — — — 5 mL — Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL 4 mL Water a sufficient a sufficient a sufficient a sufficient a sufficient amount amount amount amount amount Total amount 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL Switching of mobile phase: Switch the mobile phases A, B, C, D and E sequentially so that when proceed with 20 mL of the standard solution under the above conditions, aspartic acid, threonine, serine, glutamic acid, proline, glycine, ala-nine, cystine, valine, methionine, isoleucine, leucine, tyro-sine, phenylalanine, lysine, ammonia, histidine and arginine are eluted in this order with the resolution between the peaks of isoleucine and leucine being not less than 1.2. Reaction reagent: Dissolve 204 g of lithium acetate dihy-drate in an appropriate amount of water, add 123 mL of acetic acid (100), 401 mL of 1-methoxy-2-propanol and water to make 1000 mL, pass nitrogen for 10 minutes, and use this solution as Solution (I). Separately, to 979 mL of 1-methoxy-2-propanol add 39 g of ninhydrin, pass nitrogen for 5 minutes, add 81 mg of sodium borohydride, pass nitrogen for 30 minutes, and use this solution as Solution (II). Pre-pare a mixture with an equal volume of the Solution (I) and (Prepare before use). Flow rate of mobile phase: 0.20 mL per minute. Flow rate of reaction regent: 0.24 mL per minute. System suitability— System performance: When the test is run with 20 mL of the standard solution under the above operating conditions, the resolution between the peaks of glycine and alanine is not less than 1.2. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak height of each amino acid other than proline in the standard solution is not more than 5.0z, and the relative standard deviation of the retention time is not more than 1.0z. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.12 g of L-Proline, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.51 mg of C5H9NO2
L-Serine, when dried, contains not less than 98.
L-Serine occurs as white, crystals or a crystal-line powder. It has a slight sweet taste. It is freely soluble in water and in formic acid, and practi-cally insoluble in ethanol (99.5). It dissolves in 2 mol/L hydrochloric acid TS.
Determine the infrared absorption spectrum of L-Serine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋14.0 – ＋16.09(After dry-ing, 2.5 g, 2 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of L-Serine in 10 mL of water is between 5.2 and 6.2. actly 10 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Then develop with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of methanol and acetic acid (100) (97:3) (1 in 100) on the plate, and heat the plate at 809C for 10 minutes: the spot other than the princi-pal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on Ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.11 g of L-Serine, previ-ously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 10.51 mg of C3H7NO3
L-Threonine, when dried, contains not less than 98.
L-Threonine occurs as white, crystals or crys-talline powder. It is odorless or has a slight, characteristic odor, and has a slightly sweet taste. It is freely soluble in formic acid, soluble in water, and practically insoluble in ethanol (95).
Determine the infrared absorption spectrum of L-Threonine, previously dried, as directed in the potas-sium bromide disk method under the Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －26.0 – －29.09(after dry-ing, 1.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.20 g of L-Threonine in 20 mL of water: the pH of this solution is between 5.2 and 6.2.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Threonine in 20 mL of water: the solution is clear and colorless. ‌Chloride <1.03>—Perform the test with 0.5 g of L-Threonine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Threonine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Threonine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Threo-nine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of L-Threonine in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 2 ppm). Related substances—Dissolve 0.30 g of L-Threonine in 50 mL of water, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly the plate with a solution of ninhydrin in acetone (1 in 50), and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.12 g of L-Threonine, pre-viously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.91 mg of C4H9NO3
L-Tryptophan, when dried, contains not less than 98.
L-Tryptophan occurs as white to yellowish white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in formic acid, slightly soluble in water, and very slightly soluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Tryptophan, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: －30.0 – －33.09Weigh accu- Amount (mg) of troxipide (C ＝ MS × QT/QS × 40 15H22N2O4) rately about 0.25 g of L-Tryptophan, previously dried, and dissolve in 20 mL of water by warming. After cooling, add water to make exactly 25 mL, and determine the optical rota- MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 258 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: To 1500 mL of diluted phosphoric acid (1 in 500) add diethylamine to adjust the pH to 3.0. To 1500 mL of this solution add 100 mL of methanol and 50 mL of tetrahydrofuran. Flow rate: Adjust so that the retention time of troxipide is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, troxipide and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of troxipide to that of the internal standard is not more than 1.0z.
(1) Clarity of solution—Dissolve 0.20 g of L-Tryp-tophan in 10 mL of 2 mol/L hydrochloric acid TS: the solu-tion is clear. Chloride <1.03>—Dissolve 0.5 g of L-Tryptophan in 6 mL of dilute nitric acid, and add water to make 50 mL. Per- form the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochlo-ric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 0.6 g of L-Tryptophan in 40 mL of water and 1 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Tryptophan. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Tryp-tophan according to Method 4, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of L-Tryptophan in 3 mL of 1 mol/L hydrochloric acid TS and 2 mL of water by heating, and perform the test with this solution as the test so-lution (not more than 2 ppm). Related substances—Dissolve 0.30 g of L-Tryptophan in 1 mL of 1 mol/L hydrochloric acid TS, add water to make 50 mL, and use this solution as the sample solution. Pipet 1 ‌mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of L-Tryptophan, pre-viously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.42 mg of C11H12N2O2
L-Tyrosine, when dried, contains not less than 99.
L-Tyrosine occurs as white, crystals or a crys-talline powder. It is freely soluble in formic acid, and practically insoluble in water and in ethanol (99.5). It dissolves in dilute hydrochloric acid and in ammonia TS. ‌Identification (1) Determine the absorption spectrum of a solution of L-Tyrosine in 0.1 mol/L hydrochloric acid (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-‌ acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of ＝ 18.12 mg of C9H 11NO3 L-Tyrosine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －10.5 – －12.59(after dry-ing, 2.5 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of L-Tyrosine in 20 mL of 1 mol/L hydrochloric acid TS by warming: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of L-Tyrosine in 12 mL of dilute nitric acid and 20 mL of water, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 12 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate <1.14>—Dissolve 0.6 g of L-Tyrosine in 5 mL of dilute hydrochloric acid, and add water to make 45 mL. Perform the test using this solution as the test solution. Pre-pare the control solution as follows: To 0.35 mL of 0.005 mol/L sulfuric acid VS add 5 mL of dilute hydrochloric acid and water to make 45 mL. To the test solution and the con-trol solution add 5 mL of barium chloride TS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Tyrosine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Tyro-sine according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Tyrosine according to Method 3, and perform the test ac- cording to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of L-Tyrosine in 10 mL of diluted ammonia solution (28) (1 in 2), add water to make 20 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 10 mL, pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Then develop with a mixture of 1-propanol and ammonia solution (28) (67:33) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of metha-nol and acetic acid (100) (97:3) (1 in 100) on the plate, and then heat the plate at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.18 g of L-Tyrosine previ-ously dried, dissolve in 6 mL of formic acid, add 50 mL of
L-Valine, when dried, contains not less than 98.
L-Valine occurs as white, crystals or crystalline powder. It is odorless or has a faint characteristic odor, and has a slightly sweet taste, which becomes bitter. It is freely soluble in formic acid, soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid.
Determine the infrared absorption spectrum of L-Valine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋26.5 – ＋29.09(after dry-ing, 2 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 0.5 g of L-Valine in 20 mL of water: the pH of this solution is between 5.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.5 g of L-Valine in 20 mL of water: the solution is clear and col-orless. Chloride <1.03>—Perform the test with 0.5 g of L-Valine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Valine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Valine. Prepare the control solution with 5.0 mL of Stand-ard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Valine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Proceed with 1.0 g of L-Valine, pre-pare the test solution according to Method 2, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Valine in 25 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot obtained from the ‌sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.12 g of L-Valine, previ-ously dried, and dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.72 mg of C5H11NO2
fibrous. A transverse section, with fine dots
solution in a test tube, and add carefully 0.5 mL
a saccharized substance obtained by hydrolysis
a clear, colorless liquid, having the odor
order with the resolution between the peaks
Adsorbed Diphtheria-Purified Pertussis-Tetanus Combined Vaccine
沈降精製百日せきジフテリア破傷風混合ワクチン
Adsorbed Diphtheria-Purified Pertussis-Teta-nus Combined Vaccine becomes a homogeneous, white tur-bid liquid on shaking. Adsorbed Diphtheria-Tetanus Combined Toxoid
Adsorbed Hepatitis B Vaccine becomes a ho-mogeneous, whitish turbid liquid on shaking. L-Histidine
Adsorbed Purified Pertussis Vaccine forms a homogeneous, white turbidity on shaking. Pethidine Hydrochloride
Albumin Tannate occurs as a light brown powder. It is odorless, or has a faint, characteristic odor. It is practically insoluble in water and in ethanol (95). It dissolves in sodium hydroxide TS with turbidity.
(1) To 0.1 g of Albumin Tannate add 10 amount amount amount amount amount mL of ethanol (95), and heat in a water bath for 3 minutes Total volume 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL Changing mobile phases: When the procedure is run with 20 mL of the standard solution under the above operating conditions, switchover in sequence to mobile phases A, B, C, D and E so that aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionine, isoleu-cine, leucine, tyrosine, phenylalanine, lysine, ammonia, histidine, and arginine are eluted in this order with the reso-lution between the peaks of isoleucine and leucine being not less than 1.2. Reaction reagents: Dissolve 204 g of lithium acetate dihy-drate in water, add 123 mL of acetic acid (100) and 401 mL of 1-methoxy-2-propanol, and water to make 1000 mL, introduce nitrogen for 10 minutes, and use this solution as solution (I). Separately, add 39 g of ninhydrin to 979 mL of 1-methoxy-2-propanol, introduce nitrogen for 5 minutes, add 81 mg of sodium borohydride, introduce nitrogen for 30 minutes, and use this solution as solution (II). To 1 volume of solution (I) add 1 volume of solution (II). Prepare before use. Flow rate of mobile phase: 0.20 mL per minute. Flow rate of reaction reagent: 0.24 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating conditions, the resolution between the peaks of glycine and alanine is not less than 1.2. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak height and retention time of each amino acid obtained from the standard solution are not more than 5.0z and not more than 1.0z, respectively. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Acidity—Shake 1.0 g of Albumin Tannate with 50 mL of water for 5 minutes, and filter. To 25 mL of the fil-trate add 1.0 mL of 0.1 mol/L sodium hydroxide VS and 2 drops of phenolphthalein TS: a red color develops. (2) Fats—To 2.0 g of Albumin Tannate add 20 mL of petroleum benzine, shake vigorously for 15 minutes, and filter. Evaporate 10 mL of the filtrate on a water bath: the mass of the residue is not more than 50 mg. Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (0.5 g). Digestion test To 1.00 g of Albumin Tannate add 0.25 g of saccharated pepsin and 100 mL of water, shake well, and allow to stand for 20 minutes at 40 ± 19C in a water bath. Add 1.0 mL of dilute hydrochloric acid, shake, and allow to stand for 3 hours at 40 ± 19C. Cool rapidly to ordinary tem-perature, and filter. Wash the residue with three 10-mL por-tions of water, dry in a desiccator (silica gel) for 18 hours, and dry at 1059C for 5 hours: the mass of the residue is 0.50 to 0.58 g.
Weigh accurately about 90 mg of L-Alanine, previ-ously dried, dissolve in 3 mL of formic acid, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 8.909 mg of C3H7NO2
ジヒドロエルゴトキシンメシル酸塩
Dihydroergotoxine Mesilate contains not less than 97.
Dihydroergotoxine Mesilate occurs as a white to pale yellow powder. It is soluble in methanol, sparingly soluble in ethanol (95), slightly soluble in water, in acetonitrile and in chloroform, and practically insoluble in diethyl ether. JP XVIII Official Monographs / Dihydroergotoxine Mesilate 861
Determine the infrared absorption spectrum of Dihydroergotoxine Mesilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios of the peak areas of dihydroergocornine, dihydro-a-ergocryptine, dihydroergocristine and dihydro-b-ergocryptine to the peak area of the internal standard of these solutions. Amount (mg) of dihydroergotoxine mesilate D Optical rotation <2.49> [a]20: ＋11.0 – ＋15.09(0.2 g calcu- ＝ MS × (QTA ＋ QTB ＋ QTC ＋ QTD)/ lated on the anhydrous basis, dilute ethanol, 20 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.10 g of Dihydroergotoxine Mesilate in 20 mL of water: the solu-tion is clear and the color of the solution is not more intense than that of the following control solution. Control solution: To a mixture of 1.0 mL of Cobalt (II) Chloride CS, 0.4 mL of Copper (II) Sulfate CS and 2.4 mL of Iron (III) Chloride CS add diluted hydrochloric acid (1 in 40) to make exactly 200 mL. Heavy metals <1.07>—Proceed with 1.0 g of Di- hydroergotoxine Mesilate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Weigh accurately 0.100 g of Di-hydroergotoxine Mesilate, dissolve it in a mixture of chlo-roform and methanol (9:1) to make exactly 5 mL, and use this solution as the sample solution. Separately, weigh accu-rately 10 mg of dihydroergocristine mesilate for thin-layer chromatography, and dissolve in a mixture of chloroform and methanol (9:1) to make exactly 100 mL. Pipet 6 mL, 4 mL and 2 mL of this solution, add a mixture of chloroform and methanol (9:1) to make exactly 10 mL, respectively, and use these solutions as the standard solutions (1), (2) and (3), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03> without putting the filter paper in the developing vessel. Spot 5 mL each of the sample solution and the standard solutions (1), (2) and on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, ethyl acetate, methanol and ammonia solution (28) (50:50:3:1) to a distance of about 15 cm, and dry the plate with the aid of a cool air stream. Immediately after that, develop the plate again with a newly prepared mixture of dichloromethane, ethyl acetate, methanol and ammonia solution (28) (50:50:3:1) to a distance of about 15 cm, and dry the plate within 1 minute with the aid of a cool air stream. Spray evenly p-dimethylaminobenzal-dehyde-hydrochloric acid TS on the plate, dry the plate within 2 minutes with the aid of a cool air stream, and heat it at 409C for 15 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution (1), not more than 2 spots are more intense than that from the standard solution (2), and not more than 4 spots are more intense than that from the standard solution (3). Water <2.48> Not more than 5.0z (0.2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Dihydroergotoxine mesilate—Weigh accurately about 30 mg each of Dihydroergotoxine Mesilate and Di-hydroergotoxine Mesilate RS, and dissolve them separately in a suitable amount of a mixture of water and acetonitrile (3:1). To these solutions add exactly 10 mL of the internal standard solution and an amount of a mixture of water and acetonitrile (3:1) to make 50 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 20 mL of the sample solution and standard solution (QSA ＋ QSB ＋ QSC ＋ QSD) MS: Amount (mg) of Dihydroergotoxine Mesilate RS taken, calculated on the anhydrous basis QTA: Ratio of the peak area of dihydroergocornine to that of the internal standard of the sample solution × 659.80 QTB: Ratio of the peak area of dihydro-a-ergocryptine to that of the internal standard of the sample × 673.83 QTC: Ratio of the peak area of dihydroergocristine to that of the internal standard of the sample solution × 707.85 QTD: Ratio of the peak area of dihydro-b-ergocryptine to that of the internal standard of the sample solution × 673.83 QSA: Ratio of the peak area of dihydroergocornine to that of the internal standard of the standard solution × 659.80 QSB: Ratio of the peak area of dihydro-a-ergocryptine to that of the internal standard of the standard solution × 673.83 QSC: Ratio of the peak area of dihydroergocristine to that of the internal standard of the standard solution × 707.85 QSD: Ratio of the peak area of dihydro-b-ergocryptine to that of the internal standard of the standard solution × 673.83 Internal standard solution—Dissolve 0.04 g of chloram-phenicol in a mixture of water and acetonitrile (3:1) to make 250 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and triethylamine (30:10:1). Flow rate: Adjust so that the retention time of chloram-phenicol is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard, dihydroergocornine, dihydro-a-ergocryptine, dihydroergocristine and dihydro-b-ergocryp-tine are eluted in this order with the resolution between the peaks of dihydro-a-ergocryptine and dihydroergocristine being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of dihydroergocornine, dihydro-a-ergocryp-tine, dihydroergocristine and dihydro-b-ergocryptine to that of the internal standard is not more than 0.5z. Relative contents of dihydroergocornine mesilate, dihydroergocryptine mesilate and dihydroergocristine mesilate—Calculate the relative amounts of dihydroergocor- ‌862 nine mesilate, dihydroergocryptine mesilate (dihydro-a-ergocryptine mesilate and dihydro-b-ergocryptine mesilate) and dihydroergocristine mesilate from the chromatogram obtained in Assay (1) for the sample solution using the fol-lowing equations: Relative amount (z) of dihydroergocornine mesilate To 5 mL of a solution of Dilazep Hydrochloride Hy-drate (3 in 500) add 0.3 mL of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Dilazep Hydrochloride Hydrate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and ＝ QTA/(QTA ＋ QTB ＋ QTC ＋ QTD ) × 100 compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same Relative amount (z) of dihydroergocryptine mesilate ＝ (QTB ＋ QTD)/(QTA ＋ QTB ＋ QTC ＋ QTD) × 100 Relative amount (z) of dihydroergocristine mesilate wavelengths. Determine the infrared absorption spectrum of Dilazep Hydrochloride Hydrate as directed in the potassium ＝ QTC/(QTA ＋ QTB ＋ QTC ＋ QTD) × 100 chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Ratio of the content of dihydro-a-ergocryptine mesi-late to dihydro-b-ergocryptine mesilate—Calculate the ratio of the amount of dihydro-a-ergocryptine mesilate to di-hydro-b-ergocryptine mesilate from the chromatogram ob-tained in the Assay (1) for the sample solution using the fol-lowing equations: Ratio of the content of dihydro-a-ergocryptine mesilate to dihydro-b-ergocryptine mesilate ＝ QTB/QTD
Dried Thyroid is the fresh thyroid gland, previously deprived of connective tissue and fat, minced, dried rapidly at a temperature not above 509C, and pow-dered, or diluted with suitable diluents. It is obtained from domesticated animals that are used for food by man. It contains not less than 0.
Dried Thyroid occurs as a light yellow to grayish brown powder. It has a slight, characteristic, meat-like odor.
Mount Dried Thyroid in diluted formalde-hyde solution (1 in 10), stain in hematoxylin TS for 10 to 30 minutes, wash with water, soak in a mixture of 1 mL of hy-drochloric acid and 99 mL of diluted ethanol (7 in 10) for 5 to 10 seconds, and again wash with water for about 1 hour. Stain in a solution of eosin Y (1 in 100) for 1 to 5 minutes, wash with water, dehydrate, and soak successively in diluted ethanol (7 in 10) for 5 to 10 seconds, in diluted ethanol (4 in 5) for 5 to 10 seconds, in diluted ethanol (9 in 10) for 1 to 2 minutes, in ethanol (95) for 1 to 5 minutes then in ethanol (99.5) for 1 to 5 minutes. Interpenetrate in xylene, seal with balsam, and examine under a microscope: epithelial nuclei forming follicles peculiar to the thyroid gland are observed.
(1) Inorganic iodides—Mix 1.0 g of Dried Thyroid with 10 mL of a saturated solution of zinc sulfate, shake for 5 minutes, and filter. To 5 mL of the filtrate add 0.5 mL of starch TS, 4 drops of sodium nitrite TS and 4 drops of dilute sulfuric acid with thorough shaking: no blue color is pro-duced. (2) Fat—Extract 1.0 g of Dried Thyroid with diethyl ether for 2 hours using a Soxhlet extractor. Evaporate the diethyl ether extract, and dry the residue at 1059C to con-stant mass: the mass of the residue is not more than 30 mg. Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, constant mass). Total ash <5.01> Not more than 5.0z (0.5 g).
Transfer about 1 g of Dried Thyroid, accurately weighed, to a crucible, add 7 g of potassium carbonate, mix carefully, and gently tap the crucible on the table to compact the mixture. Overlay with 10 g of potassium carbonate, and compact again thoroughly by tapping. Place the crucible in a muffle furnace preheated to a temperature between 6009C and 7009C, and ignite the mixture for 25 minutes. Cool, add 20 mL of water, heat gently to boiling, and filter into a flask. To the residue add 20 mL of water, boil, and filter into the same flask. Rinse the crucible and the char on the funnel with boiling water until the filtrate measures 200 mL. Add slowly 7 mL of freshly prepared bromine TS, 40 mL of di-luted phosphoric acid (1 in 2), and boil until starch iodide paper is no longer colored blue by the evolved gas. Wash down inside of the flask with water, and continue boiling for 5 minutes. During the boiling add water from time to time to maintain a volume at not less than 200 mL. Cool, add 5 mL of a solution of phenol (1 in 20), again rinse inside of the flask with water, and allow to stand for 5 minutes. Add 2 mL of diluted phosphoric acid (1 in 2) and 5 mL of potas- ‌sium iodide TS, and titrate <2.50> immediately the liberated iodine with 0.01 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction.‌ Each mL of 0.01 mol/L sodium thiosulfate VS ＝ 0.2115 mg of I
Freeze-dried BCG Vaccine (for Percutaneous Use)
Freeze-dried BCG Vaccine (for Percutaneous Use) is a preparation for injection which is dissolved before use. It contains live bacteria derived from a culture of the bacillus of Calmette and Gueárin.
Freeze-dried BCG Vaccine (for Percutaneous Use) becomes a white to light yellow, turbid liquid on addi-tion of solvent. Beclometasone Dipropionate
Freeze-dried Inactivated Tissue Culture Rabies Vaccine
乾燥組織培養不活化狂犬病ワクチン
Freeze-dried Inactivated Tissue Culture Rabies Vaccine becomes a colorless or light yellow-red clear liquid on addition of solvent. Ranitidine Hydrochloride
Freeze-dried Live Attenuated Measles Vaccine
Freeze-dried Live Attenuated Measles Vaccine is a preparation for injection which is dissolved before use. It contains live attenuated measles virus.
Freeze-dried Live Attenuated Measles Vaccine becomes a colorless, yellowish or reddish clear liquid on ad-dition of solvent. Meclofenoxate Hydrochloride
Freeze-dried Live Attenuated Mumps Vaccine
乾燥弱毒生おたふくかぜワクチン
Freeze-dried Live Attenuated Mumps Vaccine becomes a clear, colorless, yellowish or reddish liquid on ad-dition of solvent. Mupirocin Calcium Hydrate
Freeze-dried Live Attenuated Rubella Vaccine
Freeze-dried Live Attenuated Rubella Vaccine is a preparation for injection which is dissolved before use. It contains live attenuated rubella virus.
Freeze-dried Live Attenuated Rubella Vaccine becomes a colorless, yellowish or reddish, clear liquid on ad-dition of solvent. Saccharated Pepsin
Freeze-dried Smallpox Vaccine Prepared in Cell Culture
Freeze-dried Smallpox Vaccine Prepared in Cell Culture is a preparation for injection which is dis-solved before use. It contains live vaccinia virus.
Freeze-dried Smallpox Vaccine Prepared in Cell Culture becomes a reddish clear liquid on addition of solvent. Sodium Acetate Hydrate
Freeze-dried Smallpox Vaccine is a preparation for injection which is dissolved before use. It contains live vaccinia virus.
Freeze-dried Smallpox Vaccine becomes a white to gray, turbid liquid on addition of solvent. Freeze-dried Smallpox Vaccine Prepared in Cell Culture
Heparin Calcium is the calcium salt of sulfated glycosaminoglycans composed of disaccharide units of D-glucosamine and uronic acid (L-iduronic acid or D- glucuronic acid) obtained from the intestinal mucosa of healthy edible swine. It contains not less than 180 Heparin Units (anti-factor IIa activity) per mg and not less than 8.
Heparin Calcium occurs as a white to grayish brown, powder or grains. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic.
(1) Dissolve 10 mg of Heparin Calcium in 5 mL of water, and add 0.1 mL of 1 mol/L hydrochloric acid TS and 5 mL of toluidine blue O solution (1 in 20,000): a purple to red-purple color develops. Dissolve 1 mg each of Heparin Calcium and Heparin Sodium for Identification RS in 1 mL of water, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions: the retention time for the major peak from the sam-ple solution and standard solution is identical. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase and flow rate: Proceed as di-rected under the operating conditions in Purity (9). System suitability— System performance: Dissolve 1.0 mg of Heparin Sodium for Identification RS in 0.60 mL of water. Dissolve 0.10 mg of Over-sulfated Chondroitin Sulfate for System Suitability RS in 0.20 mL of water. Dissolve 1.0 mg of dermatan sulfate in 2.0 mL of water. To 90 mL of the solution of Heparin So-dium for Identification RS add 30 mL each of the solutions of Over-sulfated Chondroitin Sulfate for System Suitability RS and dermatan sulfate, and mix. When the procedure is run with 20 mL of the mixture under the above operating conditions, dermatan sulfate, heparin and over-sulfated chondroitin sulfate are eluted in this order with the resolu-tion between the peaks of dermatan sulfate and heparin being not less than 1.0 and that between the peaks of heparin and over-sulfated chondroitin sulfate being not less than 1.5. A solution of 50 mg of Heparin Calcium in 5 mL of water responds to Qualitative Tests <1.09> for calcium salt. pH <2.54> Dissolve 1.0 g of Heparin Calcium in 100 mL of water: the pH of the solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Heparin Calcium in 20 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.05. Chloride <1.03>—Perform the test with 0.5 g of Heparin Calcium. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Heavy metals <1.07>—Proceed with 0.5 g of Heparin Calcium according to Method 2, and perform the test. Pre-pare the control solution with 1.5 mL of Standard Lead So-lution (not more than 30 ppm). Barium—Dissolve 30 mg of Heparin Calcium in 3.0 mL of water, and use this solution as the sample solution. To 1.0 mL of the sample solution add 3 drops of dilute sul-furic acid, and allow to stand for 10 minutes: no turbidity is produced. Total nitrogen—Weigh accurately about 0.1 g of Heparin Calcium, previously dried, and perform the test as directed under Nitrogen Determination <1.08>: the amount of nitrogen (N: 14.01) is not more than 3.0z. Protein—(i) Sodium carbonate solution: To 4 volumes of a mixture of sodium hydroxide solution (1 in 100) and anhydrous sodium carbonate solution (1 in 20) (1:1) add 1 volume of water. Copper sulfate solution: To 4 volumes of a mixture of copper (II) sulfate pentahydrate solution (1 in 80) and sodium tartrate dihydrate solution (149 in 5000) (1:1) add 1 volume of water. Alkaline copper solution for heparin: Mix 50 volumes of the sodium carbonate solution and 1 volume of the copper sulfate solution. Prepare before using. Procedure: Use a solution of Heparin Calcium (1 in 200) as the sample solution. Use a solution of bovine serum albumin (1 in 40,000) as the standard solution. To exactly 1 mL each of the sample solution and standard solution add exactly 5 mL of the alkaline copper solution for heparin, mix, and allow them to stand at room temperature for 10 minutes. To each of these solutions add exactly 0.5 mL of diluted Folin's TS (1 in 2), shake, allow them to stand at room temperature for 30 minutes, and centrifuge at room temperature. Determine the absorbances at 750 nm of the su-pernatant liquids as directed under Ultraviolet-visible Spec-trophotometry <2.24> using water as the blank: the absor-bance of the solution obtained from the sample solution is not more than that of the solution from the standard solu-tion. Nucleic acid—Dissolve 40 mg of Heparin Calcium in 10 mL of a solution of disodium dihydrogen ethylene-diamine tetraacetate dihydrate (93 in 50,000), and determine JP XVIII Official Monographs / Heparin Calcium 1097 the absorbance of this solution at 260 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absor-bance is not more than 0.15. Over-sulfated chondroitin sulfate—Dissolve 20 mg of phoric acid (1 in 10). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Heparin Calcium in 0.60 mL of a solution of sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) spectroscopy (1 in 10,000). Determine the spectrum of this solution as directed under Nuclear Magnetic Resonance Spectroscopy <2.21> (1H) in accordance with the following conditions, using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an internal ref-erence compound: it exhibits no signal corresponding to N-acetyl proton of over-sulfated chondroitin sulfate at d 2.18 ± 0.05 ppm, or the signal disappears when determining the spectrum of the sample solutions as directed under 1H with 13C-decoupling. Operating conditions— Spectrometer: 1.1. FT-NMR, Not less than 400 MHz. Temperature: 259C. Spinning: off. Number of data points: 32,768. Spectral range: Signal of DHO ± 6.0 ppm. Flip angle: 909. Delay time: 20 seconds. Dummy scans: 4. Number of scans: SN ratio of the signal of N-acetyl pro-ton of heparin is not less than 1000. Window function: Exponential function (Line broadening factor ＝ 0.2 Hz). System suitability— System performance: Dissolve 20 mg of Heparin Calcium in 0.40 mL of a solution of sodium 3-trimethylsilyl-propionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance spectroscopy (1 in 10,000). Dissolve 0.10 mg of Over-sulfated Chondroitin Sulfate for System Suitability RS in 1.0 mL of a solution of sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance spectroscopy (1 in 10,000). To the solution of heparin calcium add 0.20 mL of the solution of Over-sulfat-ed Chondroitin Sulfate for System Suitability RS. When de-termining the spectrum of this solution under the above op-erating conditions, it exhibits the signal of N-acetyl proton of heparin and the signal of N-acetyl proton of over-sulfated chondroitin sulfate at d 2.04 ± 0.02 ppm and d 2.18 ± 0.05 ppm, respectively. Related substances—Dissolve 2.0 mg of Heparin Cal-cium in 0.1 mL of water, and perform the test with exactly 20 mL of this solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions: it exhib-its no peaks after the heparin peak. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 202 nm). Column: A stainless steel column 2.0 mm in inside diame-ter and 7.5 cm in length, packed with synthetic polymer for liquid chromatography to which diethylaminoethyl group binds (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Dissolve 0.4 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (1 in 10). Mobile phase B: Dissolve 0.4 g of sodium dihydrogen phosphate dihydrate and 106.4 g of lithium perchlorate in 1000 mL of water, and adjust to pH 3.0 with diluted phos- 0 – 3 90 10 3 – 15 90 → 0 10 → 100 Flow rate: 0.2 mL per minute. Time span of measurement: About 2 times as long as the retention time of heparin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 10 mg of Heparin Sodium for Identification RS in 0.40 mL of water, and use this solution as the heparin sodium standard stock solution. Separately, dissolve 0.10 mg of Over-sulfated Chondroitin Sulfate for System Suitability RS in 0.20 mL of water, and use this solution as the over-sulfated chondroitin sulfate standard solution. To 60 mL of the heparin sodium standard stock solution add 3 mL of the over-sulfated chondroitin sul-fate standard solution and 12 mL of water, and mix. When the procedure is run with 20 mL of the mixture under the above operating conditions, it exhibits an over-sulfated chondroitin sulfate peak. System performance: To 120 mL of the heparin sodium standard stock solution add 30 mL of the over-sulfated chon-droitin sulfate standard solution, mix and use this solution as the solution for system suitability test. When the proce-dure is run with 20 mL of the solution for system suitability test under the above operating conditions, heparin and over-sulfated chondroitin sulfate are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of over-sulfated chondroitin sulfate is not more than 2.0z. Loss on drying <2.41> Not more than 8z (50 mg, in vacu-um, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.0030 EU/heparin Unit. Anti-factor Xa activity to anti-factor IIa activity ratio The ratio of the anti-factor Xa activity determined by the follow-ing method to the anti-factor IIa activity obtained in the Assay, calculated by dividing the former with the later, is 0.9 – 1.1. Anti-factor Xa activity determination Substrate solution: Dissolve 25 mg of N-benzoyl-L-isoleucyl-L-glutamyl(g-OR)-glycyl-L-arginyl-p-nitroanilide hydrochloride in 33.3 mL of water. Anti-thrombin solution: Dissolve human anti-throm-bin in water so that each mL contains 1 IU. To 150 mL of this solution add 2250 mL of buffer solution. Factor Xa solution: To 1200 mL of factor Xa TS add 1200 mL of buffer solution. Buffer solution: Proceed as directed in the Assay (1). Stopping solution: Proceed as directed in the Assay (1). Heparin standard solutions: Proceed as directed in the Assay (1). However, the standard solutions are prepared based on anti-factor Xa activity Unit instead of Heparin Unit. Heparin sample solutions: Proceed as directed in the Assay (1). However, the sample solutions are prepared based on anti-factor Xa activity Unit instead of Heparin Unit. Procedure: Transfer separately two 50-mL portions of each different dilution of the heparin standard solutions and the heparin sample solutions and five 50-mL portions of buffer solution as the blank to 1.5-mL tubes. Warm these 21 tubes, anti-thrombin solution, factor Xa solution and sub-strate solution at 379C all together. Start the following procedure at 2 minutes after warming in the order: buffer so-lution, S1, S2, S3, S4, buffer solution, T1, T2, T3, T4, buffer solution, T1, T2, T3, T4, buffer solution, S1, S2, S3, S4, and buffer solution. To each tube add 50 mL of anti-thrombin so-lution, mix, and warm at 379C for exactly 4 minutes, add 100 mL of factor Xa solution, mix, and incubate for exactly 12 minutes. Then, add 100 mL of substrate solution, mix, incubate for exactly 4 minutes, add 50 mL of stopping solu-tion to each tube, and mix immediately. Separately, to 50 mL of stopping solution add 100 mL of substrate solution, 100 mL of factor Xa solution, 50 mL of anti-thrombin solution and 50 mL of buffer solution, mix, and use this solution as a control. Determine the absorbance of each solution at 405 nm against the control. Confirm that the relative standard deviation of the reading of the blank is not more than 10z. Calculations: When the regression expression, y ＝ Ic ＋ AXs ＋ BXt, is obtained using y as log of the absorbance values, Xs as the concentration of the heparin standard solu-tions and Xt as the concentration of the heparin sample solutions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate anti-factor Xa activity per mg of Heparin Cal-cium by the following formula. Anti-factor Xa activity per mg of Heparin Calcium ＝ 100 × R × V/M V: Total volume (mL) of the solution (the sample stock solution) prepared as containing about 100 anti-factor Xa activity Units per mL M: Amount (mg) of Heparin Calcium taken for the sam-ple stock solution However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?Xs ＋ B?Xt ＋ D, where D is a constant term showing the difference between the blank and the intercept assumed from the two lines, is not in the range of between －0.2 and 0.2, analyze by excluding the measurements of the blank. The criteria for the test suitability are performed as di-rected in the Assay (1). When these criteria are not satisfied, repeat the test after changing the dilution rate so that the po-tency ratio becomes about 1 using the obtained potency as reference.
(1) Heparin Substrate solution: Dissolve 25 mg of H-D-pheny-lalanyl-L-pipecolyl-L-arginyl-p-nitroanilide dihydrochloride in 32.0 mL of water. Anti-thrombin solution (for heparin assay): Dissolve human anti-thrombin in water so that each mL contains 1 IU. Dilute this solution to an appropriate dilution factor of approximately more than 16 times with the buffer solution, and designate this solution as the anti-thrombin solution (for heparin assay). The dilution factor with the buffer solution is adjusted so that the absorbance of reaction solution with the blank solution (average of five tubes) is not more than 2.0, and that of reaction solution with S4 (0.020 Unit/mL heparin standard solution) (average of two tubes) is not less than 0.2 and not more than 1.0 when the test is performed according to the Assay. The absorbance is measured with 1 cm light path in length. Factor IIa solution: Add an equivalent volume of water to the buffer solution, and use this solution as the fac-tor IIa diluent. Dissolve factor IIa in the factor IIa diluent to make a solution so that each mL contains 20 IU. Dilute this solution to an appropriate dilution factor of approximately less than 4 times with the factor IIa diluent, and designate this solution as the factor IIa solution. Adjust the dilution factor with the factor IIa diluent so that the absorbance of reaction solution with the blank solution (average of five tubes) is not more than 2.0, and that of reaction solution with S4 (0.020 Unit/mL heparin standard solution) (average of two tubes) is not less than 0.2 and not more than 1.0 when the test is performed according to the Assay. The absorbance is measured with 1 cm light path in length. Buffer solution: Dissolve 6.1 g of 2-amino-2-hydrox-ymethyl-1,3-propanediol, 10.2 g of sodium chloride, 2.8 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate and 1.0 g of polyethylene glycol 6000 in 800 mL of water, adjust to pH 8.4 with 1 mol/L hydrochloric acid TS, and add water to make 1000 mL. Stopping solution: To 2 mL of acetic acid (100) add water to make 10 mL. Heparin standard solutions: Dissolve Heparin So-dium RS in water so that each mL contains 100 Heparin Units, and use this solution as the standard stock solution. Dilute the standard stock solution with buffer solution so that each mL contains exactly 0.1 Heparin Units, and use this solution as the standard solution. Make heparin stand-ard solutions S1, S2, S3 and S4 respectively by adding the standard solution to buffer solution as directed in the fol-lowing table. Heparin standard solution Buffer solution (mL) Standard solution (mL) No. Heparin concentration (Unit/mL) S1 0.005 S2 0.010 S3 0.015 S4 0.020 Heparin sample solutions: Weigh accurately an ap-propriate amount of Heparin Calcium, dissolve in water so that each mL contains about 100 Heparin Units, and use this solution as the sample stock solution. Dilute exactly the sample stock solution with buffer solution so that each mL contains 0.1 Heparin Units, and use this solution as the sam-ple solution. Make heparin sample solutions T1, T2, T3 and T4 respectively by adding the sample solution to buffer solution as directed in the following table. JP XVIII Official Monographs / Heparin Calcium 1099 Heparin sample solution Buffer solution (mL) Sample solution (mL) No. Heparin concentration (Unit/mL) T1 0.005 T2 0.010 T3 0.015 T4 0.020 Procedure: Transfer separately two 50-mL portions of each dilution of the heparin standard solutions and the heparin sample solutions and five 50-mL portions of buffer solution as the blank to 1.5-mL tubes. Warm these 21 tubes, anti-thrombin solution (for heparin assay), factor IIa solu-tion and substrate solution at 379C all together. Start the fol-lowing procedure at 2 minutes after warming in the order: buffer solution, S , S , S , S , buffer solution, T , T , T , T , from the two lines When the regression expression, y ＝ Is ＋ A!Xs ＋ B!Xt ＋ It-s, is obtained from the data of the heparin standard solu-tions and the heparin sample solutions except of the blank solution, a 90z confidence interval of the constant term, It-s, is between －0.2 and 0.2. Is: Intercept of the regression expression of the heparin standard solution It-s: Difference of the intercepts assumed from the two lines (2) Judgment on linearity When the regression expression, y ＝ Ic ＋ A?X?s ＋ B?X?t ＋ QsXs2 ＋ QtXt2, is obtained from the data of the heparin standard solutions and the heparin sample solutions, a 90z confidence interval of the secondary coefficients, Qs and Qt, is between －1000 and 1000. Qs: Secondary coefficient of the regression expression of the heparin standard solution Qt: Secondary coefficient of the regression expression of 1 2 3 4 1 2 3 4 the heparin sample solution buffer solution, T1, T2, T3, T4, buffer solution, S1, S2, S3, S4, and buffer solution. To each tube add 100 mL of anti-throm-bin solution (for heparin assay), mix, and warm at 379C for exactly 4 minutes, add 25 mL of factor IIa solution, mix, and incubate for exactly 4 minutes. Then, add 50 mL of substrate solution, mix, incubate for exactly 4 minutes, add 50 mL of stopping solution to each tube, and mix. Separately, to 50 mL of stopping solution add 50 mL of substrate solution, 25 mL of factor IIa solution, 100 mL of anti-thrombin solution (for heparin assay) and 50 mL of buffer solution, mix, and use this solution as a control. Determine the absorbance of each solution at 405 nm against the control. Confirm that the relative standard deviation of the reading of the blank is not more than 10z. Calculations: When the regression expression, y ＝ Ic ＋ AXs ＋ BXt, is obtained using y as log of the absorbance values, Xs as the concentration of the heparin standard solutions and Xt as the concentration of the heparin sample solutions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate Heparin Unit (anti-factor IIa activity) per mg of Heparin Calcium by the following formula. Heparin Unit (anti-factor IIa activity) per mg of Heparin Calcium ＝ 100 × R × V/M V: Total volume (mL) of the solution (the sample stock solution) prepared as containing about 100 Heparin Units (anti-factor IIa activity) per mL M: Amount (mg) of Heparin Calcium taken for the sam-ple stock solution However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?Xs ＋ B?Xt ＋ D, where D is a constant term showing the difference between the blank and the intercept assumed from the two lines, is not in the range of between －0.2 and 0.2, analyze by excluding the measurements of the blank. The criteria for the test suitability are the following 3 items, (1), (2) and (3). (1) Judgment on consistence of the intercept assumed (3) Judgment by checking if the relative potency ob-tained is within the range previously validated on this test method The potency ratio obtained is not less than 0.8 and not more than 1.2. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the potency ratio becomes about 1 using the obtained potency as reference. (2) Calcium: Weigh accurately about 50 mg of Heparin Calcium, dissolve in 20 mL of water, add 2 mL of 8 mol/L potassium hydroxide TS, allow to stand for 3 to 5 minutes with occasional shaking, add 0.1 g of NN indicator, and im-mediately titrate <2.50> with 0.01 mol/L disodium dihydro-gen ethylenediamine tetraacetate VS until the color of the so-lution changes from red-purple to blue. Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 0.4008 mg of Ca
Heparin Sodium is a sodium salt of sulfated glycosaminoglycans composed of disaccharide units of D-glucosamine and uronic acid (L-iduronic acid or D- glucuronic acid) obtained from the intestinal mucosa of healthy edible swine. It contains not less than 180 Heparin Units (anti-factor IIa activity) per mg, calculated on the dried basis.
Heparin Sodium occurs as a white to grayish brown, powder or grains. It is odorless. It is soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic.
Dissolve 1 mg each of Heparin Sodium and Heparin Sodium for Identification RS in 1 mL of water, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: the retention time for the major peak from the sample solution and standard solution is identical. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase and flow rate: Proceed as di-rected under the operating conditions in Purity (7). System suitability— System performance: Dissolve 1.0 mg of Heparin Sodium for Identification RS in 0.60 mL of water. Dissolve 0.10 mg of Over-sulfated Chondroitin Sulfate for System Suitability RS in 0.20 mL of water. Dissolve 1.0 mg of dermatan sulfate in 2.0 mL of water. To 90 mL of the solution of Heparin So-dium for Identification RS add 30 mL each of the solutions of Over-sulfated Chondroitin Sulfate for System Suitability RS and dermatan sulfate, and mix. When the procedure is run with 20 mL of the mixture under the above operating conditions, dermatan sulfate, heparin and over-sulfated chondroitin sulfate are eluted in this order with the resolu-tion between the peaks of dermatan sulfate and heparin being not less than 1.0 and that between the peaks of heparin and over-sulfated chondroitin sulfate being not less than 1.5. pH <2.54> The pH of a solution of 1.0 g of Heparin So-dium in 100 mL of water is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Heparin Sodium in 20 mL of water: the solution is clear and colorless to light yellow. Barium—Dissolve 30 mg of Heparin Sodium in 3.0 mL of water, and use this solution as the sample solution. To 1.0 mL of the sample solution add 3 drops of dilute sul-furic acid, and allow to stand for 10 minutes: no turbidity is produced. Total nitrogen—Weigh accurately about 0.1 g of Heparin Sodium, previously dried at 609C for 3 hours under reduced pressure, and perform the test as directed under Nitrogen Determination <1.08>: the amount of nitrogen (N: 14.01) is not more than 3.0z. Protein—(i) Sodium carbonate solution: To 4 volumes of a mixture of sodium hydroxide solution (1 in 100) and anhydrous sodium carbonate solution (1 in 20) (1:1) add 1 volume of water. Copper sulfate solution: To 4 volumes of a mixture of copper (II) sulfate pentahydrate solution (1 in 80) and sodium tartrate dihydrate solution (149 in 5000) (1:1) add 1 volume of water. Alkaline copper solution for heparin: Mix 50 volumes of the sodium carbonate solution and 1 volume of the copper sulfate solution. Prepare before using. Procedure: Use a solution of Heparin Sodium (1 in 200) as the sample solution. Use a solution of bovine serum albumin (1 in 40,000) as the standard solution. To exactly 1 mL each of the sample solution and standard solution add exactly 5 mL of the alkaline copper solution for heparin, mix, and allow them to stand at room temperature for 10 minutes. To each of these solutions add exactly 0.5 mL of diluted Folin's TS (1 in 2), shake, and allow them to stand at room temperature for 30 minutes. Determine the absor-bances at 750 nm of these solutions as directed under Ultra-violet-visible Spectrophotometry <2.24> using water as the blank: the absorbance of the solution obtained from the sample solution is not more than that of the solution from the standard solution. Nucleic acid—Dissolve 40 mg of Heparin Sodium in 10 mL of water, and determine the absorbance of this solu-tion at 260 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>: the absorbance is not more than 0.15. Over-sulfated chondroitin sulfate—Dissolve 20 mg of Heparin Sodium in 0.60 mL of a solution of sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance spectroscopy (1 in 10,000). Determine the spectrum of this solution as directed under Nuclear Magnetic Resonance Spectroscopy <2.21> (1H) in accordance with the following conditions, using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an internal ref-erence compound: it exhibits no signal corresponding to N-acetyl proton of over-sulfated chondroitin sulfate at d 2.15 ± 0.02 ppm, or the signal disappears when determining the spectrum of the sample solutions as directed under 1H with 13C-decoupling. Operating conditions— Spectrometer: 1.1. FT-NMR, Not less than 400 MHz. Temperature: 259C. Spinning: off. Number of data points: 32,768. Spectral range: Signal of DHO ± 6.0 ppm. Flip angle: 909. Delay time: 20 seconds. Dummy scans: 4. Number of scans: SN ratio of the signal of N-acetyl pro-ton of heparin is not less 1000. JP XVIII Official Monographs / Heparin Sodium 1101 Window function: Exponential function (Line broadening factor ＝ 0.2 Hz). System suitability— System performance: Dissolve 20 mg of Heparin Sodium for Identification RS in 0.40 mL of a solution of sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance spectroscopy (1 in 10,000). Dissolve 0.10 mg of Over-sulfat-ed Chondroitin Sulfate for System Suitability RS in 1.0 mL of a solution of sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy in heavy water for nuclear magnetic resonance spectroscopy (1 in 10,000). To the solution of Heparin Sodium for Identification RS add 0.20 mL of the solution of Over-sulfated Chondroitin Sul-fate for System Suitability RS. When determining the spec-trum of this solution under the above operating conditions, it exhibits the signal of N-acetyl proton of heparin and the signal of N-acetyl proton of over-sulfated chondroitin sul-fate at d 2.04 ± 0.02 ppm and d 2.15 ± 0.02 ppm, respec-tively. Related substances—Dissolve 2.0 mg of Heparin So-dium in 0.1 mL of water and perform the test with exactly 20 mL of this solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions: it exhibits no peaks after the heparin peak. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 202 nm). Column: A stainless steel column 2.0 mm in inside diame-ter and 7.5 cm in length, packed with synthetic polymer for liquid chromatography to which diethylaminoethyl group binds (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Dissolve 0.4 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (1 in 10). Mobile phase B: Dissolve 0.4 g of sodium dihydrogen phosphate dihydrate and 106.4 g of lithium perchlorate in 1000 mL of water, and adjust to a pH of 3.0 with diluted phosphoric acid (1 in 10). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. System performance: To 120 mL of the heparin sodium standard stock solution add 30 mL of the over-sulfated chon-droitin sulfate standard solution, mix and use this solution as the solution for system suitability test. When the proce-dure is run with 20 mL of the solution for system suitability test under the above operating conditions, heparin and over-sulfated chondroitin sulfate are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of over-sulfated chondroitin sulfate is not more than 2.0z. Galactosamine—Dissolve 2.4 mg of Heparin Sodium in 1.0 mL of a mixture of water and hydrochloric acid (7:5), and use this solution as the heparin sodium stock solution. Dissolve 8.0 mg of D-glucosamine hydrochloride in a mixture of water and hydrochloric acid (7:5) to make exactly 10 mL. Dissolve 8.0 mg of D-galactosamine hydrochloride in a mix-ture of water and hydrochloric acid (7:5) to make exactly 10 mL. To 99 volumes of the solution of D-glucosamine add 1 volume of the solution of D-galactosamine, and use this solu-tion as the standard stock solution. Transfer 500 mL each of the heparin sodium stock solution and the standard stock solution to a glass-stoppered test tube, stopper tightly, and heat at 1009C for 6 hours. After cooling to room tempera-ture, evaporate 100 mL each of the reaction solutions to dry-ness. Add 50 mL of methanol to each of the residues and evaporate to dryness at room temperature. Dissolve each of the residues in 10 mL of water, add 40 mL of aminobenzoate derivatization TS, and heat at 809C for 1 hour. After cooling to room temperature, evaporate the reaction solutions to dryness. Add 200 mL of each of water and ethyl acetate to each of the residues, shake vigorously, and then centrifuge. After remove the upper layers, add 200 mL of ethyl acetate to each of the lower layers, shake vigorously, and then centri-fuge. These lower layers are used as the sample solution and the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions: the peak area ratio of galactosamine to glucosamine of the sample solution is not larger than that of the standard solution. Operating conditions— Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Detector: A fluorescence photometer (excitation wave-length: 305 nm; emission wavelength: 360 nm). Column: A stainless steel column 4.6 mm in inside diame- 0 – 3 90 10 3 – 15 90 → 0 10 → 100 Flow rate: 0.2 mL per minute. Time span of measurement: About 2 times as long as the retention time of heparin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 10 mg of Heparin Sodium for Identification RS in 0.40 mL of water, and use this solution as the heparin sodium standard stock solution. Separately, dissolve 0.10 mg of Over-sulfated Chondroitin Sulfate for System Suitability RS in 0.20 mL of water, and use this solution as the over-sulfated chondroitin sulfate standard solution. To 60 mL of the heparin sodium standard stock solution add 3 mL of the over-sulfated chondroitin sul-fate standard solution and 12 mL of water, and mix. When the procedure is run with 20 mL of the mixture under the above operating conditions, it exhibits a peak for over-sul-fated chondroitin sulfate. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: To 100 mL of a mixture of water and trifluoroacetic acid (1000:1) add 100 mL of acetonitrile. Add 140 mL of the solution to 860 mL of a mixture of water and trifluoroacetic acid (1000:1). Flow rate: 1.0 mL per minute. Time span of measurement: About 50 minutes after inject-ed. System suitability— Test for required detectability: Dissolve 8.0 mg of D-man-nosamine hydrochloride in 10 mL of a mixture of water and hydrochloric acid (7:5), and use this solution as the man-nosamine standard solution. Transfer 500 mL of a mixture of the standard stock solution and the mannosamine standard solution (100:1) to a glass-stoppered test tube, stopper tight-ly, and heat at 1009C for 6 hours. After cooling this solution to room temperature, evaporate 100 mL of the reaction solu- tion to dryness. Add 50 mL of methanol to the residue and evaporate to dryness at room temperature. Dissolve the residue in 10 mL of water, add 40 mL of aminobenzoate derivatization TS, and heat at 809C for 1 hour. After cooling to room temperature, evaporate the reaction solution to dry-ness. Add 200 mL each of water and ethyl acetate to the residue, shake vigorously, and then centrifuge. After remov-ing the upper layer, add 200 mL of ethyl acetate to the lower layer, shake vigorously, and then centrifuge. The lower layer is used as the solution for system suitability test. When the procedure is run with 5 mL of the solution for system suitability test under the above operating conditions, the ratio of the peak area of galactosamine to that of glucosa-mine is 0.7 – 2.0z. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above operating conditions, glucosamine, mannosamine and galac-tosamine are eluted in this order with the resolutions be-tween the peaks of glucosamine and mannosamine and be-tween the peaks of mannosamine and galactosamine being not less than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the ratio of the peak area of galactosamine to that of glucosamine is not more than 4.0z. Loss on drying <2.41> Not more than 10z (20 mg, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 40z (after dry-ing, 20 mg). Bacterial endotoxins <4.01> Less than 0.0030 EU/Heparin Unit. Anti-factor Xa activity to anti-factor IIa activity ratio The ratio of the anti-factor Xa activity determined by the follow-ing method to the anti-factor IIa activity obtained in the Assay, calculated by dividing the former with the later, is 0.9 – 1.1. Anti-factor Xa activity determination Substrate solution: Dissolve 25 mg of N-benzoyl-L-isoleucyl-L-glutamyl(g-OR)-glycyl-L-arginyl-p-nitroanilide hydrochloride in 33.3 mL of water. Anti-thrombin solution: Dissolve human anti-throm-bin in water so that each mL contains 1 IU. To 150 mL of this solution add 2250 mL of buffer solution Factor Xa solution: To 1200 mL of factor Xa TS add 1200 mL of buffer solution. Buffer solution: Proceed as directed in the Assay. Stopping solution: Proceed as directed in the Assay. Heparin standard solutions: Proceed as directed in the Assay. However, the standard solutions are prepared based on anti-factor Xa activity Unit instead of Heparin Unit. Heparin sample solutions: Proceed as directed in the Assay. However, the sample solutions are prepared based on anti-factor Xa activity Unit instead of Heparin Unit. Procedure: Transfer separately two 50-mL portions of each dilution of the heparin standard solutions and the heparin sample solutions and five 50-mL portions of buffer solution as the blank to 1.5 mL-tubes. Warm these 21 tubes, anti-thrombin solution, factor Xa solution and substrate so-lution at 379C all together. Start the following procedure at 2 minutes after warming in the order: buffer solution, S1, S2, S3, S4, buffer solution, T1, T2, T3, T4, buffer solution, T1, T2, T3, T4, buffer solution, S1, S2, S3, S4, and buffer solu- tion. To each tube add 50 mL of anti-thrombin solution, mix, and warm at 379C for exactly 4 minutes, add 100 mL of fac-tor Xa solution, mix, and incubate for exactly 12 minutes. Then, add 100 mL of substrate solution, mix, incubate for exactly 4 minutes, add 50 mL of stopping solution to each tube, and mix immediately. Separately, to 50 mL of stopping solution add 100 mL of substrate solution, 100 mL of factor Xa solution, 50 mL of anti-thrombin solution and 50 mL of buffer solution, mix, and use this solution as a control. De-termine the absorbance of each solution at 405 nm against the control. Confirm that the relative standard deviation of the reading of the blank is not more than 10z. Calculations: When the regression expression, y ＝ Ic ＋ AXs ＋ BXt, is obtained using y as log of the absorbance values, Xs as the concentration of the heparin standard solutions and Xt as the concentration of the heparin sample solutions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate anti-factor Xa activity per mg of Heparin Sodium by the following formula. Anti-factor Xa activity per mg of Heparin Sodium ＝ 100 × R × V/M V: Total volume (mL) of the solution (the sample stock solution) prepared as containing about 100 anti-factor Xa activity Units per mL M: Amount (mg) of Heparin Sodium taken for the sample stock solution However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?Xs ＋ B?Xt ＋ D, where D is a constant term showing the difference between the blank and the intercept assumed from the two lines, is not in a range of between －0.2 and 0.2, analyze by excluding the measurements of the blank. The criteria for the test suitability are performed as di-rected in the Assay. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the po-tency ratio becomes about 1 using the obtained potency as reference.
Substrate solution: Dissolve 25 mg of H-D-phenylalanyl-L-pipecolyl-L-arginyl-p-nitroanilide dihydro-chloride in 32.0 mL of water. Anti-thrombin solution (for heparin assay): Dissolve human anti-thrombin in water so that each mL contains 1 IU. Dilute this solution to an appropriate dilution factor of approximately more than 16 times with the buffer solution, and designate this solution as the anti-thrombin solution (for heparin assay). The dilution factor with the buffer solution is adjusted so that the absorbance of reaction solution with the blank solution (average of five tubes) is not more than 2.0, and that of reaction solution with S4 (0.020 Unit/mL heparin standard solution) (average of two tubes) is not less than 0.2 and not more than 1.0 when the test is performed according to the Assay. The absorbance is measured with 1 cm light path in length. Factor IIa solution: Add an equivalent volume of water to the buffer solution, and use this solution as the fac-tor IIa diluent. Dissolve factor IIa in the factor IIa diluent to make a solution so that each mL contains 20 IU. Dilute this solution to an appropriate dilution factor of approximately JP XVIII Official Monographs / Heparin Sodium 1103 less than 4 times with the factor IIa diluent, and designate this solution as the factor IIa solution. Adjust the dilution factor with the factor IIa diluent so that the absorbance of reaction solution with the blank solution (average of five tubes) is not more than 2.0, and that of reaction solution with S4 (0.020 Unit/mL heparin standard solution) (average of two tubes) is not less than 0.2 and not more than 1.0 when the test is performed according to the Assay. The absorbance is measured with 1 cm light path in length. Buffer solution: Dissolve 6.1 g of 2-amino-2-hydrox-ymethyl-1,3-propanediol, 10.2 g of sodium chloride, 2.8 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate and 1.0 g of polyethylene glycol 6000 in 800 mL of water, adjust to pH 8.4 with 1 mol/L hydrochloric acid TS, and add water to make 1000 mL. Stopping solution: To 2 mL of acetic acid (100) add water to make 10 mL. Heparin standard solutions: Dissolve Heparin So-dium RS in water so that each mL contains 100 Heparin Units, and use this solution as the standard stock solution. Dilute the standard stock solution with buffer solution so that each mL contains exactly 0.1 Heparin Units, and use this solution as the standard solution. Make heparin stand-ard solutions S1, S2, S3 and S4 respectively by adding the standard solution to buffer solution as directed in the fol-lowing table. Heparin standard solution Buffer solution (mL) Standard solution (mL) No. Heparin concentration (Unit/mL) S1 0.005 S2 0.010 S3 0.015 S4 0.020 Heparin sample solutions: Weigh accurately an ap-propriate amount of Heparin Sodium, dissolve in water so that each mL contains about 100 Heparin Units, and use this solution as the sample stock solution. Dilute exactly the sam-ple stock solution with buffer solution so that each mL con-tains 0.1 Heparin Units, and use this solution as the sample solution. Make heparin sample solutions T1, T2, T3 and T4 respectively by adding the sample solution to buffer solution as directed in the following table. Heparin sample solution Buffer solution (mL) Sample solution (mL) No. Heparin concentration (Unit/mL) T1 0.005 T2 0.010 T3 0.015 T4 0.020 Procedure: Transfer separately two 50-mL portions of each dilution of the heparin standard solutions and the heparin sample solutions and five 50-mL portions of buffer solution as the blank to 1.5 mL-tubes. Warm these 21 tubes, anti-thrombin solution (for heparin assay), factor IIa solu- tion and substrate solution at 379C all together. Start the fol-lowing procedure at 2 minutes after warming in the order: buffer solution, S1, S2, S3, S4, buffer solution, T1, T2, T3, T4, buffer solution, T1, T2, T3, T4, buffer solution, S1, S2, S3, S4, and buffer solution. To each tube add 100 mL of anti-throm-bin solution (for heparin assay), mix, and warm at 379C for exactly 4 minutes, add 25 mL of factor IIa solution, mix, and incubate for exactly 4 minutes. Then, add 50 mL of substrate solution, mix, incubate for exactly 4 minutes, add 50 mL of stopping solution to each tube, and mix. Separately, to 50 mL of stopping solution add 50 mL of substrate solution, 25 mL of factor IIa solution, 100 mL of anti-thrombin solution (for heparin assay) and 50 mL of buffer solution, mix, and use this solution as a control. Determine the absorbance of each solution at 405 nm against the control. Confirm that the relative standard deviation of the reading of the blank is not more than 10z. Calculations: When the regression expression, y ＝ Ic ＋ AXs ＋ BXt, is obtained using y as log of the absorbance values, Xs as the concentration of the heparin standard solutions and Xt as the concentration of the heparin sample solutions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate Heparin Unit (anti-factor IIa activity) per mg of Heparin Sodium by the following formula. Heparin Unit (anti-factor IIa activity) per mg of Heparin So-dium ＝ 100 × R × V/M V: Total volume (mL) of the solution (the sample stock solution) prepared as containing about 100 Heparin Units (anti-factor IIa activity) per mL M: Amount (mg) of Heparin Sodium taken for the sample stock solution However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?Xs ＋ B?Xt ＋ D, where D is a constant term showing the difference between the blank and the intercept assumed from the two lines, is not in the range of between －0.2 and 0.2, analyze by excluding the measurements of the blank. The criteria for the test suitability are the following 3 items, (1), (2) and (3). Judgment on consistence of the intercept assumed from the two lines When the regression expression, y ＝ Is ＋ A!Xs ＋ B!Xt ＋ It-s, is obtained from the data of the heparin standard solu-tion and the heparin sample solution except of the blank so-lution, a 90z confidence interval of the constant term, It-s, is between －0.2 and 0.2. Is: Intercept of the regression expression of the heparin standard solution It-s: Difference of the intercepts assumed from the two lines Judgment on linearity When the regression expression, y ＝ Ic ＋ A?X?s ＋ B?X?t ＋ QsXs2 ＋ QtXt2, is obtained from the data of the heparin standard solution and the heparin sample solution, a 90z confidence interval of the secondary coefficients, Qs and Qt, is between －1000 and 1000. ‌1104 Qs: Secondary coefficient of the regression expression of the heparin standard solution Qt: Secondary coefficient of the regression expression of the heparin sample solution Judgment by checking if the relative potency ob-tained is within the range previously validated on this test method ‌The potency ratio obtained is not less than 0.8 and not more than 1.2. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the potency ratio becomes about 1 using the obtained potency as reference.
Human Normal Immunoglobulin is a clear, colorless or yellow-brown liquid. Hydralazine Hydrochloride
Weigh accurately about 0.15 g of Hydralazine Hy-drochloride, previously dried, transfer it to a glass-stoppered flask, dissolve in 25 mL of water, add 25 mL of hydrochloric acid, cool to room temperature, add 5 mL of chloroform, and titrate <2.50> with 0.05 mol/L potassium iodate VS while shaking until the purple color of the chloroform layer disappears. The end point is reached when the red-purple color no more reappears in the chloroform layer within 5 minutes after the layer has been decolorized. Each mL of 0.05 mol/L potassium iodate VS ＝ 9.832 mg of C H N .HCl 8 8 4 Description Human Normal Immunoglobulin is a clear, colorless or yellow-brown liquid. Hydralazine Hydrochloride
Influenza HA Vaccine is a liquid for injection con-taining hemagglutinin of influenza virus. It conforms to the requirements of Influenza HA Vaccine in the Minimum Requirements for Biological Products. Description Influenza HA Vaccine is a clear liquid or a slightly whitish turbid liquid. A chain C99H153N25O35S4: 2381.68 B chain C158H234N40O42S2: 3429.92 [11061-68-0] Insulin Human (Genetical Recombination) is a recombinant human insulin. It is a peptide composed of A chain consisting of 21 amino acid residues and B chain consisting of 30 amino acid residues. It contains not less than 27.
Influenza HA Vaccine is a clear liquid or a slightly whitish turbid liquid. A chain C99H153N25O35S4: 2381.68 B chain C158H234N40O42S2: 3429.92 [11061-68-0] Insulin Human (Genetical Recombination) is a recombinant human insulin. It is a peptide composed of A chain consisting of 21 amino acid residues and B chain consisting of 30 amino acid residues. It contains not less than 27.5 Insulin Units per mg, calculated on the dried basis. Description Insulin Human (Genetical Recombination) occurs as a white powder. It is practically insoluble in water and in ethanol (95). It dissolves in 0.01 mol/L hydrochloric acid TS and in sodium hydroxide TS with decomposition. It is hygroscopic.
Weigh accurately a suitable amount of Insu-lin Human (Genetical Recombination), and dissolve in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains 2.0 mg. Transfer 500 mL of this solution into a clean test tube, add 2.0 mL of HEPES buffer solution (pH 7.5) and 400 mL of V8-protease TS, incubate at 259C for 6 hours, then add 2.9 mL of ammonium sulfate buffer solu-tion to stop the reaction, and use this solution as the sample solution. Separately, proceed with Insulin Human RS in the same manner as above, and use this solution as the standard solution. Perform the test with 50 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and compare the chromatograms obtained from these solu-tions: a similar peak is observed at the same retention time in the both chromatograms. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, ammonium sulfate buffer solution and acetonitrile (7:2:1). Mobile phase B: A mixture of water, acetonitrile and am-monium sulfate buffer solution (2:2:1). Flowing of mobile phase: Change the mixing ratio of the mobile phase A and B linearly from 9:1 to 3:7 in 60 minutes after sample injection, further change to 0:10 linearly in 5 minutes, and then flow the mobile phase B only for 5 minutes. Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con- ditions, the symmetry factor of the two larger peaks which appear next to the first peak just after the solvent peak are not more than 1.5 respectively, and the resolution between these peaks is not less than 3.4.
(1) Related substances—Perform this procedure rapidly. Dissolve 7.5 mg of Insulin Human (Genetical Recombination) in 2 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak area of human insulin, A1, the peak area of the desamido substance having the relative retention time of about 1.3 to human insulin, AD, and the total area of the peaks other than the solvent peak, AT: the amounts of the desamido substance and related substances other than the desamido substance are not more than 2.0z, respectively. Previously, perform the test with 0.01 mol/L hydrochloric acid TS in the same manner to confirm the sol-vent peak. Amount (z) of the desamido substance ＝ AD/AT × 100 Amount (z) of related substances other than the desamido substance ＝ [{AT － (A1 ＋ AD)}/AT] × 100 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of phosphoric acid-sodium sulfate buffer solution (pH 2.3) and acetonitrile for liquid chromatography (41:9). Mobile phase B: A mixture of phosphoric acid-sodium sul-fate buffer solution (pH 2.3) and acetonitrile for liquid chro-matography (1:1). Flowing of mobile phase: Flow a mixture of the mobile phase A and B (78:22) for 36 minutes before and after the sample injection, then change the mixing ratio to 33:67 linearly in 25 minutes, and maintain this ratio for 6 minutes. Then flow the first mixture (78:22) for the next 15 minutes. Adjust the mixing ratio of the first mixture so that the reten-tion time of human insulin is about 25 minutes. Flow rate: 1.0 mL per minute. Time span of measurement: For about 75 minutes after the sample is injected. System suitability— Test for required detectability: Confirm that the peak height of the desamido substance obtained with 20 mL of human insulin desamido substance-containing TS is between 30z and 70z of the full scale. System performance: When the procedure is run with 20 mL of human insulin desamido substance-containing TS under the above operating conditions, human insulin and human insulin desamido substance are eluted in this order with the resolution between these peaks being not less than 2.0, and the symmetry factor of the peak of human insulin is not more than 1.8. High-molecular mass proteins—Dissolve 4 mg of In-sulin Human (Genetical Recombination) in 1 mL of 0.01 mol/L hydrochloric acid TS. Perform the test with 100 mL of this solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate each peak area: the total of areas of the peaks having smaller retention time than human insulin is not more than 1.0z of the total area of all peaks. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 276 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of L-arginine (1 in 1000), acetonitrile and acetic acid (100) (13:4:3). Flow rate: Adjust so that the retention time of human in-sulin is about 20 minutes. Time span of measurement: Until the peak of human insu-lin monomer has appeared. System suitability— Test for required detectability: Confirm that the peak height of the dimer obtained with 100 mL of human insulin dimer-containing TS is between 10z and 50z of the full scale. System performance: When the procedure is run with 100 mL of human insulin dimer-containing TS under the above operating conditions, polymer, dimer and monomer are eluted in this order, and the ratio, H1/H2, of the peak height of the dimer H1 to the height of the bottom between the peaks of the dimer and the monomer H2 is not less than 2.0. Product related impurities—Being specified sepa-rately when the drug is granted approval based on the Law. Process related impurities—Being specified sepa-rately when the drug is granted approval based on the Law. Loss on drying <2.41> Not more than 10.0z (0.2 g, 1059C, 24 hours). Bacterial endotoxins <4.01> Less than 10 EU/mg. Zinc content Weigh accurately about 50 mg of Insulin Hu-man (Genetical Recombination), and dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL. If necessary, dilute with 0.01 mol/L hydrochloric acid TS to make a solu-tion so that each mL contains between 0.4 mg and 1.6 mg of zinc (Zn: 65.38), and use this solution as the sample solution. Separately, take exactly a suitable amount of Standard Zinc Solution for Atomic Absorption Spectrophotometry, dilute with 0.01 mol/L hydrochloric acid TS to make solutions containing 0.40 mg, 0.80 mg, 1.20 mg and 1.60 mg of zinc (Zn: 65.38) in each mL, respectively, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23>, and calculate the amount of zinc (Zn: 65.38) in the sample solution by using a calibration curve obtained from the absorbances of the standard solutions: not more than 1.0z, calculated on the dried basis. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm.
Perform this procedure quickly. Weigh accurately about 7.5 mg of Insulin Human (Genetical Recombination), dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 5 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately a suitable amount of Insulin Human RS, dissolve exactly in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains about 40 Insulin Units, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution JP XVIII Official Monographs / Insulin Human (Genetical Recombination) Injection 1157 and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas of human insulin, ATI and ASI, and the peak areas of the desamido substance having the relative retention time of 1.3 to human insulin, ATD and ASD, respec-tively, of these solutions. ‌Amount (Insulin Unit/mg) of human insulin (C257H383N65O77S6) ＝ (MS × F )/D × (ATI ＋ ATD)/(ASI ＋ ASD) × 5/MT F: Labeled unit (Insulin Unit/mg) of Insulin Human RS D: Volume (mL) of 0.01 mol/L hydrochloric acid TS used to dissolve the reference standard MT: Amount (mg) of Insulin Human (Genetical Recombi-nation) taken, calculated on the dried basis MS: Amount (mg) of Insulin Human RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphoric acid-sodium sul-fate buffer solution (pH 2.3) and acetonitrile for liquid chro-matography (3:1). Adjust the mixing ratio of the component of the mobile phase so that the retention time of human in-sulin is between 10 minutes and 17 minutes. Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of human insulin desamido substance-containing TS under the above operating conditions, human insulin and human insulin desamido substance are eluted in this order with the resolution between these peaks being not less than 2.0, and the symmetry factor of the peak of human insulin is not more than 1.8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of human insulin is not more than 1.6z.
Containers—Tight containers. Storage—Not exceeding －209C.
Insulin Aspart (Genetical Recombination)
インスリン アスパルト(遺伝子組換え)
Insulin Aspart (Genetical Recombination) oc-curs as a white powder. It is practically insoluble in water and in ethanol (95). It dissolves in 0.01 mol/L hydrochloric acid TS. It is hygroscopic.
Weigh a suitable amount of Insulin Aspart (Genetical Recombination), and dissolve in 0.01 mol/L hy-drochloric acid TS to make a solution so that each mL con-tains 2.0 mg. Separately, dissolve Insulin Aspart RS in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains 2.0 mg. Transfer 25 mL each of these solutions into clean test tubes, add 100 mL of HEPES buffer solution (pH 7.5) and 20 mL of V8-protease TS, and allow to react at 259C for 6 hours. Then add 145 mL of ammonium sulfate buffer solution to stop the reaction, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 50 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and compare the peak (peak 1) eluted just after the peak of the solvent and the succeeding three peaks (peaks 2, 3 and 4) with apparently higher peak height in the chromato-grams obtained from these solutions: the similar peaks are observed at the same retention times. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (not exceeding 5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, ammonium sulfate buffer solution and acetonitrile for liquid chromatography (7:2:1). Mobile phase B: A mixture of water, acetonitrile for liquid chromatography and ammonium sulfate buffer solution (2:2:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 60 90 → 30 10 → 70 60 – 65 30 → 0 70 → 100 65 – 70 Flow rate: 1 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con- ditions, the symmetry factors of the peaks 2 and 3 are not more than 1.5, respectively, and the resolution between these peaks is not less than 8.
(1) Related substances—Perform the test with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amounts of them by the area percentage method: the amount of the peak of B28isoAsp insulin aspart, having the relative retention time of about 0.9 to insulin aspart, is not more than 0.3z, the total amount of the peak of A21Asp insulin aspart and B3Asp insulin aspart, having the relative retention times of about 1.3, and the peak of B3isoAsp insulin aspart, having the relative retention time of about 1.5, is not more than 1.0z, and the total amount of the peaks other than the peaks mentioned above is not more than 0.5z. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase and flow rate: Pro-ceed as directed in the operating conditions in the Assay. Time span of measurement: From 4 minutes to 50 minutes after injection of the sample solution. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the solution for system suitability test obtained in the Assay, add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL. Con-firm that the area percentage of the peak of B28isoAsp insu-lin aspart obtained with 10 mL of this solution is equivalent to 80 to 120z of that with 10 mL of the solution for system suitability test. High-molecular proteins—Store the sample solution at a temperature between 29C and 89C, and use within 48 hours after preparation. Dissolve 4 mg of Insulin Aspart (Genetical Recombination) in 1 mL of 0.01 mol/L hydro-chloric acid TS, and use this solution as the sample solution. Perform the test with 100 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the total amount of the peaks other than insulin aspart is not more than 0.3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 276 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of a solution of L-arginine (1 in 1000), acetonitrile for liquid chromatography and acetic acid (100) (13:4:3). Flow rate: 0.5 mL per minute. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the completion of the elution of insulin aspart. System suitability— Test for required detectability: Allow Insulin Aspart (Genetic Recombination) to stand at ordinary temperature for about 10 days, which results in containing about 0.4z of high-molecular proteins, dissolve in 0.01 mol/L hydrochlo-ride TS so that each mL contains about 4 mg of insulin aspart, and use this solution as the solution for system ‌suitability test. Store the solution for system suitability test at a temperature between 29C and 89C, and use within 7 days. Pipet 5 mL of the solution for system suitability test, and add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL. Confirm that the area percentage of the peak of insulin aspart dimer obtained with 100 mL of this solution is equiva-lent to 80 to 120z of that with 100 mL of the solution for system suitability test. System performance: When the procedure is run with 100 mL of the solution for system suitability test under the above Mobile phase A: Dissolve 142.0 g of anhydrous sodium sulfate in water, add 13.5 mL of phosphoric acid, and add water to make 5 L. Adjust to pH 3.6 with sodium hydroxide TS. To 4500 mL of this solution add 500 mL of acetonitrile for liquid chromatography. Mobile phase B: A mixture of water and acetonitrile for liquid chromatography (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. operating conditions, insulin aspart polymer (retention time: 13 to 17 minutes), insulin aspart dimer (retention time: about 17.5 minutes) and insulin aspart (retention time: 18 to 20 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) minutes) are eluted in this order, and determine the peak height of the dimer and the height of the bottom between the peaks of the dimer and the monomer: the peak-valley ratio is not less than 2.0. System repeatability: When the test is repeated 6 times with 100 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of insulin aspart is not more than 2.0z. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Loss on drying <2.41> Not more than 10.0z (0.2 g, 1059C, 24 hours). Residue on ignition <2.44> Not more than 6.0z (0.2 g).
Store the sample solution and the standard solution at a temperature between 29C and 89C, use the sample solu-tion within 24 hours after preparation, and use the standard solution within 48 hours after preparation. Weigh accurately a suitable amount of Insulin Aspart (Genetical Recombina-tion), dissolve in 0.01 mol/L hydrochloric acid TS so that each mL contains 4.0 mg, and use this solution as the sample solution. Separately, dissolve Insulin Aspart RS in 0.01 mol/L hydrochloric acid TS so that each mL contains 4.0 mg, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the total areas, AT and AS, of the peak of B28isoAsp insulin aspart (relative retention time to insulin aspart: about 0.9), the peak of insulin aspart (retention time: 20 to 24 minutes), the peak of A21Asp insulin aspart and B3Asp insulin aspart (usually eluted together having the relative retention time of about 1.3) and the peak of B3isoAsp insulin aspart (relative retention time: about 1.5) in each solution. Amount (mg) of insulin aspart (C256H381N65O79S6) ＝ MS × AT/AS MS: Total amount (mg) of insulin aspart, B28isoAsp insu-lin aspart, A21Asp insulin aspart and B3Asp insulin aspart, and B3isoAsp insulin aspart in 1 mL of the standard solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (not exceeding 5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. 0 – 35 58 42 35 – 40 58 → 20 42 → 80 40 – 45 20 80 45 – 46 20 → 58 80 → 42 46 – 60 58 42 Flow rate: 1 mL per minute. System suitability— System performance: Dissolve Insulin Aspart (Genetical Recombination) in 0.01 mol/L sodium dihydrogen phos-phate TS (pH 7.5) so that each mL contains 8 mg, and allow to stand at ordinary temperature for 10 to 15 days. To 1 mL of this solution add 1 mL of 0.01 mol/L hydrochloric acid TS, allow to stand at ordinary temperature for 1 to 3 days, and use this solution as the solution for system suitability test. The solution for system suitability test contains 0.1 to 2.2z of B28isoAsp insulin aspart, and not less than 1z of B3Asp insulin aspart and A21Asp insulin aspart. Store the solution for system suitability test at a temperature between 29C and 89C, and use within 72 hours. When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, B28isoAsp insulin aspart, insulin aspart, A21Asp insulin aspart and B3Asp in-sulin aspart, and B3isoAsp insulin aspart are eluted in this order with the resolution between the peak of insulin aspart and the peak of A21Asp insulin aspart and B3Asp insulin aspart being not less than 2.0. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of AS is not more than 1.5z.
Containers—Tight containers. Storage—Not exceeding －189C.
Insulin Glargine (Genetical Recombination)
インスリン グラルギン（遺伝子組換え）
Insulin Glargine (Genetical Recombination) is a recombinant human insulin analogue, being substitut- JP XVIII Official Monographs / Insulin Glargine (Genetical Recombination) 1163 ed asparagine residue with glycine residue at 21st of A chain and added two arginine residues at C-terminal of B chain. It is a peptide composed with A chain consist-ing of 21 amino acid residues and B chain consisting of 32 amino acid residues. It contains not less than 94.
Insulin Glargine (Genetical Recombination) occurs as a white powder. It is practically insoluble in water and in ethanol (99.5). It is sparingly soluble in 0.01 mol/L hydrochloric acid TS. It is hygroscopic. It is gradually decomposed by light.
Keep the sample solution and standard solu-tion at 2 – 89C. Weigh a suitable amount of Insulin Glargine (Genetical Recombination) and Insulin Glargine RS, and dis-solve separately in 0.01 mol/L hydrochloric acid TS so that each mL contains 10.0 mg. Transfer 5 mL of these solutions into clean test tubes, add 1 mL of 1 mol/L tris buffer solu-tion (pH 7.5) and 100 mL of a solution of V8 protease for in-sulin glargine in 1 mol/L tris buffer solution (pH 7.5) (20 units/mL), allow to react at 35 – 379C for 3 hours, then add 2 mL of phosphoric acid to stop the reaction, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and compare the chromatograms obtained from these solutions: The similar peaks appear at the same reten-tion times. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 3 mm in inside diameter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: To 930 mL of a solution, prepared by dissolving 11.6 g of phosphoric acid and 42.1 g of sodium perchloric acid in 1600 mL of water, adjusting to pH 2.3 with triethylamine and adding water to make 2000 mL, add 70 mL of acetonitrile for liquid chromatography. Mobile phase B: To 430 mL of a solution, prepared by dis-solving 11.6 g of phosphoric acid and 42.1 g of sodium per-chloric acid in 1600 mL of water, adjusting to pH 2.3 with triethylamine and adding water to make 2000 mL, add 570 mL of acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as follows. appear next to the first peak just after the solvent peak, are not more than 1.5, respectively, and the resolution between these peaks is not less than 3.4.
(1) Related substances—Perform the test with 5 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amount of these peaks by the area percentage method: the amount of the peak other than insulin glargine is not more than 0.4z, and the total amount of the peaks other than insulin glargine is not more than 1.0z. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL. Confirm that the peak area of insulin glargine obtained with 5 mL of this solution is equivalent to 5 to 15z of that with 5 mL of the solution for system suitabil-ity test. System performance: When the procedure is run with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of insulin glargine are not less than 20,000 and not more than 1.8, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the relative standard deviation of the peak area of insulin glargine is not more than 2.0z. High-molecular mass proteins—Keep the sample so-lution at 2 – 89C. Dissolve 15 mg of Insulin Glargine (Genet-ical Recombination) in 1.5 mL of 0.01 mol/L hydrochloric acid TS, add water to make 10 mL, and use this solution as the sample solution. Perform the test with 100 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late the amount of them by the area percentage method: the total amount of the peaks other than insulin glargine is not more than 0.3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 276 nm). Column: Two stainless steel columns connected in series of 8 mm in inside diameter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography (5 mm in Time after injection 0 – 30 90 → 20 10 → 80 30 – 35 of sample (min) Mobile phase A (volz) Mobile phase B (volz) particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 400 mL of water add 300 mL of aceto-nitrile for liquid chromatography and 200 mL of acetic acid (100), adjust to pH 3.0 with ammonia solution (28), and add Flow rate: 0.55 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the symmetry factors of the two larger peaks, which water to make 1000 mL. Flow rate: Adjust so that the retention time of insulin glar-gine is about 35 minutes. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the completion of the elution of insulin glargine. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.01 mol/L hydrochloric acid TS to make 50 mL, the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of insulin glargine in each solution. and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add 0.01 mol/L hydrochloric acid TS to make exactly 10 Amount (mg) of insulin glargine (C ＝ MS × AT/AS H404N 72O78 S6) mL. Confirm that the peak area of insulin glargine obtained with 100 mL of this solution is equivalent to 5 to 15z of that with 100 mL of the solution for system suitability test. ‌System performance: Heat 15 mg of Insulin Glargine (Genetical Recombination) at 1009C for 1.5 – 3 hours, then dissolve in 1.5 mL of 0.01 mol/L hydrochloric acid TS, and add water to make exactly 10 mL. When the procedure is run with 100 mL of this solution under the above operating con-ditions, the high-molecular mass protein and insulin glargine are eluted in this order with the resolution between these peaks is not less than 1.5. System repeatability: When the test is repeated 6 times with 100 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of insulin glargine is not more than 2.0z. Other product-related impurities—Being specified separately when the drug is granted approval based on the Law. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 8.0z (90 mg, coulometric titration). MS: Amount (mg) of insulin glargine in 1 mL of the stand-ard solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 3 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Dissolve 20.7 g of anhydrous sodium di-hydrogen phosphate in 900 mL of water, adjust to pH 2.5 with phosphoric acid, and add water to make 1000 mL. To 250 mL of this solution add 250 mL of acetonitrile for liquid chromatography, dissolve 18.4 g of sodium chloride in this solution, and add water to make 1000 mL. Mobile phase B: Dissolve 20.7 g of anhydrous sodium di-hydrogen phosphate in 900 mL of water, adjust to pH 2.5 with phosphoric acid, and add water to make 1000 mL. To 250 mL of this solution add 650 mL of acetonitrile for liquid chromatography, dissolve 3.2 g of sodium chloride in this solution, and add water to make 1000 mL. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Bacterial endotoxins <4.01> Less than 10 EU/mg. Zinc content Weigh accurately about 45 mg of Insulin Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Glargine (Genetical Recombination), dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 10 mL of this solution, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, take exactly a suitable amount of Standard Zinc Solution for Atomic Absorption Spectrophotometry, dilute with 0.01 mol/L hydrochloric acid TS to make three solu-tions containing 0.20 mg, 0.40 mg and 0.60 mg of zinc (Zn: 65.38) in each mL, respectively, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and calculate the amount of zinc in the sample solution using a calibration curve obtained from the absorbances of the standard solutions: not more than 0.80z of zinc (Zn: 65.38), calculated on the anhydrous basis. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow-cathode lamp. Wavelength: 213.9 nm.
Keep the sample solution and standard solution at 2 – 89C. Weigh accurately about 15 mg of Insulin Glargine (Genetical Recombination), dissolve in 1.5 mL of 0.01 mol/L hydrochloric acid TS, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve Insulin Glargine RS in 0.01 mol/L hydrochloric acid TS so that each mL contains about 10 mg of insulin glargine, then exactly dilute with water so that each mL contains about 1.5 mg of insulin glargine, and use this solution as the standard solution. Perform the test with exactly 5 mL each of 0 – 20 96 → 83 4 → 17 20 – 30 83 → 63 17 → 37 30 – 40 63 → 96 37 → 4 Flow rate: 0.55 mL per minute (the retention time of insu-lin glargine is about 21 minutes). System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of insulin glargine are not less than 20,000 and not more than 1.8, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of insulin glargine is not more than 2.0z.
Containers—Tight containers. Storage—Not exceeding －159C.
Interferon Alfa (NAMALWA) is essentially a hu-man interferon alfa, which is a glycoprotein (molecu-lar mass: 17,000 – 30,000) produced by the human lymphoblast NAMALWA cell induced by Sendai vi-rus. It is an aqueous solution. It possesses the antiviral activity. It contains not less than 50 mg and not more than 500 mg of protein per mL, and not less than 1.
Interferon Alfa (NAMALWA) occurs as a clear and colorless liquid.
(1) To Interferon Alfa (NAMALWA) add Eagle's minimum essential medium containing bovine serum so that each mL contains 5000 Units, and use this solution as the sample stock solution. To anti-interferon alfa antiserum add an amount of Eagle's minimum essential medium con-taining bovine serum so that each mL contains an amount of anti-interferon alfa antiserum which neutralizes 10,000 Units of interferon alfa. To this solution add an equal volume of the sample stock solution, stir, and use this solution as the sample solution. Separately, to the sample stock solution add an equal volume of Eagle's minimum essential medium containing bovine serum, stir, and use this solution as the control solution. Determine the remained potency of the sample solution and control solution after allowing to stand at 37 ± 19C for 1 hour, according to the Assay. When the antiviral activity of Interferon Alfa (NAMALWA) is neu-tralized by anti-interferon alfa antiserum, it meets the requirement. Not detection of the remaining potency of the sample solution is a criterion of neutralization. (2) Soak polyvinylidene fluoride membrane in methanol for 10 – 20 seconds, then soak additionally in phosphate-buffered sodium chloride TS for more than 30 minutes. To the well in the dot blot apparatus mounted the polyvinyli- dene fluoride membrane, add a volume of Interferon Alfa (NAMALWA), corresponding to about 20 mg protein, allow to stand for 15 minutes, and aspirate. After repeating twice to aspirate with a 0.2-mL portion of phosphate-buffered so-dium chloride TS, take out the polyvinylidene fluoride mem-brane, soak in 0.01 mol/L tris buffer solution-sodium chlo-ride TS (pH 7.4), and stir gently for 10 minutes. Replace the liquid, and repeat this operation two more times. Remove 0.01 mol/L tris buffer solution-sodium chloride TS (pH 7.4), add elderberry lectin TS, and stir gently for 2 hours. Remove the elderberry lectin TS, add 0.01 mol/L tris buffer solution-sodium chloride TS (pH 7.4), and stir gently for 10 minutes. Replace the liquid, and repeat this operation two more times. Remove 0.01 mol/L tris buffer solution-sodium chlo-ride TS (pH 7.4), add the peroxidase-labeled avidin TS, and stir gently for 15 minutes. Remove the peroxidase-labeled avidin TS, add 0.01 mol/L tris buffer solution-sodium chlo-ride TS (pH 7.4), and stir gently for 10 minutes. Replace the liquid, and repeat this operation two more times. Remove 0.01 mol/L tris buffer solution-sodium chloride TS (pH 7.4), add substrate TS for interferon alfa identification, and allow to develop the color: a brown dot is observed. Constituent amino acids When perform the test by Method 2 of 2. Methodologies of Amino Acid Analysis after hydrolyzing by Method 1 (but not containing phenol) of 1. Hydrolysis of Protein and Peptide under Amino Acid Analy-sis of Proteins <2.04>, the molar ratios of each constituent amino acid are 8 – 11 for aspartic acid, 4 – 7 for threonine, 7 – 10 for serine, 16 – 19 for glutamic acid, 2 – 4 for glycine and tyrosine, 5 – 7 for alanine, phenylalanine and lysine, 3 – 6 for valine, 2 – 5 for methionine, 4 – 6 for isoleucine, 12 – 15 for leucine, 1 – 3 for histidine and 6 – 9 for arginine. Hydrolysis—To Interferon Alfa (NAMALWA) add tris-glycine buffer solution (pH 6.8) so that each mL con-tains 6,000,000 Units. Pass 3 mL of the solution through a column 4 mm in internal diameter, packed with 0.145 g of ethylsilanized silica gel for column chromatography and pre-viously washed with 5 mL of a mixture of water, acetonitrile and diluted trifluoroacetic acid (1 in 50) (13:6:1). Then, after washing with not less than 10 mL of a mixture of water, acetonitrile and diluted trifluoroacetic acid (1 in 50) (13:6:1), elute interferon alfa with 0.5 mL of a mixture of acetonitrile and diluted trifluoroacetic acid (1 in 50) (19:1), and use the phenylalanine and L-norleucine, dissolve in diluted 6 mol/L hydrochloric acid TS for amino acid automatic analysis (31 in 10,000) so that each mL contains a certain concentration of about 20 nmol for each amino acid, and use this solution as the standard solution. Amino acid analysis—When perform the test with 15 mL each of the sample solutions (1) and (2) and 10 mL of the standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, either chro- matogram obtained from the sample solutions shows the peaks corresponding to the peaks obtained from the stand-ard solution. The molar ratios of each constituent amino acids are calculated. When calculate the molar ratios of each constituent amino acid, for threonine and serine the molar value is corrected by extrapolation to 0 hour-heating based on the values obtained from the sample solutions (1) and (2), for isoleucine and valine use the value from the sample solu-tion (2), and for the other amino acids use the value from the sample solution (1). The molar ratios of cystine, proline and tryptophan are excluded from calculation. Internal standard solution—To exactly 32.81 mg of L-norleucine add diluted 6 mol/L hydrochloric acid TS for amino acid automatic analysis (31 in 10,000) to make exactly 100 mL. Pipet 4 mL of this solution, add diluted 6 mol/L hydrochloric acid TS for amino acid automatic analysis (31 in 10,000) to make exactly 100 mL. Operating conditions— Detector: A fluorophotometer (excitation wavelength: 340 nm, fluorescence wavelength: 450 nm). Column: A stainless steel column 5 mm in inside diameter and 8 cm in length, packed with strongly acidic ion-exchange resin (Na type) for liquid chromatography composed with a sulfonated styrene-divinylbenzene copolymer (3 mm in parti-cle diameter). Column temperature: Inject the sample at 50 ± 19C, maintain the temperature for 11 minutes, change to 40 ± 19 C and maintain for 23 minutes, then change to 65 ± 19C and maintain for 56 minutes, and change to 45 ± 19C. Reaction vessel temperature: A constant temperature of about 519C. Mobile phase: Prepare the mobile phases A, B, C and D according to the following table. eluate as the sample stock solution. To 0.45 mL of the sam-ple stock solution add 50 mL of the internal standard solu- Mobile phase A Mobile phase B Mobile phase C Mobile phase D tion, and stir. Transfer 0.1 mL each of this solution into two glass vessels for hydrolysis, and evaporate to dryness under reduced pressure. Add 20 mL of a solution which is prepared Citric acid monohydrate Sodium citrate hydrate 15.93 g 8.40 g 6.10 g — 6.97 g 10.00 g 26.67 g — by adding 10 mL of mercapto acetic acid to 1 mL of 6 mol/L hydrochloric acid TS for amino acid automatic analysis, and 0.18 mL of 6 mol/L hydrochloric acid TS for amino acid au- Sodium chloride 6.36 g 2.34 g 54.35 g — Sodium hydroxide — — 2.0 g 8.0 g Ethanol (99.5) 54 mL — — — tomatic analysis to the bottom of the glass vessels, replace the air in the vessels with nitrogen, close the vessels tightly Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL under reduced pressure, and heat at 110 ± 29C for 24 hours for one of the vessels and for 72 hours for another. After cooling, open the vessels, evaporate the hydrolyzate to dry-ness under reduced pressure, dissolve the residue in 20 mL of water, and evaporate to dryness under reduced pressure. Dis-solve the residues with 0.1 mL each of diluted 6 mol/L hy-drochloric acid TS for amino acid automatic analysis (31 in 10,000), and use these solutions as the sample solutions (1) and (2), respectively. Separately, weigh exactly a suitable amount each of L-lysine hydrochloride, L-histidine hydro-chloride monohydrate, L-arginine, L-aspartic acid, L-threo-nine, L-serine, L-glutamic acid, glycine, L-alanine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L- Benzyl alcohol — 2 mL 5 mL — Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL Water a suitable a suitable a suitable a suitable quantity quantity quantity quantity Total amount 1000 mL 1000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B, C and D as directed in the following table. JP XVIII Official Monographs / Interferon Alfa (NAMALWA) 1167 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) Mobile phase D (volz) 0 – 11 11 – 12 100 → 0 0 → 100 12 – 34 34 – 39.1 100 → 0 0 → 100 39.1 – 71 71 – 86 Reaction reagent: Prepare the reaction reagents A, B and C according to the following table. Reaction reagent A Reaction reagent B Reaction reagent C Sodium hydroxide 24.0 g — — Boric acid — 21.60 g 21.60 g logarithm of molecular mass. Determine the relative mobili-ty of the center of the main band obtained from the sample solution, and calculate the molecular mass of Interferon Alfa (NAMALWA) from the calibration curve: at least 4 bands are observed between 17,000 and 30,000 of molecular mass.
(1) Egg albumin, Sendai virus coat protein, other foreign proteins, and other process-related impurities—Being specified separately when the drug is granted approval based on the Law. Nucleic acids—Perform the test according to the fol-lowing method: the amount of nucleic acids is not more than 1.0 pg as DNA per 1,000,000 Units of interferon alfa (NAMALWA). DNA standard solutions: To the DNA standard stock solution for interferon alfa (NAMALWA) add salmon sperm DNA solution (1 in 10,000,000) so that each mL con-tains exactly 20 ng DNA. Hereinafter, the concentration of Phthalaldehyde in ethanol (99.5) solution (2 in 25) — — 10 mL DNA is the concentration of DNA for interferon alfa Lauromacrogol solution (1 in 4) — — 4 mL 2-Mercaptoethanol — — 2 mL 10z Sodium hypochlorite TS — 0.1 mL — (NAMALWA). To this solution add tris-glycine buffer solu- tion (pH 6.8) exactly so that each mL contains 10 ng DNA. Then, dilute serially by adding tris-glycine buffer solution Water a suitable quantity a suitable quantity a suitable quantity (pH 6.8). Dilute exactly with a mixture of tris-glycine buffer solution (pH 6.8) and 1 mol/L tris buffer solution (pH 8.0) Total amount 1000 mL 1000 mL 1000 mL Flow rate of mobile phase: Adjust so that the retention times of aspartic acid, glutamic acid and methionine are about 12, 20 and 42 minutes, respectively. Flow rate of reaction reagent: About 0.2 mL per minute for each of reagent A, B, and C. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the resolutions between the peaks of threonine and serine, glycine and alanine, and isoleucine and leucine are not less than 0.6, not less than 0.8 and not less than 1.2, re-spectively. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of aspartic acid, proline, valine, and arginine are not more than 2.5z, respectively. Molecular mass To a suitable amount of Interferon Alfa (NAMALWA) add tris-glycine buffer solution (pH 6.8) so that each mL contains 6,000,000 Units. To 3 volumes of this solution add 1 volume of reduction liquid for molecular mass determination, heat on a water bath for 90 seconds, and use this solution as the sample solution. Separately, to 3 volumes of molecular mass marker for interferon alfa add 1 volume of reduction liquid for molecular mass determina-tion, heat on a water bath for 90 seconds, and use this solu-tion as the standard solution. After performing the elec-trophoresis with 40 mL of the sample solution and 15 mL of the standard solution using tris buffer solution (pH 8.3) and polyacrylamide gel for interferon alfa, fix the gel by immers-ing for 1 hour in a solution of trichloroacetic acid (3 in 20). Then, stain the gel by immersing for more than 2 hours in a solution, prepared by dissolving 1.0 g of Coomassie brilliant blue R-250 in 450 mL of methanol and 100 mL of acetic acid (100) and adding water to make 1000 mL, and destain by im-mersing the gel in 1000 mL of a mixture of water, methanol and acetic acid (100) (33:4:3). Determine the relative mobili-ty of each band obtained from the standard solution, and prepare a calibration curve by linear regression against the (40:1) so that each mL contains 128, 64, 32, 16, 8, and 4 pg of DNA, respectively, and use these solutions as DNA stand-ard solutions. Procedure: Use Interferon Alfa (NAMALWA) as the sample solution. Place 0.11 mL each of DNA standard solu-tions, a mixture of tris-glycine buffer solution (pH 6.8) and 1 mol/L tris buffer solution (pH 8.0) (43:1), and the sample solution into tubes separately. Heat these solutions in an alu-minum block thermostat bath at 989C for 10 minutes. After ice-cooling, centrifuge, and transfer 50 mL each of the super-natants to new tubes. In separate wells of a PCR microplate place 6 mL each of DNA standard solutions which have been treated by heating for DNA extraction, a mixture of tris-glycine buffer solution (pH 6.8) and 1 mol/L tris buffer so-lution (pH 8.0) (43:1), and the sample solution. Then, add 20 mL each of a mixture of 2-fold PCR reaction solution con-taining SYBR Green, nuclease free water, primer F TS and primer R TS (167:70:10:10) into each well. Seal with plate film, and centrifuge. After centrifugation, attach the plate to a real-time PCR system, repeat 40 cycles of 15 seconds at 959C and 1 minute at 609C, and measure the fluorescence in-tensity of each well in every PCR cycle. Plot the fluorescent amount on the vertical axis and the PCR cycle number on the horizontal axis, and determine the PCR cycle number at which the fluorescence of each well is greater than a certain value. Further, make a calibration curve by plotting the number of PCR cycles on the vertical axis and the logarithm of the concentration of DNA standard solution on the horizontal axis to calculate the concentration of DNA in the sample solution. System suitability— Test for required detectability: The PCR cycle number ob-tained with 4 pg/mL DNA standard solution is not greater than that obtained with a mixture of tris-glycine buffer solu-tion (pH 6.8) and tris buffer solution (pH 8.0) (43:1). System performance: When the procedure is run with each DNA standard solution under the above conditions, the cor-relation coefficient of the calibration curve obtained is 0.990 or more. Infective virus test—Inject 0.2 mL each of Interferon Alfa (NAMALWA) into the allantoic cavity of not less than 6 embryonated eggs, allow them to stand at 36 ± 19C for 3 ‌1168 days, and then allow to stand at 49C for a night. Collect more than 1 mL of the allantoic fluid from each egg. To 50 mL of the allantoic fluid add 50 mL of 0.5 volz chicken erythrocyte suspension, mix, and allow to stand at room temperature for 1 hour. Examine the presence of the aggre-gation. When the aggregation is not found, inject 0.2 mL each of this allantoic fluid into the allantoic cavity of the embryonated eggs, and repeat the same procedure as above: the test is met when the aggregation is not found. As a positive control, inoculate the Sendai virus 1.6 × 10－4 to 6.4 × 10－4 HA value per embryonated chicken egg into the allantoic cavity, and perform the test at the same time.
(1) Protein content— Sample solution: Dilute Interferon Alfa (NAMAL-WA) with isotonic sodium chloride solution so that each mL contains 3,000,000 to 4,000,000 Units, and use this solution as the sample solution. 1 cm Standard solution: Weigh accurately about 50 mg of bovine serum albumin, and dissolve in isotonic sodium chlo-ride solution to make exactly 50 mL. Determine the absor-bance of this solution at 280 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Calculate the protein concentration based on E1z (280 nm) ＝ 6.6. To this solu-tion add isotonic sodium chloride solution so that each mL contains exactly 50, 25, 12.5, 6.25, and 3.13 mg of the bovine serum albumin, and use these solutions as the standard solu-tions. Procedure: To exactly 0.25 mL each of the sample solution and the standard solutions add exactly 0.25 mL of Coomassie brilliant blue TS for interferon alfa, and allow to stand at room temperature for exactly 30 seconds. Determine the absorbance of these solutions at 614 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the blank. Plot the absorbance of the standard solu-tions on the vertical axis and their protein concentrations on the horizontal axis to prepare a calibration curve. Determine the protein content of the sample solution from its absorbance using the calibration curve, and calculate the amount of protein per mL of the sample solution. Perform a blank determination in the same manner with isotonic so-dium chloride solution, and make any necessary correction. (2) Specific activity—To each well of a flat-bottom microplate add 45,000 to 60,000 cells of FL cell, prepared with Eagle's minimum essential medium containing bovine serum, and incubate at 37 ± 19C for 18 to 22 hours in an in-cubator filled with 5z carbon dioxide. Dilute Interferon Alfa (NAMALWA) and Interferon Alfa RS separately with Eagle's minimum essential medium containing bovine serum so that each mL contains about 30 Units, and use these solu-tions as the sample solution (1) and the standard solution (1), respectively. To 200 mL each of these solutions add 117 mL of Eagle's minimum essential medium containing bovine se-rum, and use these solutions as the sample solution (2) and the standard solution (2), respectively. Repeat this opera-tion, and prepare the sample solutions and standard solu-tions with log dilutions of 8 serials (dilution ratio per stage is 0.2 log10 fold). Repeat to prepare the sample solutions three or more times. Add each sample solution or standard solu-tion into each well of the cell culture, and incubate at 37 ± 19C for 6 hours. Discard the culture medium, add 1 × 105 to × 106 PFU of Sindbis virus per well, and incubate at 37 ± 19C for 38 to 42 hours. Discard the culture medium, add neutral red-Eagle's minimum essential medium containing bovine serum, and incubate at 37 ± 19C for 45 to 75 minutes. Discard the culture medium, and add 0.01 mol/L phosphate buffer solution. Discard the liquid. Repeat this operation. Elute the neutral red that is taken up by the cells by adding sodium dihydrogen phosphate-ethanol TS. Deter-mine the absorbance at 540 nm, prepare the dose-response curves by plotting the absorbances on the vertical axis and the logarithm of the dilution ratio on the horizontal axis with the absorbances obtained from the sample solution and standard solution. On the dose-response curves of the sam-ple solution and standard solution, calculate the relative potency of the sample solution (n ＝ 3 or more), obtained in-dependently, to the standard solution by comparing the points where the intermediate of absorbances in cells infect-ed with virus and cells not infected with virus, and calculate the average value of them as the potency of Interferon Alfa (NAMALWA) in 1 mL. Calculate the specific activity by dividing the obtained potency by the amount of protein con-tent. When all of the following conditions are satisfied, the test is valid. Absorbance obtained from cells not infected with virus is 0.8 to 1.2. Absorbance obtained from the cells infected with virus is not more than 0.1. Standard deviation of the (log) potency of Interferon Alfa (NAMALWA) in 1 mL obtained from the sample solution prepared three or more times independently is not more than 0.06.
Pancreatin is a substance containing enzymes pre-pared from the pancreas of edible animals, mostly the hog, and has amylolytic, proteolytic and lipolytic ac-tivities. It contains not less than 2800 starch saccharifying activity units, not less than 28,000 proteolytic activity units, and not less than 960 lipolytic activity units per g.
Pancreatin occurs as a white to light yellow powder. It has a characteristic odor.
(1) Rancidity—Pancreatin has no unpleasant or rancid odor and is tasteless. Fat—Add 20 mL of diethyl ether to 1.0 g of Pancrea-tin, extract with occasional shaking for 30 minutes, and filter. Wash the residue with 10 mL of diethyl ether, combine the washing with the filtrate, evaporate the diethyl ether, and dry the residue at 1059C for 2 hours: the mass of the residue does not exceed 20 mg. Loss on drying <2.41> Not more than 4.0z (1 g, in vacu-um, phosphorus (V) oxide, 24 hours). Residue on ignition <2.44> Not more than 5z (1 g).
(1) Starch digestive activity <4.03> Substrate solution—Use potato starch TS for amylo-lytic activity test, prepared by adding 10 mL of phosphate buffer solution for pancreatin instead of 10 mL of 1 mol/L acetic acid-sodium acetate buffer solution (pH 5.0). Sample solution—Weigh accurately about 0.1 g of Pancreatin, add a suitable amount of ice-cold water, stir, and add ice-cold water to make exactly 100 mL. Pipet 10 mL of this solution, and add ice-cold water to make exactly 100 mL. Procedure—Proceed as directed in 1.1. Measure-ment of starch saccharifying activity of 1. Assay for starch digestive activity under Digestion Test. Protein digestive activity <4.03> Substrate solution—Use the substrate solution 2 described in 2.3. (ii) of 2. Assay for protein digestive activity under Digestion Test after adjusting the pH to 8.5. Sample solution—Weigh accurately about 0.1 g of Pancreatin, add a suitable amount of ice-cold water, stir, and add ice-cold water to make exactly 200 mL. Procedure—Proceed as directed in 2. Assay for pro-tein digestive activity under Digestion Test, using trichloroa-cetic acid TS B as the precipitation reagent. Fat digestive activity <4.03> Emulsifier—Prepare with 18 g of polyvinyl alcohol I and 2 g of polyvinyl alcohol II as directed in 3. Assay for fat digestive activity under Digestion Test. Substrate solution—Use the substrate solution Amount (mg) of ozagrel sodium (C H N NaO ) described in 3. Assay for fat digestive activity under the ＝ MS × QT/QS × 16 13 11 2 2 Digestion Test. Sample solution—Weigh accurately about 0.1 g of MS: Amount (mg) of Ozagrel Sodium RS taken Internal standard solution—A solution of benzoic acid in methanol (1 in 100).
Pepsin A
Gastric Protease
Pepsin is a proteolytic enzyme obtained from the gastric mucosa of the pig, Sus scrofa Linné var. domesticus Gray (Suidae), or of the cow, Bos taurus Linné var. domesticus Gmelin (Bovidae), or from the gastric juice of the hog, having not less than 3000 Units per g when determined as directed in the Assay.
Gastric mucosa of pig (Sus scrofa domesticus) or cow (Bos taurus), or gastric juice of hog
Sus scrofa Linné var. domesticus Gray (Suidae) or Bos taurus Linné var. domesticus Gmelin (Bovidae)
One unit of pepsin activity is the amount of enzyme which liberates 1 μmol of tyrosine per minute at 37°C and pH 1.7 using hemoglobin as substrate
White to light yellow powder or flakes, having a characteristic, slightly acid odor
Soluble in water, practically insoluble in ethanol (95) and in diethyl ether. The solution in water is usually slightly turbid.
Place 10 mL of a solution of egg white in a test tube, add 2 mL of dilute hydrochloric acid and 0.1 g of Pepsin, and keep at 38-40°C: the mixture becomes turbid, then gradually clarifies.
To 5 mL of a solution of Pepsin (1 in 100) add 2 mL of ninhydrin TS, and heat: a blue-purple color is produced.
Dissolve 0.5 g of Pepsin in 10 mL of water: the solution is slightly turbid and light yellow to yellow-brown in color.
To 0.5 g of Pepsin add 10 mL of water, shake well, filter, and to the filtrate add 2 drops of dilute acetic acid: no precipitate is formed.
Place 1.0 g of Pepsin in a glass-stoppered bottle, add 30 mL of petroleum ether, shake for 15 minutes, and filter through a dry filter paper. Evaporate 10 mL of the filtrate on a water bath, and dry the residue at 105°C for 2 hours: the mass of the residue is not more than 10 mg.
Proceed according to Method 2 using 1.0 g of Pepsin. Prepare the control solution with 3.0 mL of Standard Lead Solution.
Prepare the test solution with 1.0 g of Pepsin according to Method 1, and perform the test.
Dry 0.5 g of Pepsin at 105°C for 4 hours
Proceed as directed under Residue on Ignition Test, using 0.5 g of Pepsin
Weigh accurately about 0.05 g of Pepsin, dissolve in dilute hydrochloric acid (1 in 100) to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve 0.05 g of hemoglobin in 5 mL of water, add 10 mL of 0.1 mol/L hydrochloric acid TS, and add water to make exactly 50 mL. Pipet 5 mL of this solution, warm to 37°C, add exactly 1 mL of the sample solution previously warmed to 37°C, shake immediately, and allow to stand at 37°C for exactly 10 minutes. Then add exactly 10 mL of trichloroacetic acid TS, shake, allow to stand for 30 minutes, and filter through a dry filter paper. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the test solution. Prepare a blank solution in the same manner, but add 1 mL of dilute hydrochloric acid (1 in 100) instead of the sample solution. Determine the absorbance of the test solution at 275 nm, taking the blank solution as the blank. Calculate the activity using the formula: Activity (Units/g) = (A × 50 × V) / (W × 1.05 × 10), where A is the absorbance of the test solution, V is the dilution factor of the sample solution, and W is the mass (g) of the sample.
Store in a cool place, protect from moisture
Saccharated Pepsin is a mixture of pepsin obtained from the gastric mucosa of hog or cattle and Lactose Hydrate, and it is an enzyme drug having a proteolytic activity. It contains not less than 3800 units and not more ‌than 6000 units per g.
Saccharated Pepsin occurs as a white powder. It has a characteristic odor, and has a slightly sweet taste. It dissolves in water to give a slightly turbid liquid, and does not dissolve in ethanol (95) and in diethyl ether. It is slightly hygroscopic.
(1) Rancidity—Saccharated Pepsin has no un-pleasant or rancid odor. (2) Acidity—Dissolve 0.5 g of Saccharated Pepsin in 50 mL of water, and add 0.50 mL of 0.1 mol/L sodium hydrox-ide VS and 2 drops of phenolphthalein TS: the solution is red in color. Loss on drying <2.41> Not more than 1.0z (1 g, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
(i) Substrate solution—Use the substrate solution 1 described in Assay for protein digestive activity under Diges-tion Test <4.03> after adjusting the pH to 2.0. Sample solution—Weigh accurately an amount of Saccharated Pepsin equivalent to about 1250 units, dissolve in ice-cold 0.01 mol/L hydrochloric acid TS to make exactly 50 ml. Standard solution—Weigh accurately a suitable amount of Saccharated Pepsin RS, and dissolve in ice-cold 0.01 mol/L hydrochloric acid TS to make a solution contain-ing about 25 units per mL. Procedure—Proceed as directed in Assay for protein digestive activity under Digestion Test <4.03>, and determine the absorbances, AT and ATB, of the sample solution, using trichloroacetic acid TS A as the precipitation reagent. Sepa-rately, determine the absorbances, AS and ASB, of the stand-ard solution in the same manner as the sample solution. Units in 1 g of Saccharated Pepsin ＝ US × (AT － ATB)/(AS － ASB) × 1/M US: Units per mL of the standard solution M: Amount (g) of Saccharated Pepsin per mL of the sam-ple solution taken
Containers—Tight containers. Storage—Not exceeding 309C.
Whole Human Blood is a deep red liquid from which the erythrocytes settle upon standing, leaving a yellow supernatant layer. A gray layer which mainly consists of leu-cocytes may appear on the surface of the settled erythrocyte layer. The supernatant layer may become turbid in the presence of fat, or may show the faint color of hemoglobin. Wine
Nux Vomica Extract Powder occurs as a yel-low-brown to grayish brown powder. It has a slight, charac-teristic odor and a bitter taste.
(1) To 3 g of Nux Vomica Extract Powder add 3 mL of ammonia TS and 20 mL of chloroform, macer-ate for 30 minutes with occasional shaking, and filter. Re-move most of the chloroform from the filtrate by warming on a water bath, add 5 mL of diluted sulfuric acid (1 in 10), and warm on a water bath while shaking well until the odor of chloroform is no longer perceptible. After cooling, filter through a pledget of absorbent cotton, and add 2 mL of nitric acid to 1 mL of the filtrate: a red color develops. (2) To the remaining filtrate obtained in (1) add 1 mL of potassium dichromate TS, and allow to stand for 1 hour: a yellow-red precipitate is produced. Collect the precipitate by filtration, and wash with 1 mL of water. Transfer a part of the precipitate to a small test tube, add 1 mL of water, dis-solve by warming, cool, and add 5 drops of sulfuric acid dropwise carefully along the wall of the test tube: the layer of sulfuric acid shows a purple color which turns immedi-ately red to red-brown.
Weigh accurately about 2.0 g of Nux Vomica Ex-tract Powder, place in a glass-stoppered centrifuge tube, add 15 mL of ammonia TS, and shake. Add 20 mL of diethyl ether, stopper tightly, shake for 15 minutes, centrifuge to separate the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, proceed in the same manner, and repeat this procedure three times. Combine the extracts, and evapo-rate the solvent on a water bath. Dissolve the residue in 10 mL of the mobile phase, add exactly 10 mL of the internal standard solution, and add the mobile phase to make 100 mL. Filter this solution through a membrane filter with a porosity not more than 0.8 mm, discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 75 mg of strychnine nitrate for assay (separately determine the loss on drying), and dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the inter-nal standard solution, then add the mobile phase to make 100 mL, and use this solution as the standard solution. Per-form the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Calcu-late the ratio, QT and QS, of the peak area of strychnine to MS: Amount (mg) of strychnine nitrate for assay taken, calculated on the dried basis Internal standard solution—A solution of barbital sodium in the mobile phase (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: Room temperature. Mobile phase: A mixture of a solution of potassium dihy-drogenphosphate (6.8 in 1000), acetonitrile and triethyla-mine (45:5:1), adjusted the pH to 3.0 with phosphoric acid. Flow rate: Adjust so that the retention time of strychnine is about 17 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and strychnine are eluted in this order with the resolution between these peaks being not less than 1.5.
Containers—Tight containers. Storage—Light-resistant.
Acebutolol Hydrochloride, when dried, contains not less than 98.
Acebutolol Hydrochloride occurs as white to pale yellow-white, crystals or crystalline powder. It is freely soluble in water, in methanol, in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether. A solution of Acebutolol Hydrochloride (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Acebutolol Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Acebutolol Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Acebutolol Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Acebutolol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Acebutolol Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 40 mg of Acebutolol Hydrochloride in 2 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 25 mL, and pipet 1 mL of this so-lution, add methanol to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with the upper layer of a mixture of water, 1-butanol and acetic acid (100) (5:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 365 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.25 g of Acebutolol Hydro-chloride, previously dried, dissolve in 20 mL of acetic acid (100), add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.29 mg of C18H28N2O4.HCl
Acemetacin, when dried, contains not less than 99.
Acemetacin occurs as a light yellow crystalline powder. It is soluble in acetone, sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water.
(1) To 1 mg of Acemetacin add 1 mL of concentrated chromotropic acid TS, and heat in a water bath for 5 minutes: a red-purple color develops. Determine the absorption spectrum of a solution of Acemetacin in methanol (1 in 50,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Acemetacin as directed in the potassium bromide disk method under Infrared Spectrometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave ‌400 numbers. Perform the test with Acemetacin as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Acemetacin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.40 g of Acemetacin in 10 mL of acetone, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add acetone to make exactly 50 mL. Pipet 1 mL of this solution, add ace-tone to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin Layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin layer chromatography. Develop the plate with a mixture of hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the num-ber of spots other than the principal spot obtained from the sample solution is not more than 2, and these spots are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Acemetacin, previ-ously dried, dissolve in 20 mL of acetone, add 10 mL of water, and then titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS (main wavelength: 254 nm): the spots obtained from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Acemetacin Cap-sules, add 40 mL of methanol, shake well, and add methanol to make exactly V mL so that each mL contains about 0.6 mg of acemetacin (C21H18ClNO6). Filter this solution, dis-card the first 10 mL of the filtrate, pipet 5 mL of the subse-quent filtrate, add exactly 2 mL of the internal standard so-lution, add methanol to make 50 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of acemetacin (C21H18ClNO6) ＝ MS × QT/QS × V/50 MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Acemetacin Capsules is not less than 70z. Start the test with 1 capsule of Acemetacin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 33 mg of acemetacin (C21H18ClNO6), and use this solution as the sample solution. Separately, weigh accurately about 17 mg of acemetacin for assay, previously dried at 1059C for 2 hours, dissolve in the ＝ 41.58 mg of C 21H 18ClNO6 dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 20
Paracetamol
N-Acetyl-p-aminophenol
APAP
Acetaminophen, when dried, contains not less than 98.0% of C8H9NO2.
White crystals or crystalline powder, odorless, slightly bitter taste
Freely soluble in ethanol (95), sparingly soluble in water, slightly soluble in diethyl ether. Dissolves in sodium hydroxide TS.
Determine the infrared absorption spectrum of the sample as directed in the potassium bromide disk method under Infrared Spectrophotometry, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
To 0.1 g of Acetaminophen add 10 mL of dilute hydrochloric acid, boil for 1 minute, cool, add 1 drop of sodium nitrite TS, allow to stand for 2 minutes, then add 2 mL of ammonium sulfamate TS, mix, and allow to stand for 10 minutes. Add 2 mL of N-(1-naphthyl)-N'-diethyl ethylenediamine oxalate TS: a red-purple color develops.
Dissolve 1.0 g of Acetaminophen in 20 mL of ethanol (95): the solution is clear and colorless to pale yellow.
p-Aminophenol and other related substances
Dissolve 0.10 g of Acetaminophen in 10 mL of a mixture of water and methanol (1:1), and use this solution as the sample solution. Perform the test with 10 μL of this solution as directed under Liquid Chromatography according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak appearing at the retention time of p-aminophenol is not more than 0.05% of the peak area of acetaminophen, and the area of any peak other than acetaminophen and p-aminophenol is not more than 0.1% of the peak area of acetaminophen.
Proceed according to Method 1 using 2.0 g of Acetaminophen. Prepare the control solution with 2.0 mL of Standard Lead Solution (for the test, use a quartz or porcelain crucible).
Prepare the test solution with 1.0 g of Acetaminophen according to Method 1, and perform the test (not more than 2 ppm).
Weigh accurately about 0.5 g of Acetaminophen, add exactly 5 mL of the internal standard solution, and dissolve by warming if necessary. Cool, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of p-Chloroacetanilide for thin-layer chromatography, add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 1 μL each of the sample solution and standard solution as directed under Gas Chromatography according to the following conditions. Calculate the ratios of the peak area of p-chloroacetanilide to that of the internal standard from the sample solution and the standard solution.
Proceed as directed under Residue on Ignition Test
Weigh accurately about 0.3 g of Acetaminophen, previously dried, add 40 mL of water and 40 mL of hydrochloric acid, and dissolve by heating. After cooling, add 0.2 g of potassium bromide, titrate with 0.1 mol/L sodium nitrite VS (potentiometric titration) while cooling the solution in ice water and stirring. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium nitrite VS = 15.12 mg of C8H9NO2.
Acetazolamide contains not less than 98.
Acetazolamide occurs as a white to pale yellow-white crystalline powder. It is odorless, and has a slight bitter taste. It is slightly soluble in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether. Melting point: about 2559C (with decomposition).
(1) To 0.1 g of Acetazolamide add 5 mL of sodium hydroxide TS, then add 5 mL of a solution of 0.1 g Chloride <1.03>—To 1.5 g of Acetazolamide add 75 mL of water, and warm at 709C for 20 minutes with occa-sional shaking. After cooling, filter, and to 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Sulfate <1.14>—To 25 mL of the filtrate obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 1.0 g of Aceta- zolamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Silver-reducing substances—Wet 5 g of Acetazola-mide with 5 mL of aldehyde-free ethanol, and add 125 mL of water, 10 mL of nitric acid and exactly 5 mL of 0.1 mol/L silver nitrate VS. Stir for 30 minutes by protecting from light, filter through a glass filter (G3), and wash the residue on the glass filter with two 10-mL portions of water. Com-bine the filtrate with the washings, to the solution add 5 mL of ammonium iron (III) sulfate TS, and titrate <2.50> with 0.1 mol/L ammonium thiocyanate VS: not less than 4.8 mL of 0.1 mol/L ammonium thiocyanate VS is consumed. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.15 g of Acetazolamide, and dissolve in 400 mL of water in a water bath by heating. After cooling, add water to make exactly 1000 mL. Pipet 5 mL of the solution, add 10 mL of 1 mol/L hydrochloric acid TS, and then add water to make exactly 100 mL. Determine the absorbance A of this solution at the wavelength of maxi-mum absorption at about 265 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of acetazolamide (C4H6N4O3S2) ＝ A/474 × 200,000
Acetic Acid is a clear, colorless liquid. It has a pungent, characteristic odor and an acid taste. It is miscible with water, with ethanol (95) and with glycerin. Specific gravity d 20: about 1.04
Acetic Acid changes blue litmus paper to red, and responds to Qualitative Tests <1.09> for acetate.
(1) Clarity and color of solution—Dissolve 1.0 g of Acetazolamide in 10 mL of sodium hydroxide TS: the solution is clear and colorless to pale yellow. Description Acetic Acid is a clear, colorless liquid. It has a pungent, characteristic odor and an acid taste. It is miscible with water, with ethanol (95) and with glycerin. Specific gravity d 20: about 1.04 Identification Acetic Acid changes blue litmus paper to red, and responds to Qualitative Tests <1.09> for acetate. Purity (1) Chloride—To 20 mL of Acetic Acid add 40 mL of water, and use this solution as the sample solution. To 10 mL of the sample solution add 5 drops of silver nitrate TS: no opalescence is produced. ‌404‌ Sulfate—To 10 mL of the sample solution obtained in (1) add 1 mL of barium chloride TS: no turbidity is pro-duced. Heavy metals <1.07>—Evaporate 10 mL of Acetic Acid on a water bath to dryness, and to the residue add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test with this solution as the test solution. Prepare the con-trol solution with 3.0 mL of Standard Lead Solution by add-ing 2 mL of dilute acetic acid and water to make 50 mL (not more than 3 ppm). Potassium permanganate-reducing substances—To 20 mL of the sample solution obtained in (1) add 0.10 mL of 0.02 mol/L potassium permanganate VS: the red color does not disappear within 30 minutes. Non-volatile residue—Evaporate 30 mL of Acetic Acid on a water bath to dryness, and dry at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Measure exactly 5 mL of Acetic Acid, add 30 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 drops of phenolphthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 60.05 mg of C2H4O2
Acetohexamide, when dried, contains not less than 98.
Acetohexamide occurs as a white to yellowish white powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in acetone, slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water. Melting point: about 1859C (with decomposition).
(1) Dissolve 0.10 g of Acetohexamide in 100 mL of methanol. To 5 mL of the solution add 20 mL of 0.5 mol/L hydrochloric acid TS and 75 mL of methanol, and use the solution as the sample solution (1). Determine the absorption spectrum of the sample solution (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>, using methanol as the blank, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, to exactly 10 mL of the sample solution (1) add methanol to make ex-actly 50 mL, and use the solution as the sample solution (2). Determine the absorption spectrum of the sample solution (2) as directed under Ultraviolet-visible Spectrophotometry <2.24>, using methanol as the blank, and compare the spec-trum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Acetohexamide, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.5 g of Aceto-hexamide in 40 mL of N,N-dimethylformamide, add 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL. Perform the test using this solution as the test solution. JP XVIII Official Monographs / Acetohexamide 405 Prepare the control solution as follows: to 0.45 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL (not more than 0.011z). Sulfate <1.14>—Dissolve 2.0 g of Acetohexamide in 40 mL of N,N-dimethylformamide, and add 1 mL of dilute hydrochloric acid and N,N-dimethylformamide to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid and N,N-dimethylformamide to make 50 mL (not more than 0.010z). Heavy metals <1.07>—Proceed with 1.0 g of Aceto- hexamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances (i) Cyclohexylamine—Dissolve exactly 1.0 g of Acetohexamide in exactly 30 mL of 0.5 mol/L sodium hydroxide TS, add exactly 5 mL of hexane, shake vigorously for 60 minutes, allow to stand for 5 minutes, and use the upper layer as the sample solution. Separately, dissolve exactly 50 mg of cyclohexylamine in 0.5 mol/L sodium hydroxide TS to make exactly 50 mL. Pipet 2 mL of this solution, and add 0.5 mol/L sodium hydroxide TS to make exactly 300 mL. Pipet 30 mL of this solution, add exactly 5 mL of hexane, shake vigorously for 60 minutes, allow to stand for 5 minutes, and use the upper layer as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and determine the peak area of cyclo-hexylamine by the automatic integration method: the peak area of cyclohexylamine obtained from the sample solution is not more than that from the standard solution. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused-silica column 0.53 mm in inside diame- ter and 30 m in length, coated the inner surface with methyl-silicone polymer for gas chromatography 1.5 mm in thick-ness. Column temperature: A constant temperature of about 909C. Injection port temperature: A constant temperature of about 1509C. Detector temperature: A constant temperature of about 2109C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of cyclohex-ylamine is about 4 minutes. Split ratio: 1:1. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the number of theoretical plates of the peak of cyclo-hexylamine is not less than 8000. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of cyclohexylamine is not more than 5z. Dicyclohexylurea—Dissolve exactly 1.0 g of Aceto-hexamide in exactly 10 mL of 0.5 mol/L sodium hydroxide TS, add exactly 20 mL of methanol, shake, then add exactly 5 mL of diluted hydrochloric acid (1 in 10), shake vigorously for 15 minutes, and centrifuge. Filter 10 mL or more of the supernatant liquid through a membrane filter with pore size of not larger than 0.5 mm. Discard the first 5 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, dissolve exactly 50 mg of dicyclohexylurea in methanol to make exactly 100 mL. Pipet 2 mL of this solu-tion, and add methanol to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 10 mL of 0.5 mol/L sodium hydroxide TS, shake, then add exactly 5 mL of diluted hy-drochloric acid (1 in 10), shake, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak area of dicyclo-hexylurea by the automatic integration method: the peak area of dicyclohexylurea obtained from the sample solution is not more than that from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.5 g of sodium hydroxide in 1000 mL of 0.05 mol/L sodium dihydrogen phosphate TS, and adjust the pH to 6.5 with 0.5 mol/L sodium hydroxide TS. To 500 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of dicyclo-hexylurea is about 10 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating conditions, the number of theoretical plates of the peak of dicyclohexylurea is not less than 10,000. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of dicyclohexylurea is not more than 2.0z. Other related substances—Dissolve 0.10 g of Aceto-hexamide in 10 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add acetone to make exactly 20 mL. Pipet two 1 mL portions of this solution, add acetone to make exactly 10 mL and 25 mL, respectively, and use these solutions as the standard solution (1) and the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, ammonia solution (28) and cyclohexane (6:2:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and the number of them which are more intense than the spot from the standard solution (2) is not more than 4. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Acetohexamide, previously dried, dissolve in 30 mL of N,N-dimethylfor-mamide, add 10 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner using a solution prepared by adding 19 mL of water to 30 mL of N,N-dimethylformamide, and make any necessary correc- ‌406‌ tion. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 32.44 mg of C15H20N2O4S of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement.
Aspirin occurs as white, crystals, granules or powder. It is odorless, and has a slight acid taste. It is freely soluble in ethanol (95) and in acetone, soluble in diethyl ether, and slightly soluble in water. It dissolves in sodium hydroxide TS and in sodium car-bonate TS. In moist air, it gradually hydrolyzes to salicylic acid and acetic acid. Melting point: about 1369C (bath fluid is heated at 1309C previously).
(1) Boil 0.1 g of Aspirin in 5 mL of water for 5 to 6 minutes, cool, and add 1 to 2 drops of iron (III) chloride TS: a red-purple color is produced. Boil 0.5 g of Aspirin in 10 mL of sodium carbonate TS for 5 minutes, and add 10 mL of dilute sulfuric acid: the odor of acetic acid is perceptible, and a white precipitate is produced. Filter the precipitate, add 3 mL of ethanol (95) and 3 mL of sulfuric acid to the filtrate, and heat: the odor of ethyl acetate is perceptible.
(1) Clarity of solution—Dissolve 0.5 g of Aspirin in 10 mL of warm sodium carbonate TS: the solution is clear. Salicylic acid—Dissolve 2.5 g of Aspirin in 25 mL of ethanol (95), and add 1.0 mL of this solution to a solution which is prepared by transferring 1 mL of a freshly prepared dilute ammonium iron (III) sulfate TS to a Nessler tube and diluting with water to 50 mL. Allow to stand for 30 seconds: the solution has no more color than the following control so-lution. Control solution: Dissolve 0.100 g of salicylic acid in water, and add 1 mL of acetic acid (100) and water to make 1000 mL. Add 1.0 mL of this solution to a solution which is prepared by transferring 1 mL of freshly prepared dilute am-monium iron (III) sulfate TS and 1 mL of ethanol (95) to a Nessler tube and diluting with water to 50 mL. Allow to stand for 30 seconds. Chloride <1.03>—Boil 1.8 g of Aspirin in 75 mL of water for 5 minutes, cool, add water to make 75 mL, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.015z). Sulfate <1.14>—To 25 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.040z). Heavy metals <1.07>—Dissolve 2.5 g of Aspirin in 30 mL of acetone, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 2.5 mL of Standard Lead Solution, 30 mL of acetone, 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Readily carbonizable substances <1.15>—Weigh 0.5 g of Aspirin, and perform the test. The solution has no more color than Matching Fluid Q. Loss on drying <2.41> Not more than 0.5z (3 g, silica gel, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1.5 g of Aspirin, previously dried, add exactly 50 mL of 0.5 mol/L sodium hydroxide VS, and boil gently for 10 minutes under a reflux condenser with a carbon dioxide-absorbing tube (soda lime). Cool, and titrate <2.50> immediately the excess sodium hydroxide with 0.25 mol/L sulfuric acid VS (indicator: 3 drops of phenol-phthalein TS). Perform a blank determination in the same manner. Each mL of 0.5 mol/L sodium hydroxide VS ＝ 45.04 mg of C9H8O4
Aciclovir occurs as a white to pale yellow-white crystalline powder. It is slightly soluble in water and very slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS and in dilute sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Aciclovir in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Aciclovir RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Aciclovir as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Aciclovir RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Aciclovir in 20 mL of dilute sodium hydroxide TS: the so-lution is clear and is not more colored than the following control solution. Control solution: To 2.5 mL of Matching Fluid F add diluted dilute hydrochloric acid (1 in 10) to make 100 mL. ‌Heavy metals <1.07>—Proceed with 1.0 g of Aciclovir according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Separately, weigh accurately about 25 mg of guanine, dissolve in 50 mL of dilute sodium hydroxide TS, and add the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas of guanine, AT and AS, and calculate the amount of guanine by the follow-ing equation: it is not more than 0.7z. Determine each peak area from the sample solution by the automatic integration method, and calculate the amount of each related substance other than aciclovir and guanine by the area percentage method: it is not more than 0.2z. Furthermore, the sum of the amount of guanine calculated above and the amounts of related substances determined by the area percentage method is not more than 1.5z. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × AT/AS MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 1.0 g of sodium 1-decanesulfonate and 6.0 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust the pH to 3.0 with phosphoric acid. To this solution add 40 mL of acetonitrile. Flow rate: Adjust so that the retention time of aciclovir is about 3 minutes. System suitability— System performance: Dissolve 0.1 g of Aciclovir in 5 mL of dilute sodium hydroxide TS, add 2 mL of a solution of guanine in dilute sodium hydroxide TS (1 in 4000), and add the mobile phase to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, aciclovir and guanine are eluted in this order with the resolution between these peaks being not less than Amount (z) of guanine ＝ MS/MT MS: Amount (mg) of guanine taken × AT/AS × 2/5 17. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- MT: Amount (mg) of Aciclovir taken Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 8 times as long as the retention time of aciclovir, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the sample solution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of aciclovir obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of guanine is not more than 2.0z. Water <2.48> Not more than 6.0z (50 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Aciclovir and Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), dissolve each in 1 mL of dilute sodium hydroxide TS, add the mobile phase to make exactly 20 mL each, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of aciclovir in each solution. ing conditions, the relative standard deviation of the peak area of aciclovir is not more than 1.0z.
Aclarubicin Hydrochloride is the hydrochloride of an anthracycline substance having antitumor activity produced by the growth of Streptomyces galilaeus. It contains not less than 920 mg (potency) and not more than 975 mg (potency) per mg, calculated on the anhydrous basis.
Aclarubicin Hydrochloride occurs as a yellow to pale orange-yellow powder. It is very soluble in chloroform and in methanol, freely soluble in water, and slightly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Aclarubicin Hydrochloride in diluted methanol (4 in 5) (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi- ties of absorption at the same wavelengths. ‌Determine the infrared absorption spectrum of Aclarubicin Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Aclarubicin Hydrochloride in methanol (1 in 200) responds to Qualitative Tests <1.09> (2) for chlo-ride. D Optical rotation <2.49> [a]20: －146 – －1629(50 mg calcu-lated on the anhydrous basis, water, 10 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.05 g of Aclarubicin Hydrochloride in 10 mL of water is between 5.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.10 g of Aclarubicin Hydrochloride in 10 mL of water: the solu-tion is clear and yellow to pale orange-yellow. Heavy metals <1.07>—Proceed with 1.0 g of Aclaru- bicin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 10 mg of Aclarubicin Hydrochloride in 10 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method. Calculate the amount of the related substances by the area and allow to stand for 60 minutes. To 1.0 mL of this solu-tion add 1.0 mL of 0.2 mol/L sodium hydroxide TS, 1.0 mL of phosphate buffer solution (pH 8.0) and 1.0 mL of chlo-roform, shake vigorously, and take the chloroform layer. When the procedure is run with 20 mL of the chloroform under the above operating conditions, aclarubicin and 1-deoxypyrromycin are eluted in this order with the resolution between these peaks being not less than 3.0. System repeatability: When the test is repeated 5 times with 20 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of aclarubicin is not more than 2.0z. Water <2.48> Not more than 3.5z (0.1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Aclarubicin Hydro-chloride, equivalent to about 20 mg (potency), and dissolve in diluted methanol (4 in 5) to make exactly 100 mL. Pipet 15 mL of this solution, add diluted methanol (4 in 5) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Aclarubicin RS, equivalent to about 20 mg (potency), add 0.6 mL of diluted hydrochloric acid (1 in 250) and diluted methanol (4 in 5) to make exactly 100 mL. Pipet 15 mL of this solution, add diluted methanol (4 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 433 nm. percentage method: the amount of aklavinone having the relative retention time of about 0.6 to aclarubicin is not more than 0.2z, aclacinomycin L1 having the relative retention Amount [ mg (potency)] of aclarubicin (C ＝ MS × AT/AS × 1000 42H53NO15) time of about 0.75 is not more than 0.5z, 1-deoxypyrromy-cin having the relative retention time of about 1.7 is not more than 1.5z and aclacinomycin S1 having the relative retention time of about 2.3 is not more than 0.5z, and the total amount of the peaks other than aclarubicin and the peaks mentioned above is not more than 1.0z of the peak area of aclarubicin. Operating conditions— Detector: A visible absorption photometer (wavelength: 436 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of chloroform, methanol, acetic acid (100), water and triethylamine (6800:2000:1000:200:1). Flow rate: Adjust so that the retention time of aclarubicin is about 5 minutes. Time span of measurement: As long as about 4 times of the retention time of aclarubicin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of aclarubicin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: Dissolve 5 mg of Aclarubicin Hy-drochloride in 10 mL of 0.1 mol/L hydrochloric acid TS, MS: Amount [mg (potency)] of Aclarubicin RS taken
Containers—Tight containers. Storage—Light-resistant and at 59C or below.
Purified Lanolin is a light yellow to yellowish brown, viscous, ointment-like substance, and has a faint, ‌2062 Lard / Crude Drugs and Related Drugs characteristic but not rancid odor. It is very soluble in diethyl ether and in cyclohexane, freely soluble in tetrahydrofuran and in toluene, and very slightly soluble in ethanol (95). It is practically insoluble in water, but miscible without separation with about twice its mass of water, retaining ointment-like viscosity. Melting point: 37 – 439C
Superimpose carefully 1 mL of a solution of Purified Lanolin in cyclohexane (1 in 50) on 2 mL of sulfuric acid: a red-brown color develops at the zone of contact, and the sulfuric acid layer shows a green fluorescence. Acid value <1.13> Not more than 1.0. Iodine value 18 – 36 Weigh accurately about 0.8 g of Purified Lanolin in a glass-stoppered 500-mL flask, add 20 mL of cyclohexane to dissolve, and add exactly 25 mL of Hanus' TS, and mix well. If a clear solution is not obtained, add more cyclohexane to make clear, and allow the mixture to stand for 1 hour between 209C and 309C in light-resistant, well-closed containers, with occasional shaking. Add 20 mL of a solution of potassium iodide (1 in 10) and 100 mL of water, shake, and titrate the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Per-form a blank determination in the same manner. Iodine value ＝ (a － b) × 1.269/M M: amount (g) of Purified Lanolin taken a: Volume (mL) of 0.1 mol/L sodium thiosulfate VS used in the blank determination b: Volume (mL) of 0.1 mol/L sodium thiosulfate VS used in the titration of the sample
(1) Acid or alkali—To 5 g of Purified Lanolin add 25 mL of water, boil for 10 minutes, and cool. Add water to restore the previous mass, and separate the aqueous layer: the aqueous layer is neutral. Chloride <1.03>—To 2.0 g of Purified Lanolin add 40 mL of water, boil for 10 minutes, and cool. Add water to re-store the previous mass, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Use this solution as the test solution, and perform the test. Pre-pare the control solution with 1.0 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.036z). Ammonia—To 10 mL of the aqueous layer obtained in (1) add 1 mL of sodium hydroxide TS, and boil: the gas evolved does not turn moistened red litmus paper to blue. Water-soluble organic substances—To 5 mL of the aqueous layer obtained in (1) add 0.25 mL of 0.002 mol/L potassium permanganate VS, and allow to stand for 5 minutes: the red color of the solution does not disappear. Petrolatum—Dissolve 1.0 g of Purified Lanolin in 10 mL of a mixture of tetrahydrofuran and isooctane (1:1), and use this solution as the sample solution. And dissolve 20 mg of vaseline in 10 mL of a mixture of tetrahydrofuran and isooctane (1:1), and use this solution as the standard solu-tion. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with isooctane to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 2) on the plate, heat the plate at 809C for 5 minutes, cool, and examine under ultraviolet light (main wavelength: 365 nm): no fluo-rescent spot is observable same level of the spot of standard solution. Use a thin-layer plate previously developed with isooctane to the upper end, dried in air, and heated at 1109C for 60 minutes. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Total ash <5.01> Not more than 0.1z.
Lard occurs as a white, soft, unctuous mass, and has a faint, characteristic odor and a bland taste. It is freely soluble in diethyl ether and in petroleum ether, very slightly soluble in ethanol (95), and practically insoluble in water. Melting point: 36 – 429C Congealing point of the fatty acids: 36 – 429C Acid value <1.13> Not more than 2.0. Saponification value <1.13> 195 – 203 Iodine value <1.13> 46 – 70
(1) Moisture and coloration—Melt 5 g of Lard by heating on a water bath: it forms a clear liquid, from which no water separates. Observe the liquid in a layer 10 mm thick: the liquid is colorless to slightly yellow. Alkalinity—To 2.0 g of Lard add 10 mL of water, melt by warming on a water bath, and shake vigorously. After cooling, add 1 drop of phenolphthalein TS to the sepa-rated aqueous layer: the layer is colorless. Chloride <1.03>—To 1.5 g of Lard add 30 mL of ethanol (95), heat under a reflux condenser for 10 minutes, and filter after cooling. To 20 mL of the filtrate add 5 drops of a solution of silver nitrate in ethanol (95) (1 in 50): the opalescence of the mixture does not exceed that of the fol-lowing control solution. Control solution: To 1.0 mL of 0.01 mol/L hydrochloric acid VS add ethanol (95) to make 20 mL, and add 5 drops of a solution of silver nitrate in ethanol (95) (1 in 50). Beef tallow—Dissolve 5 g of Lard in 20 mL of diethyl ether, stopper lightly with absorbent cotton, and allow to stand at 209C for 18 hours. Collect the separated crystals, moisten them with ethanol (95), and examine under a micro-scope of 200 magnifications: the crystals are in the form of rhomboidal plates grouped irregularly, and do not contain prisms or needles grouped in fan-shaped clusters.
Adsorbed Diphtheria-Tetanus Combined Toxoid
沈降ジフテリア破傷風混合トキソイド
Adsorbed Diphtheria-Tetanus Combined Tox-oid becomes a homogeneous, whitish turbid liquid on shak-ing. ‌874 Dipyridamole
Adsorbed Diphtheria Toxoid for Adult Use
成人用沈降ジフテリアトキソイド
Adsorbed Diphtheria Toxoid for Adult Use becomes a homogeneous, whitish turbid liquid on shaking. Adsorbed Diphtheria-Purified Pertussis-Tetanus Combined Vaccine
Adsorbed Tetanus Toxoid becomes a uniform white-turbid liquid on shaking. Tetracaine Hydrochloride
Afloqualone, when dried, contains not less than 98.
Afloqualone occurs as white to light yellow, crystals or crystalline powder. It is soluble in acetonitrile, sparingly soluble in ethanol (99.5), and practically insoluble in water. It is gradually colored by light. Melting point: about 1979C (with decomposition).
(1) Conduct this procedure without ex-posure to light, using light-resistant containers. Determine the absorption spectrum of a solution of Afloqualone in ethanol (99.5) (1 in 150,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Afloqualone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Acidity or alkalinity—Take 1.0 g of Afloqua-lone in a light-resistant vessel, add 20 mL of freshly boiled and cooled water, shake well, and filter. To 10 mL of the fil-trate add 2 drops of bromothymol blue TS: a yellow color develops. The color changes to blue by adding 0.20 mL of 0.01 mol/L sodium hydroxide TS. Heavy metals <1.07>—Proceed with 2.0 g of Afloqua-lone in a platinum crucible according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 10 mg of Afloqualone in 25 mL of the mobile phase, and use this solution as the sample solution. Pipet 3 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the total area of the peaks other than afloqualone from the sample solution is not larger than the peak area of afloqua-lone from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.2 g of disodium hydrogen phosphate dodecahydrate in 1000 mL of water, adjust to pH 5.5 with diluted phosphoric acid (1 in 10). To 600 mL of this solution add 400 mL of acetonitrile. Flow rate: Adjust so that the retention time of afloqua-lone is about 5.5 minutes. Time span of measurement: About 4 times as long as the retention time of afloqualone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 25 mL, and confirm that the peak area of afloqualone obtained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: Dissolve 0.01 g of Afloqualone in a suitable amount of the mobile phase, add 5 mL of a solution of propyl parahydroxybenzoate in the mobile phase (1 in 2000) and the mobile phase to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, afloqualone and propyl parahydroxy-benzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of afloqualone is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1.0 g, platinum crucible).
Weigh accurately about 0.4 g of Afloqualone, previ-ously dried, dissolve in 10 mL of hydrochloric acid and 40 mL of water, and add 10 mL of a solution of potassium bro-mide (3 in 10). After cooling at 159C or below, titrate <2.50> with 0.1 mol/L sodium nitrite VS according to the potentio-metric titration or amperometric titration under the Electro-metric Titration method. Each mL of 0.1 mol/L sodium nitrite ＝ 28.33 mg of C16H14FN3O
Powdered Agar appears as a white powder, is odorless, and is tasteless and mucilaginous. Under a microscope <5.01>, Powdered Agar, immersed in olive oil or liquid paraffin, reveals angular granules with stri-ations or nearly spheroidal granules 5 to 60 mm in diameter. It becomes transparent in chloral hydrate TS. It is practically insoluble in organic solvents. A boiling solution of Powdered Agar (1 in 100) is neutral.
(1) To a part of Powdered Agar add drop-wise iodine TS: a dark blue to reddish purple color develops. (2) Dissolve 1 g of Powdered Agar in 65 mL of water by boiling for 10 minutes with constant stirring, and add a sufficient amount of hot water to maintain the original volume lost by evaporation: the solution is clear. Cool the solution between 309C and 399C: the solution forms a firm, resilient gel, which does not melt below 859C.
(1) Sulfuric acid—Dissolve 1.0 g of Powdered Agar in 100 mL of water by boiling: the solution is not acid. Sulfurous acid and starch—To 5 mL of the solution obtained in (1) add 2 drops of iodine TS: the solution is not decolorized immediately, and does not show a blue color. Insoluble matter—To 7.5 g of Powdered Agar add 500 mL of water, boil for 15 minutes, and add water to make exactly 500 mL. Take exactly 100 mL of the solution, add 100 mL of hot water, heat to boiling, filter while hot through a tared glass filter (G3), wash the residue with a small amount of hot water, and dry the residue at 1059C for 3 hours: the mass of the residue is not more than 15.0 mg. Water absorption—To 5.0 g of Powdered Agar add water to make 100 mL, shake well, allow to stand at 259C for 24 hours, and filter through moistened glass wool in a 100-mL graduated cylinder: the volume of the filtrate is not more than 75 mL. Loss on drying <5.01> Not more than 22.0z (6 hours). Total ash <5.01> Not more than 4.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
White, translucent rectangular column, string or flakes. Rectangular column about 26 cm in length, 4 cm square in cross section; a string of about 35 cm in length and about 3 mm in width; flakes about 3 mm in length; externally, with wrinkles and somewhat lustrous, light and pliable. Odorless; tasteless and mucilaginous. It is practically insoluble in organic solvents. A boiling solution of Agar (1 in 100) is neutral.
(1) To a fragment of Agar add dropwise iodine TS: a dark blue to reddish purple color develops. (2) Dissolve 1 g of Agar in 65 mL of water by boiling for 10 minutes with constant stirring, and add a sufficient amount of hot water to make up the water lost by evapora-tion: the solution is clear. Cool the solution between 309C and 399C: the solution forms a firm, resilient gel, which does not melt below 859C.
(1) Sulfuric acid—Dissolve 1.0 g of Agar in 100 mL of water by boiling: the solution is not acidic. Sulfurous acid and starch—To 5 mL of the solution obtained in (1) add 2 drops of iodine TS: the solution is not decolorized immediately, and does not show a blue color. Insoluble matter—To 7.5 g of Agar add 500 mL of water, boil for 15 minutes, and add water to make exactly 500 mL. Measure exactly 100 mL of the solution, add 100 mL of hot water, heat to boiling, filter while hot through a tared glass filter (G3), wash the residue with a small amount of hot water, and dry the residue at 1059C for 3 hours: the mass of the residue is not more than 15.0 mg. Water absorption—To 5.0 g of Agar add water to make 100 mL, shake well, allow to stand at 259C for 24 hours, and filter through moistened glass wool in a 100-mL graduated cylinder: the volume of the filtrate is not more than 75 mL. Loss on drying <5.01> Not more than 22.0z (6 hours). Total ash <5.01> Not more than 4.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Ajmaline, when dried, contains not less than 96.
Ajmaline occurs as a white to pale yellow crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in acetic anhydride and in chloroform, sparingly soluble in methanol, in ethanol (95), in acetone and in diethyl ether, and very slightly soluble in water. It dissolves in dilute hydrochloric acid. Melting point: about 1959C (with decomposition).
(1) Dissolve 0.05 g of Ajmaline in 5 mL of methanol, and use this solution as the sample solution. Add 3 mL of nitric acid to 1 mL of the sample solution: a deep red color develops. (2) Spot the sample solution of (1) on filter paper, and spray Dragendorff's TS: an orange color develops.
Alacepril, when dried, contains not less than 98.
Alacepril occurs as white, crystals or crystal-line powder. It is freely soluble in methanol, soluble in ethanol (95), and slightly soluble in water. It dissolves in sodium hydroxide TS.
(1) To 20 mg of Alacepril add 0.1 g of so-dium hydroxide, and heat gradually to melt: the gas evolved changes the color of a moisten red litmus paper to blue. After cooling, to the melted substance add 2 mL of water, shake, and add 1 mL of lead (II) acetate TS: a brown to black precipitate is formed. (2) Determine the infrared absorption spectrum of Alacepril, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －81 – －859(after drying, 0.25 g, ethanol (95), 25 mL, 100 mm).
(1) Chloride <1.03>—Dissolve 0.5 g of Alacepril in 30 mL of methanol, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test with this solution as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of meth-anol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate <1.14>—Dissolve 0.5 g of Alacepril in 30 mL of methanol, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.50 mL of 0.005 mol/L sulfuric acid VS add 30 mL of methanol, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Alacepril according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Alacepril in 5 mL of ethanol (95), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak other than alacepril from the sample solution is not larger than 2/5 times the peak area of alacepril from the standard solution, and the total area of the peaks other than alacepril from the sample solution is not larger than the peak area of alacepril from the standard solution. For the areas of the peaks, having the relative retention times of about 2.3 and about 2.6 to alacepril, multiply their correction factors, 1.5 and 1.9, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100), acetonitrile, methanol and tetrahydrofuran (6:2:1:1). Flow rate: Adjust so that the retention time of alacepril is about 5 minutes. Time span of measurement: About 3 times as long as the retention time of alacepril, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 4 mL of the standard solution add ethanol (95) to make exactly 10 mL. Confirm that the peak area of alacepril obtained with 10 mL of this solution is equivalent to 30 to 50z of that obtained with 10 mL of the standard solution. System performance: Dissolve 20 mg of Alacepril in 50 mL of a solution of propyl parahydroxybenzoate in ethanol (95) (1 in 80,000). When the procedure is run with 10 mL of this solution under the above operating conditions, alacepril ‌422 and propyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of alacepril is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Alacepril, previ-ously dried, dissolve in 75 mL of a mixture of methanol and water (2:1), and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 40.65 mg of C20H26N2O5S
Ethanol for Disinfection is a colorless, clear liquid. It is miscible with water. It burns with a light blue flame on ignition. It is volatile.
(1) To 1 mL of Ethanol for Disinfection add 2 mL of iodine TS and 1 mL of sodium hydroxide TS, and mix: light yellow precipitates appear. (2) To 1 mL of Ethanol for Disinfection add 1 mL of acetic acid (100) and 3 drops of sulfuric acid, and heat: the odor of ethyl acetate is produced. Specific gravity <2.56> d 15: 0.86027 – 0.87264
Proceed as directed in the Purity under Ethanol, with the exeption of (4), which is changed as follows. (4) Other impurities (absorbance)—Perform the test with Ethanol for Disinfection as directed under Ultraviolet-visible Spectorophotometry <2.24>: the absorbances at 240 nm, be-tween 250 nm and 260 nm and between 270 nm and 340 nm are not more than 0.40, 0.30, and 0.10, respectively. The ab-sorption spectrum determined in a 5-cm cell using water as a ‌JP XVIII Official Monographs / Ether 967‌ blank shows a smooth absorption curve between 235 nm and 340 nm.
Ethanol is a clear, colorless liquid. It is miscible with water. It is flammable and burns with a light blue flame on igni-tion. It is volatile.◆
Determine the infrared absorption spectrum of Ethanol as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Specific gravity <2.56> d 15: 0.80872 – 0.81601
(1) Clarity and color of solution—Dissolve 1.0 g of Ethambutol Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g Etham- butol Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand- Ethanol [64-17-5] This monograph is harmonized with the European Phar-macopoeia and the U.S. Pharmacopeia. The parts of the text that are not harmonized are marked with symbols (◆ ◆). Information on the harmonization with the European Pharmacopoeia and the U.S. Pharmacopeia is available on the website of the Pharmaceuticals and Medical Devices Agency. Ethanol contains not less than 95.1 volz and not more than 96.9 volz (by specific gravity) of ethanol (C H O) at 159C. ard Lead Solution (not more than 10 ppm). 2 6 Arsenic <1.11>—Prepare the test solution with 1.0 g of Ethambutol Hydrochloride according to Method 1, and perform the test (not more than 2 ppm). 2-Aminobutanol—Dissolve 5.0 g of Ethambutol Hy-drochloride in methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 0.05 g of 2-amino-1-butanol in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetic acid (100), hydrochloric acid and water (11:7:1:1) to a distance of about 10 cm, air-dry the plate, and heat at 1059C for 5 minutes. Cool, spray evenly ninhydrin-L-ascorbic acid TS upon the plate, air-dry the plate, and heat at 1059C for 5 minutes: the spot obtained from the sample Description Ethanol is a clear, colorless liquid. It is miscible with water. It is flammable and burns with a light blue flame on igni-tion. It is volatile.◆ Identification Determine the infrared absorption spectrum of Ethanol as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Specific gravity <2.56> d 15: 0.80872 – 0.81601 Purity (1) Clarity and color of solution—Ethanol is clear and colorless. To 1.0 mL of Ethanol add water to make 20 mL, and allow to stand for 5 minutes: the resulting liquid is clear. ‌JP XVIII Official Monographs / Anhydrous Ethanol 965 Control solution: water. Acidity or alkalinity—To 20 mL of Ethanol add 20 mL of freshly boiled and cooled water and 0.1 mL of a solu-tion prepared by addition of 7.0 mL of ethanol (95) and 2.0 mL of water to 1.0 mL of phenolphthalein TS : no color de-velops. Add 1.0 mL of 0.01 mol/L sodium hydroxide VS to this solution: a pink color develops. Volatile impurities—Pipet 500 mL of Ethanol, add 150 mL of 4-methylpentan-2-ol, and use this solution as the sample solution. Separately, to 100 mL of anhydrous metha-nol add Ethanol to make exactly 50 mL. Pipet 5 mL of this solution, add Ethanol to make exactly 50 mL, and use this solution as the standard solution (1). Separately, to 50 mL each of anhydrous methanol and acetaldehyde add Ethanol to make exactly 50 mL. To 100 mL of this solution add Ethanol to make exactly 10 mL, and use this solution as the standard solution (2). Separately, to 150 mL of acetal add Ethanol to make exactly 50 mL. To 100 mL of this solution add Ethanol to make exactly 10 mL, and use this solution as the standard solution (3). Separately, to 100 mL of benzene add Ethanol to make exactly 100 mL. To 100 mL of this solu-tion add Ethanol to make exactly 50 mL, and use this solu-tion as the standard solution (4). Perform the test with ex-actly 1 mL each of Ethanol, the sample solution and standard solutions (1), (2), (3) and (4) as directed under Gas Chroma-tography <2.02> according to the following conditions, and determine the peak areas of acetaldehyde, AE, benzene, BE and acetal, CE obtained with Ethanol, and the peak area of methanol with the standard solution (1), the peak area of acetaldehyde, AT with the standard solution (2), the peak area of acetal, CT with the standard solution (3) and the peak area of benzene, BT with the standard solution (4) by the au-tomatic integration method: the peak area of methanol ob- System suitability— System performance: When the procedure is run with 1 mL of the standard solution (2) under the above operating condi-tions, acetaldehyde and methanol are eluted in this order with the resolution between these peaks being not less than 1.5. Other impurities (absorbance)—Determine the ab-sorption spectrum of Ethanol between 235 nm and 340 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, in a 5-cm cell using water as a blank: the absorbances at 240 nm, between 250 nm and 260 nm and between 270 nm and 340 nm are not more than 0.40, 0.30, and 0.10, respectively, and the spectrum shows a steadily descending curve with no observable peaks or shoulders. Residue on evaporation—Evaporate 100 mL of Ethanol, exactly measured, in a tared dish on a water bath, and dry at 1059C for 1 hour: the mass of the residue does not exceed 2.5 mg.
◆Containers—Tight containers.◆ Storage—Without exposure to light.
Aldioxa is a condensation product of allantoin and aluminum hydroxide. When dried, it contains not less than 65.
Aldioxa occurs as a white powder. It is practically insoluble in water and in ethanol (99.5). It dissolves in dilute hydrochloric acid. A solution of Aldioxa in sodium fluoride-hydrochloric acid TS (1 in 100) shows no optical rotation. Melting point: about 2309C (with decomposition).
(1) Determine the infrared absorption spec-trum of Aldioxa, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. (2) To 0.2 g of Aldioxa add 10 mL of dilute hydrochloric acid, dissolve by warming, and cool: the solution responds to Qualitative Tests <1.09> for aluminum salt.
(1) Chloride <1.03>—To 0.10 g of Aldioxa add 6 mL of dilute nitric acid, boil to dissolve with shaking for 5 minutes, cool, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.142z). (2) Heavy metals <1.07>—To 1.0 g of Aldioxa add 3 mL of hydrochloric acid and 3 mL of water, heat gently to boil with shaking, and evaporate on a water bath to dryness. To the residue add 30 mL of water, shake under warming, cool, filter, and to the filtrate add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 3 mL of hydrochloric acid on a water bath to dry-ness, and add 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Loss on drying <2.41> Not more than 4.0z (1 g, 1059C, 2 hours).
(1) Allantoin—Weigh accurately about 0.1 g of Aldioxa, previously dried, dissolve in 50 mL of dilute sulfu-ric acid by heating, cool, and add water to make exactly 100 mL. Pipet 10 mL of this solution, and perform the test as directed under Nitrogen Determination <1.08>. Each mL of 0.005 mol/L sulfuric acid VS ＝ 0.3953 mg of C4H6N4O3 (2) Aluminum—Weigh accurately about 0.2 g of Aldioxa, previously dried, dissolve carefully in 50 mL of dilute hydrochloric acid by heating, cool, and add dilute hydrochloric acid to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 25 mL, and use this solution as the sample solution. Separately, pipet a suitable quantity of Standard Aluminum Stock Solution, dilute with water so that each mL of the solution contains not less than 16.0 mg and not more than 64.0 mg of aluminum (Al: 26.98), and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and calculate the alu-minum content of the sample solution from the calibration curve obtained from the absorbance of the standard solu-tion. Gas: Combustible gas—Acetylene. Supporting gas—Nitrous oxide. Lamp: An aluminum hollow cathode lamp. Wavelength: 309.2 nm.
Alimemazine Tartrate, when dried, contains not less than 98.
Alimemazine Tartrate occurs as a white pow-der. It is odorless, and has a bitter taste. It is freely soluble in water and in acetic acid (100), spar-ingly soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Alimemazine Tartrate in 50 mL of water is between 5.0 and 6.5. It is gradually colored by light.
(1) To 2 mL of a solution of Alimemazine Tartrate (1 in 100) add 1 drop of iron (III) chloride TS: a red-brown color is produced, and immediately a yellow pre-cipitate is formed. Dissolve 1 g of Alimemazine Tartrate in 5 mL of water, add 3 mL of sodium hydroxide TS, extract with two 10-mL portions of diethyl ether [use the aqueous layer ob-tained in the Identification (4)]. Shake the combined diethyl ether extracts with 3 g of anhydrous sodium sulfate, filter, and evaporate the diethyl ether with the aid of a current of air. Dry the residue in a desiccator (in vacuum, phosphorus (V) oxide) for 16 hours: it melts <2.60> between 669C and 709C. Determine the absorption spectrum of a solution of Alimemazine Tartrate (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. The aqueous layer, obtained in the identification (2), when neutralized with dilute acetic acid, responds to Qualita-tive Tests <1.09> (1) and (2) for tartrate.
(1) Clarity and color of solution—Dissolve 1.0 g of Alimemazine Tartrate in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Alimemazine Tartrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Alimemazine Tartrate according to Method 3, and per-form the test. Use a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 5) (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.8 g of Alimemazine Tar-trate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from red through brown to green-brown (indicator: 2 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.35 mg of (C18H22N2S)2.C4H6O6
Allopurinol, when dried, contains not less than 98.
Allopurinol occurs as white to pale yellow-white, crystals or crystalline powder. It is slightly soluble in N,N-dimethylformamide, and very slightly soluble in water and in ethanol (99.5). It dissolves in ammonia TS.
(1) Determine the absorption spectrum of a JP XVIII Official Monographs / Allopurinol Tablets 431 ‌solution of Allopurinol (1 in 200,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Allopurinol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Allopurinol according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Allopurinol according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Allopurinol in 10 mL of ammonia TS, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ammonia TS to make exactly 500 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose with fluorescent indicator for thin-layer chromatography. Develop the plate with ammonia TS-satu-rated 1-butanol to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.16 g of Allopurinol, previ-ously dried, dissolve in 70 mL of N,N-dimethylformamide by warming. Cool, and titrate <2.50> with 0.1 mol/L tetra-methylammonium hydroxide VS (potentiometric titration). To 70 mL of N,N-dimethylformamide add 12 mL of water, perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 13.61 mg of C5H4N4O
Alminoprofen, when dried, contains not less than 99.
Alminoprofen occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in ethanol (99.5) and in acetic acid (100), and very slightly soluble in water. It gradually turns brown on exposure to light. A solution of Alminoprofen in ethanol (99.5) (1 in 10) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Alminoprofen in ethanol (99.5) (3 in 500,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Alminoprofen as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
Weigh accurately about 0.3 g of Alminoprofen, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.93 mg of C13H17NO2 ‌JP XVIII Official Monographs / Alminoprofen Tablets 433
Alprazolam occurs as white, crystals or crys-talline powder. It is freely soluble in chloroform, soluble in methanol and in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in water. It dissolves in dilute nitric acid.
(1) Determine the absorption spectrum of a solution of Alprazolam in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Dissolve 0.05 g of Alprazolam in 0.7 mL of deu-terochloroform for nuclear magnetic resonance spectros-copy, and determine the spectrum of this solution using of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.67 kPa, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Alprazolam, previously dried, dissolve in 100 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 15.44 mg of C17H13ClN4
Alprenolol Hydrochloride, when dried, contains not less than 99.
Alprenolol Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water, in ethanol (95) and in acetic acid (100), slightly soluble in acetic anhydride, and practi-cally insoluble in diethyl ether.
(1) To 2 mL of a solution of Alprenolol Hydrochloride (1 in 100) add 0.05 mL of copper (II) sulfate TS and 2 mL of sodium hydroxide TS: a blue-purple color develops. To this solution add 1 mL of diethyl ether, shake well, and allow to stand: a red-purple color develops in the diethyl ether layer. Dissolve 0.05 g of Alprenolol Hydrochloride in 5 mL of water, add 1 to 2 drops of bromine TS, and shake: the color of the test solution disappears. Determine the absorption spectrum of a solution of Alprenolol Hydrochloride in ethanol (95) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Alprenolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Alprenolol Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. JP XVIII Official Monographs / Alprostadil 435 pH <2.54> Dissolve 1.0 g of Alprenolol Hydrochloride in 10 mL of water: the pH of this solution is between 4.5 and 6.0.
(1) Chloride <1.03>—Dissolve 0.5 g of Alprazolam in 10 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.20 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 2.0 g of Alprazolam according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Alprazolam in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, then pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop with a mixture of acetone, hexane, ethyl acetate and ethanol (95) (4:2:2:1) to a distance Description Alprenolol Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water, in ethanol (95) and in acetic acid (100), slightly soluble in acetic anhydride, and practi-cally insoluble in diethyl ether. Identification (1) To 2 mL of a solution of Alprenolol Hydrochloride (1 in 100) add 0.05 mL of copper (II) sulfate TS and 2 mL of sodium hydroxide TS: a blue-purple color develops. To this solution add 1 mL of diethyl ether, shake well, and allow to stand: a red-purple color develops in the diethyl ether layer. Dissolve 0.05 g of Alprenolol Hydrochloride in 5 mL of water, add 1 to 2 drops of bromine TS, and shake: the color of the test solution disappears. Determine the absorption spectrum of a solution of Alprenolol Hydrochloride in ethanol (95) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Alprenolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Alprenolol Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. JP XVIII Official Monographs / Alprostadil 435 pH <2.54> Dissolve 1.0 g of Alprenolol Hydrochloride in 10 mL of water: the pH of this solution is between 4.5 and 6.0. Melting point <2.60> 108 – 1129C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Alprenolol Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Alprenolol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Alprenolol Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). ‌Related substances—Dissolve 0.10 g of Alprenolol Hydrochloride in 10 mL of ethanol (95), and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add ethanol (95) to make exactly 100 mL. Pipet 2.5 mL of this solution, add ethanol (95) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, acetone, acetic acid (100) and water (60:42:5:3) to a distance of about 10 cm, air-dry the plate, and then dry at 809C for 30 minutes. After cooling, allow the plate to stand in iodine vapor for 30 minutes: the spots other than the principal spot and the spot on the starting point obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Alprenolol Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.58 mg of C15H23NO2.HCl
アルプロスタジル アルファデクス
Alprostadil Alfadex is a a-cyclodextrin clathrate compound of alprostadil. It contains not less than 2.
Alprostadil Alfadex occurs as a white powder. It is freely soluble in water, and practically insoluble in ethanol (95), in ethyl acetate and in diethyl ether. It is hygroscopic.
(1) Dissolve 0.02 g of Alprostadil Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, and centrifuge. Use the supernatant liquid as the sample solution (1). Separately, to 0.02 g of Alprostadil Alfadex add 5 mL of ethyl acetate, shake, and centrifuge. Use the supernatant liquid as the sample solution (2). Evaporate the solvent from these solutions under reduced pressure, add 2 mL of sulfuric acid to the residue, and shake for 5 minutes: the liquid obtained from the sample solution (1) shows an orange-yellow color, while the liquid obtained from the sample solu-tion (2) does not show that color. Dissolve 0.02 g of Alprostadil Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, and evaporate the solvent from the supernatant liquid under reduced pressure. Dissolve the residue in 2 mL of ethanol (95), add 5 mL of 1,3-dinitrobenzene TS, then add 5 mL of a solution of potassium hydroxide in ethanol (95) (17 in 100) under ice-cooling, and allow to stand for 20 minutes in a dark place under ice-cooling: a purple color develops. Dissolve 0.05 g of Alprostadil Alfadex in 1 mL of iodine TS, by heating on a water bath, and allow to stand: a dark blue precipitate is formed. Determine the absorption spectrum of a solution of Alprostadil Alfadex in dilute ethanol (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits no absorption between 220 nm and 400 nm. Separately, to 10 mL of the solution add 1 mL of potassium hydroxide-ethanol TS, allow to stand for 15 minutes, and determine the absorption spectrum as directed under Ultravi-olet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]20: ＋126 – ＋1389(0.1 g calcu-lated on the anhydrous basis, dilute ethanol, 20 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Alprostadil Alfadex in 20 mL of water: the pH of this solution is between 4.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Alprostadil Alfadex in 10 mL of water: the solution is col- ‌JP XVIII Official Monographs / Alum Solution 439 orless. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry within 30 minutes after preparation of the solution: the absorbance at 450 nm is not larger than 0.10. Prostaglandin A1—Dissolve 0.10 g of Alprostadil Alfadex in 5 mL of water, add exactly 5 mL of the internal standard solution and ethanol (95) to make 15 mL, and use this solution as the sample solution. Separately, dissolve 1.5 mg of prostaglandin A1 in ethanol (95) to make exactly 100 mL. Pipet 3 mL of this solution, add exactly 5 mL of the in-ternal standard solution, 2 mL of ethanol (95) and water to make 15 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the operating conditions described in the Assay, and calculate the ratios, QT and QS, of the peak area of prostaglandin A1 to that of the internal standard: QT is not larger than QS. Internal standard solution—A solution of propyl parahy-droxybenzoate in dilute ethanol (1 in 15,000). Related substances—Dissolve 0.10 g of Alprostadil Alfadex in 3 mL of water, add exactly 3 mL of ethyl acetate, shake, centrifuge, and use the supernatant liquid obtained as the sample solution. Separately, dissolve 1.0 mg of prostaglandin A1 in ethyl acetate to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of phosphomolybdic acid n-hydrate in ethanol (95) (1 in 4) on the plate, and heat the plate at 1009C for 5 minutes: the spots other than the principal spot obtained from the sample solution, and the spots other than the spot corresponding to the spot from the standard solution are all not more intense than the spot from the standard solution. Water <2.48> Not more than 6.0z (0.2 g, direct titration).
Weigh accurately about 0.1 g of Alprostadil Alfadex, dissolve in 5 mL of water, add exactly 5 mL of the internal standard solution and water to make 15 mL, and use this solution as the sample solution. Separately, weigh accurately about 3 mg of Alprostadil RS, dissolve in 5 mL of ethanol (95), add exactly 5 mL of the internal standard solu-tion and water to make 15 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of alprostadil to that of the internal standard. Amount (mg) of alprostadil (C20H34O5) ＝ MS × QT/QS MS: Amount (mg) of Alprostadil RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in dilute ethanol (1 in 15,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column about 5 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogenphosphate and acetonitrile (3:2). Flow rate: Adjust so that the retention time of alprostadil is about 6 minutes. Selection of column: Dissolve about 0.1 g of Alprostadil Alfadex in 5 mL of water, add 5 mL of a solution of prostaglandin A1 in ethanol (95) (3 in 200,000) and 5 mL of the internal standard solution. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of alprostadil, the internal standard and prostaglandin A1 in this order and complete separation of these peaks.
Containers—Tight containers. Storage—Light-resistant, at a temperature not exceeding
Alprostadil, when dried, contains not less than 97.
Alprostadil occurs as white, crystals or crystal-line powder. It is freely soluble in ethanol (99.5) and in tetrahydrofu-ran, slightly soluble in acetonitrile, and practically insoluble in water.
(1) The absorption spectrum of a solution of Alprostadil in ethanol (99.5) (1 in 100,000) determined as directed under Ultraviolet-visible Spectrophotometry <2.24> shows no absorption between 210 nm and 350 nm. Sepa-rately, to 10 mL of this solution add 1 mL of potassium hy-droxide-ethanol TS, allow to stand for 15 minutes, and de-termine the absorption spectrum in the same way. Compare the spectrum so obtained with the Reference Spectrum or the spectrum of a solution of Alprostadil RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Alprostadil, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Alprostadil RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －53 – －619(after drying, 25 mg, tetrahydrofuran, 5 mL, 100 mm).
Related substances—Dissolve 4 mg of Alprostadil in 2 mL of a mixture of acetonitrile for liquid chromatography and water (9:1), and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, and add the mixture of acetonitrile for liquid chromatography and water (9:1) to make exactly 10 mL. Pipet 2 mL of this solution, add the mixture of acetonitrile for liquid chromatography and water (9:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks, having the relative retention time of about 0.70 and 1.26 to alprostadil, ob-tained from the sample solution is not larger than 1/2 times the peak area of alprostadil from the standard solution, the area of the peaks, having the relative retention time of about 0.88 and 1.18 to alprostadil, is not larger than the peak area of alprostadil from the standard solution, the area of the peaks other than alprostadil and the peaks mentioned above is not larger than 1/10 times the peak area of alprostadil from the standard solution and the total area of the peaks other than alprostadil is not larger than 2 times the peak area of alprostadil from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of alprostadil, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 2 mL of the standard solution, add the mixture of acetonitrile for liq-uid chromatography and water (9:1) to make exactly 20 mL. Confirm that the peak area of alprostadil obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of ‌436 the standard solution. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of alprostadil is not more than 1.5z. Loss on drying <2.41> Not more than 1.0z (0.1 g, in vacu-um, phosphorus (V) oxide, 4 hours).
Weigh accurately about 5 mg each of Alprostadil and Alprostadil RS, previously dried, dissolve in exactly 5 mL of the internal standard solution, add a mixture of aceto-nitrile for liquid chromatography and water (9:1) to make 50 mL, and use these solutions as the sample solution and standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of alprostadil to that of the internal standard. Alprostadil Injection
硫酸アルミニウムカリウム水和物
Aluminum Potassium Sulfate Hydrate contains not less than 99.
Aluminum Potassium Sulfate Hydrate occurs as colorless or white, crystals or powder. It is odorless. It has a slightly sweet, strongly astringent taste. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. A solution of Aluminum Potassium Sulfate Hydrate (1 in 20) is acid.
A solution of Aluminum Potassium Sulfate Hydrate (1 in 10) responds to Qualitative Tests <1.09> for aluminum salt, to Qualitative Tests <1.09> (1), (3) and (4) for potassium salt, and to the Qualitative Tests <1.09> (1) and (3) for sulfate.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Aluminum Potassium Sulfate Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Aluminum Potassium Sulfate Hydrate according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.6 g of Aluminum Potassium Sulfate Hydrate, according to Method 1, and perform the test (not more than 3.3 ppm).
Weigh accurately about 4.5 g of Aluminum Potas-sium Sulfate Hydrate, and dissolve in water to make exactly 200 mL. Take exactly 20 mL of this solution, and add ex-actly 30 mL of 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS and 20 mL of acetic acid-ammo-nium acetate buffer solution (pH 4.8), boil for 5 minutes, and cool. Add 55 mL of ethanol (95), and titrate <2.50> with 0.05 mol/L zinc acetate VS (indicator: 2 mL of dithizone TS), until the color of the solution changes from light dark green to light red. Perform a blank determination in the same manner. Each mL of 0.05 mol/L disodium dihydorogen ethylenediamine tetraacetate VS ＝ 23.72 mg of AlK(SO4)2.12H2O
Amantadine Hydrochloride, when dried, contains not less than 99.
Amantadine Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is very soluble in formic acid, freely soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether.
(1) To 0.1 g of Amantadine Hydrochloride add 1 mL of pyridine and 0.1 mL of acetic anhydride, dis-solve by boiling for 1 minute, add 10 mL of dilute hydro-chloric acid, and cool in ice water. Filter the crystals sepa-rated, wash with water, and dry at 1059C for 1 hour: the residue melts <2.60> between 1479C and 1519C. Determine the infrared absorption spectrum of Amantadine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Amantadine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Amantadine Hydrochloride in 5 mL of water: the pH of this solution is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Amantadine Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Amanta- dine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Amantadine Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Amantadine Hydrochloride in 10 mL of water, add 10 mL of sodium hy-droxide TS and 10 mL of chloroform, and shake. Filter the chloroform layer through absorbent cotton with 3 g of anhy-drous sodium sulfate on a funnel, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions. Determine each peak area of these solutions by the automatic integration method: each peak area other than amantadine obtained from the sample solution is not larger than 1/3 times the peak area of amantadine from the standard solution, and the total area of each peak is not larger than the peak area of amantadine from the standard solution. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column about 3 mm in inside diameter and about 2 m in length, packed with siliceous earth for gas chromatography (150 to 180 mm in particle diameter) coated with a mixture (L) of branched hydrocarbon of petroleum hexamethyltetracosane group for gas chromatography and potassium hydroxide at the ratios of 2z and 1z, respec-tively. Column temperature: Inject at a constant temperature of about 1259C, maintain the temperature for 5 minutes, raise at the rate of 59C per minute to 1509C, and maintain at a constant temperature of about 1509C for 15 minutes. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of amantadine is about 11 minutes. Selection of column: Dissolve 0.15 g of naphthalene in 5 mL of the sample solution, and add chloroform to make 100 mL. Proceed with 2 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of naphthalene and amantadine in this order with the resolution between these peaks being not less than 2.5. Detection sensitivity: Adjust the detection sensitivity so that the peak height of amantadine obtained from 2 mL of the standard solution composes about 10z of the full scale. Time span of measurement: About twice as long as the retention time of amantadine, beginning after the solvent ‌JP XVIII Official Monographs / Amidotrizoic Acid 445‌ peak. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.2 g of Amantadine Hy-drochloride, previously dried, dissolve in 2 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat on a water bath for 30 minutes. After cooling, add acetic acid (100) to make 70 mL, and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.77 mg of C10H17N.HCl
Ambenonium Chloride contains not less than 98.
Ambenonium Chloride occurs as a white pow-der. It is freely soluble in water, in methanol and in acetic acid (100), soluble in ethanol (95), and slightly soluble in acetic anhydride. It is hygroscopic. Melting point: about 2059C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ambenonium Chloride in methanol (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ambenonium Chloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Ambenonium Chloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Ambenonium Chloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Am- benonium Chloride according to Method 4, and perform the test. Use a solution of magnesium nitrate in ethanol (95) (1 in 5). Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Ambenonium Chloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 1 mL of this solu-tion, add methanol to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, formic acid and water (12:6:5) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 11.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Ambenonium Chlo-ride, and dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3). Titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.42 mg of C28H42Cl4N4O2
Amidotrizoic Acid, calculated on the dried basis, contains not less than 98.
Amidotrizoic Acid occurs as a white crystalline powder. It is odorless. It is slightly soluble in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether. It dissolves in sodium hydroxide TS.
(1) Heat 0.1 g of Amidotrizoic Acid over a flame: a purple gas is evolved. (2) Determine the infrared absorption spectrum of Amidotrizoic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g ‌446 of Amidotrizoic Acid in 10 mL of 0.2 mol/L sodium hydrox-ide TS: the solution is clear and colorless. Primary aromatic amines—Dissolve 0.20 g of Amidotrizoic Acid in 5 mL of water and 1 mL of sodium hydroxide TS, add 4 mL of a solution of sodium nitrite (1 in 100) and 10 mL of 1 mol/L hydrochloric acid TS, shake, and allow to stand for 2 minutes. Add 5 mL of ammonium amidosulfate TS, shake well, allow to stand for 1 minute, and add 0.4 mL of a solution of 1-naphthol in ethanol (95) (1 in 10), 15 mL of sodium hydroxide TS and water to make exactly 50 mL. Determine the absorbance of this solution at 485 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24> using a solution, prepared in the same man-ner, as the blank: the absorbance is not more than 0.15. Soluble halides—Dissolve 2.5 g of Amidotrizoic Acid in 20 mL of water and 2.5 mL of ammonia TS, add 20 mL of dilute nitric acid and water to make 100 mL, allow to stand for 15 minutes with occasional shaking, and filter. Discard the first 10 mL of the filtrate, transfer the subsequent 25 mL of the filtrate to a Nessler tube, and add ethanol (95) to make 50 mL. Proceed as directed under Chloride Limit Test <1.03> using this solution as the test solution. Prepare the control solution as follows: to 0.10 mL of 0.01 mol/L hydro-chloric acid VS, add 6 mL of dilute nitric acid and water to make 25 mL, then ethanol (95) to make 50 mL. Iodine—Dissolve 0.20 g of Amidotrizoic Acid in 2.0 mL of sodium hydroxide TS, add 2.5 mL of 0.5 mol/L sul-furic acid TS, allow to stand for 10 minutes with occasional shaking, add 5 mL of chloroform, shake well, and allow to stand: the solution is colorless in the chloroform layer. Heavy metals <1.07>—Proceed with 2.0 g of Amidotrizoic Acid according to Method 2, and perform the Amikacin Sulfate
Amikacin Sulfate is the sulfate of a derivative of kanamycin. It contains not less than 691 mg (potency) and not more than 791 mg (potency) per mg, calculated on the dried basis.
Amikacin Sulfate occurs as a white to yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (95).
(1) Determine the infrared absorption spec-trum of Amikacin Sulfate, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of Amikacin Sulfate RS previously dried: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.1 g each of Amikacin Sulfate and Amika-cin Sulfate RS in 4 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer chromatog-raphy <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of water, ammonia water (28), methanol and tetrahydrofuran (1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin-citric acid-acetic acid TS on the plate, and heat at 1009C for 10 minutes: the principal spot obtained from the sample solution and the spot from the standard so-lution show a red-purple color and the same Rf value. A solution of Amikacin Sulfate (1 in 100) responds to Qualitative Tests <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: ＋76 – ＋849(1 g, water, 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Amikacin Sulfate in 100 mL of water: the pH of the solution is between 6.0 and 7.5.
(1) Heavy metals <1.07>—Proceed with 1.0 g of ‌JP XVIII Official Monographs / Amikacin Sulfate Injection 447 Amikacin Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Amikacin Sulfate in 4 mL of a water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of water, ammonia water (28), methanol and tetra-hydrofuran (1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin-citric acid-acetic acid TS on the plate, and heat the plate at 1009C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 4.0z (1 g, in vacu-um, 609C, 3 hours).
Transfer about 0.5 g of Amidotrizoic Acid, accu-rately weighed, to a saponification flask, dissolve in 40 mL of sodium hydroxide TS, add 1 g of zinc powder, connect to a reflux condenser, boil for 30 minutes, cool, and filter. Wash the flask and the filter paper with 50 mL of water, and combine the washings and the filtrate. Add 5 mL of acetic acid (100) to this solution, and titrate <2.50> with 0.1 mol/L silver nitrate VS until the color of the precipitate changes from yellow to green (indicator: 1 mL of tetrabromophenol-phthalein ethyl ester TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 20.46 mg of C11H9I3N2O4
Aminophylline Hydrate occurs as white to pale yellow, granules or powder. It is odorless or slightly ammo-nia-like odor, and has a bitter taste. It is soluble in water, slightly soluble in methanol, and practically insoluble in ethanol (95) and in diethyl ether. To 1 g of Aminophylline Hydrate add 5 mL of water, and shake: it dissolves almost completely. Separation of crystals begins in 2 to 3 minutes, and these crystals dissolve on the addition of a small amount of ethylenediamine. It is gradually affected by light, and gradually loses ethyl-enediamine in air.
(1) Dissolve 0.75 g of Aminophylline Hy-drate in 30 mL of water, and use this solution as the sample solution. To 20 mL of the sample solution add 1 mL of dilute hydrochloric acid: a precipitate is gradually formed. Filter the precipitate, recrystallize from water, and dry at 1059C for 1 hour: the crystals so obtained melt <2.60> between 2719C and 2759C. Dissolve 0.1 g of the crystals obtained in (1) in 50 mL of water, and to 2 mL of this solution add tannic acid TS dropwise: a white precipitate is produced, and this precipi-tate dissolves upon dropwise addition of tannic acid TS. To 0.01 g of the crystals obtained in (1) add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, and evaporate on a water bath to dryness: the residue shows a yellow-red color. Invert the dish containing the residue over a vessel containing 2 to 3 drops of ammonia TS: the color of the residue changes to red-purple, which is des-troyed on the addition of 2 to 3 drops of sodium hydroxide TS. Dissolve 0.01 g of the crystals obtained in (1) in 5 mL of water, add 3 mL of ammonia-ammonium chloride buffer solution (pH 8.0) and 1 mL of copper (II) sulfate-pyridine TS, and mix. Add 5 mL of chloroform to the mixture, and shake: the chloroform layer develops a green color. To 5 mL of the sample solution obtained in (1) add 2 drops of copper (II) sulfate TS: a purple color develops. Add Amount [mg (potency)] of amikacin (C ＝ MS × HT/HS 22H43N5O13) 1 mL of copper (II) sulfate TS: the color changes to blue, and green precipitates are formed on standing. MS: Amount [mg (potency)] of Amikacin Sulfate RS taken pH <2.54> Dissolve 1.0 g of Aminophylline Hydrate in 25 mL of water: the pH of the solution is between 8.0 and 9.5.
(1) Clarity and color of solution—Dissolve 1.0 g ‌JP XVIII Official Monographs / Aminophylline Injection 449 of Aminophylline Hydrate in 10 mL of hot water: the solu-tion is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Aminophylline Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Water <2.48> Not more than 7.9z (0.3 g, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Theophylline—Weigh accurately about 0.25 g of Aminophylline Hydrate, and dissolve in 50 mL of water and 8 mL of ammonia TS by gentle warming on a water bath. Add exactly 20 mL of 0.1 mol/L silver nitrate VS, warm on a water bath for 15 minutes, allow to stand be-tween 59C and 109C for 20 minutes, collect the precipitate by suction, and wash with three 10-mL portions of water. Combine the filtrate and washings, and add dilute nitric acid to make neutral. Add 3 mL of dilute nitric acid, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 18.02 mg of C7H8N4O2 (2) Ethylenediamine—Weigh accurately about 0.5 g of Aminophylline Hydrate, dissolve in 30 mL of water, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (indica-tor: 3 drops of bromophenol blue TS). Each mL of 0.1 mol/L hydrochloric acid VS ＝ 3.005 mg of C2H8N2
Amiodarone Hydrochloride, when dried, contains not less than 98.
Amiodarone Hydrochloride occurs as a white to pale yellow-white crystalline powder. It is very soluble in water at 809C, freely soluble in dichloromethane, soluble in methanol, sparingly soluble in ethanol (95), and very slightly soluble in water. Melting point: about 1619C (with decomposition).
(1) Determine the absorption spectrum of a solution of Amiodarone Hydrochloride in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Amiodarone Hydrochloride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 0.1 g of Amiodarone Hydrochloride add 10 mL of water, dissolve by warming at 809C, and cool: the solu-tion responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> To 1.0 g of Amiodarone Hydrochloride add 20 mL of freshly boiled and cooled water, dissolve by warming at 809C, and cool: the pH of this solution is between 3.2 and 3.8.
(1) Clarity and color of solution—Dissolve 0.5 g of Amiodarone Hydrochloride in 10 mL of methanol: the solution is clear, and is not more colored than the following control solutions (1) and (2). Control solution (1): To a mixture of 1.0 mL of Cobalt Chloride CS, 2.4 mL of Iron (III) Chloride CS and 0.4 mL of Copper (II) Sulfate CS, add diluted hydrochloric acid (1 in 40) to make 10.0 mL. To 2.5 mL of this solution add diluted hydrochloric acid (1 in 40) to make 20 mL. Control solution (2): To 3.0 mL of a mixture of 0.2 mL of Cobalt (II) Chloride CS, 9.6 mL of Iron (III) Chloride CS and 0.2 mL of Copper (II) Sulfate CS, add diluted hydro-chloric acid (1 in 40) to make 100 mL. Iodine—To 1.50 g of Amiodarone Hydrochloride add 40 mL of water, dissolve by warming at 809C, cool, add water to make exactly 50 mL, and use this solution as the sample stock solution. Pipet 15 mL of this solution, add exactly 1 mL of 0.1 mol/L hydrochloric acid TS and exactly 1 mL of a solution of potassium iodate (107 in 10,000), add water to make exactly 20 mL, and use this solution as the sample solution. Separately, pipet 15 mL of the sample stock solution, add exactly 1 mL of 0.1 mol/L hydrochloric acid TS, exactly 1 mL of a solution of potassium iodide (441 in 5,000,000) and exactly 1 mL of a solution of potassium iodate (107 in 10,000), add water to make exactly 20 mL, and use this solution as the standard solution. Separately, pipet 15 mL of the sample stock solution, add exactly 1 mL of 0.1 mol/L hydrochloric acid TS, add water to make ex-actly 20 mL, and use this solution as the control solution. Allow the sample solution, standard solution and control so-lution to stand in a dark place for 4 hours. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the control solution as the blank: the absorbance of the sam-ple solution at 420 nm is not larger than 1/2 times the absor-bance of the standard solution. Heavy metals <1.07>—Proceed with 1.0 g of Amioda- rone Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substance 1—Dissolve 0.5 g of Amiodarone Hydrochloride in 5 mL of dichloromethane, and use this solution as the sample solution. Separately, dissolve 10 mg of 2-chloroethyl diethylamine hydrochloride in 50 mL of dichloromethane, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, meth-anol and formic acid (17:2:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly bismuth subnitrate TS and then hydrogen peroxide TS: the spot obtained from the sample solution corresponding to the spot from the standard solution is not more intense than the spot from the standard solution. Related substance 2—Dissolve 0.125 g of Amioda-rone Hydrochloride in 25 mL of a mixture of water and acetonitrile for liquid chromatography (1:1), and use this so-lution as the sample solution. Pipet 2 mL of the sample solu-tion, and add a mixture of water and acetonitrile for liquid chromatography (1:1) to make exactly 50 mL. Pipet 1 mL of this solution, add a mixture of water and acetonitrile for liq-uid chromatography (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than amiodarone obtained from the sample solu-tion is not larger than the peak area of amiodarone from the standard solution, and the total area of the peaks other than amiodarone from the sample solution is not larger than 2.5 times the peak area of amiodarone from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: To 800 mL of water add 3.0 mL of acetic acid (100), adjust the pH to 4.95 with ammonia solution (28), and add water to make 1000 mL. To 300 mL of this so-lution add 400 mL of acetonitrile for liquid chromatography and 300 mL of methanol for liquid chromatography. ‌JP XVIII Official Monographs / Amiodarone Hydrochloride Tablets 451 Flow rate: Adjust so that the retention time of amioda-rone is about 24 minutes. Time span of measurement: About 2 times as long as the retention time of amiodarone. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water and acetonitrile for liq-uid chromatography (1:1) to make exactly 25 mL. Confirm that the peak area of amiodarone obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amiodarone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amiodarone is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.3 kPa, 509C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Amiodarone Hy-drochloride, previously dried, dissolve in 40 mL of a mixture of acetic anhydride and acetic acid (100) (3:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 68.18 mg of C25H29I2NO3.HCl
Amitriptyline Hydrochloride, when dried, contains not less than 99.
Amitriptyline Hydrochloride occurs as color-less crystals or a white to pale yellow crystalline powder. It has a bitter taste and a numbing effect. It is freely soluble in water, in ethanol (95) and in acetic acid (100), soluble in acetic anhydride, and practically in-soluble in diethyl ether. The pH of a solution of 1.0 g of Amitriptyline Hydrochlo-ride in 20 mL of water is between 4.0 and 5.0.
(1) Dissolve 5 mg of Amitriptyline Hydro-chloride in 3 mL of sulfuric acid: a red color develops. Add 5 drops of potassium dichromate TS to this solution: it turns dark brown. Acidify 1 mL of a solution of Amitriptyline Hydro-chloride (1 in 500) with 0.5 mL of dilute nitric acid, and add 1 drop of silver nitrate TS: a white, opalescent precipitate is produced. Determine the absorption spectrum of a solution of Amitriptyline Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Amitriptyline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 1.0 g of Amitriptyline Hydrochloride in 20 mL of water: the solu-tion is clear and colorless. (2) Heavy metals <1.07>—Proceed with 2.0 g of Amitrip- tyline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Amitriptyline Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.39 mg of C20H23N.HCl
Amlexanox, when dried, contains not less than 98.
Amlexanox occurs as white to yellowish white, crystals or crystalline powder. It is very slightly soluble in ethanol (99.5), and practically insoluble in water. It dissolves in diluted sodium hydroxide TS (1 in 3).
(1) Determine the absorption spectrum of a solution of Amlexanox in ethanol (99.5) (1 in 250,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Amlexanox RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Am-lexanox as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Amlexanox RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Amlexanox in 20 mL of water and 10 mL of sodium hydroxide TS, add 15 mL of dilute nitric acid and water to make 50 mL, centri-fuge, and then filter the supernatant liquid. To 25 mL of this filtrate add water to make 50 mL. Perform the test using this solution as the test solution. The control solution consists of 5 mL of sodium hydroxide TS, 7.5 mL of dilute nitric acid, 0.30 mL of 0.01 mol/L hydrochloric acid VS, and water ad-ded to make 50 mL (not more than 0.021z). Heavy metals <1.07>—Proceed with 1.0 g of Amlexa- nox according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—(i) Dissolve 30 mg of Amlexa-nox in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the area of the peak other than amlexanox obtained from the sample solution is not larger than 2 times the peak area of amlexanox from the standard solution. Operating conditions— The detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating condi-tions in the Assay. Mobile phase: Dissolve 7.2 g of disodium hydrogen phos-phate dodecahydrate in water to make 1000 mL. Adjust the pH of this solution to 8.0 by adding a solution prepared by dissolving 3.1 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water. To 400 mL of this solution add 600 mL of acetonitrile. Flow rate: To 15 mL of a solution of benzophenone in the mobile phase (3 in 1,000,000) add the mobile phase to make 20 mL. Adjust so that the retention time of benzophenone is about 6.5 minutes when perform the test with 10 mL of this solution under the conditions described above. Time span of measurement: About 3 times as long as the retention time of benzophenone, beginning after the peak of amlexanox. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of amlexanox obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Pipet 1 mL of the sample solution, and add the mobile phase to make 100 mL. To 5 mL of this solution add 15 mL of the solution of benzophenone in the mobile phase (3 in 1,000,000). When perform the test with 10 mL of this solution according to the above conditions, amlexanox and benzophenone are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlexanox is not more than 2.0z. (iii) The total amount of related substances, when calcu-lated according to the following formula, is not more than 0.5z. Total amount (z) of related substances ＝ {(AT1/AS1) ＋ (AT2/AS2)}× 1/10 Time span of measurement: Until completion of the elution of amlexanox, beginning after the solvent peak. System suitability— AT1 : Total area of the peaks other than amlexanox from the sample solution obtained in (i) System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of amlexanox obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlexanox is not more than 2.0z. (ii) Dissolve 30 mg of Amlexanox in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area of these so-lutions by the automatic integration method: the area of the AT2: Total area of the peaks other than amlexanox from the sample solution obtained in (ii) AS1: Peak area of amlexanox from the standard solution obtained in (i) AS2: Peak area of amlexanox from the standard solution obtained in (ii) Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 30 mg each of Amlexanox and Amlexanox RS, both dried, and dissolve them separately in the mobile phase to make exactly 50 mL. Pipet 5 mL each of these solutions, and add exactly 15 mL of the internal standard solution, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of amlexanox to that of the in-ternal standard, respectively. peak other than amlexanox obtained from the sample solu-tion is not larger than 2 times the peak area of amlexanox Amount (mg) of amlexanox (C 16H14N2O4) ＝ M S × QT/QS from the standard solution. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. MS: Amount (mg) of Amlexanox RS taken Internal standard solution—A solution of 3-nitroaniline in the mobile phase (1 in 4000). ‌JP XVIII Official Monographs / Amlexanox Tablets 455 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame- 50 mL. Pipet 25 mL of this solution, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Amlexanox. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Amount (mg) of amlexanox (C ＝ MS × QT/QS × V/200 16H14N2O4) 259C. Mobile phase: Dissolve 17.9 g of disodium hydrogen phos-phate dodecahydrate in water to make 1000 mL. Adjust the pH of this solution to 8.0 by adding a solution prepared by dissolving 7.8 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water. To 760 mL of this solution add 240 mL of acetonitrile. Flow rate: Adjust so that the retention time of amlexanox is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution according to the above condi-tions, amlexanox and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of amlexanox to that of the internal standard is not more than 1.0z.
Amlodipine Besilate contains not less than 98.
Amlodipine Besilate occurs as a white to yel-lowish white crystalline powder. It is freely soluble in methanol, sparingly soluble in ethanol (99.5), and slightly soluble in water. A solution of Amlodipine Besilate in methanol (1 in 100) shows no optical rotation. Melting point: about 1989C (with decomposition).
(1) Determine the absorption spectrum of a solution of Amlodipine Besilate in 0.01 mol/L hydrochloric acid-methanol TS (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Amlodipine Besilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Am-lodipine Besilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Amlodipine Besilate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. To 30 mg of Amlodipine Besilate add 0.1 g of sodium nitrate and 0.1 g of anhydrous sodium carbonate, mix, and gradually ignite. After cooling, dissolve the residue in 2 mL of dilute hydrochloric acid and 10 mL of water, filter if nec-essary, and add barium chloride TS: a white precipitate is formed.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Amlodipine Besilate according to Method 4, and perform the test. Prepare the control solution with 2.5 mL of Stand-ard Lead Solution (not more than 25 ppm). Related substances—Dissolve 0.10 g of Amlodipine Besilate in 50 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mixture of water and acetonitrile (1:1) to make exactly 100 mL. Pipet 3 mL of this solution, add the mixture of water and acetonitrile (1:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak having the relative Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 30 80 → 20 20 → 80 30 – 45 Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of amlodipine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and acetonitrile (1:1) to make exactly 10 mL. Confirm that the peak area of amlodi-pine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amlodipine are not less than 70,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlodipine is not more than 2.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 35 mg each of Amlodipine Besilate and Amlodipine Besilate RS (separately determine the water <2.48> using the same manner as Amlodipine Besi-late), dissolve them separately in the mobile phase to make exactly 250 mL. Pipet 5 mL each of these solutions, add ex-actly 5 mL each of the internal standard solution, add the mobile phase to make 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and ‌JP XVIII Official Monographs / Amlodipine Besilate Orally Disintegrating Tablets 457 QS, of the peak area of amlodipine to that of the internal standard. Amount (mg) of amlodipine besilate (C H ClN O .C H O S) lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak having the relative retention time of about 0.45 to amlodipine obtained from the sample solution is not larger than the peak area of 20 25 2 5 6 6 3 amlodipine from the standard solution, the area of the peak ＝ MS × QT/QS MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of isobutyl para-hydroxybenzoate in the mobile phase (3 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and a solution of potassium dihydrogen phosphate (41 in 10,000) (13:7). Flow rate: Adjust so that the retention time of amlodipine is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, amlodipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of amlodipine to that of the internal standard is not more than 1.0z.
Ammonia Water occurs as a clear, colorless liquid, having a very pungent, characteristic odor. It is alkaline. Specific gravity d 20: 0.95 – 0.96
(1) Hold a glass rod moistened with hydro-chloric acid near the surface of Ammonia Water: dense white fumes are produced. (2) Hold moistened red litmus paper near the surface of Ammonia Water: it turns blue.
(1) Residue on evaporation—Evaporate 10.0 mL of Ammonia Water to dryness, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 2.0 mg. (2) Heavy metals <1.07>—Evaporate 5.0 mL of Ammo- nia Water to dryness on a water bath, add 1 mL of dilute hydrochloric acid to the residue, and evaporate to dryness. Dissolve the residue in 2 mL of dilute acetic acid, add water
Pipet 5 mL of Ammonia Water, add 25 mL of water, and titrate <2.50> with 0.5 mol/L sulfuric acid VS (in-dicator: 2 drops of methyl red TS). Each mL of 0.5 mol/L sulfuric acid VS ＝ 17.03 mg of NH3
Amobarbital, when dried, contains not less than 99.
Amobarbital occurs as white, crystals or crys-talline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in ethanol (95), in acetone and in diethyl ether, sparingly soluble in chloroform, and practically in- soluble in water. It dissolves in sodium hydroxide TS and in sodium car-bonate TS. The pH of a saturated solution of Amobarbital is between 5.0 and 5.6.
(1) Boil 0.2 g of Amobarbital with 10 mL of sodium hydroxide TS: the gas evolved changes moistened red litmus paper to blue. Dissolve 0.05 g of Amobarbital in 2 to 3 drops of am-monia-ammonium chloride buffer solution (pH 10.7) and 5 mL of diluted pyridine (1 in 10). Add 5 mL of chloroform and 0.3 mL of copper (II) sulfate TS to the solution: a red-purple precipitate is produced in the aqueous layer. Shake the mixture: a red-purple color is produced in the chlo-roform layer. To 0.4 g of Amobarbital add 0.1 g of anhydrous sodium carbonate and 4 mL of water, shake, and add a solu-tion of 0.3 g of 4-nitrobenzyl chloride in 7 mL of ethanol (95). Heat the mixture on a water bath for 30 minutes under a reflux condenser, and allow to stand for 1 hour. Filter the crystals produced, wash with 7 mL of sodium hydroxide TS and a small portion of water, recrystallize from ethanol, and dry at 1059C for 30 minutes: the crystals so obtained melt <2.60> between 1689C and 1739C or between 1509C and 1549C.
(1) Clarity and color of solution—Dissolve 0.5 g of Amobarbital in 5 mL of sodium hydroxide TS: the solu-tion is clear and colorless. Chloride <1.03>—Dissolve 0.30 g of Amobarbital in 20 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: take 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of acetone and 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.035z). Sulfate <1.14>—Dissolve 0.40 g of Amobarbital in 20 mL of acetone, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: take 0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of acetone, and 1 mL of dilute hydrochloric acid, and add water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Amobar- bital according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Readily carbonizable substances <1.15>—Perform the test with 0.5 g of Amobarbital. The solution is not more colored than Matching Fluid A. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Amobarbital, previ-ously dried, and dissolve in 5 mL of ethanol (95) and 50 mL of chloroform. Titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until the color of the solution changes from yellow through light blue to purple (indicator: 1 mL of alizarin yellow GG-thymolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 22.63 mg of C11H18N2O3
Amosulalol Hydrochloride contains not less than 98.
Amosulalol Hydrochloride occurs as white crystals or a white crystalline powder. It has a bitter taste. It is very soluble in formic acid, freely soluble in metha- nol, and sparingly soluble in water and in ethanol (99.5). It is hygroscopic. A solution of Amosulalol Hydrochloride in methanol (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Amosulalol Hydrochloride (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Amosulalol Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Amosulalol Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Place 1.0 g of Amosulalol Hydrochloride in a porcelain crucible, add 1.5 mL of sulfuric acid, cover loosely, and heat gently to car-bonize. After cooling, add 2 mL of nitric acid, heat carefully until white fumes no longer are evolved, and then heat inten-sely to 500 – 6009C to incinerate. After cooling, add 2 mL of hydrochloric acid, proceed according to Method 2, and per-form the test. The control solution, processed in the same manner as the test solution using the same amounts of rea-gents, is prepared by combining 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Amosulalol Hydrochloride in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the total area of the peaks other than amosulalol obtained from the sample solution is not larger than 2/5 times the peak area of amosulalol from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.7 by adding a solution prepared by dissolving 3.58 g of disodi-um hydrogen phosphate dodecahydrate in water to make 1000 mL. To 670 mL of this solution add 330 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of amosulalol is about 7 minutes. Time span of measurement: About 2 times as long as the retention time of amosulalol, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. ‌JP XVIII Official Monographs / Amosulalol Hydrochloride Tablets 461 Confirm that the peak area of amosulalol obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amosulalol are not less than 4000 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amosulalol is not more than 1.0z. Water <2.48> Not more than 4.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Amosulalol Hydro-chloride, dissolve in 3 mL of formic acid, add 80 mL of a mixture of acetic acid (100) and acetic anhydride (3:2), and titrate <2.50> within 5 minutes with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.69 mg of C18H24N2O5S.HCl
Amoxapine, when dried, contains not less than ard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution.
Amoxapine occurs as white to light yellow- white, crystals or crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in ethanol (95) and in diethyl ether, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Amoxapine in 0.1 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption as the same wavelengths. Determine the infrared absorption spectrum of Amoxapine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Amoxapine as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Amoxapine according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). (2) Related substances—Dissolve 0.5 g of Amoxapine in 10 mL of a mixture of ethanol (95) and acetic acid (100) (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of ethanol (95) and acetic acid (100) (9:1) to make exactly 10 mL. Pipet 1 mL of this solution, add a mixture of ethanol (95) and acetic acid (100) (9:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethanol (95) and acetic acid (100) (9:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.4z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Amoxapine, previ- ‌JP XVIII Official Monographs / Amoxicillin Hydrate 463 ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to greenish blue (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 15.69 mg of C17H16ClN3O
Amoxicillin Hydrate contains not less than 950 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the anhydrous basis.
Amoxicillin Hydrate occurs as white to light yellow-white, crystals or crystalline powder. It is slightly soluble in water and in methanol, and very slightly soluble in ethanol (95).
Determine the infrared absorption spectrum of Amoxicillin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Amoxicillin RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋290 – ＋3159(0.1 g calcu-lated on the anhydrous basis, water, 100 mL, 100 mm).
(1) Heavy metals <1.07>—To 1.0 g of Amoxicillin Hydrate add 2 mL of a solution of magnesium sulfate hepta-hydrate (1 in 4), mix, and heat on a water bath to dryness. Carbonize the residue by gently heating. After cooling, add 1 mL of sulfuric acid, heat carefully, then heat at 500 – 6009C to incinerate. After cooling, add 1 mL of hydrochloric acid to the residue, and heat on a water bath to dryness. Then add 10 mL of water to the residue, and heat on a water bath to dissolve. After cooling, add ammonia TS to adjust the pH to 3 – 4, and add 2 mL of dilute acetic acid. If necessary, filter, wash the residue on the filter with 10 mL of water, transfer the filtrate and washings into a Nessler tube, add water to make 50 mL, and use this solution as the test solu-tion. Prepare the control solution as follows: To 2.0 mL of Standard Lead Solution add 2 mL of a solution of magne-sium sulfate heptahydrate (1 in 4), then proceed in the same manner as for preparation of the test solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Amoxicillin Hydrate according to Method 4, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Amoxicillin Hydrate in 50 mL of a solution of boric acid (1 in 200), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a solution of boric acid (1 in 200) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than amoxicillin obtained from the sample solution is not larger than the peak area of amoxicillin from the standard solution. Furthermore, the total area of the peaks other than amoxicillin from the sample solution is not larger than 3 times the peak area of amoxicillin from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate in 750 mL of water, adjust to pH 4.5 with acetic acid (31), and add water to make 1000 mL. To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of amoxicillin is about 8 minutes. Time span of measurement: About 4 times as long as the retention time of amoxicillin, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a solution of boric acid (1 in 200) to make exactly 10 mL. Confirm that the peak area of amoxi-cillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amoxicillin are not less than 2500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amoxicillin is not more than 1.0z. Water <2.48> Not less than 11.0z and not more than 15.0z (0.1 g, volumetric titration, direct titration).
Weigh accurately an amount of Amoxicillin Hydrate and Amoxicillin RS, equivalent to about 30 mg (potency), dissolve each in a solution of boric acid (1 in 200) to make exactly 100 mL, and use these solutions as the sample solu-tion and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the peak areas, AT and AS, of amoxicillin in each solution. Amount [ mg (potency)] of amoxicillin (C16H19N3O5S) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Amoxicillin RS taken ‌464 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.361 g of sodium acetate trihy-drate in 750 mL of water, adjust to pH 4.5 with acetic acid (31), and add water to make 1000 mL. To 950 mL of this so-lution add 50 mL of methanol. Flow rate: Adjust so that the retention time of amoxicillin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the number of theoretical plates of the peak of amoxicillin is not less than 2500. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amoxicillin is not more than 1.0z.
Amphotericin B is a polyene macrolide substance having antifungal activity produced by the growth of Streptomyces nodosus. dimethylsulfoxide, and add methanol to make 50 mL. To 1 mL of this solution add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Amphotericin B RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Purity Amphotericin A—Weigh accurately about 50 mg each of Amphotericin B and Amphotericin B RS, add ex-actly 10 mL each of dimethylsulfoxide to dissolve, and add methanol to make exactly 50 mL. Pipet 4 mL each of these solutions, add methanol to make exactly 50 mL, and use these solutions as the sample solution and standard solution (1), respectively. Separately, weigh accurately about 20 mg of Nystatin RS, add exactly 40 mL of dimethylsulfoxide to dissolve, then add methanol to make exactly 200 mL. Pipet 4 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution obtained in the same manner as the sample solution as the blank, and determine the absorbances at 282 nm and at 304 nm. Calcu-late the amount of amphotericin A by the following equa-tion: not more than 5z for Amphotericin B used for injec-tions, and not more than 15z for Amphotericin B not used for injections. Amount (z) of amphotericin A ＝ MS ×{(ASa1 × AT2) － (ASa2 × AT1)} × 25 MT ×{(ASa1 × ASb2) － (ASa2 × ASb1)} MS: Amount (mg) of Nystatin RS taken MT: Amount (mg) of Amphotericin B taken ASa1: Absorbance at 282 nm of the standard solution (1) ASb1: Absorbance at 282 nm of the standard solution (2) ASa2: Absorbance at 304 nm of the standard solution (1) ASb2: Absorbance at 304 nm of the standard solution (2) AT1: Absorbance at 282 nm of the sample solution AT2: Absorbance at 304 nm of the sample solution Loss on drying <2.41> Not more than 5.0z (0.1 g, in vacu-um, 609C, 3 hours). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Saccharomyces cerevisiae ATCC 9763 Culture medium—Use the medium 2) under (1) Agar media for seed and base layer. Preparation of cylinder-agar plate—Proceed as di- ‌466‌ rected in 1.5 Preparation of agar base layer plates under the Cylinder plate method, using Petri dish plates not dispensing the agar medium for base layer and dispensing 8.0 mL of the seeded agar medium. Standard solution—Use light-resistant vessels. Weigh accurately an amount of Amphotericin B RS equiva-lent to about 20 mg (potency), dissolve in dimethylsulfoxide to make exactly 20 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 24 hours. Take exactly a suitable amount of the standard stock solution before use, and add dimethylsulfoxide to make solutions so that each mL con-tains 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these solutions, add 0.2 mol/L phosphate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration standard solution and low concentra-tion standard solution, respectively. Sample solution—Use light-resistant vessels. Weigh accurately an amount of Amphotericin B equivalent to about 20 mg (potency), dissolve in dimethylsulfoxide to make exactly 20 mL, and use this solution as the sample stock so-lution. Take exactly a suitable amount of the sample stock solution, add dimethylsulfoxide to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency).
Amphotericin A—Weigh accurately about 50 mg each of Amphotericin B and Amphotericin B RS, add ex-actly 10 mL each of dimethylsulfoxide to dissolve, and add methanol to make exactly 50 mL. Pipet 4 mL each of these solutions, add methanol to make exactly 50 mL, and use these solutions as the sample solution and standard solution (1), respectively. Separately, weigh accurately about 20 mg of Nystatin RS, add exactly 40 mL of dimethylsulfoxide to dissolve, then add methanol to make exactly 200 mL. Pipet 4 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution obtained in the same manner as the sample solution as the blank, and determine the absorbances at 282 nm and at 304 nm. Calcu-late the amount of amphotericin A by the following equa-tion: not more than 5z for Amphotericin B used for injec-tions, and not more than 15z for Amphotericin B not used for injections. Amount (z) of amphotericin A ＝ MS ×{(ASa1 × AT2) － (ASa2 × AT1)} × 25 MT ×{(ASa1 × ASb2) － (ASa2 × ASb1)} MS: Amount (mg) of Nystatin RS taken MT: Amount (mg) of Amphotericin B taken ASa1: Absorbance at 282 nm of the standard solution (1) ASb1: Absorbance at 282 nm of the standard solution (2) ASa2: Absorbance at 304 nm of the standard solution (1) ASb2: Absorbance at 304 nm of the standard solution (2) AT1: Absorbance at 282 nm of the sample solution AT2: Absorbance at 304 nm of the sample solution Loss on drying <2.41> Not more than 5.0z (0.1 g, in vacu-um, 609C, 3 hours).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Saccharomyces cerevisiae ATCC 9763 Culture medium—Use the medium 2) under (1) Agar media for seed and base layer. Preparation of cylinder-agar plate—Proceed as di- ‌466‌ rected in 1.5 Preparation of agar base layer plates under the Cylinder plate method, using Petri dish plates not dispensing the agar medium for base layer and dispensing 8.0 mL of the seeded agar medium. Standard solution—Use light-resistant vessels. Weigh accurately an amount of Amphotericin B RS equiva-lent to about 20 mg (potency), dissolve in dimethylsulfoxide to make exactly 20 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 24 hours. Take exactly a suitable amount of the standard stock solution before use, and add dimethylsulfoxide to make solutions so that each mL con-tains 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these solutions, add 0.2 mol/L phosphate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration standard solution and low concentra-tion standard solution, respectively. Sample solution—Use light-resistant vessels. Weigh accurately an amount of Amphotericin B equivalent to about 20 mg (potency), dissolve in dimethylsulfoxide to make exactly 20 mL, and use this solution as the sample stock so-lution. Take exactly a suitable amount of the sample stock solution, add dimethylsulfoxide to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these solutions, add 0.2 mol/L phos-phate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Containers—Tight containers. Storage—Light-resistant, and in a cold place.
Ampicillin Hydrate contains not less than 960 mg (potency) and not more than 1005 mg (potency) per mg, calculated on the anhydrous basis.
Ampicillin Hydrate occurs as a white to light ＝ MS × QT/QS × 1000 16 19 3 4 yellow-white, crystals or crystalline powder. It is sparingly soluble in water, slightly soluble in metha- MS: Amount [mg (potency)] of Ampicillin RS taken Internal standard solution—A solution of guaifenesin in the mobile phase (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.94 g of diammonium hydrogen phosphate in 850 mL of water, add 100 mL of acetonitrile, adjust the pH to 5.0 with phosphoric acid, and add water to make exactly 1000 mL. Flow rate: Adjust so that the retention time of ampicillin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 40. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ampicillin to that of the internal standard is not more than 1.0z.
Determine the infrared absorption spectrum of Ampicillin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Ampicillin RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋280 – ＋3059(0.5 g calcu-lated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Ampicillin Hydrate in 400 mL of water is between 3.5 and 5.5.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ampicillin Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ampicillin hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Ampicillin Hydrate in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ampicillin obtained from the sample so-lution is not larger than the peak area of ampicillin from the standard solution, and the total area of the peaks other than ampicillin from the sample solution is not larger than 2 times the peak area of ampicillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 10 times as long as the retention time of ampicillin. JP XVIII Official Monographs / Ampicillin Hydrate 469 System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of ampicillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ampicillin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ampicillin is not more than 1.0z. N,N-Dimethylaniline—Weigh accurately about 1 g of Ampicillin Hydrate, dissolve in 5 mL of sodium hydroxide TS, add exactly 1 mL of the internal standard solution, shake vigorously for 1 minute, and use the upper layer liquid obtained after allowing it to stand as the sample solution. Separately, weigh accurately about 50 mg of N,N-dimethylaniline, dissolve in 2 mL of hydrochloric acid and 20 mL of water, add water to make exactly 50 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, and add water to make exactly 250 mL. Pipet 1 mL of this solution, add 5 mL of sodium hydroxide TS and exactly 1 mL of the internal standard solu-tion, shake vigorously for 1 minute, and use the upper layer liquid obtained after allowing it to stand as the standard peak area of N,N-dimethylaniline to that of the internal standard with the standard solution. System performance: Dissolve 50 mg of N,N-dimethylani-line in cyclohexane to make 50 mL. To 1 mL of this solution add the internal standard solution to make 50 mL, and use this solution as the solution for system suitability test. When the procedure is run with 1 mL of the solution for system suitability test under the above operating conditions, N,N-dimethylaniline and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 1 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the ratios of the peak area of N,N-dimethylaniline to that of the internal standard is not more than 2.0z. Water <2.48> 12.0 – 15.0z (0.1 g, volumetric titration, direct titration).
Weigh accurately an amount of Ampicillin Hydrate and Ampicillin RS, equivalent to about 50 mg (potency), dis-solve in a suitable volume of the mobile phase, add exactly 5 mL each of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of ampicillin to that of the internal standard. solution. Perform the test with 1 mL each of the sample Amount [ mg (potency)] of ampicillin (C H N O S) solution and standard solution as directed under Gas Chro-matography <2.02> according to the following conditions, ＝ MS × QT/QS × 1000 16 19 3 4 calculate the ratios, QT and QS, of the peak area of N,N-dimethylaniline to that of the internal standard, and calcu-late the amount of N,N-dimethylaniline by the following equation: not more than 20 ppm. Amount (ppm) of N,N-dimethylaniline ＝ MS/MT × QT/QS × 400 MS: Amount (g) of N,N-dimethylaniline taken MT: Amount (g) of Ampicillin Hydrate taken Internal standard solution—A solution of naphthalene in cyclohexane (1 in 20,000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 2.6 mm in inside diameter and 2 m in length, packed with siliceous earth for gas chromatog-raphy (180 – 250 mm in particle diameter) coated with 50z phenyl-50z methyl polysiloxane for gas chromatography at the ratio of 3z. Column temperature: A constant temperature of about 1209C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of N,N-dimethylaniline is about 5 minutes. System suitability— Test for required detectability: Measure exactly 1 mL of the standard stock solution, and add water to make exactly 250 mL. Pipet 1 mL of this solution, add 5 mL of sodium hydroxide TS and exactly 1 mL of the internal standard solu-tion, shake vigorously for 1 minute, and use the upper layer liquid obtained after allowing it to stand for the test. Con-firm that when the procedure is run with 1 mL of the upper layer liquid under the above operating conditions, the ratio of the peak area of N,N-dimethylaniline to that of the inter-nal standard is equivalent to 15 to 25z of the ratio of the MS: Amount [mg (potency)] of Ampicillin RS taken Internal standard solution—A solution of guaifenesin in the mobile phase (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.94 g of diammonium hydrogen-phosphate in 850 mL of water, add 100 mL of acetonitrile, adjust the pH to 5.0 with phosphoric acid, and add water to make exactly 1000 mL. Flow rate: Adjust so that the retention time of ampicillin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 40. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ampicillin to that of the internal standard is not more than 1.0z.
Ampiroxicam, when dried, contains not less than 99.
Ampiroxicam occurs as a white to yellowish white crystalline powder. It is freely soluble in acetic acid (100), soluble in aceto-nitrile, very slightly soluble in ethanol (99.5), and practically insoluble in water. A solution of Ampiroxicam in acetonitrile (1 in 20) shows no optical rotation. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Ampiroxicam in 0.01 mol/L hydrochloric acid-methanol TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Am-piroxicam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. icam from the standard solution. For the area of the peaks, having the relative retention time of about 0.17 and about 0.46 to ampiroxicam, multiply the correction factor, 0.37 and 0.60, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (3 in 500), methanol and acetonitrile (5:3:2). Flow rate: Adjust so that the retention time of ampiroxic-am is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of ampiroxicam, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add acetonitrile to make exactly 50 mL. Confirm that the peak area of ampiroxicam obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ampiroxicam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ampiroxicam is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ampiroxicam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Ampiroxicam in 50 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 200 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having a relative retention time of about 0.17 to ampiroxicam, obtained from the sample solution is not larger than 1/2 times the peak area of ampiroxicam from the standard solution, the area of the peak other than ampiroxicam and the peak mentioned above from the sample solution is not larger than 2/5 times the peak area of ampiroxicam from the standard solution, and the total area of the peaks other than ampiroxicam from the sample solution is not larger than the peak area of ampirox-
Weigh accurately about 0.22 g of Ampiroxicam, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.75 mg of C H N O S 20 21 3 7 Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Ampiroxicam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Ampiroxicam in 50 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 200 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having a relative retention time of about 0.17 to ampiroxicam, obtained from the sample solution is not larger than 1/2 times the peak area of ampiroxicam from the standard solution, the area of the peak other than ampiroxicam and the peak mentioned above from the sample solution is not larger than 2/5 times the peak area of ampiroxicam from the standard solution, and the total area of the peaks other than ampiroxicam from the sample solution is not larger than the peak area of ampirox-
Amyl Nitrite is the nitrous acid ester of 3-methyl-butanol-1 and contains a small quantity of 2-methyl-butanol-1 and the nitrous acid esters of other homo-logues.
Amyl Nitrite is a clear, light yellow-liquid, and has a characteristic, fruity odor. It is miscible with ethanol (95), and with diethyl ether. It is practically insoluble in water. It is affected by light and by heat. It is volatile at ordinary temperature and flammable even at a low temperature. Boiling point: about 979C
Determine the infrared spectrum of Amyl Nitrite as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. Specific gravity <2.56> d 20: 0.871 – 0.880
(1) Acidity—To 5 mL of Amyl Nitrite add a mix-ture of 1.0 mL of 1 mol/L sodium hydroxide VS, 10 mL of water and 1 drop of phenolphthalein TS, shake, and allow to stand for 1 minute: the light red color of the water layer does not disappear. Water—Allow 2.0 mL of Amyl Nitrite to stand in ice water: no turbidity is produced. Aldehyde—To 3 mL of a mixture of equal volumes of silver nitrate TS and aldehyde free-ethanol add ammonia TS dropwise until the precipitate first formed is redissolved. Add 1.0 mL of Amyl Nitrite, and warm between 609C and 709C for 1 minute: a brown to black color is not produced. Residue on evaporation—Evaporate 10.0 mL of ‌JP XVIII Official Monographs / Antipyrine 475‌ Amyl Nitrite on a water bath in a draft chamber, carefully protecting from flame, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Weigh accurately a volumetric flask containing 10 mL of ethanol (95), add about 0.5 g of Amyl Nitrite, and weigh accurately again. Add exactly 25 mL of 0.1 mol/L silver nitrate VS, then add 15 mL of potassium chlorate solu-tion (1 in 20) and 10 mL of dilute nitric acid, stopper the flask immediately, and shake it vigorously for 5 minutes. Dilute with water to make exactly 100 mL, shake, and filter through dry filter paper. Discard the first 20 mL of the fil-trate, measure exactly 50 mL of the subsequent filtrate, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammo-nium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 35.15 mg of C5H11NO2 ‌Containers and storage Containers—Hermetic containers not exceeding 10–ml capacity. Storage—Light-resistant, in a cold place, and remote from fire. Dental Antiformin Dental Sodium Hypochlorite Solution
Ethyl Aminobenzoate occurs as white, crystals or crystalline powder. It is odorless. It has a slightly bitter taste, numbing the tongue. It is freely soluble in ethanol (95) and in diethyl ether, and very slightly soluble in water. It dissolves in dilute hydrochloric acid.
(1) Dissolve 10 mg of Ethyl Aminobenzo-ate in 1 mL of dilute hydrochloric acid and 4 mL of water. This solution responds to Qualitative Tests <1.09> for prima-ry aromatic amines. Dissolve 0.1 g of Ethyl Aminobenzoate in 5 mL of water with the aid of dilute hydrochloric acid added drop-wise, and add iodine TS dropwise: a brown precipitate is produced. Warm 50 mg of Ethyl Aminobenzoate with 2 drops of acetic acid (31) and 5 drops of sulfuric acid: the odor of ethyl acetate is perceptible.
(1) Acidity—Dissolve 1.0 g of Ethyl Aminobenzo-ate in 10 mL of neutralized ethanol, and add 10 mL of water, 2 drops of phenolphthalein TS and 0.50 mL of 0.01 mol/L sodium hydroxide VS: a red color is produced. Chloride—Dissolve 0.20 g of Ethyl Aminobenzoate in 5 mL of ethanol (95), add 2 to 3 drops each of dilute nitric acid and of silver nitrate TS: no change occurs immediately. Heavy metals <1.07>—Dissolve 2.0 g of Ethyl Amino- benzoate in 20 mL of ethanol (95), add 2 mL of dilute acetic acid and ethanol (95) to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and sufficient ethanol (95) to make 50 mL (not more than 10 ppm). Readily carbonizable substances <1.15>—Perform the test with 0.5 g of Ethyl Aminobenzoate: the solution has no more color than Matching Fluid A. Loss on drying <2.41> Not more than 1.0z (1 g, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Ethyl Aminoben-zoate, previously dried, dissolve in 10 mL of hydrochloric acid and 70 mL of water, add 10 mL of a solution of potas-sium bromide (3 in 10), and cool to a temperature below 159C. Then titrate <2.50> with 0.1 mol/L sodium nitrite VS by the potentiometric titration or the amperometric titration. Each mL of 0.1 mol/L sodium nitrite VS ＝ 16.52 mg of C9H11NO2
Anesthetic Ether contains not less than 96z and not more than 98z (by specific gravity) of ether (C4H10O).
Anesthetic Ether occurs as a colorless, clear, mobile liquid, having a characteristic odor. It is miscible with ethanol (95). It is soluble in water. It is highly volatile and flammable. It is slowly oxidized by the action of air and light, with the formation of peroxides. Its vapor, when mixed with air and ignited, may explode violently. Boiling point: 35 – 379C Specific gravity <2.56> d 20: 0.718 – 0.721
(1) Foreign odor—Place 10 mL of Anesthetic Ether in an evaporating dish, and allow it to evaporate spon-taneously to a volume of about 1 mL: no foreign odor is per-ceptible. Drop this residue onto a piece of clean, odorless filter paper to evaporate the ether: no foreign odor is percep-tible. Acidity—Place 10 mL of diluted ethanol (95) (4 in 5) and 0.5 mL of phenolphthalein TS in a 50-mL glass-stoppered flask, and add 0.02 mol/L sodium hydroxide VS dropwise to produce a red color which persists after shaking for 30 seconds. Add 25 mL of Anesthetic Ether, stopper the flask, shake gently, and add 0.40 mL of 0.02 mol/L sodium hydroxide VS with shaking: a red color develops. Aldehyde—To 100 mL of water in a 200-mL glass-stoppered flask add 10 mL of Anesthetic Ether and 1 mL of a solution of sodium hydrogen sulfite (1 in 1000), stopper tightly, shake vigorously for 10 seconds, and allow the mix-ture to stand in a cool place for 30 minutes, protected from light. Add 2 mL of starch TS, and add dropwise 0.01 mol/L iodine VS until a pale blue color develops. Shake with about 2 g of sodium hydrogen carbonate to decolorize the solution, and add 1 mL of diluted 0.01 mol/L iodine VS (9 in 40): a blue color develops. Keep the temperature of the solution below 189C during the procedure. Peroxide—Place 10 mL of Anesthetic Ether in a Nessler tube, add 1 mL of a freshly prepared solution of po-tassium iodide (1 in 10), shake occasionally for 1 hour, pro-tecting from light, then add 1 mL of starch TS, and shake well: no color is produced and in the aqueous layer and in the ether layer. Residue on evaporation—Evaporate 50 mL of Anesthetic Ether, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Anhydrous Ampicillin contains not less than 960 mg (potency) and not more than 1005 mg (potency) per mg, calculated on the anhydrous basis.
Anhydrous Ampicillin occurs as white to light yellow-white, crystals or crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, very slightly soluble in ethanol (95), and practically in-soluble in acetonitrile.
Determine the infrared absorption spectrum of Anhydrous Ampicillin as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋280 – ＋3059(0.5 g calcu-lated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Anhydrous Ampicillin in 100 mL of water is be-tween 4.0 and 5.5.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Anhydrous Ampicillin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Anhydrous Ampicillin according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.05 g of Anhydrous Ampicillin in the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of each peak other than ampicillin obtained from the sample solution is not larger than the peak area of ampicillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: As long as about 10 times of ‌468 the retention time of ampicillin. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of ampicillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. Water <2.48> Not more than 2.0z (2.5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Anhydrous Ampicil-lin and Ampicillin RS, equivalent to about 50 mg (potency), add exactly 5 mL each of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ampicillin to that of the internal standard. Ampicillin Hydrate
Anhydrous Caffeine, when dried, contains not less than 98.
Anhydrous Caffeine occurs as white, crystals or powder. It is odorless, and has a bitter taste. It is freely soluble in chloroform, sparingly soluble in water, in acetic acid (100) and in acetic anhydride, and slightly soluble in ethanol (95) and in diethyl ether. The pH of a solution of 1.0 g of Anhydrous Caffeine in 100 mL of water is between 5.5 and 6.5.
(1) To 2 mL of a solution of Anhydrous Caffeine (1 in 500) add tannic acid TS dropwise: a white ‌JP XVIII Official Monographs / Caffeine Hydrate 589 precipitate, which dissolves upon the dropwise addition of tannic acid TS, is produced. To 0.01 g of Anhydrous Caffeine add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, and evaporate on a water bath to dryness: the residue acquires a yellow-red color. Invert the residue over a vessel containing 2 to 3 drops of ammonia TS: the color turns red-purple, and disappears upon the addition of 2 to 3 drops of sodium hy-droxide TS. Dissolve 0.01 g of Anhydrous Caffeine in water to make 50 mL. To 5 mL of this solution add 3 mL of diluted acetic acid (31) (3 in 100) and 5 mL of pyridine (1 in 10), mix, add 2 mL of diluted sodium hypochlorite TS (1 in 5), and allow to stand for 1 minute. Add 2 mL of sodium thio-sulfate TS and 5 mL of sodium hydroxide TS to the solution: a yellow color develops.
(1) Chloride <1.03>—Dissolve 2.0 g of Anhydrous Caffeine in 80 mL of hot water, cool rapidly to 209C, add water to make 100 mL, and use this solution as the sample solution. To 40 mL of the sample solution add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Sulfate <1.14>—To 40 mL of the sample solution ob- tained in (1) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Anhy-drous Caffeine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Anhydrous Caffeine in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 100 mL. Pipet 1 mL of this solution, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (9:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Readily carbonizable substances <1.15>—Perform the test using 0.5 g of Anhydrous Caffeine: the solution is not more colored than Matching Fluid D. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.4 g of Anhydrous Caffeine, previously dried, dissolve in 70 mL of a mixture of acetic anhydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the solution changes from purple through green to yellow (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.42 mg of C8H10N4O2
Anhydrous Citric Acid contains not less than 99.
Anhydrous Citric Acid occurs as colorless crystals, white granules or crystalline powder. It is very soluble in water, and freely soluble in ethanol (99.5).◆
Determine the infrared absorption spectrum of Anhydrous Citric Acid, previously dried at 1059C for 2 hours, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 2.0 g of Anhydrous Citric Acid in water to make 10 mL: the solu-tion is clear and colorless or has no more color than the fol-lowing control solutions (1), (2) or (3). Control solution (1): To 1.5 mL of Cobalt (II) Chloride CS and 6.0 mL of Iron (III) Chloride CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Control solution (2): To 2.5 mL of Cobalt (II) Chloride CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Control solution (3): To 0.15 mL of Cobalt (II) Chloride CS, 7.2 mL of Iron (III) Chloride CS and 0.15 mL of Cop-per (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Sulfates—Dissolve 2.0 g of Anhydrous Citric Acid in water to make 30 mL, and use this solution as the sample so-lution. Separately, dissolve 0.181 g of potassium sulfate in diluted ethanol (3 in 10) to make exactly 500 mL. Pipet 5 mL of this solution, and add diluted ethanol (3 in 10) to make exactly 100 mL. To 4.5 mL of this solution add 3 mL of a so-lution of barium chloride dihydrate (1 in 4), shake, and allow to stand for 1 minute. To 2.5 mL of this solution add 15 mL of the sample solution and 0.5 mL of acetic acid (31), and allow to stand for 5 minutes: the solution has no more turbidity than the following control solution (not more than 150 ppm). Control solution: Dissolve 0.181 g of potassium sulfate in water to make exactly 500 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and proceed in the same manner as above using this solution instead of the sample so-lution. ‌JP XVIII Official Monographs / Citric Acid Hydrate 769 Oxalic acid—Dissolve 0.80 g of Anhydrous Citric Acid in 4 mL of water, add 3 mL of hydrochloric acid and 1 g of zinc, and boil for 1 minute. After allowing to stand for 2 minutes, take the supernatant liquid, add 0.25 mL of a solution of phenylhydrazinium chloride (1 in 100), heat to boil, and then cool quickly. To this solution add the equal volume of hydrochloric acid and 0.25 mL of a solution of potassium hexacyanoferrate (III) (1 in 20), mix, and allow to stand for 30 minutes: the solution has no more color than the following control solution prepared at the same time (not more than 360 ppm expressed as oxalic anhydride). Control solution: To 4 mL of a solution of oxalic acid di-hydrate (1 in 10,000) add 3 mL of hydrochloric acid and 1 g of zinc, and proceed in the same manner as the test solution. (4) Heavy metals <1.07>—Proceed with 2.0 g of Anhy- drous Citric Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Readily carbonizable substances—Place 1.0 g of An-hydrous Citric Acid in a Nessler tube, add 10 mL of sulfuric acid, immediately heat in a 90 ± 19C water bath for 60 minutes, and cool quickly. Compare the color of 2.0 mL each of this solution and Matching Fluid K, using test tubes 12 mm in outside diameter, from a side against white back-ground: the solution is not more colored than the matching fluid. Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.55 g of Anhydrous Citric Acid, dissolve in 50 mL of water, and titrate with 1 mol/L sodium hydroxide VS (indicator: 1 drop of phenolphthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 64.04 mg of C6H8O7 ◆Containers and storage Containers—Tight containers.◆ Citric Acid Hydrate
Anhydrous Lactose occurs as white, crystals or powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).◆
Determine the infrared absorption spectrum of Anhydrous Lactose, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Anhydrous Lactose for Identification RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋54.4 – ＋55.99 Weigh ac-curately about 10 g of Anhydrous Lactose, calculated on the anhydrous basis, dissolve in 80 mL of water warmed to 509C, and add 0.2 mL of ammonia TS after cooling. After standing for 30 minutes, add water to make exactly 100 mL, and determine the optical rotation of this solution in a 100-mm cell.
(1) Clarity and color of solution—Dissolve 1.0 g of Anhydrous Lactose in 10 mL of boiling water, and allow to cool: the solution is clear, and colorless or nearly colorless and has no more color than the following control solution. Determine the absorbance at 400 nm of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, ‌1234 using water as the control solution: not more than 0.04. Control solution: To a mixture of 2.5 mL of Cobalt (II) Chloride CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II) Sulfate CS, add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity or alkalinity—Dissolve 6 g of Anhydrous Lactose by heating in 25 mL of freshly boiled and cooled water, and after cooling, add 0.3 mL of phenolphthalein TS: the solution is colorless, and not more than 0.4 mL of 0.1 mol/L sodium hydroxide VS is required to produce a pink or red color. (3) Heavy metals <1.07>—Proceed with 4.0 g of Anhy- drous Lactose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 5 ppm).◇ Proteins and light absorbing substances—Dissolve 1.0 g of Anhydrous Lactose in water to make 100 mL, and use this solution as the sample solution. Determine the ab-sorbances of the sample solution as directed under Ultravio-let-visible Spectrophotometry <2.24>, using water as the con-trol solution: not more than 0.25 at between 210 nm and 220 nm, and not more than 0.07 at between 270 nm and 300 nm. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 2 hours). Water <2.48> Not more than 1.0z (1 g, volumetric titra-tion, direct titration. Use a mixture of methanol for water determination and formamide for water determination (2:1) instead of methanol for water determination). Residue on ignition <2.44> Not more than 0.1z (1 g). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/g and 5 × 101 CFU/g, respectively, and ◇Salmonella and◇ Escherichia coli are not observed. Isomer ratio Place 10 mg of Anhydrous Lactose in a screw capped reaction vial for gas chromatography, add 4 mL of a mixture of pyridine, trimethylsilylimidazole and dimethylsul-foxide (117:44:39), stopper, and sonicate at room tempera-ture for 20 minutes. After cooling, transfer 400 mL of this solution into a vial for injection, add 1 mL of pyridine, stop-per tightly, mix, and use this fluid as the sample solution. Perform the test with 0.5 mL of the sample solution as di-rected under Gas Chromatography <2.02> according to the following conditions. Determine the peak areas of a-lactose and b-lactose, Aa and Ab, and calculate the contents (z) of a-lactose and b-lactose in Anhydrous Lactose by the follow-ing equations. Content (z) of a-lactose ＝ Aa/(Aa ＋ Ab) × 100 Flow rate: 2.8 mL per minute (Retention time of b-lactose is about 12 minutes). Splitless. System suitability— System performance: Prepare a solution with 10 mg of a mixture of a-lactose and b-lactose (1:1) in the same manner as for preparing the sample solution, and proceed with 0.5 mL of this solution under the above operating conditions, and determine the retention times of the peaks of a-lactose and b-lactose: the relative retention time of a-lactose to b-lactose is about 0.9 with the resolution between these peaks being not less than 3.0. System repeatability: When the test is repeated 6 times with 0.5 mL of the solution used in the system performance under the above operating conditions, the relative standard deviation of the peak area of b-lactose is not more than 5.0z.◇
Weigh accurately about 3 g of L-Lactic Acid, trans-fer in a conical flask, add accurately measured 40 mL of 1 mol/L sodium hydroxide VS, invert a watch glass over the flask, and heat on a water bath for 10 minutes. Titrate <2.50> the excess sodium hydroxide with 0.5 mol/L sulfuric acid VS immediately (indicator: 2 drops of phenolphthalein TS). Per-form a blank determination in the same manner. Each mL of 1 mol/L sodium hydroxide VS ＝ 90.08 mg of C3H6O3
Slightly viscous liquid or powder, milky white to light yellow in color. Odor, characteristic; taste, astrin-gent and acid.
To a portion of Royal Jelly, equivalent to 0.2 g of dried substance, add 5 mL of water, 1 mL of dilute hydrochloric acid and 10 mL of diethyl ether, shake for 15 minutes, and centrifuge. Take the diethyl ether layer, evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in 5 mL of methanol, and use this solution as the sample solution. Separately, dissolve 2 mg of 10-hydroxy-2-(E)-decenoic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, de-velop the plate with a mixture of 1-propanol and ammonia solution (28) (7:3) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution has the same color tone and Rf value with the spot from the stand-ard solution.
(1) Heavy metals <1.07>—Proceed with a portion of Royal Jelly, equivalent to 1.0 g of the dried substance, according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with an amount of Royal Jelly, equivalent to 0.40 g of the dried sub-stance according to Method 3, and perform the test (not more than 5 ppm). Loss on drying <5.01> The slightly viscous liquid: Not less than 57.0z and not more than 77.0z (6 hours). The powder: Not less than 7.0z and not more than 13.0z (6 hours). Total ash <5.01> Not more than 4.0z, calculated on the dried basis. Acid-insoluble ash <5.01> Not more than 0.5z, calculated on the dried basis.
Weigh accurately a portion of Royal Jelly, equiva-lent to 0.2 g of the dried substance, add 20 mL of methanol, sonicate for 30 minutes, and add methanol to make exactly 50 mL. Centrifuge this solution, pipet 2 mL of the superna-tant liquid, add exactly 2 mL of the internal standard solu-tion, then add 25 mL of water and methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of 10-hydroxy-2-(E)-decenoic acid for assay, dissolve in methanol to make exactly 100 mL. Pipet 3 mL of this solution, add exactly 2 mL of the internal standard solution, then add 25 mL of water and methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of 10-hydroxy-2-(E)-decenoic acid to that of the internal standard. Amount (mg) of 10-hydroxy-2-(E)-decenoic acid ＝ MS × QT/QS × 3/4 MS: Amount (mg) of 10-hydroxy-2-(E)-decenoic acid for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in methanol (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, methanol for liquid chromatography and phosphoric acid (550:450:1). Flow rate: Adjust so that the retention time of 10-hydroxy-2-(E)-decenoic acid is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, 10-hydroxy-2-(E)-decenoic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of 10-hydroxy-2-(E)-decenoic acid to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage—At not exceeding 109C.
Apricot Kernel Water contains not less than 0.
(1) Sulfate <1.14>—Add a few drops of 0.1 mol/L sodium hydroxide VS to 5.0 mL of Apricot Kernel Water to make slightly alkaline, evaporate on a water bath to dryness, and ignite between 4509C and 5509C. Dissolve the residue in 1.0 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.005z). Heavy metals <1.07>—Evaporate 50 mL of Apricot Kernel Water on a water bath to dryness, ignite between 4509C and 5509C, dissolve the residue in 5 mL of dilute acetic acid with warming, add water to make exactly 50 mL, and filter. Remove the first 10 mL of the filtrate, dilute the subsequent 20 mL to 50 mL with water, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 1 ppm). Free hydrogen cyanide—To 10 mL of Apricot Kernel Water add 0.8 mL of 0.1 mol/L silver nitrate VS and 2 to 3 drops of nitric acid at 159C, filter, and add 0.1 mol/L silver nitrate VS to the filtrate: no change occurs. Residue on evaporation—Evaporate 5.0 mL of Apricot Kernel Water to dryness, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Measure exactly 25 mL of Apricot Kernel Water, add 100 mL of water, 2 mL of potassium iodide TS and 1 mL of ammonia TS, and titrate <2.50> with 0.1 mol/L silver nitrate VS until a yellow turbidity persists. Each mL of 0.1 mol/L silver nitrate VS ＝ 5.405 mg of HCN
Aprindine Hydrochloride, when dried, contains not less than 98.
Aprindine Hydrochloride occurs as a white to pale yellow-white crystalline powder. It has a bitter taste, numbing the tongue. It is very soluble in water, in methanol and in acetic acid (100), and freely soluble in ethanol (99.5). It gradually turns brown on exposure to light.
(1) Dissolve 10 mg of Aprindine Hydro-chloride in a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Aprindine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Aprindine Hydrochloride (1 in 50) add 1 mL of dilute nitric acid: this solution re-sponds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Aprindine Hydrochloride in 50 mL of water: the pH of the solution is between 6.4 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Aprindine Hydrochloride in 10 mL of methanol: the solu-tion is clear, and its absorbance at 420 nm determined as di-rected under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.10. Heavy metals <1.07>—Proceed with 1.0 g of Aprin- dine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 25 mg of Aprindine Hydrochloride in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than aprindine obtained from the sample solution is not larger than 1/10 times the peak area of aprindine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.40 g of potassium dihydrogen phosphate in 500 mL of water, and adjust the pH to 3.0 with hydrochloric acid. To 500 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of aprindine is about 6 minutes. Time span of measurement: About 4 times as long as the retention time of aprindine. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of aprindine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of aprindine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of aprindine is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Aprindine Hydro-chloride, previously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.90 mg of C22H30N2.HCl
Arbekacin Sulfate is the sulfate of a derivative of dibekacin. It contains not less than 670 mg (potency) and not more than 750 mg (potency) per mg, calculated on the dried basis.
Arbekacin Sulfate occurs as a white powder. It is very soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 10 mg each of Arbekacin Sul-fate and Arbekacin Sulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia solution (28), methanol, chloroform and ethanol (95) (7:6:4:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the principal spot obtained from the sample solu-tion and the spot from the standard solution are purple-brown in color and their Rf values are the same. (2) A solution of Arbekacin Sulfate (1 in 50) responds to Qualitative Tests <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: ＋69 – ＋799 (0.25 g after drying, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.75 g of Arbekacin Sulfate in 10 mL of water is between 6.0 and 8.0.
(1) Clarity and color of solution—A solution obtained by dissolving 1.0 g of Arbekacin Sulfate in 5 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Arbeka- cin Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Dibekacin—Weigh accurately about 20 mg of Ar-bekacin Sulfate, add exactly 10 mL of the internal standard solution to dissolve, add water to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Dibekacin Sulfate RS, equivalent to about 10 mg (potency), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 10 mL of the inter-nal standard solution and water to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of dibekacin to that of the internal standard. Calculate the amount of dibekacin by the following equa-tion: not more than 2.0z. Amount (z) of dibekacin ＝ MS/MT × QT/QS × 1/10 × 100 MS: Amount [mg (potency)] of Dibekacin Sulfate RS taken MT: Amount (mg) of Arbekacin Sulfate taken Internal standard solution—A solution of bekanamycin sul-fate (1 in 2000). Operating conditions— Detector: Fluorometric detector (excitation wavelength: 340 nm, detection wavelength: 460 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Reaction coil: A column about 0.3 mm in inside diameter and about 3 m in length. Reaction coil temperature: A constant temperature of about 509C. Mobile phase: Dissolve 8.70 g of sodium 1-pentane sul-fonate and 8.52 g of anhydrous sodium sulfate in 980 mL of water, adjust the pH to 4.0 with acetic acid (100), and add water to make 1000 mL. To 230 mL of this solution add 20 mL of methanol. Reagent: Dissolve 12.36 g of boric acid in 960 mL of water, add 10 mL of a solution of o-phthalaldehyde in ethanol (99.5) (1 in 25), adjust the pH to 10.5 with 8 mol/L potassium hydroxide TS, and add water to make 1000 mL. To this solution add 1 mL of 2-mercaptoethanol. Reaction temperature: A constant temperature of about 509C. Flow rate of mobile phase: 0.5 mL per minute. Flow rate of reagent: 1 mL per minute. System suitability— System performance: Dissolve 20 mg each of Arbekacin Sulfate, becanamycin sulfate and dibekacin sulfate in 200 mL of water. When the procedure is run with 5 mL of this so-lution under the above operating conditions, becanamycin, arbekacin and dibekacin are eluted in this order, and the resolution between the peaks, becanamycin and arbekacin is not less than 5 and arbekacin and dibekacin is not less than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of dibekacin to that of the internal standard is not more than 2.0z. Related substances—Dissolve 20 mg of Arbekacin Sulfate in 20 mL of water, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add water to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the area of each peak by the automatic integration method: the total area of the peaks other than ar-bekacin and dibekacin obtained from the sample solution is not larger than the peak area of arbekacin from the standard solution. Operating conditions— Detector, column, column temperature, reaction coil, reaction coil temperature, mobile phase, reagent, reaction temperature, flow rate of mobile phase, and flow rate of rea-gent: Proceed as directed in the operating conditions in the Purity (3). Time span of measurement: About 1.5 times as long as the retention time of arbekacin. System suitability— System performance: Dissolve 10 mg each of Arbekacin Sulfate and dibekacin sulfate in 200 mL of water. When the procedure is run with 5 mL of this solution under the above operating conditions, arbekacin and dibekacin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of arbekacin is not more than 5.0z. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). ‌JP XVIII Official Monographs / Argatroban Hydrate 479‌
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer, having pH 7.8 – 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Arbekacin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solu-tion (pH 6.0) (1 in 2) to make exactly 50 mL, and use this so-lution as the standard stock solution. Keep the standard stock solution at 5 to 159C and use within 30 days. Take ex-actly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentra-tion standard solution and low concentration standard solu-tion, respectively. Sample solutions—Weigh accurately an amount of Arbekacin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Argatroban Hydrate occurs as white, crystals or crystalline powder. It has a bitter taste. It is freely soluble in acetic acid (100), sparingly soluble in methanol, slightly soluble in ethanol (99.5), and very slightly soluble in water. It is gradually decomposed on exposure to light.
(1) Determine the absorption spectrum of a solution of Argatroban Hydrate in ethanol (99.5) (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ar-gatroban Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋175 – ＋1859(0.2 g calcu-lated on the anhydrous basis, methanol, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Argatroban Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Incinerate 2.0 g of Argatroban Hy- drate according to Method 4. After cooling, add 10 mL of dilute hydrochloric acid to the residue, dissolve by warming on a water bath, and perform the test using this solution as the test solution. Add 10 mL of a solution of magnesium ni-trate hexahydrate in ethanol (95) (1 in 10), then add 1.5 mL of hydrogen peroxide (30), and fire to burn (not more than 1 ppm). Related substance 1—Dissolve 50 mg of Argatroban Hydrate in 40 mL of methanol, add water to make 100 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area from the sample solution by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of each peak other than argatroban is not more than 0.1z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase A: To 2.5 mL of acetic acid (100) add water to make 1000 mL, and adjust the pH to 5.0 with ammonia TS. To 500 mL of this solution add 500 mL of methanol. Mobile phase B: To 2.5 mL of acetic acid (100) add water to make 1000 mL, and adjust the pH to 5.0 with ammonia TS. To 200 mL of this solution add 800 mL of methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. the procedure is run with 10 mL of this solution under the above operating conditions, methyl benzoate and argatroban are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of argatroban is not more than 2.0z. Related substance 2—Dissolve 0.10 g of Argatroban Hydrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 3 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol, ethyl acetate and water (10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the number of spots other than the principal spot obtained from the sample solu-tion is not more than 2, and they are not more intense than the spot from the standard solution. Water <2.48> 2.5 – 4.5z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 50 mg of Argatroban Hydrate in 50 mL of methanol, add water to make 100 mL, and use this so-lution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine the areas of two adjacent peaks, Aa and Ab, having the reten-tion times of about 40 minutes, where Aa is the peak area of shorter retention time and Ab is the peak area of longer retention time: Ab/(Aa ＋ Ab) is between 0.30 and 0.40. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Time after injection 0 – 5 5 – 35 100 → 5 0 → 95 of sample (min) Mobile phase A (volz) Mobile phase B (volz) Column temperature: A constant temperature of about 409C. Mobile phase: To 500 mL of water add 500 mL of metha-nol, 13 mL of diluted 40z tetrabutylammonium hydroxide TS (1 in 4) and 0.68 mL of phosphoric acid, and adjust the pH to 6.8 with ammonia TS and diluted ammonia solution Flow rate: About 1.0 mL per minute. Time span of measurement: About 1.5 times as long as the retention time of argatroban, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase A to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of argatroban obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: Dissolve 5 mg of Argatroban Hy-drate and 5 mL of methyl benzoate in 40 mL of methanol, and add water to make 100 mL. To 5 mL of this solution add 40 mL of methanol and water to make 100 mL. When (28) (1 in 20). Flow rate: Adjust so that the retention time of the peak having the shorter retention time of the two peaks of ar-gatroban is about 40 minutes. System suitability— System performance: When the procedure is run with 10 mL of the sample solution under the above operating condi-tions, the resolution between the two peaks is not less than 1.2. System repeatability: When the test is repeated 6 times with 10 mL of the sample solution under the above operating conditions, the relative standard deviation of the total area of the two separate peaks of argatroban is not more than 2.0z.
Weigh accurately about 0.5 g of Argatroban Hy-drate, dissolve in 20 mL of acetic acid for nonaqueous titra-tion, add 40 mL of acetone for nonaqueous titration, and ‌JP XVIII Official Monographs / L-Arginine Hydrochloride 481‌ titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 50.86 mg of C23H36N6O5S
Aromatic Castor Oil is a colorless or yellow-ish, clear, viscous liquid. It has an aromatic odor.
To 3 g of Aromatic Castor Oil add 1 g of po-tassium hydroxide, and heat the mixture carefully to fuse: a characteristic odor is perceptible. Dissolve the fused matter in 30 mL of water, add an excess of magnesium oxide, and filter. Acidify the filtrate with hydrochloric acid: white crys-tals are produced.
Arotinolol Hydrochloride, when dried, contains not less than 99.
Arotinolol Hydrochloride occurs as a white to light yellow crystalline powder. It is freely soluble in dimethylsulfoxide, slightly soluble in methanol and in water, very slightly soluble in ethanol (99.5), and practically insoluble in diethyl ether. ‌JP XVIII Official Monographs / Arsenical Paste 483 A solution of Arotinolol Hydrochloride in methanol (1 in 125) does not show optical rotation.
(1) Determine the absorption spectrum of a solution of Arotinolol Hydrochloride in methanol (1 in 75,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Arotinolol Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Arotinolol Hydrochloride (1 in 200) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Arotinolol Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.05 g of Arotinolol Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 40 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol, acetone and ammonia solution (28) (30:10:10:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, in vacu-um, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1.5 g of Arotinolol Hydro-chloride, previously dried, dissolve in dimethylsulfoxide to make exactly 25 mL. Pipet 5 mL of this solution, add 100 mL of water and 5 mL of sodium hydroxide TS, and extract with three 50-mL portions of dichloromethane. Filter each dichloromethane extract through a pledget of absorbent cot-ton with anhydrous sodium sulfate on it. Evaporate com-bined filtrate to dryness in vacuum. Dissolve the residue in 70 mL of acetic acid (100), and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 20.40 mg of C15H21N3O2S3.HCl
Arsenic Trioxide, when dried, contains not less than 99.
Arsenic Trioxide occurs as a white powder. It is odorless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in sodium hydroxide TS.
Dissolve 0.2 g of Arsenic Trioxide in 40 mL of water by heating on a water bath: the solution responds to Qualitative Tests <1.09> for arsenite.
Clarity of solution—To 1.0 g of Arsenic Trioxide add 10 mL of ammonia TS, and heat gently: the solution is clear. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately about 0.15 g of Arsenic Trioxide, previously dried, dissolve in 20 mL of a solution of sodium hydroxide (1 in 25), by warming, if necessary. Add 40 mL of water and 2 drops of methyl orange TS, then add dilute hydrochloric acid until the color of the solution becomes light red. Add 2 g of sodium hydrogen carbonate and 50 mL of water to this solution, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 3 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 4.946 mg of As2O3
Arsenical Paste contains not less than 36.
Arsenical Paste is grayish black and has the odor of clove oil.
(1) Place 0.1 g of Arsenical Paste in a small flask, add 5 mL of fuming nitric acid and 5 mL of sulfuric acid, and heat over a flame until the reacting liquid becomes colorless and white fumes begin to evolve. After cooling, add the reacting liquid to 20 mL of water cautiously, and add 10 mL of hydrogen sulfide TS while warming: a yellow precipitate is produced (arsenic trioxide). Shake thoroughly 0.5 g of Arsenical Paste with 25 mL of diethyl ether, 5 mL of dilute hydrochloric acid and 20 mL of water, separate the water layer, and filter: 5 mL of the filtrate responds to Qualitative Tests <1.09> for primary aro-matic amines (procaine hydrochloride). Shake thoroughly 0.5 g of Arsenical Paste with 25 mL of diethyl ether and 25 mL of water, separate the water layer, filter, and use the filtrate as the sample solution. Dis-solve 0.01 g of procaine hydrochloride in 5 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia solution (28) (50:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 mm): the spots obtained from the sample solu-tion and standard solution exhibit the same Rf value.
Weigh accurately about 0.3 g of Arsenical Paste into a 150-mL Kjeldahl flask, add 5 mL of fuming nitric acid and 10 mL of sulfuric acid, and shake thoroughly. Heat cau-tiously the mixture, gently at first, and then continue strong heating, until red fumes of nitrogen oxide are sparingly evolved. After cooling, add 5 mL of fuming nitric acid, heat again until red fumes of nitrogen oxide are no longer evolved and the reacting liquid becomes clear, and cool. Add 30 mL of a saturated solution of ammonium oxalate monohydrate, heat again until white fumes of sulfuric acid are evolved, and continue the heating for 10 minutes. Decompose completely oxalic acid, cool, transfer cautiously the colorless reacting liquid to a glass-stoppered flask, containing 40 mL of water. Wash thoroughly the Kjeldahl flask with 60 mL of water, add the washings to the content of the glass-stoppered flask, and cool. Dissolve 3 g of potassium iodide in this solution, ‌484‌‌ allow to stand in a dark place at room temperature for 45 minutes, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 5 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 4.946 mg of As2O3
Aspoxicillin Hydrate contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Aspoxicillin Hydrate occurs as a white, crys-tals or crystalline powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in water, and practically insoluble in acetonitrile, in methanol and in ethanol (95).
(1) Determine the absorption spectrum of a solution of Aspoxicillin Hydrate (1 in 4000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Aspoxicillin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. pH <2.54> Dissolve 1.0 g of Aspoxicillin Hydrate in 50 mL of water: the pH of the solution is between 4.2 and 5.2.
(1) Clarity and color of solution—Dissolve 1.0 g of Aspoxicillin Hydrate in 50 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Aspoxi- cillin Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Aspoxicillin Hydrate according to Method 5, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.05 g of Aspoxicillin Hydrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas of each peak by the au-tomatic integration method: the area of each peak other than aspoxicillin obtained from the sample solution is not larger than 3/10 times the peak area of aspoxicillin from the stand-ard solution, and the total of peak areas other than aspox-icillin from the sample solution is not larger than the peak area of aspoxicillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 6 times as long as the retention time of aspoxicillin. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, add the mobile phase to make exactly 10 mL. Con-firm that the peak area of aspoxicillin obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of aspoxicillin is not more than 5z. Water <2.48> Not less than 9.5z and not more than 13.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately an amount of Aspoxicillin Hy-drate and Aspoxicillin RS, equivalent to about 0.1 g (po-tency), dissolve each in a suitable amount of water, add exactly 10 mL of the internal standard solution, 6.5 mL of acetonitrile and water to make 50 mL, and use these solu-tions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of these solu-tions as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of aspoxicillin to that of the in- ‌JP XVIII Official Monographs / Atenolol 491 ternal standard. Amount [ mg (potency)] of aspoxicillin (C21H27N5O7S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Aspoxicillin RS taken Internal standard solution—A solution of N-(3-hydroxy-phenyl)acetamide (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 130 mL of acetonitrile add potassium dihydrogenphosphate TS (pH 3.0) to make 1000 mL. Flow rate: Adjust so that the retention time of aspoxicillin is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, aspoxicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of aspoxicillin to that of the internal stand-ard is not more than 0.8z.
Atenolol, when dried, contains not less than 99.
Atenolol occurs as a white to pale yellow crys-talline powder. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (99.5), and slightly soluble in water. A solution of Atenolol in methanol (1 in 25) shows no op-tical rotation.
(1) Determine the absorption spectrum of a solution of Atenolol in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Atenolol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting Pint <2.60> 152 – 1569C
(1) Heavy metals <1.07>—Proceed with 1.0 g of Atenolol according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Atenolol in 25 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the area of the peak other than atenolol obtained from the sample solution is not larger than 1/2 times the peak area of atenolol from the standard solution, and the total area of the peaks other than atenolol from the sample solution is not larger than the peak area of atenolol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 226 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.0 with phosphoric acid. To 40 volume of this solution add 9 volume of methanol and 1 volume of tetrahydrofuran. Dissolve 1 g of sodium 1-octanesulfonate and 0.4 g of tetrabutylammo-nium hydrogensulfate in 1000 mL of this solution. Flow rate: Adjust so that the retention time of atenolol is about 8 minutes. Time span of measurement: About 4 times as long as the retention time of atenolol. System suitability— Test for required detectability: To exactly 10 mL of the standard solution add the mobile phase to make exactly 50 mL. Confirm that the peak area of atenolol obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of atenolol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of atenolol is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Atenolol, previ-ously dried, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.63 mg of C14H22N2O3
Atropine Sulfate Hydrate, when dried, contains not rately about 44 mg of Atorvastatin Calcium RS (separately determine the water <2.
Atropine Sulfate Hydrate occurs as colorless crystals or a white crystalline powder. It is odorless. It is very soluble in water and in acetic acid (100), freely soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: 188 – 1949C (with decomposition). Intro-duce a capillary tube charged with dried sample into a bath previously heated to 1809C, and continue to heat at a rate of rise of about 39C per minute. It is affected by light.
(1) To 1 mg of Atropine Sulfate Hydrate add 3 drops of fuming nitric acid, and evaporate the mixture on a water bath to dryness. Dissolve the residue in 1 mL of N,N-dimethylformamide, and add 5 to 6 drops of tetraethyl-ammonium hydroxide TS: a red-purple color develops. To 2 mL of a solution of Atropine Sulfate Hydrate (1 in 50) add 4 to 5 drops of hydrogen tetrachloroaurate (III) TS: a lusterless, yellow-white precipitate is formed. To 5 mL of a solution of Atropine Sulfate Hydrate (1 in 25) add 2 mL of ammonia TS, and allow to stand for 2 to 3 minutes. Collect the precipitate, wash with water, and dry in a desiccator (in vacuum, silica gel) for 4 hours: it melts <2.60> between 1159C and 1189C. A solution of Atropine Sulfate Hydrate (1 in 20) re-sponds to Qualitative Tests <1.09> for sulfate.
(1) Clarity and color of solution —Dissolve 0.5 g JP XVIII Official Monographs / Atropine Sulfate Injection 495 of Atropine Sulfate Hydrate in 10 mL of water: the solution is clear and colorless. ‌Acidity—Dissolve 1.0 g of Atropine Sulfate Hydrate in 20 mL of water, and add 0.30 mL of 0.02 mol/L sodium hydroxide VS and 1 drop of methyl red-methylene blue TS: a green color develops. Related substances—Dissolve 0.25 g of Atropine Sul-fate Hydrate in 1 mL of diluted hydrochloric acid (1 in 10), add water to make 15 mL, and use this solution as the sam-ple solution. To 5 mL of the sample solution add 2 to 3 drops of hydrogen hexachloroplatinate (IV) TS: no precipitate is formed. To 5 mL of the sample solution add 2 mL of ammo-nia TS, and shake vigorously: the turbidity of the solution is not greater than that of the following control solution. Control solution: To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL. To this solution add 1 mL of silver nitrate TS, and allow 7 mL of the mixture to stand for 5 minutes. D Hyoscyamine—Weigh accurately about 1 g of Atro-pine Sulfate Hydrate, previously dried, and dissolve in water to make exactly 10 mL: the specific optical rotation [a]20 <2.49> of this solution in a 100-mm cell is between －0.609 and ＋0.109. Readily carbonizable substances <1.15>—Take 0.20 g of Atropine Sulfate Hydrate, and perform the test: the solu-tion has no more color than Matching Fluid A. Loss on drying <2.41> Not more than 4.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 1109C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Dissolve about 0.25 g of Atropine Sulfate Hydrate, previously dried and accurately weighed, in 30 mL of acetic acid (100). If necessary, dissolve it by warming, and cool. Titrate <2.50> with 0.05 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 33.84 mg of (C17H23NO3)2.H2SO4
Auranofin, when dried, contains not less than 98.
Auranofin occurs as a white crystalline pow-der. It is very soluble in chloroform, freely soluble in metha-nol, sparingly soluble in ethanol (99.5), and practically in-soluble in water. It shows crystal polymorphism.
(1) To 50 mg of Auranofin add 3 mL of water, 3 mL of nitric acid and 3 mL of sulfuric acid, shake, and allow to stand: golden colored suspended matters are produced. Determine the infrared absorption spectrum of Auranofin as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Auranofin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Prepare the test solution with 1 mg of Auranofin as directed under Oxygen Flask Combustion Method <1.06>, using 10 mL of water as the absorbing liquid. Wash out the test solution into a Nessler tube with water to make 30 mL. Add 10 mL of dilute sulfuric acid, 3 mL of hexaammonium heptamolybdate-sulfuric acid TS and 0.1 mL of tin (II) chlo-ride TS, shake, and allow to stand for 10 to 15 minutes: a blue color is developed. D Optical rotation <2.49> [a]20: －54.0 – －62.09(after dry-ing, 0.2 g, methanol, 20 mL, 100 mm).
(1) Chloride <1.03>—Put 0.5 g of Auranofin in a porcelain crucible, add 0.25 g of anhydrous sodium carbon-ate, mix well, and ignite until the carbonized substance is dis-appeared. After cooling, add 20 mL of water, heat, and filter after cooling. Wash the residue with 20 mL of water, combine the filtrate and the washings, neutralize with dilute nitric acid, then add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: Dissolve 0.25 g of anhydrous sodium carbonate in 20 mL of water, neutralize with dilute nitric acid, add 0.50 mL of 0.01 mol/L hydrochloric acid, 6 mL of dilute nitric acid, and water to make 50 mL (not more than 0.036z). Heavy metals <1.07>—Proceed with 1.0 g of Auranofin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Put 0.5 g of Auranofin in a Kjeldahl flask, add cautiously 2 mL of sulfuric acid and 5 mL of nitric acid, and heat until the solution becomes almost color-less. After cooling, add 15 mL of a saturated solution of am-monium oxalate monohydrate, heat until white fumes are evolved, and concentrate to 1 to 2 mL. Then, add 3 mL of water and 1 drop of methyl orange TS, neutralize with am-monia solution (28), filter, and perform the test using the fil-trate as the test solution: the color is not darker than that of the following control solution. Control solution: Heat a mixture of 2 mL of sulfuric acid and 5 mL of nitric acid until white fumes are no longer evolved. After cooling, add 15 mL of a saturated solution of ammonium oxalate monohydrate, heat until white fumes are evolved, and concentrate to 1 to 2 mL. Add 3 mL of water and 1 drop of methyl orange TS, neutralize with ammonia solution (28), and filter. To the filtrate add 2.0 mL of Stand-ard Arsenic Solution, then proceed in the same manner as for the test solution (not more than 4 ppm). Related substances—Dissolve 50 mg of Auranofin in 5 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add chlo-roform to make exactly 100 mL. To exactly 3 mL of this so-lution add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform and acetone (4:1) to a distance of about 10 cm, and air-dry the plate. Dry the plate, furthermore, at 809C for 30 minutes. After cooling, allow the plate to stand in a iodine vapor for 30 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately about 20 mg each of Auranofin and Auranofin RS, both previously dried, dissolve each in 10 mL of a mixture of water and acetonitrile (1:1), and add exactly 5 mL each of the internal standard solution. Then add a mixture of water and acetonitrile (1:1) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of JP XVIII Official Monographs / Auranofin Tablets 497 ‌the peak area of auranofin to that of the internal standard. Amount (mg) of auranofin (C20H34AuO9PS) ＝ MS × QT/QS MS: Amount (mg) of Auranofin RS taken Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of water and acetonitrile (1:1) (3 in 1250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of sodium dihydrogen phosphate dihydrate solution (1 in 100), tetrahydrofuran and aceto-nitrile (12:5:3). Flow rate: Adjust so that the retention time of auranofin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, auranofin and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of auranofin to that of the internal standard is not more than 1.0z.
Usually quartered sections of a sphere, some-times warped or flattened, 4 – 8 cm in length, 2.5 – 4.5 cm in width and 0.5 – 0.8 cm in thickness; the outer surface is dark red-brown to grayish yellow-brown, with numerous small dents associated with oil sacs; the inner surface is white to light grayish yellow-red, with irregular indented reticulation left by vascular bundles; light and brittle in texture. Odor, characteristic aroma; taste, bitter, somewhat mucilaginous and slightly pungent.
To 1.0 g of Bitter Orange Peel add 10 mL of ethanol (95), allow to stand for 30 minutes with occasional shaking, filter, and use the filtrate as the sample solution. Separately, dissolve 10 mg of naringin for thin-layer chroma-tography in 10 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute 2,6-dibromo-N-chloro-1,4-benzo-quinone monoimine TS on the plate, and allow to stand in ammonia gas: one of the several spots obtained from the sample solution and a grayish green spot from the standard solution show the same color tone and the same Rf value. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 5.5z. Acid-insoluble ash <5.01> Not more than 0.5z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Bitter Orange Peel provided that 1 mL of silicon resin is previously added to the test sample in the flask: the volume of essential oil is not less than 0.2 mL.
Azathioprine, when dried, contains not less than 98.
Azathioprine is light yellow, crystals or crystal-line powder. It is odorless. It is sparingly soluble in N,N-dimethylformamide and in pyridine, very slightly soluble in water and in ethanol (99.5), and practically insoluble in chloroform and in diethyl ether. It dissolves in sodium hydroxide TS and in ammonia TS. It is gradually colored by light. Melting point: about 2409C (with decomposition).
(1) Dissolve 0.01 g of Azathioprine in 50 mL of water by warming. To 5 mL of this solution add 1 mL of dilute hydrochloric acid and 0.01 g of zinc powder, and allow to stand for 5 minutes: a yellow color is produced. Filter this solution: the filtrate responds to Qualitative Tests <1.09> for primary aromatic amines, and a red color is pro- duced. Dissolve 0.01 g of Azathioprine in 50 mL of water by warming. To 1 mL of this solution add 0.5 mL of phos-photungstic acid TS and 0.5 mL of dilute hydrochloric acid: a white precipitate is formed. Prepare the test solution by proceeding with 0.03 g of Azathioprine according to Oxygen Flask Combustion Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Azathioprine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths.
(1) Clarity and color of solution—Dissolve 0.5 g of Azathioprine in 50 mL of N,N-dimethylformamide: the solution is clear and shows a light yellow color. Acidity or alkalinity—Add 100 mL of water to 2.0 g of Azathioprine, shake well for 15 minutes, centrifuge for 5 minutes at 10,000 revolutions per minute, and filter. Discard the first 20 mL of the filtrate, add 2 drops of methyl red TS to 40 mL of the subsequent filtrate, and use this solution as the sample solution. Add 0.10 mL of 0.02 mol/L hydrochloric acid VS to 20 mL of the sample solution: a red color develops. Add 0.10 mL of 0.02 mol/L sodium hydroxide VS to 20 mL of the sample solution: a yellow color develops. Sulfate <1.14>—To 25 mL of the filtrate obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Azathioprine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Azathioprine, according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 10 mg of Azathioprine in 80 mL of the mobile phase by warming, cool, add the mobile phase to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the total area of the peaks other than azathioprine obtained from the sample solution is not larger than 1/2 times the peak area of azathioprine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 296 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 2.5 of a solution of 0.05 mol/L potassium dihydrogenphosphate TS (1 in 2) with diluted phosphoric acid (3 in 2000). To 800 mL of this solu-tion add 200 mL of methanol. Flow rate: Adjust so that the retention time of azathioprine is about 8 minutes. Time span of measurement: About three times as long as the retention time of azathioprine, beginning after the sol- ‌JP XVIII Official Monographs / Azathioprine Tablets 499 vent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add water to make exactly 50 mL. Con-firm that the peak area of azathioprine obtained with 20 mL of this solution is equivalent to 8 to 12z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg of Azathioprine in 80 mL of water by warming, cool, and add water to make 100 mL. To 2 mL of this solution add 2 mL of a solution, separately prepared by dissolving 0.06 g of benzoic acid in 3 mL of methanol and diluting with water to make 10 mL, and add the mobile phase to make 25 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, azathioprine and benzoic acid are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of azathioprine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Azathioprine, pre- as the sample solution. Separately, dissolve 0.1 g of Azathioprine RS in 10 mL of a solution of ammonia solution (28) in ethanol (95) (1 in 10), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethanol (95), ethyl acetate, and ammonia solution (28) (5:5:1) to a distance of about 7 cm, and air-dry the plate. Ex-amine under ultraviolet light (main wavelength: 254 nm): the spots obtained from the sample solution and the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Azathioprine Tablets add 1 mL of dimethyl-sulfoxide for ultraviolet-visible spectrophotometry per 5 mg of azathioprine (C9H7N7O2S), shake well, add 0.1 mol/L hy-drochloric acid TS to make exactly V mL so that each mL contains about 0.2 mg of azathioprine (C9H7N7O2S), and filter. Discard the first 20 mL of the filtrate, pipet 3 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of azathioprine (C H N O S) viously dried, add 80 mL of N,N-dimethylformamide, and warm to dissolve. After cooling, titrate <2.50> with 0.1 ＝ MS × AT/AS × V/500 9 7 7 2 mol/L tetramethylammonium hydroxide VS until the color of the solution changes from yellow through yellow-green to blue-green (indicator: 1 mL of thymol blue-dimethylfor-mamide TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 27.73 mg of C9H7O7O2S
Azelastine Hydrochloride occurs as a white crystalline powder. It is freely soluble in formic acid, and slightly soluble in water and in ethanol (99.5). Melting point: about 2259C (with decomposition). A solution of Azelastine Hydrochloride (1 in 200) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Azelastine Hydrochloride (3 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Azelastine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>: both spectra exhibit similar intensities of absorption at the same wave numbers. To 10 mL of a saturated solution of Azelastine Hydrochloride add 1 mL of dilute nitric acid, and filter to separate formed crystals: the filtrate responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Azelastine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). each peak area other than azelastine obtained from the sam-ple solution is not larger than 1/10 times the peak area of azelastine from the standard solution, and the total area of the peaks other than the peak of azelastine from the sample solution is not larger than 1/2 times the peak area of azelastine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of water, acetonitrile and per-chloric acid (660:340:1). Flow rate: Adjust so that the retention time of azelastine is about 10 minutes. Time span of measurement: About 2 times as long as the retention time of azelastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of azelastine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of azelastine is not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of azelastine is not more than 1.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of previously dried Azelastine Hydrochloride, dissolve in 5 mL of formic acid, add 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.84 mg of C22H24ClN3O.HCl
Azelnidipine occurs as a light yellow to yellow, crystalline powder or powder containing masses. It is freely soluble in ethanol (99.5) and in acetic acid (100), and practically insoluble in water. A solution of Azelnidipine in ethanol (99.5) (1 in 100) shows no optical rotation. Azelnidipine shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Azelnidipine in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Azel-nidipine as directed in the paste method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Azelnidipine according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Azelnidipine in a mixture of acetonitrile and water (4:1) to make 100 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add a mixture of acetonitrile and water (4:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the areas of the peak, having the relative retention time of about 0.50 and about 1.42 to azel-nidipine, obtained from the sample solution are not larger than 1/5 times and 3/10 times the peak area of azelnidipine from the standard solution, respectively, the area of the peak other than azelnidipine and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of azelnidipine from the standard solution, and the total area of the peaks other than azelnidipine from the sam-ple solution is not larger than 7/10 times the peak area of azelnidipine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.05 g of potassium dihydrogen phosphate in 350 mL of water, add 650 mL of a mixture of acetonitrile and methanol (7:3), and adjust to pH 5.5 with diluted phosphoric acid (1 in 10). Flow rate: Adjust so that the retention time of azelnidi-pine is about 36 minutes. Time span of measurement: About 2 times as long as the retention time of azelnidipine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (4:1) to make exactly 20 mL. Confirm that the peak area of azelnidipine obtained with 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 10 mL of the standard solu-tion. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of azelnidipine are not less than 15,000 and 0.8 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of azelnidipine is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 709C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Azelnidipine, dis-solve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.13 mg of C33H34N4O6
Azithromycin Hydrate occurs as a white crys-talline powder. It is freely soluble in methanol and in ethanol (99.5), and practically insoluble in water.
Determine the infrared absorption spectrum of Azithromycin Hydrate as directed in the potassium bro-mide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Azithromycin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －45 – －499(0.4 g calculated on the anhydrous basis, ethanol (99.5), 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Azithromycin Hydrate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not less than 4.0z and not more than 5.0z (0.4 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Azithromycin Hy-drate and Azithromycin RS, equivalent to about 50 mg (po-tency), dissolve each in an adequate amount of a mixture of acetonitrile and water (3:2), add exactly 2 mL of the internal standard solution and the mixture of acetonitrile and water (3:2) to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of azithromycin to that of the internal stand-ard. Amount [ mg (potency)] of azithromycin (C38H72N2O12) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Azithromycin RS taken Internal standard solution—A solution of 4,4?-bis(diethylamino)benzophenone in acetonitrile (3 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized poly-vinyl alcohol gel polymer for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.97 g of dipotassium hydrogen phosphate in about 750 mL of water, adjust the pH to 11.0 with potassium hydroxide TS, and add water to make 1000 mL. To 400 mL of this solution add 600 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of azithromy-cin is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, azithromycin and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of azithromycin to that of the internal stand-ard is not more than 1.0z.
Azosemide, when dried, contains not less than 99.
Azosemide occurs as a white to yellow-white crystalline powder. It is freely soluble in N,N-dimethylformamide, slightly soluble in methanol and in ethanol (99.5), and practically in-soluble in water. It dissolves in dilute sodium hydroxide TS. It is gradually colored to yellow by light. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 37.08 mg of C12H11ClN6O2S2
Weigh accurately about 0.6 g of Azosemide, previ-ously dried, dissolve in 50 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until the color of the solution changes from yel-low to yellow-green (indicator: 10 drops of thymol blue-N,N-dimethylformamide TS). Perform a blank determina-tion in the same manner with a solution prepared by adding 15 mL of ethanol (95) to 50 mL of N,N-dimethylformamide, 12 11 6 2 2 and make any necessary correction. 2-Chloro-5-(1H-tetrazol-5-yl)-4-[(thien-2-ylmethyl)amino] benzenesulfonamide [27589-33-9] Azosemide, when dried, contains not less than 99.0z and not more than 101.0z of azosemide (C12H11ClN6O2S2). Description Azosemide occurs as a white to yellow-white crystalline powder. It is freely soluble in N,N-dimethylformamide, slightly soluble in methanol and in ethanol (99.5), and practically in-soluble in water. It dissolves in dilute sodium hydroxide TS. It is gradually colored to yellow by light. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 37.08 mg of C12H11ClN6O2S2
Aztreonam contains not less than 920 mg (potency) and not more than 1030 mg (potency) per mg, calcu-lated on the anhydrous basis.
Aztreonam occurs as a white to yellowish white crystalline powder. It is freely soluble in dimethylsulfoxide, slightly soluble in water and in methanol, and very slightly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Aztreonam (3 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the JP XVIII Official Monographs / Aztreonam for Injection 507 spectrum with the Reference Spectrum or the spectrum of a solution of Aztreonam RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the spectrum of a solution of Aztreonam in deuterated dimethylsulfoxide for nuclear magnetic reso-nance spectroscopy (1 in 10), using a light hydrogen sub-stance existing in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy as an internal reference compound and 2.50 ppm for its chemical shift, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21> (1H): it exhibits a multiple signal at around d 1.5 ppm, and a single signal at around d 7.0 ppm. The ratio of integrated in-tensity of each signal is 9:1. D Optical rotation <2.49> [a]20: －26 – －329(0.25 g calcu-lated on the anhydrous bases, water, 50 mL, 100 mm). pH <2.54> Dissolve 0.05 g of Aztreonam in 10 mL of water: the pH of this solution is between 2.2 and 2.8. ‌Purity (1) Clarity and color of solution—Dissolve 1.0 g of Aztreonam in 20 mL of dimethylsulfoxide: the solution is clear, and its absorbance at 420 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.06. Heavy metals <1.07>—Proceed with 2.0 g of Aztreo- nam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 40 mg of Aztreonam in 100 mL of water, and use this solution as the sample solu-tion. Pipet 2 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area of both solutions by the automatic integration method: the area of the peak other than aztreonam obtained from the sample solution is not larger than the peak area of aztreonam from the standard solution, and the total area of peaks other than aztreonam from the sample solution is not larger than 2.5 times the peak area of aztreonam from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Time span of measurement: About 4 times as long as the retention time of aztreonam, beginning after the solvent peak. System suitability— Test for required detectability: To 5 mL of the standard solution add water to make 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the so-lution for system suitability test, and add water to make ex-actly 10 mL. Confirm that the peak area of aztreonam ob-tained with 25 mL of this solution is equivalent to 7 to 13z of that with 25 mL of the solution for system suitability test. System performance: When the procedure is run under the above operating conditions with 25 mL of the standard solu-tion obtained in the Assay, the internal standard and aztreo-nam are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak areas of aztreonam is not more than 2.0z. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Aztreonam and Az-treonam RS, equivalent to about 20 mg (potency), dissolve each in 70 mL of water, add exactly 10 mL of the internal standard solution and water to make 100 mL, and use these solutions as the sample solution and standard solution, re-spectively. Perform the test with 25 mL each of these solu-tions as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of aztreonam to that of the in-ternal standard. Amount [ mg (potency)] of aztreonam (C13H17N5O8S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Aztreonam RS taken Internal standard solution—A solution of 4-aminobenzoic acid (1 in 6250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.7 g of tetrabutylammonium hydrogensulfate in 300 mL of water, adjust to pH 3.0 with 0.5 mol/L disodium hydrogenphosphate TS, and add water to make 1000 mL. To 650 mL of this solution add 350 mL of methanol. Flow rate: Adjust so that the retention time of aztreonam is about 8 minutes. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the internal standard and aztreonam are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of aztreonam to that of the internal stand-ard is not more than 1.5z.
b-ガラクトシダーゼ（アスペルギルス）
b-Galactosidase (Aspergillus) contains an enzyme produced by Aspergillus oryzae.
b-Galactosidase (Aspergillus) occurs as a white to light yellow powder. It is slightly soluble in water with a turbidity, and practi-cally insoluble in ethanol (95) and in diethyl ether.
(1) Dissolve 25 mg of b-Galactosidase (Aspergillus) in 100 mL of water, then to 1 mL of this solu-tion add 9 mL of lactose substrate TS, and stand at 309C for 10 minutes. To 1 mL of this solution add 6 mL of glucose detection TS, and stand at 309C for 10 minutes: a red to red-purple color develops. (2) Dissolve 0.1 g of b-Galactosidase (Asperigillus) in 100 mL of water, and filter the solution if necessary. Deter-mine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Odor—b-Galactosidase (Aspergillus) has no ‌1054 any rancid odor. Heavy metals <1.07>—Proceed with 1.0 g of b-Galac-tosidase (Aspergillus) according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of b-Galactosidase (Aspergillus) according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 9.0z (0.5 g, in vacu-um, 809C, 4 hours). Residue on ignition <2.44> Not more than 3z (0.5 g). Nitrogen content Weigh accurately about 70 mg of b-Galactosidase (Aspergillus), and perform the test as directed under Nitrogen Determination <1.08>: the amount of nitro-gen (N: 14.01) is between 0.5z and 5.0z, calculated on the dried basis.
(i) Substrate solution—Dissolve 0.172 g of 2-nitrophenyl-b-D-galactopyranoside in disodium hydrogen-phosphate-citric acid buffer solution (pH 4.5) to make 100 mL. (ii) Procedure—Weigh accurately about 25 mg of b-Galactosidase (Aspergillus), dissolve in water to make ex-actly 100 mL, then pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the sample so-lution. Take exactly 3.5 mL of the substrate solution, stand at 30 ± 0.19C for 5 minutes, add exactly 0.5 mL of the sam-ple solution, immediately mix, and stand at 30 ± 0.19C for exactly 10 minutes, then add exactly 1 mL of sodium carbon-ate TS and mix immediately. Perfoum the test as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbance, A1, of this solution at 420 nm using water as the control. Separately, take exactly 3.5 mL of the substrate solution, add exactly 1 mL of sodium carbonate TS and mix, then add exactly 0.5 mL of the sample solution and mix. Determine the absorbance, A2, of this solution in the same manner as above. Units per g of b-Galactosidase (Aspergillus) ＝ 1/M × (A1 － A2)/0.917 × 1/0.5 × 1/10 0.917: Absorbance of 1 mmol/5 mL of o-nitrophenol M: Amount (g) of b-Galactosidase (Aspergillus) in the sample solution per mL Unit: One unit indicates an amount of the enzyme which decomposes 1 mmol of 2-nitrophenyl-b-D-galac-topyranoside in 1 minute under the above condi-tions.
Containers—Tight containers. Storage—In a cold place.
b-Galactosidase (Penicillium) contains an enzyme, having lactose decomposition activity, produced by Penicillium multicolor.
b-Galactosidase (Penicillium) occurs as a white to pale yellow-white, crystalline powder or powder. It is soluble in water with a turbidity, and practically in-soluble in ethanol (95). It is hygroscopic.
(1) Dissolve 0.05 g of b-Galactosidase (Penicillium) in 100 mL of water, then to 0.2 mL of this so-lution add 0.2 mL of lactose substrate TS for b-galactosidase (penicillium), and allow to stand at 309C for 10 minutes. To this solution add 3 mL of glucose detection TS for penicilli-um origin b-galactosidase, and allow to stand at 309C for 10 minutes: a red to red-purple color develops. (2) Dissolve 0.15 g of b-Galactosidase (Penicillium) in 100 mL of water, filter if necessary, and determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 278 nm and 282 nm.
(1) Odor—b-Galactosidase (Penicillium) has no any rancid odor. Heavy metals <1.07>—Proceed with 1.0 g of b-Galac- tosidase (Penicillium) according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of b-Galactosidase (Penicillium) according to Method 3, and perform the test (not more than 2 ppm). Nitrogen—Weigh accurately about 0.1 g of b-Galac-tosidase (Penicillium), and perform the test as directed under Nitrogen Determination <1.08>: not more than 3 mg of nitro-gen (N: 14.01) is found for each labeled 1000 Units. Protein contaminants—Dissolve 0.15 g of b-Galac-tosidase (Penicillium) in 4 mL of water, and use this solution as the sample solution. Perform the test with 15 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than the peak having retention time of about 19 minutes is not more than 75z of the total area of all peaks, and the areas of peaks other than the peaks having retention times of about 3, 16 and 19 minutes are not more than 15z of the total area of all peaks. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column about 7.5 mm in inside diameter and about 75 mm in length, packed with strongly acidic ion-exchange resin for liquid chromatography of sul-fopropyl group-binding hydrophilic polymer (10 mm in parti-cle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A solution obtained by dissolving 2.83 g of sodium acetate in 1000 mL of water, and adjusting to pH 4.5 with acetic acid (100) (mobile phase A), and a solution ob-tained by dissolving 29.2 g of sodium chloride in 1000 mL of mobile phase A (mobile phase B). Flow system: Adjust a linear concentration gradient from the mobile phase A to the mobile phase B immediately after injection of the sample so that the retention times of non-retaining protein and the enzyme protein are about 3 minutes and 19 minutes, respectively, when the flow runs 0.8 mL per minute, and then continue the running of the mobile phase B. Selection of column: Dissolve 15 mg of b-lactoglobulin in 4.5 mL of water, add 0.5 mL of a solution of cytosine (1 in 5000), and use this solution as the column-selecting solution. Proceed with 15 mL of the column-selecting solution under ‌JP XVIII Official Monographs / Gati‰oxacin Hydrate 1055‌ the above operating conditions, and calculate the resolution. Use a column giving elution of cytosine and b-lactoglobulin in this order with the resolution between these peaks being not less than 4. Detection sensitivity: Adjust the detection sensitivity so that the peak height of b-lactoglobulin from 15 mL of the column-selecting solution is between 5 cm and 14 cm. Time span of measurement: About 1.4 times as long as the retention time of b-lactoglobulin. Loss on drying <2.41> Not more than 5.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 2z (1 g).
(i) Substrate solution—Dissolve 0.603 g of 2-nitrophenyl-b-D-galactopyranoside in disodium hydrogen phosphate-citric acid buffer solution for penicillium origin b-galactosidase (pH 4.5) to make 100 mL. (ii) Procedure—Weigh accurately about 0.15 g of b-Galactosidase (Penicillium), dissolve in water with thorough shaking to make exactly 100 mL, and allow to stand at room temperature for an hour. Pipet 2 mL of this solution, add disodium hydrogen phosphate-citric acid buffer solution for penicillium origin b-galactosidase (pH 4.5) to make exactly 100 mL, and use this solution as the sample solution. Trans-fer exactly 0.5 mL of the sample solution to a test tube, stand at 30 ± 0.19C for 10 minutes, add exactly 0.5 mL of the substrate solution previously kept at 30 ± 0.19C, then mix immediately, and stand at 30 ± 0.19C for exactly 10 minutes. Then add exactly 1 mL of sodium carbonate TS, mix immediately to stop the reaction. To this solution add exactly 8 mL of water, mix, and use as the colored sample solution. Separately, pipet 0.5 mL of disodium hydrogen phosphate-citric acid buffer solution for penicillium origin b-galactosidase (pH 4.5), then proceed in the same manner as the sample solution, and use the solution so obtained as the colored blank solution. Perform the test with the colored sample solution and the colored blank solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and determine the absorbances, AT and AB, at 420 nm. Units per g of b-Galactosidase (Penicillium) ＝ 1/M × (AT － AB)/0.459 × 1/10 0.459: Absorbance of 1 mmol/10 mL of o-nitrophenol M: Amount (g) of b-Galactosidase (Penicillium) in 0.5 mL of the sample solution Unit: One unit indicates an amount of the enzyme which decomposes 1 mmol of 2-nitrophenyl-b-D-galac-topyranoside in 1 minute under the above condi-tions.
Bacampicillin Hydrochloride is a hydrochloride of ampicillin ethoxycarbonyloxyethyl ester. It contains not less than 626 mg (potency) and not more than 710 mg (potency) per mg, calculated on the anhydrous basis.
Bacampicillin Hydrochloride occurs as a white to pale yellow crystalline powder. It is freely soluble in methanol and in ethanol (95), and soluble in water.
(1) Determine the absorption spectrum of a solution of Bacampicillin Hydrochloride in methanol (1 in 1000) as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Bacampicillin Hy-drochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Bacampicillin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Bacampicillin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Bacampicillin Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: ＋140 – ＋1709(0.1 g calcu-lated on the anhydrous basis, ethanol (95), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Amount [mg (potency)] of aztreonam (C ＝ MS × QT/QS × 250 13H17N5O8S2) Bacampicillin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). MS: Amount [mg (potency)] of Aztreonam RS taken Internal standard solution—A solution of 4-aminobenzoic acid (1 in 6250).
Containers—Hermetic containers. Storage—Light-resistant.
Bacitracin A System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- C66 H103 N17 O16S: 1422.69 ditions, ampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ampicillin to that of the internal standard is not more than 2.0z. Water <2.48> Not more than 1.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 1.5z (1 g). Assay Weigh accurately an amount of Bacampicillin Hydrochloride and Bacampicillin Hydrochloride RS, equiva-lent to about 40 mg (potency), dissolve each in water to make exactly 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of bacampicillin in each solution. [22601-59-8] [1405-87-4, Bacitracin] Bacitracin is a mixture of peptide substances having antibacterial activity including bacitracin A as the main component produced by the growth of Bacillus subtilis or Bacillus licheniformis. It contains not less than 60 Units per mg, calculated on the dried basis.
Bacitracin occurs as a white to light brown powder. It is freely soluble in water, and slightly soluble in ethanol (99.5).
(1) To 3 mL of a solution of Bacitracin (1 in 100) add 3 mL of 4-dimethylaminobenzaldehyde TS, shake until red-rosy to red-purple color appears, then add several drops of a solution of sodium nitrite (1 in 100), and Amount [ mg (potency)] of ampicillin (C H N O S) shake: a green to dark green color is produced. ＝ MS × AT/AS × 1000 16 19 3 4 (2) Dissolve 60 mg each of Bacitracin and Bacitracin RS in 10 mL of water, and use these solutions as the sample so- MS: Amount [mg (potency)] of Bacampicillin Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 500 mL of diluted 2 mol/L sodium dihy-drogen phosphate TS (1 in 100), add diluted 0.05 mol/L dis- lution and standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, acetic acid (100), water, pyridine and ethanol (99.5) (30:15:10:6:5) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 1109C for 5 minutes: the spots obtained from the sample solution and standard solution show the same Rf value.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bacitracin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.15 g of Bacitracin in 0.05 mol/L sulfuric acid TS to make 100 mL. To 2 mL of this solution add 0.05 mol/L sulfuric acid TS to make 10 mL, and determine the absorbances of this solution, A1 and A2, at 252 nm and 290 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: A2/A1 is not more than 0.20. ‌Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
Weigh accurately an amount of Bacampicillin Hydrochloride and Bacampicillin Hydrochloride RS, equiva-lent to about 40 mg (potency), dissolve each in water to make exactly 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of bacampicillin in each solution. [22601-59-8] [1405-87-4, Bacitracin] Bacitracin is a mixture of peptide substances having antibacterial activity including bacitracin A as the main component produced by the growth of Bacillus subtilis or Bacillus licheniformis. It contains not less than 60 Units per mg, calculated on the dried basis. The potency of Bacitracin is expressed as unit calculated from the amount of bacitracin A (C66H103N17O16S: 1422.69). One unit of Bacitracin is equivalent to 23.8 mg of bacitracin A (C66H103N17O16S). Description Bacitracin occurs as a white to light brown powder. It is freely soluble in water, and slightly soluble in ethanol (99.5). Identification (1) To 3 mL of a solution of Bacitracin (1 in 100) add 3 mL of 4-dimethylaminobenzaldehyde TS, shake until red-rosy to red-purple color appears, then add several drops of a solution of sodium nitrite (1 in 100), and Amount [ mg (potency)] of ampicillin (C H N O S) shake: a green to dark green color is produced. ＝ MS × AT/AS × 1000 16 19 3 4 (2) Dissolve 60 mg each of Bacitracin and Bacitracin RS in 10 mL of water, and use these solutions as the sample so- MS: Amount [mg (potency)] of Bacampicillin Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 500 mL of diluted 2 mol/L sodium dihy-drogen phosphate TS (1 in 100), add diluted 0.05 mol/L dis- lution and standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, acetic acid (100), water, pyridine and ethanol (99.5) (30:15:10:6:5) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 1109C for 5 minutes: the spots obtained from the sample solution and standard solution show the same Rf value. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Bacitracin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.15 g of Bacitracin in 0.05 mol/L sulfuric acid TS to make 100 mL. To 2 mL of this solution add 0.05 mol/L sulfuric acid TS to make 10 mL, and determine the absorbances of this solution, A1 and A2, at 252 nm and 290 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: A2/A1 is not more than 0.20. ‌Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Micrococcus luteus ATCC 10240. Culture medium—Use the medium iii in 3) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Bacitracin RS, equivalent to about 400 units, dissolve in phosphate buffer solution (pH 6.0) to make exactly 20 mL, and use this solution as the standard stock solution. Keep the standard stock solution at not exceeding 109C and use within 2 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 2 units and 0.5 units, and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Bacitracin, equivalent about 400 units, dissolve in phosphate buffer solution (pH 6.0) to make exactly 20 mL. Take ex-actly a suitable amount of this solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 2 units and 0.5 units, and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Baclofen occurs as a white to pale yellow-white crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in water, very slightly soluble in methanol and in ethanol (95), and practically insoluble in diethyl ether. It dissolves in dilute hydrochloric acid.
(1) To 5 mL of a solution of Baclofen (1 in 1000) add 1 mL of ninhydrin TS, and heat on a water bath for 3 minutes: a blue-purple color develops. Determine the absorption spectrum of a solution of Baclofen in 0.1 mol/L hydrochloric acid TS (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Baclofen RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Perform the test with Baclofen as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—Dissolve 0.5 g of Baclofen in 50 mL of acetic acid (100), and add water to make 100 mL. To 10 mL of this solution add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.21z). Heavy metals <1.07>—Proceed with 2.0 g of Baclofen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Baclofen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Baclofen in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1.0 mL and 1.5 mL of the sample solu-tion, to each add the mobile phase to make exactly 100 mL, and use these solutions as the standard solutions (1) and (2), respectively. Perform the test with exactly 25 mL each of the sample solution and standard solutions (1) and (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak height of these solutions: each height of the peaks other than the peak of baclofen obtained from the sample solution is not larger than the peak height of baclofen from the standard solution (1), and the total height of these peaks is not larger than the peak height of baclofen from the standard solution (2). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and diluted acetic acid (100) (1 in 900) (3:2). Flow rate: Adjust so that the retention time of baclofen is about 4 minutes. Time span of measurement: About 3 times as long as the retention time of baclofen, beginning after the solvent peak. System suitability— Test for required detectability: Adjust the sensitivity so that the peak height of baclofen obtained with 25 mL of the standard solution (1) is between 5 and 10 mm. System performance: Dissolve 0.40 g of Baclofen and 5 mg of methyl parahydroxybenzoate in 200 mL of the mobile phase. To 10 mL of this solution add the mobile phase to make 100 mL. When the procedure is run with 25 mL of this solution under the above operating conditions, baclofen and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution (1) under the above ‌JP XVIII Official Monographs / Baclofen Tablets 511 operating conditions, the relative standard deviation of the peak heights of baclofen is not more than 3.0z. Water <2.48> Not more than 1.0z (1 g, direct titration). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.5 g of Baclofen, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to greenish blue (indica-tor: 2 drops of crystal violet TS). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.37 mg of C10H12ClNO2
Bakumondoto Extract contains not less than 1.
Bakumondoto Extract occurs as a light yellow to light brown powder or black-brown viscous extract. It has a slight odor, and a sweet taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 5 mL of 1-butanol, shake, centrifuge, remove the 1-butanol layer, and use the aqueous layer as the sample solution. Separately, heat 3.0 g of pulverized ophiopogon root in 50 mL of water under a reflux condenser for 1 hour. After cooling, shake 20 mL of the extract with 5 mL of 1-butanol, centrifuge, re-move the 1-butanol layer, and use the aqueous layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL of the sample solution and 5 mL of the standard solution as bands on the original line on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5), water and acetic acid (100) (120:80:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark blue-green spot (Rf value: about 0.3) from the standard solution (Ophiopogon Root). Shake 5.0 g of the dry extract (or 15 g of the viscous extract) with 15 mL of water, add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sam-ple solution. Separately, dissolve 1 mg of cycloartenyl feru-late for thin-layer chromatography in 1 mL of ethyl acetate, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 30 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (50:20:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution. Or examine under ultraviolet light (main wavelength: 365 nm) after spraying evenly a mixture of sulfuric acid and ethanol (99.5) (1:1) on the plate, and heating the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Brown Rice). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Bakumondoto Extract / Crude Drugs and Related Drugs
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 7.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Ginsenoside Rb1—Weigh accurately about 2 g of the dry extract (or an amount of the viscous extract, equivalent to about 2 g of dried substance), add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 15 mL of diluted methanol (3 in 5), and repeat the same proce-dure. Combine all of the supernatant liquid, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of sodium hydroxide TS, allow to stand for 30 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 20 mL. Apply exactly 5 mL of this solution to a column [about 10 mm in inside di-ameter, packed with 0.36 g of octadecylsilanized silica gel for pre-treatment (55 – 105 mm in particle size), and washed just before using with methanol and then diluted methanol (3 in 10)], and wash the column in sequence with 2 mL of diluted methanol (3 in 10), 1 mL of sodium carbonate TS and 10 mL of diluted methanol (3 in 10). Finally, elute with methanol to collect exactly 5 mL, and use this as the sample solution. Separately, weigh accurately about 10 mg of Gin-senoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solution. ditions, the number of theoretical plates and the symmetry factor of the peak of ginsenoside Rb1 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z. (2) Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about Amount (mg) of ginsenoside Rb1 ＝ MS × AT/AS × 1/5 (C54 H92O 23) 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times MS: Amount (mg) of Ginsenoside Rb1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with carbamoyl group bound silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (400:100:1). Flow rate: 1.0 mL per minute (the retention time of gin-senoside Rb1 is about 16 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Bamethan Sulfate, when dried, contains not less than 99.
Bamethan Sulfate occurs as white, crystals or crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in water and in acetic acid (100), soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 1699C (with decomposition).
(1) To 1 mL of a solution of Bamethan Sulfate (1 in 1000) add 5 mL of a solution of 4-nitroben-zenediazonium fluoroborate (1 in 2000) and 10 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.2): an orange-red color develops. Determine the absorption spectrum of a solution of Bamethan Sulfate in 0.01 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Bamethan Sulfate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>: it exhibits absorption at the wave numbers of about 1618, 1597, 1518, 1118 and 833 cm－1. A solution of Bamethan Sulfate (1 in 100) responds to Qualitative Tests <1.09> for sulfate. spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.75 g of Bamethan Sulfate, previously dried, dissolve in 80 mL of acetic acid (100), and titrate with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 51.67 mg of (C12H19NO2)2.H2SO4
Barbital, when dried, contains not less than 99.
Barbital occurs as colorless or white crystals or a white crystalline powder. It is odorless, and has a slightly bitter taste. ‌JP XVIII Official Monographs / Barium Sulfate 513 It is freely soluble in acetone and in pyridine, soluble in ethanol (95), sparingly soluble in diethyl ether, and slightly soluble in water and in chloroform. It dissolves in sodium hydroxide TS and in ammonia TS. The pH of its saturated solution is between 5.0 and 6.0.
(1) Boil 0.2 g of Barbital with 10 mL of sodium hydroxide TS: the gas evolved changes moistened red litmus paper to blue. Dissolve 0.05 g of Barbital in 5 mL of diluted pyri-dine (1 in 10), add 0.3 mL of copper (II) sulfate TS, shake, and allow to stand for 5 minutes: a red-purple precipitate is formed. Shake the mixture with 5 mL of chloroform: a red-purple color develops in the chloroform layer. Separately, dissolve 0.05 g of Barbital in 2 to 3 drops of ammonia-ammonium chloride buffer solution (pH 10.7) and 5 mL of diluted pyridine (1 in 10). Add 5 mL of chloroform and 0.3 mL of copper (II) sulfate TS to the solution: a red-purple precipitate is produced in the aqueous layer. The red-purple precipitate is not dissolved in the chloroform by shaking. To 0.4 g of Barbital add 0.1 g of anhydrous sodium carbonate and 4 mL of water, shake, and add a solution of 0.3 g of 4-nitrobenzyl chloride in 7 mL of ethanol (95). Heat the mixture on a water bath under a reflux condenser for 30 minutes, and allow to stand for 1 hour. Collect the separated crystals, wash with 7 mL of sodium hydroxide TS and a small amount of water, recrystallize from a mixture of ethanol (95) and chloroform (1:1), and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1929C and 1969C.
(1) Clarity and color of solution—Dissolve 1.0 g Description Barbital occurs as colorless or white crystals or a white crystalline powder. It is odorless, and has a slightly bitter taste. ‌JP XVIII Official Monographs / Barium Sulfate 513 It is freely soluble in acetone and in pyridine, soluble in ethanol (95), sparingly soluble in diethyl ether, and slightly soluble in water and in chloroform. It dissolves in sodium hydroxide TS and in ammonia TS. The pH of its saturated solution is between 5.0 and 6.0. Identification (1) Boil 0.2 g of Barbital with 10 mL of sodium hydroxide TS: the gas evolved changes moistened red litmus paper to blue. Dissolve 0.05 g of Barbital in 5 mL of diluted pyri-dine (1 in 10), add 0.3 mL of copper (II) sulfate TS, shake, and allow to stand for 5 minutes: a red-purple precipitate is formed. Shake the mixture with 5 mL of chloroform: a red-purple color develops in the chloroform layer. Separately, dissolve 0.05 g of Barbital in 2 to 3 drops of ammonia-ammonium chloride buffer solution (pH 10.7) and 5 mL of diluted pyridine (1 in 10). Add 5 mL of chloroform and 0.3 mL of copper (II) sulfate TS to the solution: a red-purple precipitate is produced in the aqueous layer. The red-purple precipitate is not dissolved in the chloroform by shaking. To 0.4 g of Barbital add 0.1 g of anhydrous sodium carbonate and 4 mL of water, shake, and add a solution of 0.3 g of 4-nitrobenzyl chloride in 7 mL of ethanol (95). Heat the mixture on a water bath under a reflux condenser for 30 minutes, and allow to stand for 1 hour. Collect the separated crystals, wash with 7 mL of sodium hydroxide TS and a small amount of water, recrystallize from a mixture of ethanol (95) and chloroform (1:1), and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1929C and 1969C. Melting point <2.60> 189 – 1929C Purity (1) Clarity and color of solution—Dissolve 0.5 g of Barbital in 5 mL of sodium hydroxide TS: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.30 g of Barbital in 20 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: take 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of ace-tone and 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.035z). Sulfate <1.14>—Dissolve 0.40 g of Barbital in 20 mL of acetone, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: take 0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of ace-tone, and 1 mL of dilute hydrochloric acid, and add water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Barbital according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 20 ppm). Readily carbonizable substances <1.15>—Perform the test with 0.5 g of Barbital. The solution is not more colored than Matching Fluid A. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Barbital, previously dried, and dissolve in 5 mL of ethanol (95) and 50 mL of chloroform. Titrate <2.50> with 0.1 mol/L potassium hy-droxide-ethanol VS until the color of the solution changes from yellow through light blue to purple (indicator: 1 mL of alizarin yellow GG-thymolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 18.42 mg of C8H12N2O3
Barium Sulfate occurs as a white powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It does not dissolve in hydrochloric acid, in nitric acid and in sodium hydroxide TS.
(1) Mix 0.5 g of Barium Sulfate with 2 g each of anhydrous sodium carbonate and potassium carbon-ate in a crucible, heat the mixture until fusion is complete, treat the cooled mass with hot water, and filter. The filtrate, acidified with hydrochloric acid, responds to Qualitative Tests <1.09> for sulfate. (2) Wash the hot water-insoluble residue obtained in (1) with water, dissolve in 2 mL of acetic acid (31), and filter, if necessary: the solution responds to Qualitative Tests <1.09> for barium salt.
(1) Acidity or alkalinity—Agitate 1.0 g of Barium Sulfate with 20 mL of water for 5 minutes: the solution is neutral. Phosphate—Boil 1.0 g of Barium Sulfate with 3 mL of nitric acid and 5 mL of water for 5 minutes, cool, and add water to restore the original volume. Filter through a filter paper, previously washed with dilute nitric acid, to the fil-trate add an equal volume of hexaammonium heptamolyb-date TS, and allow to stand between 509C and 609C for 1 hour: no yellow precipitate is produced. Sulfide—Place 10 g of Barium Sulfate in a 250-mL conical flask, add 10 mL of dilute hydrochloric acid and water to make 100 mL, and boil for 10 minutes: the gas evolved does not darken moistened lead (II) acetate paper. Heavy metals <1.07>—Boil 5.0 g of Barium Sulfate with 2.5 mL of acetic acid (100) and 50 mL of water for 10 minutes, cool, add 0.5 mL of ammonia TS and water to make 100 mL, and filter. Perform the test with a 50-mL por-tion of this filtrate. Prepare the control solution with 2.5 mL of Standard Lead Solution, 1.25 mL of acetic acid (100), 0.25 mL of ammonia TS and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Barium Sulfate according to Method 1, and perform the test (not more than 1 ppm). Hydrochloric acid-soluble substances and soluble barium salts—Cool the solution obtained in (3), add water to make 100 mL, and filter. Evaporate 50 mL of the filtrate on a water bath to dryness, add 2 drops of hydrochloric acid and 10 mL of warm water, filter through a filter paper for quantitative analysis, and wash with 10 mL of warm water. Evaporate the combined filtrate and washings on a water bath to dryness, and dry the residue at 1059C for 1 hour: the residue weighs not more than 15 mg. Shake the residue, if any, with 10 mL of water, and filter. To the filtrate add 0.5 mL of dilute sulfuric acid, and allow to stand for 30 minutes: no turbidity is produced. ‌514‌
ベクロメタゾンプロピオン酸エステル
Beclometasone Dipropionate, when dried, contains not less than 97.
Beclometasone Dipropionate occurs as a white to pale yellow powder. It is soluble in methanol, sparingly soluble in ethanol cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of beclometasone dipropionate to that of the internal standard, respectively. (99.5), and practically insoluble in water. Melting point: about 2089C (with decomposition). It shows crystal polymorphism. Amount (mg) of beclometasone dipropionate (C ＝ MS × QT/QS 28H 37ClO7)
(1) Dissolve 2 mg of Beclometasone Dipropionate in 2 mL of sulfuric acid: initially a yellowish color develops, and gradually changes through orange to dark red-brown. To this solution add carefully 10 mL of water: the color changes to bluish green, and a flocculent precipitate is formed. Dissolve 0.01 g of Beclometasone Dipropionate in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: a red to red-brown precipitate is formed. Perform the test with 0.02 g of Beclometasone Dipropionate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 1 mL of sodium hydrox-ide TS and 20 mL of water as an absorbing liquid: the test solution responds to Qualitative Tests <1.09> for chloride. Determine the infrared absorption spectrum of Beclometasone Dipropionate, previously dried, as directed in MS: Amount (mg) of Beclometasone Dipropionate RS taken Internal standard solution—A solution of testosterone propionate in methanol (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and water (3:2). Flow rate: Adjust so that the retention time of beclometa- sone dipropionate is about 6 minutes. ‌JP XVIII Official Monographs / Bekanamycin Sulfate 515 System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, beclometasone dipropionate and the internal stand-ard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of beclometasone dipropionate to that of the internal standard is not more than 1.0z.
(1) Heavy metals <1.07>—Proceed with 0.5 g of Beclometasone Dipropionate according to Method 2, and perform the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 30 ppm). (2) Related substances—Dissolve 20 mg of Beclometa-sone Dipropionate in 5 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chlo-roform and methanol (9:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1,2-dichloroethane, methanol and water (475:25:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly alkaline blue tetrazolium TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 20 mg each of Beclometa-sone Dipropionate and Beclometasone Dipropionate RS, previously dried, and dissolve each in methanol to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution and metha-nol to make 50 mL, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the beclometasone dipropionate (C 28H 37ClO7). test with 20 mL each of the sample solution and standard so- lution as directed under Liquid Chromatography <2.01> ac- Description Beclometasone Dipropionate occurs as a white to pale yellow powder. It is soluble in methanol, sparingly soluble in ethanol cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of beclometasone dipropionate to that of the internal standard, respectively. (99.5), and practically insoluble in water. Melting point: about 2089C (with decomposition). It shows crystal polymorphism. Amount (mg) of beclometasone dipropionate (C ＝ MS × QT/QS 28H 37ClO7) Identification (1) Dissolve 2 mg of Beclometasone Dipropionate in 2 mL of sulfuric acid: initially a yellowish color develops, and gradually changes through orange to dark red-brown. To this solution add carefully 10 mL of water: the color changes to bluish green, and a flocculent precipitate is formed. Dissolve 0.01 g of Beclometasone Dipropionate in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: a red to red-brown precipitate is formed. Perform the test with 0.02 g of Beclometasone Dipropionate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 1 mL of sodium hydrox-ide TS and 20 mL of water as an absorbing liquid: the test solution responds to Qualitative Tests <1.09> for chloride. Determine the infrared absorption spectrum of Beclometasone Dipropionate, previously dried, as directed in MS: Amount (mg) of Beclometasone Dipropionate RS taken Internal standard solution—A solution of testosterone propionate in methanol (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and water (3:2). Flow rate: Adjust so that the retention time of beclometa- sone dipropionate is about 6 minutes. ‌JP XVIII Official Monographs / Bekanamycin Sulfate 515 System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, beclometasone dipropionate and the internal stand-ard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of beclometasone dipropionate to that of the internal standard is not more than 1.0z.
Bekanamycin Sulfate occurs as a white pow-der. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 20 mg of Bekanamycin Sulfate in 2 mL of 1/15 mol/L phosphate buffer solution (pH 5.6), add 1 mL of ninhydrin TS, and boil: a blue-purple color develops. Dissolve 30 mg each of Bekanamycin Sulfate and Bekanamycin Sulfate RS in 5 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solu-tion of potassium dihydrogen phosphate (3 in 40) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the principal spots obtained from the sample solution and the standard solution show a purple-brown color and the same Rf value. To a solution of Bekanamycin Sulfate (1 in 5) add 1 drop of barium chloride TS: a white turbidity is produced. D Optical rotation <2.49> [a]20: ＋102 – ＋1169(after drying, 0.25 g, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.50 g of Bekanamycin Sulfate in 10 mL of water is between 6.0 and 8.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Bekanamycin Sulfate in 5 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Bekana- mycin Sulfate according to Method 4, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Bekanamycin Sulfate according to Method 1, and per-form the test (not more than 1 ppm). Related substances—Dissolve 60 mg of Bekanamycin Sulfate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer having pH <2.54> 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Bekanamycin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 5 to 159C and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 10 mg (po-tency) and 2.5 mg (potency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Bekanamycin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 10 mg (potency) and 2.5 mg (potency), and use these solutions as the high concentration sample solution and low concentration sample solution, respectively. ‌516
Belladonna Extract contains not less than 0.
Belladonna Extract has a dark brown color, a characteristic odor and a bitter taste.
Mix 0.5 g of Belladonna Extract with 30 mL of ammonia TS in a flask, transfer the mixture to a separa-tor, then add 40 mL of ethyl acetate, and shake the mixture. Drain off the ethyl acetate layer, add 3 g of anhydrous so-dium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent to dryness under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. Proceed as directed in the Identification under Belladonna Root.
Heavy metals <1.07>—Prepare the test solution with 1.0 g of Belladonna Extract as directed under the Extracts (4), and perform the test (not more than 30 ppm).
Weigh accurately about 0.4 g of Belladonna Extract, place in a glass-stoppered centrifuge tube, add 15 mL of am-monia TS, and shake. Add 25 mL of diethyl ether, stopper tightly, shake for 15 minutes, centrifuge, and separate the diethyl ether layer. Repeat this procedure twice with the aqueous layer, using 25 mL each of diethyl ether. Combine the extracts, and evaporate the solvent on a water bath. Dis-solve the residue in 5 mL of the mobile phase, add exactly 3 mL of the internal standard solution, and add the mobile phase to make exactly 25 mL. Proceed as directed under Bel-ladonna Root. Amount (mg) of hyoscyamine (C17H23NO3) ＝ MS × QT/QS × 1/5 × 0.855 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500).
Belladonna Total Alkaloids contains not less than 95.
Belladonna Total Alkaloids occurs as white, crystals or crystalline powder. It is very soluble in methanol, freely soluble in ethanol (99.5), and slightly soluble in water.
Dissolve 2 mg of Belladonna Total Alkaloids in 1 mL of ethanol (95), and use this solution as the sample solution. Then proceed as directed in the Identification under Belladonna Root. D Optical rotation <2.49> [a]20: －18.5 – －22.09(after dry-ing, 1 g, ethanol (99.5), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Place 1.0 g of Bella-donna Total Alkaloids in a porcelain crucible, and mix with 1.2 mL of dilute hydrochloric acid. Mix with 10 mL of a so-lution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and after evaporating the solvent on a boiling water bath, carbonize by gradual heating. Then proceed according to Method 4, and perform the test. The control solution is prepared as follows: Mix 1.2 mL of dilute hydrochloric acid with 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and evaporate the solvent on a boil-ing water bath. After cooling, add 1 mL of sulfuric acid, then proceed according to Method 4, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 2.0 g of Belladonna Total Alkaloids according to Method 4, and perform the test (not more than 1 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 6 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Weigh accurately about 25 mg of Belladonna Total Alkaloids, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 3 mL of the internal standard solution and the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Atropine Sulfate RS (separately determine the loss on drying <2.41> under the same condi-tions as Atropine Sulfate Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solution as the standard stock solution (1). Also, weigh accurately about 25 mg of Scopolamine Hydrobromide RS (separately determine the loss on drying <2.41> under the same conditions as Scopolamine Hydrobromide Hydrate), and dissolve in the mobile phase to make exactly 25 mL. Pipet 3 mL of this so-lution, add the mobile phase to make exactly 25 mL, and use this solution as the standard stock solution (2). Take exactly 5 mL of standard stock solution (1), add exactly 2 mL of the ‌JP XVIII Crude Drugs and Related Drugs / Benincasa Seed 1959 standard stock solution (2), and add exactly 3 mL of the in-ternal standard solution. To this solution add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QTA and QSA, of the peak area of hyoscyamine (atropine) to that of the internal standard and the ratios, QTS and QSS, of the peak area of scopolamine to that of the internal standard. Then calculate the amounts of hyoscyamine and scopolamine using the following equa-tions. The amount of the total alkaloids is obtained as the sum of them. Benincasa Seed Benincasae Semen
Benidipine Hydrochloride, when dried, contains not less than 99.
Benidipine Hydrochloride occurs as a yellow crystalline powder. It is very soluble in formic acid, soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Benidipine Hydrochloride in methanol (1 in 100) shows no optical rotation. Melting point: about 2009C (with decomposition).
(1) Determine the absorption spectrum of a solution of Benidipine Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Benidipine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Benidipine Hydrochloride (1 in 10) add 5 mL of ammonia TS, heat on a water bath for 5 minutes, cool, and filter. The filtrate, which is acidified with dilute nitric acid, responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Benidipine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Benidipine Hydrochloride in 100 mL of a mixture of water and metha-nol (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and methanol (1:1) to make exactly 500 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the peak areas of bisbenzyl-piperidyl ester having the relative retention time of about 0.35 to benidipine, dehydro derivative having the relative retention time of about 0.75 and other related substances obtained from the sample solution are not larger than 1/2 times the peak area of benidipine from the standard solu-tion, and the total area of the peaks other than benidipine from the sample solution is not larger than the peak area of benidipine from the standard solution. For the areas of the peaks of bisbenzylpiperidyl ester and dehydro derivative, multiply their correction factor 1.6, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0), methanol and tetrahydrofu-ran (65:27:8). Flow rate: Adjust so that the retention time of benidipine is about 20 minutes. Time span of measurement: About 2 times as long as the retention time of benidipine, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add the mixture of water and methanol (1:1) to make exactly 20 mL. Confirm that the peak area of benidipine obtained with 10 mL of this solution is equivalent to 18 to 32z of that obtained with 10 mL of the standard solution. System performance: Dissolve 6 mg of Benidipine Hydro-chloride and 5 mg of benzoin in 200 mL of the mixture of water and methanol (1:1). When the procedure is run with 10 mL of this solution under the above operating conditions, benzoin and benidipine are eluted in this order with the reso-lution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of benidipine is not more than 3.5z. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Benidipine Hydro-chloride, previously dried, dissolve in 10 mL of formic acid, add 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 54.20 mg of C28H31N3O6.HCl
Oxybuprocaine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a saline taste. It exhibits anesthetic properties when placed on the tongue. It is very soluble in water, freely soluble in ethanol (95) and in chloroform, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Oxybuprocaine Hydro- chloride in 10 mL of water is between 5.0 and 6.0. It is gradually colored by light.
(1) Dissolve 10 mg of Oxybuprocaine Hy-drochloride in 1 mL of dilute hydrochloric acid and 4 mL of water. This solution responds to Qualitative Tests <1.09> for primary aromatic amines. Dissolve 0.1 g of Oxybuprocaine Hydrochloride in 8 mL of water, and add 3 mL of ammonium thiocyanate TS: an oily substance is produced. Rub the inner surface of the container with a glass rod: white crystals are formed. Collect the crystals so obtained, recrystallize from water, and dry in a desiccator (in vacuum, phosphorus (V) oxide) for 5 hours: the crystals melt <2.60> between 1039C and 1069C. Determine the absorption spectrum of a solution of Oxybuprocaine Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Oxybuprocaine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Oxybuprocaine Hydrochloride in 10 mL of water: the so-lution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Oxybu- procaine Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.25 g of Oxybu-procaine Hydrochloride in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add chloroform to make exactly 20 mL. Pipet 1 mL of this solution, add chloroform to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, ethanol (95) and formic acid (7:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Oxybuprocaine Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.49 mg of C17H28N2O3.HCl
Benserazide Hydrochloride contains not less than 98.
Benserazide Hydrochloride occurs as a white to grayish white crystalline powder. It is freely soluble in water and in formic acid, soluble in methanol, very slightly soluble in ethanol (95). It dissolves in 0.1 mol/L hydrochloric acid TS. The pH of a solution of 1.0 g of Benserazide Hydrochlo-ride in 100 mL of water is between 4.0 and 5.0. It is hygroscopic. It is gradually colored by light. A solution of Benserazide Hydrochloride (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Benserazide Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ben-serazide Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 10 mL of a solution of Benserazide Hydrochloride (1 in 30) add silver nitrate TS: a white precipitate is formed. To a portion of this precipitate add dilute nitric acid: the precipitation does not dissolve.
(1) Clarity and color of solution—Dissolve 0.5 g of Benserazide Hydrochloride in 10 mL of water, and per-form the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance of this so-lution at 430 nm is not more than 0.10. Heavy metals <1.07>—Proceed with 1.0 g of Benser- azide Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.25 g of Benserazide Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL and 3 mL of the sample solution, add methanol to make exactly 200 mL, and use these solutions as the standard solution (1) and (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solutions (1) and (2) on a plate of cellulose for thin-layer chromatogra- ‌JP XVIII Official Monographs / Benzalkonium Chloride 519‌ phy. Develop the plate with a solution of formic acid in so-dium chloride TS (1 in 1000) to a distance of about 10 cm, and air-dry the plate. Spray evenly sodium carbonate TS, air-dry, and then spray evenly Folin's TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution (2), and the number of the spots which intense more than the spot from the standard solution (1) are not more than 2. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration). Use a solution of salicylic acid in methanol for water determination (3 in 20) instead of metha-nol for water determination. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Benserazide Hydro-chloride, dissolve in 5 mL of formic acid, add 50 mL of ace-tic acid (100), and titrate <2.50> immediately with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.37 mg of C10H15N3O5.HCl
Bentonite occurs as a very fine, white to light yellow-brown powder. It is odorless. It has a slightly earthy taste. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It swells in water.
(1) Add 0.5 g of Bentonite to 3 mL of diluted sulfuric acid (1 in 3), and heat until white fumes are evolved. Cool, add 20 mL of water, and filter. To 5 mL of the filtrate add 3 mL of ammonia TS: a white, gelatinous precipitate is produced, which turns red on the addition of 5 drops of alizarin red S TS. (2) Wash the residue obtained in (1) with water, add 2 mL of a solution of methylene blue trihydrate (1 in 10,000), and wash again with water: the residue is blue in color. pH <2.54> To 1.0 g of Bentonite add 50 mL of water, and shake: the pH of the suspension is between 9.0 and 10.5.
(1) Heavy metals <1.07>—To 1.5 g of Bentonite add 80 mL of water and 5 mL of hydrochloric acid, and boil gently for 20 minutes with thorough stirring. Cool, centri-fuge, collect the supernatant liquid, wash the residue with two 10-mL portions of water, and centrifuge each. Combine the supernatant liquid and the washings, and add dropwise ammonia solution (28). When a precipitate is produced, add dropwise dilute hydrochloric acid with vigorous stirring, and dissolve. To the solution add 0.45 g of hydroxylammonium chloride, and heat. Cool, and add 0.45 g of sodium acetate trihydrate, 6 mL of dilute acetic acid and water to make 150 mL. Pipet 50 mL of the solution, and perform the test using this solution as the test solution. Prepare the control solution as follows: mix 2.5 mL of Standard Lead Solution, 0.15 g of hydroxylammonium chloride, 0.15 g of sodium acetate trihy-drate, and 2 mL of dilute acetic acid, and add water to make 50 mL (not more than 50 ppm). Arsenic <1.11>—To 1.0 g of Bentonite add 5 mL of dilute hydrochloric acid, and gently heat to boil while stir-ring well. Cool immediately, and centrifuge. To the residue add 5 mL of dilute hydrochloric acid, shake well, and centri-fuge. To the residue add 10 mL of water, and perform the same operations. Combine all the extracts, and heat on a water bath to concentrate to 5 mL. Perform the test with this solution as the test solution (not more than 2 ppm). Foreign matter—Place 2.0 g of Bentonite in a mor-tar, add 20 mL of water to swell, disperse evenly with a pes-tle, and dilute with water to 100 mL. Pour the suspension through a No. 200 (74 mm) sieve, and wash the sieve thor-oughly with water. No grit is felt when the fingers are rubbed over the wire mesh of the sieve. Loss on drying <2.41> 5.0 – 10.0z (2 g, 1059C, 2 hours). Gel formation Mix 6.0 g of Bentonite with 0.30 g of mag-nesium oxide. Add the mixture, in several portions, to 200 mL of water contained in a glass-stoppered 500-mL cylinder. Agitate for 1 hour, transfer 100 mL of the suspension to a 100-mL graduated cylinder, and allow to stand for 24 hours: not more than 2 mL of supernatant appears on the surface. Swelling power To 100 mL of water in a glass-stoppered 100-mL cylinder add 2.0 g of Bentonite in ten portions, allowing each portion to settle before adding the next, and allow to stand for 24 hours: the apparent volume of the sedi-ment at the bottom is not less than 20 mL.
Benzalkonium Chloride is represented by the for mula [C6H5CH2N(CH3)2R]Cl, in which R extends from C8H17 to C18H37, with C12H25 and C14H29 com-prising the major portion. It contains not less than 95.
Benzalkonium Chloride occurs as a white to yellow-white powder, colorless to light yellow, gelatinous pieces, or jelly-like fluid or mass. It has a characteristic odor. It is very soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. A solution of Benzalkonium Chloride foams strongly when shaken.
(1) Dissolve 0.2 g of Benzalkonium Chlo-ride in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate, and heat for 5 minutes on a water bath. After cooling, add 10 mL of water and 0.5 g of zinc powder, heat for 5 minutes, cool, and filter: the filtrate responds to Qualitative Tests <1.09> for primary aromatic amines. The color of the solu- tion is red. To 2 mL of a solution of Benzalkonium Chloride (1 in 1000) add a mixture of 0.2 mL of a solution of bromophe-nol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS: a blue color develops. Add 4 mL of chloroform to this solu-tion, and shake vigorously: the blue color shifts to the chlo-roform layer. Collect the chloroform layer, and add drop-wise, with stirring, a solution of sodium lauryl sulfate (1 in 1000): the chloroform layer turns colorless. Determine the absorption spectrum of a solution of Benzalkonium Chloride in 0.1 mol/L hydrochloric acid TS (1 in 2000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. ‌To 1 mL of a solution of Benzalkonium Chloride (1 in 100) add 2 mL of ethanol (95), 0.5 mL of dilute nitric acid and 1 mL of silver nitrate TS: a white precipitate is pro-duced. This precipitate does not dissolve on the addition of dilute nitric acid, but dissolves on the addition of ammonia TS.‌
(1) Clarity and color of solution—Dissolve 1.0 g of Benzalkonium Chloride in 10 mL of water: the solution is clear and colorless to light yellow. (2) Petroleum ether-soluble substances—To 3.0 g of Benzalkonium Chloride add water to make 50 mL, then add 50 mL of ethanol (99.5) and 5 mL of 0.5 mol/L sodium hy- It foams strongly on shaking. Identification (1) Evaporate a volume of Benzalkonium Chloride Solution, equivalent to 0.2 g of Benzalkonium Chloride, on a water bath to dryness, and proceed with the residue as directed in the Identification (1) under Benzalko-nium Chloride. To a volume of Benzalkonium Chloride Solution, equivalent to 0.01 g of Benzalkonium Chloride, add water to make 10 mL. Proceed with 2 mL of this solution as directed in the Identification (2) under Benzalkonium Chloride. To a volume of Benzalkonium Chloride Solution, equivalent to 1 g of Benzalkonium Chloride, add water or concentrate on a water bath, if necessary, to make 10 mL. To 1 mL of this solution add 0.1 mol/L hydrochloric acid VS to make 200 mL, and proceed as directed in the Identifi-cation (3) under Benzalkonium Chloride. To a volume of Benzalkonium Chloride Solution, equivalent to 0.1 g of Benzalkonium Chloride, add water or concentrate on a water bath, if necessary, to make 10 mL. Proceed with 1 mL of this solution as directed in the Identifi-cation (4) under Benzalkonium Chloride.
Pipet a volume of Benzalkonium Chloride Solution, equivalent to about 0.15 g of benzalkonium chloride droxide TS, and extract with three 50-mL portions of petro- leum ether. Combine the petroleum ether extracts, and wash (C22H ClN), dilute with water to make 75 mL, if necessary, with three 50-mL portions of dilute ethanol. After shaking well with 10 g of anhydrous sodium sulfate, filter through a dry filter paper, and wash the filter paper with two 10-mL portions of petroleum ether. Evaporate the petroleum ether on a water bath by heating, and dry the residue at 1059C for 1 hour: the residue is not more than 1.0z. Water <2.48> Not more than 15.0z (volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 0.15 g of Benzalkonium Chloride, and dissolve in 75 mL of water. Adjust the pH be-tween 2.6 and 3.4 by adding dropwise diluted dilute hydro-chloric acid (1 in 2), add 1 drop of methyl orange TS, and titrate <2.50> with 0.02 mol/L sodium tetraphenylboron VS until the color of the solution becomes red. Each mL of 0.02 mol/L sodium tetraphenylboron VS and proceed as directed in the Assay under Benzalkonium Chloride. Each mL of 0.02 mol/L sodium tetraphenylboron VS ＝ 7.080 mg of C22H40ClN
Benzbromarone, when dried, contains not less than 98.
Benzbromarone occurs as a white to light yel- low crystalline powder. It is very soluble in N,N-dimethylformamide, freely solu-ble in acetone, sparingly soluble in ethanol (99.5), and prac-tically insoluble in water. It dissolves in dilute sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Benzbromarone in 0.01 mol/L sodium hydroxide TS (1 in 100,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Benz-bromarone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—Dissolve 1.0 g of Benzbro-marone in 40 mL of acetone, and add 1 mL of dilute hydro-chloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 40 mL of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.019z). Soluble halides—Dissolve 0.5 g of Benzbromarone in 40 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Proceed with this solution as directed under Chloride Limit Test <1.03>. Prepare the control solu-tion as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL. Heavy metals <1.07>—Proceed with 2.0 g of Benzbro- marone according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Benzbromarone according to Method 3, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Benzbro-marone in 10 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ace-tone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of cyclo-hexane, 4-methyl-2-pentanone, ethanol (99.5) and acetic acid (100) (100:20:2:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 509C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Benzbromarone, previously dried, dissolve in 30 mL of N,N-dimethylfor-mamide, and titrate <2.50> with 0.1 mol/L tetramethylam-monium hydroxide VS (indicator: 5 drops of thymol blue dimethylformamide TS). Perform a blank determination in ‌522‌ the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 42.41 mg of C17H12Br2O3
Benzethonium Chloride, when dried, contains not less than 97.
Benzethonium Chloride occurs as colorless or white crystals. It is odorless. It is very soluble in ethanol (95), freely soluble in water, and practically insoluble in diethyl ether. A solution of Benzethonium Chloride foams strongly when shaken.
(1) Dissolve 0.2 g of Benzethonium Chlo-ride in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate, and heat for 5 minutes on a water bath. After cooling, add 10 mL of water and 0.5 g of zinc powder, heat for 5 minutes, cool, and filter: the filtrate responds to Qualitative Tests <1.09> for primary aromatic amines, developing a red color. To 2 mL of a solution of Benzethonium Chloride (1 in 1000) add a mixture of 0.2 mL of a solution of bromo-phenol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS: a blue color develops. Add 4 mL of chloroform to this solu-tion, and shake vigorously: the blue color shifts to the chlo-roform layer. Collect the chloroform layer, and add drop-wise a solution of sodium lauryl sulfate (1 in 1000) with stirring: the chloroform layer turns colorless. Determine the absorption spectrum of a solution of Benzethonium Chloride in 0.1 mol/L hydrochloric acid TS (1 in 5000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. To 1 mL of a solution of Benzethonium Chloride (1 in 100) add 2 mL of ethanol (95), 0.5 mL of dilute nitric acid and 1 mL of silver nitrate TS: a white precipitate is pro-duced. This precipitate does not dissolve on addition of dilute nitric acid, but dissolves on addition of ammonia TS.
Ammonium—Dissolve 0.10 g of Benzethonium Chloride in 5 mL of water, and boil with 3 mL of sodium hy-droxide TS: the evolving gas dose not change moistened red litmus paper to blue. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Benzethonium Chloride, previously dried, dissolve in 75 mL of water, add diluted dilute hydrochloric acid (1 in 2) dropwise to adjust the pH to 2.6–3.4, then add 1 drop of methyl orange TS, and titrate <2.50> with 0.02 mol/L tetraphenylboron VS until the solution develops a red. Each mL of 0.02 mol/L sodium tetraphenylboron VS ＝ 8.962 mg of C27H42ClNO2
Benzoic acid [65-85-0] Benzoic Acid, when dried, contains not less than 99.
Benzoic Acid occurs as white, crystals or crys-talline powder. It is odorless, or has a faint, benzaldehyde-like odor. It is freely soluble in ethanol (95), in acetone and in diethyl ether, soluble in hot water, and slightly soluble in water.
Dissolve 1 g of Benzoic Acid in 8 mL of so-dium hydroxide TS, and add water to make 100 mL. This solution responds to Qualitative Tests <1.09> (2) for benzo-ate.
(1) Heavy metals <1.07>—Dissolve 1.0 g of Ben-zoic Acid in 25 mL of acetone, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid, 25 mL of acetone and water to make 50 mL (not more than 20 ppm). Chlorinated compounds—Take 0.5 g of Benzoic Acid and 0.7 g of calcium carbonate in a crucible, mix with a small amount of water, and dry. Ignite it at about 6009C, dissolve in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washing, add water to make 50 mL, and add 0.5 mL of silver nitrate TS: this solution has not more turbid than the follow-
Weigh accurately about 0.5 g of Benzoic Acid, pre-viously dried, dissolve in 25 mL of neutralized ethanol and 25 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 12.21 mg of C H O 7 6 2 Description Benzoic Acid occurs as white, crystals or crys-talline powder. It is odorless, or has a faint, benzaldehyde-like odor. It is freely soluble in ethanol (95), in acetone and in diethyl ether, soluble in hot water, and slightly soluble in water. Identification Dissolve 1 g of Benzoic Acid in 8 mL of so-dium hydroxide TS, and add water to make 100 mL. This solution responds to Qualitative Tests <1.09> (2) for benzo-ate. Melting point <2.60> 121 – 1249C Purity (1) Heavy metals <1.07>—Dissolve 1.0 g of Ben-zoic Acid in 25 mL of acetone, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid, 25 mL of acetone and water to make 50 mL (not more than 20 ppm). Chlorinated compounds—Take 0.5 g of Benzoic Acid and 0.7 g of calcium carbonate in a crucible, mix with a small amount of water, and dry. Ignite it at about 6009C, dissolve in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washing, add water to make 50 mL, and add 0.5 mL of silver nitrate TS: this solution has not more turbid than the follow-
Benzoin occurs as grayish brown to dark red-brown blocks varying in size; the fractured surface exhibiting whitish to light yellow-red grains in the matrix; hard and brittle at ordinary temperature but softened by heat. Odor, characteristic and aromatic; taste, slightly pungent and acrid. ‌Identification (1) Heat a fragment of Benzoin in a test tube: it evolves an irritating vapor, and a crystalline subli-mate is produced. (2) Digest 0.5 g of Benzoin with 10 mL of diethyl ether, decant 1 mL of the diethyl ether into a porcelain dish, and add 2 to 3 drops of sulfuric acid: a deep red-brown to deep red-purple color develops.
Ethanol-insoluble substances—Boil gently 1.0 g of Benzoin with 30 mL of ethanol (95) for 15 minutes under a reflux condenser. After cooling, collect the insoluble sub-stances through a tared glass filter (G3), and wash with three 5-mL portions of ethanol (95). Dry the residue at 1059C for 4 hours: the mass of the residue does not exceed 0.30 g. Total ash <5.01> Not more than 2.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Benzyl alcohol [100-51-6] This monograph is harmonized with the European Phar-macopoeia and the U.S. Pharmacopeia. The parts of the text that are not harmonized are marked with symbols (◆ ◆). Information on the harmonization with the European Pharmacopoeia and the U.S. Pharmacopeia is available on the website of the Pharmaceuticals and Medical Devices Agency. Benzyl Alcohol contains not less than 98.
Benzyl Alcohol is a clear, colorless oily liq-uid. It is miscible with ethanol (95), with fatty oils and with essential oils. It is soluble in water. Specific gravity d 20: 1.043 – 1.049◆
Determine the infrared absorption spec-trum of Benzyl Alcohol as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ D Refractive index <2.45> n20: 1.538 – 1.541
◆(1) Clarity and color of solution—Dissolve 2.0 mL of Benzyl Alcohol in 60 mL of water: the solution is clear and colorless.◆ Acidity—To 10 mL of Benzyl Alcohol add 10 mL of ethanol (95) and 2 drops of phenolphthalein TS, and add dropwise 0.1 mol/L sodium hydroxide VS until the solution acquires a light red color: the amount of 0.1 mol/L sodium hydroxide VS used is not more than 1.0 mL. Benzaldehyde and other related substances—Use Benzyl Alcohol as the sample solution. Separately, dissolve exactly 0.100 g of ethylbenzene in Benzyl Alcohol to make exactly 10 mL. Pipet 2 mL of this solution, add Benzyl Alco-hol to make exactly 20 mL, and use this solution as the ethyl-benzene stock solution. Separately, dissolve exactly 2.000 g of dicyclohexyl in Benzyl Alcohol to make exactly 10 mL. Pipet 2 mL of this solution, add Benzyl Alcohol to make ex-actly 20 mL, and use this solution as the dicyclohexyl stock solution. Separately, weigh exactly 0.750 g of benzaldehyde and 0.500 g of cyclohexylmethanol, and add Benzyl Alcohol to make exactly 25 mL. Pipet 1 mL of this solution, add ex-actly 2 mL of ethylbenzene stock solution and exactly 3 mL of dicyclohexyl stock solution, then add Benzyl Alcohol to make exactly 20 mL, and use this solution as the standard solution (1). Perform the test with exactly 0.1 mL each of the sample solution and standard solution (1) as directed under Gas Chromatography <2.02> according to the following con-ditions, and when the peak having the retention time cor-responding to ethylbenzene and dicyclohexyl appears on the chromatogram obtained from the sample solution, correct the peak areas of ethylbenzene and dicyclohexyl from the standard solution (1) by deducting the relevant peak area from the sample solution, the peak area of benzaldehyde from the sample solution is not more than the difference be-tween the peak areas of benzaldehyde of the sample solution and the standard solution (1) (0.15z), and the peak area of cyclohexylmethanol from the sample solution is not more than the difference between the peak areas of cyclohexyl-methanol of the sample solution and the standard solution (1) (0.10z). The total area of the peaks having smaller reten-tion time than benzyl alcohol and other than benzaldehyde and cyclohexylmethanol from the sample solution is not more than 4 times the peak area or the corrected peak area of ethylbenzene from the standard solution (1) (0.04z). The total area of the peaks having larger retention time than benzyl alcohol from the sample solution is not more than the peak area or the corrected peak area of dicyclohexyl from the standard solution (1) (0.3z). For these calculations the peak areas less than 1/100 times the peak area or the cor-rected peak area of ethylbenzene with the standard solution (1) are excluded. Benzyl Alcohol labeled that it is suitable for use in the manufacture of injection forms meets the following require-ments. Use Benzyl Alcohol as the sample solution. Separately, weigh exactly 0.250 g of benzaldehyde and 0.500 g of cyclo-hexylmethanol, and add Benzyl Alcohol to make exactly 25 mL. Pipet 1 mL of this solution, add exactly 2 mL of the ethylbenzene stock solution and exactly 2 mL of the dicyclo-hexyl stock solution, then add Benzyl Alcohol to make ex-actly 20 mL, and use this solution as the standard solution (2). Perform the test with exactly 0.1 mL each of the sample solution and standard solution (2) as directed under Gas Chromatography <2.02> according to the following condi-tions, and when the peak having the retention time cor-responding to ethylbenzene and dicyclohexyl appears on the chromatogram obtained from the sample solution, correct cyclohexylmethanol from the sample solution is not more than the difference between the peak areas of cyclohexyl-methanol of the sample solution and the standard solution (2) (0.10z). The total area of the peaks having smaller reten-tion time than benzyl alcohol and other than benzaldehyde and cyclohexylmethanol from the sample solution is not more than 2 times the peak area or the corrected peak area of ethylbenzene from the standard solution (2) (0.02z). The total area of the peaks having larger retention time than ben-zyl alcohol from the sample solution is not more than the peak area of or the corrected peak area dicyclohexyl from the standard solution (2) (0.2z). For these calculation the peak areas less than 1/100 times the peak area or the cor-rected peak area of ethylbenzene from the standard solution (2) are excluded. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.32 mm in inside diameter and 30 m in length, coated inside with polyethylene glycol 20 M for gas chromatography in 0.5 mm thickness. Column temperature: Inject at a constant temperature of about 509C, raise the temperature at a rate of 59C per minute to 2209C, and maintain at 2209C for 35 minutes. Temperature of injection port: A constant temperature of about 2009C. Temperature of detector: A constant temperature of about 3109C. Carrier gas: Helium. Flow rate: 25 cm/second. Splitless. Detection sensitivity: When 0.1 mL of the standard solu-tion (1) is injected, adjust the sensitivity of the detector so that the height of the peak of ethylbenzene is not less than 30z of the full scale of the recorder. For Benzyl Alcohol labeled to use for injection, use the standard solution (2) instead of the standard solution (1). System suitability— System performance: When the procedure is run with the standard solution (1) under the above operating conditions, the retention time of benzyl alcohol is about 26 minutes, the relative retention times of ethylbenzene, dicyclohexyl, ben-zaldehyde and cyclohexylmethanol to benzyl alcohol are about 0.28, about 0.59, about 0.68 and about 0.71, respec-tively, and the resolution between the peaks of benzaldehyde and cyclohexylmethanol is not less than 3.0. In the case of Benzyl Alcohol labeled to use for injection, proceed with the standard solution (2) instead of the standard solution (1). Peroxide value—Weigh accurately about 5 g of Ben-zyl Alcohol, and dissolve in 30 mL of a mixture of acetic acid (100) and chloroform (3:2) in a 250-mL glass-stoppered conical flask. Add 0.5 mL of saturated potassium iodide so-lution, shake for exactly 1 minute, add 30 mL of water, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS, adding the titrant slowly with continuous vigorous shaking, until the blue color of the solution disappears after addition of 5 mL of starch TS near the end point where the solution is a pale yellow color. Perform a blank determination in the same manner. Calculate the amount of peroxide by the following formula: not more than 5. In the blank determination, the required amount of 0.01 mol/L sodium thiosulfate VS must not exceed 0.1 mL. the peak areas of ethylbenzene and dicyclohexyl from the standard solution (2) by deducting the relevant peak area Amount (mEq/kg) of peroxide ＝ 10 × (V1 － V0)/M from the sample solution, the peak area of benzaldehyde from the sample solution is not more than the difference be-tween the peak areas of benzaldehyde of the sample solution and the standard solution (2) (0.05z), and the peak area of V1: Volume (mL) of 0.01 mol/L sodium thiosulfate VS consumed in the test V0: Volume (mL) of 0.01 mol/L sodium thiosulfate VS consumed in the blank determination ‌JP XVIII Official Monographs / Benzylpenicillin Benzathine Hydrate 525‌ M: Amount (g) of Benzyl Alcohol taken Residue on evaporation—Perform the test after conformation that the sample meets the requirement of the peroxide value. Transfer 10.0 g of Benzyl Alcohol to a porcelain or quartz crucible or platinum dish, previously weighed accurately, and heat on a hot-plate at not exceeding 2009C, taking care to avoid boiling, to evaporate to dryness. Dry the residue on the hot-plate for 1 hour, and allow to cool in a desiccator: not more than 5 mg.
Weigh accurately about 0.9 g of Benzyl Alcohol, add exactly 15 mL of a freshly prepared mixture of de-hydrated pyridine and acetic anhydride (7:1), and heat on a water bath under a reflux condenser for 30 minutes. Cool, add 25 mL of water, and titrate <2.50> the excess acetic acid with 1 mol/L sodium hydroxide VS (indicator: 2 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 1 mol/L sodium hydroxide VS ＝ 108.1 mg of C7H8O
Containers—Tight containers. Storage—Light-resistant.◆
ベンジルペニシリンベンザチン水和物
Benzylpenicillin Benzathine Hydrate is the N,N?-dibenzylethylenediamine salt of a penicillin compound having antibacterial activity produced by the growth of Penicillium species. It contains not less than 1213 Units and not more than 1333 Units per mg, calculated on the anhydrous basis.
Benzyl Benzoate is a colorless, clear, viscous liquid. It has a faint, aromatic odor and a pungent, burning taste. It is miscible with ethanol (95) and with diethyl ether. It is practically insoluble in water. Congealing point: about 179C Specific gravity d 20: about 1.123 Boiling point: about 3239C
(1) Heat gently 1 mL of Benzyl Benzoate with 5 mL of sodium carbonate TS and 2 mL of potassium permanganate TS: the odor of benzaldehyde is perceptible. Warm the titrated mixture obtained in the Assay on a water bath to remove ethanol, and add 0.5 mL of iron (III) chloride TS: a light yellow-red precipitate is produced, which turns white on the addition of dilute hydrochloric acid. D Refractive index <2.45> n20: 1.568 – 1.570
Acidity—Dissolve 5.0 mL of Benzyl Benzoate in 25 mL of neutralized ethanol, and add 0.50 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Residue on ignition <2.44> Not more than 0.05z (2 g).
Weigh accurately about 2 g of Benzyl Benzoate, add exactly 50 mL of 0.5 mol/L potassium hydroxide-ethanol VS, and boil gently for 1 hour under a reflux condenser with a carbon dioxide-absorbing tube (soda lime). Cool, and titrate <2.50> the excess potassium hydroxide with 0.5 mol/L Description Benzylpenicillin Benzathine Hydrate occurs as a white crystalline powder. It is slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water. Identification (1) Determine the absorption spectrum of a solution of Benzylpenicillin Benzathine Hydrate in methanol (1 in 2000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ben-zylpenicillin Benzathine Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋217 – ＋2339(0.1 g calcu-lated on the anhydrous basis, methanol, 20 mL, 100 mm). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Benzylpenicillin Benzathine Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Benzylpenicillin Benzathine Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 70 mg of Benzylpenicil-lin Benzathine Hydrate in 25 mL of methanol, add a solution prepared by dissolving 1.02 g of disodium hydrogen phos-phate and 6.80 g of potassium dihydrogen phosphate in water to make 1000 mL to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak having the relative retention time of about 2.4 to benzylpenicillin obtained from the sample solution is not larger than 2 times the total area of the peaks of benzyl-penicillin and N,N?-dibenzylethylenediamine from the standard solution, and the area of the peak other than benzylpenicillin, N,N?-dibenzylethylenediamine and the peak having the relative retention time of about 2.4 to benzylpenicillin from the sample solution is not larger than the total area of the peaks of benzylpenicillin and N,N?-dibenzylethylenediamine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Assay (1) Benzylpenicillin—Weigh accurately an amount of Benzylpenicillin Benzathine Hydrate, equivalent to about 85,000 Units, dissolve in 25 mL of methanol, and add a solu-tion containing 1.02 g of disodium hydrogen phosphate and 6.80 g of potassium dihydrogen phosphate in 1000 mL of water to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of the solution containing 1.02 g of disodium hydrogen phosphate and 6.80 g of potassium dihydrogen phosphate in 1000 mL of water and methanol (1:1) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Benzylpenicillin Potassium RS, equivalent to about 85,000 Units, and about 25 mg of N,N?-dibenzylethylenediamine diacetate, dissolve in 25 mL of methanol, and add the solution containing 1.02 g of disodium hydrogen phosphate and 6.80 g of potas-sium dihydrogen phosphate in 1000 mL of water to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of the solution containing 1.02 g of disodium hydrogen phos-phate and 6.80 g of potassium dihydrogen phosphate in 1000 mL of water and methanol (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT ands AS, of benzylpenicillin in each solution. Column: A stainless steel column 4.0 mm in inside diame- ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Amount (unit) of benzylpenicillin sodium (C ＝ MS × AT/AS 16H17N2NaO4S) Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, methanol and 0.25 mol/L potassium dihydrogen phosphate TS (pH 3.5) (6:3:1). Mobile phase B: A mixture of methanol, water and 0.25 mol/L potassium dihydrogen phosphate TS (pH 3.5) (6:3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 10 – 20 75 → 0 25 → 100 20 – 55 Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of benzylpenicillin, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase A to make exactly 20 mL. Confirm that the peak area of benzylpenicillin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, N,N?-dibenzylethylenediamine and benzylpenicillin are eluted in this order with the resolution between these peaks being not less than 25. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of benzylpenicillin is not more than 2.0z. Water <2.48> 5.0 – 8.0z (1 g, volumetric titration, direct titration). MS: Amount (unit) of Benzylpenicillin Potassium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, methanol and 0.25 mol/L potassium dihydrogen phosphate TS (pH 3.5) (11:7:2). Flow rate: Adjust so that the retention time of benzyl-penicillin is about 18 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, N,N?-dibenzylethylenediamine and benzylpenicillin are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak areas of N,N?-dibenzylethylenediamine and benzylpenicillin are not more than 2.0z, respectively. (2) N,N?-Dibenzylethylenediamine—Determine the areas, AT and AS, of the peak corresponding to N,N?- dibenzylethylenediamine on the chromatograms obtained in with the sample solution and standard solution. Amount (z) of N,N?-dibenzylethylenediamine (C16H20N2) ＝ MS/MT × AT/AS × 100 × 0.667 MS: Amount (mg) of N,N?-dibenzylethylenediamine dia-cetate taken MT: Amount (mg) of Benzylpenicillin Benzathine Hydrate taken ‌JP XVIII Official Monographs / Benzylpenicillin Potassium 527 0.667: Conversion factor for the molecular mass of N,N?-dibenzylethylenediamine diacetate (C16H20N2･ 2CH3COOH) to that of N,N?-dibenzylethylenedia-mine (benzathine, C16H20N2)
Bepotastine Besilate contains not less than 99.
Bepotastine Besilate occurs as white to pale yellow-white, crystals or crystalline powder. It is very soluble in acetic acid (100), and sparingly soluble in water and in ethanol (99.5). The pH of a solution of 1 g of Bepotastine Besilate in 100 mL of water is about 3.8.
(1) Determine the absorption spectrum of a solution of Bepotastine Besilate (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bepotastine Besilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Bepotastine Besilate as directed under Flame Coloration Test <1.04> (2): a green color ap-pears. Mix well 30 mg of Bepotastine Besilate with 0.1 g of sodium nitrate and 0.1 g of anhydrous sodium carbonate, and gradually ignite. After cooling, dissolve the residue in 2 mL of dilute hydrochloric acid and 10 mL of water, filter if necessary, and add barium chloride TS: a white precipitate is produced.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Bepotastine Besilate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 10 mg of Bepotastine Besilate in 25 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having a relative retention time of about 2.5 to bepotastine, obtained from the sample solution is not larger than the peak area of bepotastine from the standard solu-tion, and the area of the peak other than bepotastine and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of bepotastine from the stand-ard solution. Furthermore, the total area of the peaks other than bepotastine from the sample solution is not larger than the peak area of bepotastine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium 1-pentane sul-fonate in a mixture of 0.05 mol/L potassium dihydrogen phosphate TS (pH 3.0) and acetonitrile (7:3) to make 1000 mL. Flow rate: Adjust so that the retention time of bepotastine is about 6 minutes. Time span of measurement: About 5 times as long as the retention time of bepotastine, beginning after the peak of benzenesulfonic acid. System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of bepotastine obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bepotastine are not less than 3000 and 0.8 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bepotastine is not more than 2.0z. Enantiomer—Dissolve 5.0 mg of Bepotastine Besilate in 25 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the area of each peak by the automatic integration method: the peak area of the enan-tiomer having the relative retention time of about 0.9 to bepotastine obtained from the sample solution is not larger than the peak area of bepotastine from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with b-cyclodextrin binding silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogen phosphate TS and acetonitrile (3:1). Flow rate: Adjust so that the retention time of bepotastine is about 17 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- ‌530 ditions, the number of theoretical plates and the symmetry factor of the peak of bepotastine are not less than 3000 and 0.8 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bepotastine is not more than 5.0z. Water <2.48> Not more than 0.1z (0.3 g, coulometric titration). in 30 minutes of Bepotastine Besilate Tablets is not less than 85z. Start the test with 1 tablet of Bepotastine Besilate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 2.2 mg of bepotastine besilate (C21H25ClN2O3.C6H6O3S), and use this solution as the sam- Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.8 g of Bepotastine Besi-late, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 54.71 mg of C21H25ClN2O3.C6H6O3S
Berberine Chloride Hydrate contains not less than 95.
Berberine Chloride Hydrate occurs as yellow, crystals or crystalline powder. It is odorless or has a faint, characteristic odor. It has a very bitter taste. It is sparingly soluble in methanol, slightly soluble in ethanol (95), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Berberine Chloride Hydrate (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, Amount (mg) of beraprost sodium (C24 ＝ MS × QT/QS × 1/500 H29 NaO5) and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Berberine Chloride RS prepared in the same manner as the sample solution: both MS: Amount (mg) of beraprost sodium for assay taken Internal standard solution—A mixture of water and a solu-tion of 4-isopropylphenol in methanol (1 in 250,000) (1:1). Operating conditions— Detector: A fluorophotometer (excitation wavelength: 285 nm, fluorescence wavelength: 614 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ber-berine Chloride Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Berberine Chloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.1 g of Berberine Chloride Hydrate in 20 ‌534 mL of water by warming, add 0.5 mL of nitric acid, cool, and filter after allowing to stand for 10 minutes. To 3 mL of the filtrate add 1 mL of silver nitrate TS, and collect the pro-duced precipitate: the precipitate does not dissolve in dilute nitric acid, but it dissolves in an excess amount of ammonia TS.
(1) Acidity—Shake thoroughly 0.10 g of Berberine Chloride Hydrate with 30 mL of water, and filter. To the fil-trate add 2 drops of phenolphthalein TS and 0.10 mL of 0.1 mol/L sodium hydroxide VS: the yellow color changes to an orange to red color. Sulfate <1.14>—Shake 1.0 g of Berberine Chloride Hydrate with 48 mL of water and 2 mL of dilute hydrochlo-ric acid for 1 minute, and filter. Discard the first 5 mL of the filtrate, take the subsequent 25 mL of the filtrate, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS, 1 mL of dilute hydrochloric acid, 5 to 10 drops of bromophenol blue TS and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Berber- ine Chloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 30 ppm). Related substances—Dissolve 10 mg of Berberine Chloride Hydrate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 4 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of the peaks other than berberine ob-tained from the sample solution is not larger than the peak area of berberine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of berberine, beginning after the solvent peak. Detection sensitivity: Adjust so that the peak height of berberine obtained from 10 mL of the standard solution is about 10z of the full scale. Water <2.48> 8 – 12z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 10 mg of Berberine Chloride Hydrate, dissolve in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately, determine the water content <2.48> in the same manner as Berberine Chloride Hydrate), and dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS of berberine in each solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column about 4 mm in inside diameter and about 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of monobasic potassium phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. Selection of column: Dissolve each 1 mg of berberine chloride and palmatin chloride in the mobile phase to make 10 mL. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of palmatin and berberine in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with the standard solution under the above operating condi-tions, the relative standard deviation of the peak areas of berberine is not more than 1.5z.
Berberine Tannate is a compound of berberine and tannic acid. It contains not less than 27.
Berberine Tannate occurs as a yellow to light yellow-brown powder. It is odorless or has a faint, charac-teristic odor, and is tasteless. It is practically insoluble in water, in acetonitrile, in meth-anol and in ethanol (95).
(1) To 0.1 g of Berberine Tannate add 10 mL of ethanol (95), and heat in a water bath for 3 minutes with shaking. Cool, filter, and to 5 mL of the filtrate add 1 drop of iron (III) chloride TS: a blue-green color is pro-duced, and on allowing to stand, a bluish black precipitate is formed. Dissolve 0.01 g of Berberine Tannate in 10 mL of methanol and 0.4 mL of 1 mol/L hydrochloric acid TS, and add water to make 200 mL. To 8 mL of the solution add water to make 25 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Ber-berine Tannate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com- Amount (mg) of berberine chloride (C ＝ MS × AT/AS 20H18ClNO4) pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis
(1) Acidity—To 0.10 g of Berberine Tannate add ‌JP XVIII Official Monographs / Betahistine Mesilate 535 30 mL of water, and filter after shaking well. To the filtrate add 2 drops of phenolphthalein TS and 0.10 mL of 0.1 mol/L sodium hydroxide VS: the color of the solution changes from yellow to orange to red. Chloride <1.03>—Shake 1.0 g of Berberine Tannate with 38 mL of water and 12 mL of dilute nitric acid for 5 to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine the peak areas, AT and AS, of berberine in each solution. minutes, and filter. Discard the first 5 mL of the filtrate, to 25 mL of the subsequent filtrate add water to make 50 mL, and perform the test using this solution as the test solution. Amount (mg) of berberine (C ＝ MS × AT/AS × 0.950 20H19NO5) Prepare the control solution with 0.50 mL of 0.01 mol/L hy-drochloric acid VS by adding 6 mL of dilute nitric acid, 10 to 15 drops of bromophenol blue TS and water to make 50 mL (not more than 0.035z). Sulfate <1.14>—Shake 1.0 g of Berberine Tannate with 48 mL of water and 2 mL of dilute hydrochloric acid for 1 minute, and filter. Discard the first 5 mL of the filtrate, take the subsequent 25 mL of the filtrate, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS, 1 mL of dilute hydrochloric acid, 5 to 10 drops of bromophenol blue TS and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Berber- ine Tannate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Related substances—Dissolve 10 mg of Berberine Tannate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 4 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of the peaks other than berberine obtained from the sample solution is not larger than the peak area of berberine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of berberine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of berberine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 3.0z. Water <2.48> Not more than 6.0z (0.7 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 1.0z (1 g).
Weigh accurately about 30 mg of Berberine Tan-nate, dissolve in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately, determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in the mobile phase MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. System suitability— System performance: Dissolve 1 mg each of berberine chloride and palmatin chloride in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, palmatin and ber-berine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Betahistine Mesilate, when dried, contains not less than 98.
Betahistine Mesilate occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), and sparingly soluble in ethanol (99.5). It dissolves in dilute hydrochloric acid. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Betahistine Mesilate in 0.1 mol/L hydrochloric acid (1 in 50,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the ‌536 Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Beta-histine Mesilate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A 30 mg portion of Betahistine Mesilate responds to Qualitative Tests <1.09> (2) for mesilate.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Betahistine Mesilate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Betahistine Mesilate in 10 mL of a mixture of water and acetonitrile (63:37), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and acetonitrile (63:37) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 709C, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Betahistine Mesi-late, previously dried, dissolve in 1 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.42 mg of C8H12N2.2CH4O3S
ベタメタゾンジプロピオン酸エステル
Betamethasone Dipropionate, when dried, contains not less than 97.
Betamethasone Dipropionate occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in acetone and in chloroform, soluble in the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test as directed under Thin-layer Chromatography <2.03> with these solutions. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent ‌JP XVIII Official Monographs / Betamethasone Sodium Phosphate 541 indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and acetone (7:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g, platinum crucible).
(1) To 1 mL of a solution of Betametha-sone Dipropionate in methanol (1 in 10,000) add 4 mL of isoniazid TS, and heat on a water bath for 2 minutes: a yellow color develops. Proceed with 0.01 g of Betamethasone Dipropionate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solu-tion so obtained responds to the Qualitative Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Betamethasone Dipropionate in methanol (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Beta-methasone Dipropionate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋84 – ＋899(after drying, 50 mg, ethanol (99.5), 10 mL, 100 mm).
(1) Fluoride—To 0.10 g of Betamethasone Dipro-pionate add 10.0 mL of diluted 0.01 mol/L sodium hydrox-ide TS (1 in 20), shake for 10 minutes, and filter through a membrane filter (0.4-mm pore size). Place 5.0 mL of the fil-trate in a 20-mL volumetric flask, and add 10 mL of a mix-ture of alizarin complexone TS, acetic acid-potassium ace-tate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1), add water to make 20 mL, allow to stand for 1 hour, and use this solution as the sample solution. Separately, place 1.0 mL of Standard Fluorine Solution in a 20-mL volu-metric flask, add 5.0 mL of diluted 0.01 mol/L sodium hy-droxide TS (1 in 20), then 10 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solu-tion (pH 4.3) and cerium (III) nitrate TS (1:1:1), proceed in the same manner as the preparation of the sample solution, and use this solution as the standard solution. Place 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20) in a 20-mL volumetric flask, and proceed in the same manner as the preparation of the sample solution. Using this solution as the blank, determine the absorbances of the sample solution and standard solution at 600 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>: the absorbance of the sample solution is not more than that of the standard solu-tion (not more than 0.012z). Heavy metals <1.07>—Proceed with 1.0 g of Beta-methasone Dipropionate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 10 betamethasone dipropionate (C28 H37 FO7), and not less mg of Betamethasone Dipropionate in 10 mL of chloroform, and use this solution as the sample solution. Pipet 3 mL of than 3.4z and not more than 4.1z of fluorine (F:19.00). Description Betamethasone Dipropionate occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in acetone and in chloroform, soluble in the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test as directed under Thin-layer Chromatography <2.03> with these solutions. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent ‌JP XVIII Official Monographs / Betamethasone Sodium Phosphate 541 indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and acetone (7:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g, platinum crucible).
(1) Betamethasone dipropionate—Weigh accu-rately about 15 mg of Betamethasone Dipropionate, previ-ously dried, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, and dilute with methanol to exactly 50 mL. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 239 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of betamethasone dipropionate (C28H37FO7) ＝ A/312 × 10,000 (2) Fluorine—Weigh accurately about 10 mg of Beta-methasone Dipropionate, previously dried, and proceed as directed in the procedure of determination for fluorine under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid.
Betamethasone Valerate, when dried, contains not less than 97.
Betamethasone Valerate occurs as a white crys-talline powder. It is odorless. It is freely soluble in chloroform, soluble in ethanol (95), sparingly soluble in methanol, slightly soluble in diethyl ether, and practically insoluble in water. Melting point: about 1909C (with decomposition).
(1) Proceed with 0.01 g of Betamethasone Valerate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution so obtained responds to Qualitative Tests <1.09> for fluoride. (2) Determine the infrared absorption spectrum of Beta-methasone Valerate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of dried Betamethasone Valerate RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋77 – ＋839(after drying, 0.1 g, methanol, 20 mL, 100 mm).
Related substances—Conduct this procedure with-out exposure to daylight. Dissolve 0.02 g of Betamethasone Valerate in 5 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (9:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of chloroform and methanol (9:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly alkaline blue tetrazolium TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g, platinum crucible).
Dissolve about 10 mg each of Betamethasone Valer-ate and Betamethasone Valerate RS, previously dried and JP XVIII Official Monographs / Betamethasone Valerate and Gentamicin Sulfate Cream 543 ‌accurately weighed, in methanol to make exactly 100 mL. Pipet 10 mL each of these solutions, add 10 mL each of the internal standard solution, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of betamethasone valerate to that of the internal standard. these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy, develop the plate with ethyl acetate to a distance of about 10 cm, and air-dry the plate. Spray evenly alkaline blue tetrazolium TS on the plate, and heat the plate at 1009C: the principal spot obtained from the sample solution and the spot from the standard solution are purple in color, and their Rf values are the same. Amount (mg) of betamethasone valerate (C ＝ MS × QT/QS 27H37FO6) (2) To a quantity of Betamethasone Valerate and Gen-tamicin Sulfate Cream, equivalent to about 2 mg (potency) of Gentamicin Sulfate, add 20 mL of ethyl acetate and 10 MS: Amount (mg) of Betamethasone Valerate RS taken Internal standard solution—A solution of isoamyl benzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (7:3). Flow rate: Adjust so that the retention time of betametha- sone valerate is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, betamethasone valerate and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of betamethasone valerate to that of the inter-nal standard is not more than 1.0z.
Betamethasone, when dried, contains not less than 96.
Betamethasone occurs as a white to pale yel-low-white crystalline powder. It is sparingly soluble in methanol, in ethanol (95) and in acetone, and practically insoluble in water. Melting point: about 2409C (with decomposition). It shows crystal polymorphism.
(1) Proceed with 10 mg of Betamethasone as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solu-tion so obtained responds to Qualitative Tests <1.09> for fluoride. Dissolve 1.0 mg of Betamethasone in 10 mL of ethanol (95). Mix 2.0 mL of the solution with 10 mL of phenylhydrazinium hydrochloride TS, heat in a water bath at 609C for 20 minutes, and cool the solution. Determine the absorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, using as the blank the solution prepared with 2.0 mL of ethanol (95) in the same manner as the former solution, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Betamethasone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Beta-methasone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Betamethasone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Betamethasone and Betamethasone RS in acetone, respectively, then evaporate the acetone to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋118 – ＋1269(after drying, 0.1 g, methanol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 0.5 g of Betamethasone according to Method 2, and perform the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 30 ppm). (2) Related substances—Dissolve 10 mg of Betametha-sone in 5 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and meth-anol (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, diethyl ether, methanol and water (385:75:40:6) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.5z (0.1 g, platinum crucible).
Dissolve about 20 mg each of Betamethasone and Betamethasone RS, previously dried and accurately weighed, in methanol to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL each of the internal stand-ard solution, then add methanol to make 50 mL, and use these solutions as the sample solution and standard solution, respectively. Perform the test with 10 mL each of these solu-tions as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of betamethasone to that of the internal standard. Amount (mg) of betamethasone (C22H29FO5) ＝ MS × QT/QS MS: Amount (mg) of Betamethasone RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 1750). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column about 4.0 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (3:2). Flow rate: Adjust so that the retention time of betametha- sone is about 4 minutes. System suitability— System performance: When proceed the test with 10 mL of the standard solution under the above operating conditions, betamethasone and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of betamethasone to that of the internal stand-ard is not more than 1.0z.
Betamipron occurs as white, crystals or crys-talline powder. It is freely soluble in methanol, soluble in ethanol (99.5), and slightly soluble in water. It dissolves in sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Betamipron in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Beta-mipron as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 0.25 g of Betamipron in 100 mL of water by warming, and cool: the pH of this solution is be-tween 3.0 and 3.4.
(1) Clarity and color of solution—Dissolve 1.0 g of Betamipron in 10 mL of sodium hydroxide TS: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Beta- mipron according to Method 4, and perform the test. Pre-pare the control solution with 1.0 mL of Standard Lead So-lution (not more than 10 ppm). b-Alanine—Dissolve 0.25 g of Betamipron in 10 mL of methanol, and use this solution as the sample solution. Separately, dissolve 50 mg of b-alanine in methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of methanol, ethyl acetate, ammonia solution (28) and water (200:200:63:37) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin-butanol TS on the plate, and heat the plate at 1059C for 5 minutes: the spot ob-tained from the sample solution corresponding to the spot ‌546 from the standard solution is not more intense than the spot from the standard solution. Related substances—Dissolve 20 mg of Betamipron in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than betamipron from the sample solution is not larger than 2/5 times the peak area of betamipron from the standard solu-tion, and the total area of the peaks other than betamipron from the sample solution is not larger than the peak area of betamipron from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.12 g of sodium dihydrogen phos-phate dihydrate in 800 mL of water, adjust to pH 7.0 with dilute sodium hydroxide TS, and add water to make 1000 mL. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of betamipron is about 6 minutes. Time span of measurement: About 2 times as long as the retention time of betamipron, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of betamipron obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 5 mg of Betamipron and 5 mg of benzoic acid in 200 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, benzoic acid and betamipron are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betamipron is not more than 2.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Betamipron, dis-solve in 25 mL of ethanol (99.5), add 25 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (poten-tiometric titration). Perform the blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 19.32 mg of C10H11NO3
Betaxolol Hydrochloride, when dried, contains not less than 99.
Betaxolol Hydrochloride occurs as white, crys-tals or crystalline powder. It is very soluble in water, and freely soluble in methanol, in ethanol (99.5) and in acetic acid (100). Dissolve 1.0 g of Betaxolol Hydrochloride in 50 mL of water: the pH of the solution is between 4.5 and 6.5. A solution of Betaxolol Hydrochloride (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Betaxolol Hydrochloride in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Be-taxolol Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Betaxolol Hydrochloride (1 in 10) re-sponds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Betaxolol Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Betax- olol Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Betaxolol Hydrochloride according to Method 3, and per-form the test (not more than 1 ppm). Related substance I—Dissolve 0.10 g of Betaxolol Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 3 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 1 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (10:3:3) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 1 hour: the number of the spots other than the principal spot obtained from the sample solution is not more ‌JP XVIII Official Monographs / Bethanechol Chloride 547 than 3, and they are not more intense than the spot from the standard solution. Related substance II—Dissolve 0.10 g of Betaxolol Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than betaxolol obtained from the sample solution is not larger than the peak area of betaxolol from the standard solution, and the total area of the peaks other than the peak of betaxolol from the sample solution is not larger than 2 times the peak area of betaxolol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted 0.05 mol/L potassium dihydrogen phosphate TS (1 in 2) with the pH adjusted to 3.0 with 1 mol/L hydrochloric acid TS, acetonitrile and methanol (26:7:7). Flow rate: Adjust so that the retention time of betaxolol is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of betaxolol, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 4 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of betaxolol obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of Betaxolol Hydro-chloride and 5 mg of 2-naphthol in 200 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, betaxolol and 2-naphthol are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betaxolol is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Betaxolol Hydro-chloride, previously dried, dissolve in 30 mL of acetic acid (100), add 30 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.39 mg of C18H29NO3.HCl
Bethanechol Chloride, when dried, contains not less than 98.
Bethanechol Chloride occurs as colorless or white crystals or a white, crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), and sparingly soluble in ethanol (99.5). It is hygroscopic. A solution of Bethanechol Chloride (1 in 10) shows no optical rotation.
(1) To 2 mL of a solution of Bethanechol Chloride (1 in 40) add 0.1 mL of a solution of cobalt (II) chloride hexahydrate (1 in 100), then add 0.1 mL of potas-sium hexacyanoferrate (II) TS: A green color is produced, and almost entirely fades within 10 minutes. To 1 mL of a solution of Bethanechol Chloride (1 in 100) add 0.1 mL of iodine TS: a brown precipitate is pro-duced, and the solution shows a greenish brown color. Determine the infrared absorption spectrum of Bethanechol Chloride as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. A solution of Bethanechol Chloride (1 in 100) re-sponds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bethanechol Chloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 1.0 g of Bethanechol Chloride in 2.5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of a solution of ammonium acetate (1 in 100), acetone, 1-butanol and formic acid (20:20:20:1) to a distance of about 10 cm, and dry the plate at 1059C for 15 minutes. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate, and allow to stand for 30 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). ‌548
Weigh accurately about 0.4 g of Bethanechol Chlo-ride, previously dried, dissolve in 2 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.67 mg of C7H17ClN2O2
Bezafibrate, when dried, contains not less than 98.
Bezafibrate occurs as a white crystalline pow-der. It is freely soluble in N,N-dimethylformamide, soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Bezafibrate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bezafibrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Bezafibrate as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—Dissolve 3.0 g of Bezafibrate in 15 mL of N,N-dimethylformamide, add water to make 60 mL, shake well, allow to stand for more than 12 hours, and filter. To 40 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.70 mL of 0.01 mol/L hydrochloric acid VS add 10 mL of N,N-dimethylformamide, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.012z). Heavy metals <1.07>—Proceed with 2.0 g of Bezafibrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Bezafibrate in 35 mL of methanol, add diluted 0.5 mol/L ammonium ace-tate TS (1 in 50) to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add 70 mL of methanol and diluted 0.5 mol/L ammonium acetate TS (1 in 50) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the areas of the peaks having the relative retention times of about 0.65 and 1.86 to bezafibrate obtained from the sample solution are not larger than 1/2 times the peak area of bezafibrate from the stand-ard solution, the area of the peaks other bezafibrate and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of bezafibrate from the standard solution, and the total area of the peaks other than the peak of bezafibrate from the sample solution is not larger than 3/4 times the peak area of bezafibrate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and diluted acetic acid (100) (1 in 100) (9:4). Flow rate: Adjust so that the retention time of bezafibrate is about 6 minutes. Time span of measurement: About 2.5 times as long as the retention time of bezafibrate, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add a mixture of methanol and diluted 0.5 mol/L ammonium acetate TS (1 in 50) (7:3) to make exactly 50 mL. Confirm that the peak area of bezafibrate obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: Dissolve 20 mg of Bezafibrate and 10 mg of 4-chlorobenzoate in 70 mL of methanol, and add diluted 0.5 mol/L ammonium acetate TS (1 in 50) to make 100 mL. When the procedure is run with 5 mL of this solu-tion under the above operating conditions, 4-chlorobenzoate and bezafibrate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of bezafibrate is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Bezafibrate, previ-ously dried, dissolve in 50 mL of ethanol (99.5), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 36.18 mg of C19H20ClNO4
Spherical or massive stone, 1 – 4 cm in diame-ter; externally yellow-brown to red-brown; light, fragile and easily broken. Fractured surface shows yellow-brown to red-brown annular rings, often containing white granular sub-stances or thin layers in these annular rings. Odor, slight; taste, slightly bitter, followed by slight sweet-ness.
To 25 mg of pulverized Oriental Bezoar add 10 mL of methanol, shake for 5 minutes, and centrifuge. Take the supernatant liquid, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 0.5 mL of metha-nol, and use this solution as the sample solution. Separately, dissolve 5 mg each of cholic acid for thin-layer chromatogra-phy and deoxycholic acid for thin-layer chromatography in 5 mL of methanol, respectively, and use these solutions as the standard solution (1) and the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion, standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, formic acid and methanol (30:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 10 minutes: two of the sever-al spots obtained from the sample solution have the same color tone and Rf value with each spot from the standard so-lutions (1) and (2).
(1) Synthetic dye—To 2 mg of pulverized Oriental Bezoar add 1 mL dilute hydrochloric acid: no violet color de-velops. Starch—To 5 mg of pulverized Oriental Bezoar add 2 mL of water, and heat on a water bath for 5 minutes. Cool and add 2 to 3 drops of iodine TS: no blue-purple color de-velops. Sucrose—To 0.02 g of pulverized Oriental Bezoar add 10 mL of water, shake for 15 minutes, and filter. To 1 mL of the filtrate add 2 mL of anthrone TS, and shake: no deep blue-green to dark green color develops. Total ash <5.01> Not more than 10.0z.
Conduct this procedure without exposure to light us-ing light-resistant vessels. The following sample solution and standard solution should be prepared before use. Weigh ac-curately about 10 mg of pulverized Oriental Bezoar, add 10 mL of a mixture of dimethyl sulfoxide and acetic acid (100) (9:1), warm at 609C for 20 minutes, and add a mixture of dimethyl sulfoxide and acetic acid (100) (9:1) to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, ˆlter through a membrane ˆlter, and use the ˆltrate as the sample solution. Separately, weigh ac-curately about 10 mg of bilirubin for assay, add about 350 mg of L-ascorbic acid, and dissolve in a mixture of dimethyl sulfoxide and acetic acid (100) (9:1) to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. ‌JP XVIII Crude Drugs and Related Drugs / Otsujito Extract 2087 Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of bilirubin in each solution. Amount (mg) of bilirubin ＝ MS × AT/AS × 1/2 MS: Amount (mg) of bilirubin for assay taken Operating conditions— Detector: A visible absorption photometer (wavelength: 450 nm). Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of acetonitrile and diluted acetic acid (100) (1 in 100) (19:1). Flow rate: Adjust so that the retention time of bilirubin is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bilirubin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bilirubin is not more than 1.5z.
Bicalutamide occurs as a white, powder or crystalline powder. It is freely soluble in acetone, sparingly soluble in metha-nol, slightly soluble in ethanol (99.5), and practically insolu- ble in water. A solution of Bicalutamide in acetone (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Bicalutamide in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Bicalutamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Bicalutamide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Bicalutamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Alterna-tively, perform the test by the ATR method, and compare the spectrum with the spectrum of Bicalutamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize Bicalutamide and Bicalutamide RS with acetone, respectively, filter and dry the crystals, and perform the test by the potassium bromide disk method or the ATR method in the same manner.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Bicalutamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Bicalutamide in 25 mL of a mixture of water, acetonitrile and phosphoric acid (1000:1000:1), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add a mixture of water, acetonitrile and phosphoric acid (1000:1000:1) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of water, acetonitrile and phosphoric acid (1000:1000:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak areas of related sub-stance M, having the relative retention time of about 0.26 to bicalutamide, related substance N, having the relative reten-tion time of about 0.34, related substance L, having the rela-tive retention time of about 1.03 and related substance K, having the relative retention time of about 1.13, obtained from the sample solution, are not larger than the peak area of bicalutamide from the standard solution, and the area of the peak other than bicalutamide and the peaks mentioned above from the sample solution is not larger than the peak area of bicalutamide from the standard solution. Further-more, the total area of the peaks other than bicalutamide from the sample solution is not larger than 5 times the peak area of bicalutamide from the standard solution. For the areas of the peaks, related substance G, having the relative retention times of about 0.21 and about 0.25, related sub-stance I, having the relative retention time of about 0.23, related substance M, related substance N, related substance O, having the relative retention time of about 0.55, related substance A, having the relative retention time of about 0.95, and related substance L, and related substance P, having the relative retention time of about 1.09 from the sample solution, multiply their correction factors, 0.5, 0.5, 0.5, 0.4, 0.7, 0.5, 1.1, 0.9 and 0.7, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 47 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water, acetonitrile and phos-phoric acid (1000:1000:1) to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the SN ratio of the peak of bi-calutamide is not less than 10. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bicalutamide are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bicalutamide is not more than 5.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately, and powder not less than 20 more than 102.0z of bicalutamide (C calculated on the dried basis. 18H14F4N2O4S), Bezafibrate Extended-release Tablets. Weigh accurately a portion of the powder, equivalent to about 20 mg of bezafibrate (C19H20ClNO4), add 60 mL of methanol and exactly 10 mL of the internal standard solution, and shake Description Bicalutamide occurs as a white, powder or crystalline powder. It is freely soluble in acetone, sparingly soluble in metha-nol, slightly soluble in ethanol (99.5), and practically insolu- ble in water. A solution of Bicalutamide in acetone (1 in 100) shows no optical rotation. Melting point <2.60> 192 – 1979C Bicalutamide shows crystal polymorphism. Identification (1) Determine the absorption spectrum of a solution of Bicalutamide in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Bicalutamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Bicalutamide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Bicalutamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Alterna-tively, perform the test by the ATR method, and compare the spectrum with the spectrum of Bicalutamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize Bicalutamide and Bicalutamide RS with acetone, respectively, filter and dry the crystals, and perform the test by the potassium bromide disk method or the ATR method in the same manner. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Bicalutamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Bicalutamide in 25 mL of a mixture of water, acetonitrile and phosphoric acid (1000:1000:1), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add a mixture of water, acetonitrile and phosphoric acid (1000:1000:1) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of water, acetonitrile and phosphoric acid (1000:1000:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak areas of related sub-stance M, having the relative retention time of about 0.26 to bicalutamide, related substance N, having the relative reten-tion time of about 0.34, related substance L, having the rela-tive retention time of about 1.03 and related substance K, having the relative retention time of about 1.13, obtained from the sample solution, are not larger than the peak area of bicalutamide from the standard solution, and the area of the peak other than bicalutamide and the peaks mentioned above from the sample solution is not larger than the peak area of bicalutamide from the standard solution. Further-more, the total area of the peaks other than bicalutamide from the sample solution is not larger than 5 times the peak area of bicalutamide from the standard solution. For the areas of the peaks, related substance G, having the relative retention times of about 0.21 and about 0.25, related sub-stance I, having the relative retention time of about 0.23, related substance M, related substance N, related substance O, having the relative retention time of about 0.55, related substance A, having the relative retention time of about 0.95, and related substance L, and related substance P, having the relative retention time of about 1.09 from the sample solution, multiply their correction factors, 0.5, 0.5, 0.5, 0.4, 0.7, 0.5, 1.1, 0.9 and 0.7, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 47 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water, acetonitrile and phos-phoric acid (1000:1000:1) to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the SN ratio of the peak of bi-calutamide is not less than 10. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bicalutamide are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bicalutamide is not more than 5.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible). Assay Weigh accurately about 25 mg each of Bicalutamide and Bicalutamide RS (separately determine the loss on dry-ing <2.41> in the same conditions as Bicalutamide), and dis-solve each in a mixture of water, acetonitrile and phosphoric acid (1000:1000 1) to make exactly 25 mL. Pipet 5 mL each of these solutions, add a mixture of water, acetonitrile and phosphoric acid (1000:1000:1) to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of bicalutamide in each solution. Amount (mg) of bicalutamide (C18H14F4N2O4S) ＝ MS × AT/AS MS: Amount (mg) of Bicalutamide RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase A: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography and diluted phosphoric acid (1 in 1000) (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Official Monographs / Bicalutamide 551 Time after injection Mobile phase A Mobile phase B Related substance O: (2RS)-3-[(4-Fluorophenyl)sulfanyl]-2-hydroxy-2- of sample (min) (volz) (volz) methylpropanoic acid 0 – 20 92 → 67 8 → 33 20 – 40 67 → 50 33 → 50 40 – 47 Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bicalutamide are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bicalutamide is not more than 1.0z.
Bifonazole, when dried, contains not less than thin-layer chromatography.
Bifonazole occurs as a white to pale yellow powder. It is odorless and tasteless. It is freely soluble in dichloromethane, soluble in metha-nol, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. A solution of Bifonazole in methanol (1 in 100) does not show optical rotation.
(1) Determine the absorption spectrum of a solution of Bifonazole in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Bi-fonazole, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—To 2.0 g of Bifonazole add 40 mL of water, warm for 5 minutes, and after cooling, filter. To 10 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—To 10 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Bifona- zole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Bifonazole in 10 mL of methanol, and use this solution as the sample solution. Pipet 3 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 25 mL and 5 mL of this solution, add methanol to make exactly 50 mL each, and use these solutions as the standard solutions (1) and (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for solution of methyl yellow in dichloromethane (1 in 500) as indicator, and titrate <2.50>, while shaking vigorously, with 0.01 mol/L sodium lauryl sulfate VS by a buret with 0.02-mL minimum graduation. The end point is reached when the color of the dichloromethane layer changes from yellow to orange-red after dropwise addition of 0.01 mol/L sodium lauryl sulfate VS, strong shaking, and standing for a while. Each mL of 0.01 mol/L sodium lauryl sulfate VS ＝ 3.104 mg of C22H18N2
Weigh accurately about 0.15 g of Bifonazole, previ-ously dried, and dissolve in dichloromethane to make exactly 50 mL. Pipet 5 mL of this solution in a glass-stoppered coni- 98.5z of bifonazole (C 22H18 N2). cal flask, add 10 mL of water, 5 mL of dilute sulfuric acid and 25 mL of dichloromethane, and add 2 to 3 drops of a Description Bifonazole occurs as a white to pale yellow powder. It is odorless and tasteless. It is freely soluble in dichloromethane, soluble in metha-nol, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. A solution of Bifonazole in methanol (1 in 100) does not show optical rotation. Identification (1) Determine the absorption spectrum of a solution of Bifonazole in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Bi-fonazole, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 147 – 1519C Purity (1) Chloride <1.03>—To 2.0 g of Bifonazole add 40 mL of water, warm for 5 minutes, and after cooling, filter. To 10 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—To 10 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Bifona- zole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Bifonazole in 10 mL of methanol, and use this solution as the sample solution. Pipet 3 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 25 mL and 5 mL of this solution, add methanol to make exactly 50 mL each, and use these solutions as the standard solutions (1) and (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for solution of methyl yellow in dichloromethane (1 in 500) as indicator, and titrate <2.50>, while shaking vigorously, with 0.01 mol/L sodium lauryl sulfate VS by a buret with 0.02-mL minimum graduation. The end point is reached when the color of the dichloromethane layer changes from yellow to orange-red after dropwise addition of 0.01 mol/L sodium lauryl sulfate VS, strong shaking, and standing for a while. Each mL of 0.01 mol/L sodium lauryl sulfate VS ＝ 3.104 mg of C22H18N2
Biperiden Hydrochloride, when dried, contains not less than 99.
Biperiden Hydrochloride occurs as a white to brownish yellow-white crystalline powder. It is freely soluble in formic acid, slightly soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 2709C (with decomposition).
(1) Dissolve 0.02 g of Biperiden Hydro-chloride in 5 mL of phosphoric acid: a green color develops. Dissolve 0.01 g of Biperiden Hydrochloride in 5 mL of water by heating, cool, and add 5 to 6 drops of bromine TS: a yellow precipitate is formed. Determine the absorption spectrum of a solution of Biperiden Hydrochloride (1 in 2000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Biperiden Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Dissolve 0.02 g of Biperiden Hydrochloride in 10 mL of water by heating, and cool: the solution responds to Qualitative Tests <1.09> for chloride.
(1) Acidity or alkalinity—To 1.0 g of Biperiden Hydrochloride add 50 mL of water, shake vigorously, filter, and to 20 mL of the filtrate add 1 drop of methyl red TS: no red to yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of Biperiden Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Biperiden Hydrochloride according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Biperiden Hy-drochloride in 20 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and am-monia solution (28) (80:15:2) to a distance of about 15 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Biperiden Hydro-chloride, previously dried, dissolve in 5 mL of formic acid, add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.79 mg of C21H29NO.HCl
Bisacodyl Suppositories contain not less than 90.0z and not more than 110.0z of the labeled amount of bisacodyl (C22H19NO4: 361.39). Method of preparation Prepare as directed under Supposi-tories, with Bisacodyl. Identification (1) To a quantity of Bisacodyl Supposito-ries, equivalent to 6 mg of Bisacodyl, add 20 mL of ethanol (95), warm on a water bath for 10 minutes, shake vigorously for 10 minutes, and allow to stand in ice water for 1 hour. Centrifuge the solution, filter the supernatant liquid, and to 2 mL of the filtrate add ethanol (95) to make 20 mL. Deter-mine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 261 nm and 265 nm. (2) Use the filtrate obtained in (1) as the sample solution. Separately, dissolve 6 mg of Bisacodyl RS in 20 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of 2-butanone, chloroform and xylene (1:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution and that from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 suppository of Bisacodyl Suppositories add a suitable amount of tetrahydrofuran, warm to 409C, and shake to dis-solve. After cooling, add tetrahydrofuran to make exactly V mL so that each mL contains about 0.
(1) To a quantity of Bisacodyl Supposito-ries, equivalent to 6 mg of Bisacodyl, add 20 mL of ethanol (95), warm on a water bath for 10 minutes, shake vigorously for 10 minutes, and allow to stand in ice water for 1 hour. Centrifuge the solution, filter the supernatant liquid, and to 2 mL of the filtrate add ethanol (95) to make 20 mL. Deter-mine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 261 nm and 265 nm. (2) Use the filtrate obtained in (1) as the sample solution. Separately, dissolve 6 mg of Bisacodyl RS in 20 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of 2-butanone, chloroform and xylene (1:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution and that from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 suppository of Bisacodyl Suppositories add a suitable amount of tetrahydrofuran, warm to 409C, and shake to dis-solve. After cooling, add tetrahydrofuran to make exactly V mL so that each mL contains about 0.2 mg of bisacodyl (C22H19NO4). Pipet 5 mL of this solution, and proceed as directed in the Assay. Amount (mg) of bisacodyl (C22H19NO4) ＝ MS × QT/QS × V/50 JP XVIII Official Monographs / Bismuth Subgallate 555 MS: Amount (mg) of Bisacodyl RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in acetonitrile (3 in 100,000). ‌Assay Weigh accurately not less than 20 Bisacodyl Sup-positories, make them fine fragments carefully, and mix uniformly. Weigh accurately a portion of the fragments, equivalent to about 10 mg of bisacodyl (C22H19NO4), add 40 mL of tetrahydrofuran, warm to 409C, dissolve by shaking, cool, and add tetrahydrofuran to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, and add the mobile phase to make 100 mL. Cool this solution in ice for 30 minutes, centrifuge, filter the supernatant liquid through a membrane filter with pore size of 0.5 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 10 mg of Bisacodyl RS, previously dried at 1059C for 2 hours, and dissolve in tetrahydrofuran to make exactly 50 mL. Pipet 5 mL of this solution, proceed in the same manner as the sample solution, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of bisacodyl to that of the internal standard. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid, in dilute nitric acid and in dilute sulfuric acid on warming. It dissolves in sodium hydroxide TS, forming a clear, yellow solution, which turns red immediately. It is affected by light. Identification (1) Ignite 0.5 g of Bismuth Subgallate: it chars at first, and leaves finally a yellow residue. The residue responds to Qualitative Tests <1.09> for bismuth salt. (2) To 0.5 g of Bismuth Subgallate add 25 mL of water and 20 mL of hydrogen sulfide TS, and shake well. Filter off the blackish brown precipitate, and add 1 drop of iron (III) chloride TS to the filtrate: a blue-black color is produced.
(1) Clarity of solution—Dissolve 1.0 g of Bismuth Subgallate in 40 mL of diluted sodium hydroxide TS (1 in 8): the solution is clear. Sulfate—Ignite 3.0 g of Bismuth Subgallate in a por-celain crucible, and cautiously dissolve the residue in 2.5 mL of nitric acid by warming. Pour the solution into 100 mL of water, shake, and filter. Evaporate 50 mL of the filtrate on a water bath to 15 mL. Add water to make 20 mL, filter again, and use the filtrate as the sample solution. To 5 mL of the sample solution add 2 to 3 drops of barium nitrate TS: no Amount (mg) of bisacodyl (C 22H19NO4) ＝ M S × QT/QS turbidity is produced. Nitrate—To 0.5 g of Bismuth Subgallate add 5 mL of MS: Amount (mg) of Bisacodyl RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in acetonitrile (3 in 100,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.01 mol/L citric acid TS, acetonitrile and methanol (2:1:1). Flow rate: Adjust so that the retention time of bisacodyl is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and bisacodyl are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of bisacodyl to that of the internal standard is not more than 1.0z.
Bisacodyl, when dried, contains not less than 98.
Bisacodyl occurs as a white crystalline powder. It is freely soluble in acetic acid (100), soluble in acetone, slightly soluble in ethanol (95) and in diethyl ether, and prac- tically insoluble in water. It dissolves in dilute hydrochloric acid.
(1) Determine the absorption spectrum of a solution of Bisacodyl in ethanol (95) (3 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Bisacodyl RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Bi-sacodyl, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Bisacodyl RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Bisacodyl in 30 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.35 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.012z). Sulfate <1.14>—Dissolve 1.0 g of Bisacodyl in 2 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution as follows: to 0.35 mL of 0.005 mol/L sulfuric acid VS add 2 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.017z). Heavy metals <1.07>—Proceed with 2.0 g of Bi- sacodyl according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 0.20 g of Bisacodyl in 10 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 2-butanone, chlo- roform and xylene (1:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Bisacodyl, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from orange-yellow to green (indicator: 0.5 mL of p-naphtholbenzein TS). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.14 mg of C22H19NO4
Bismuth Subgallate, when dried, contains not less than 47.
Bismuth Subgallate occurs as a yellow powder. It is odorless and tasteless. dilute sulfuric acid and 25 mL of iron (II) sulfate TS, shake well, and filter. Superimpose carefully 5 mL of the filtrate on sulfuric acid: no red-brown color develops at the zone of contact. Ammonium—Dissolve 1.0 g of Bismuth Subgallate in 5 mL of sodium hydroxide TS, and heat: the gas evolved does not change moistened red litmus paper to blue. Copper—To 5 mL of the sample solution obtained in (2) add 1 mL of ammonia TS, and filter: no blue color de-velops in the filtrate. Lead—Ignite 1.0 g of Bismuth Subgallate at about 5009C in a porcelain crucible, dissolve the residue in a smallest possible amount of nitric acid added dropwise, evaporate over a low flame to dryness , and cool. Add 5 mL of a solution of potassium hydroxide (1 in 6) to the residue, boil carefully for 2 minutes, cool, and centrifuge. Take the supernatant liquid in a test tube, add 10 drops of potassium chromate TS, and acidify the solution by adding acetic acid (100) dropwise: neither turbidity nor a yellow precipitate is produced. Silver—To 5 mL of the sample solution obtained in (2) add 0.5 mL of nitric acid and 2 to 3 drops of dilute hy-drochloric acid: no turbidity is produced. Alkaline earth metals and alkali metals—Boil 1.0 g of Bismuth Subgallate with 40 mL of diluted acetic acid (31) (1 in 2) for 2 minutes, cool, add water to make 40 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute hydrochlo-ric acid, boil, immediately pass hydrogen sulfide thoroughly through the solution, filter the precipitate produced, and wash with water. Combine the filtrate and the washings, add 5 drops of sulfuric acid, and evaporate to dryness. Ignite as directed under Residue on Ignition <2.44>: the mass of the residue is not more than 5.0 mg. Arsenic <1.11>—Mix well 0.20 g of Bismuth Subgal-late with 0.20 g of calcium hydroxide, and ignite the mixture. Dissolve the residue in 5 mL of dilute hydrochloric acid, use this solution as the test solution, and perform the test (not more than 10 ppm). Gallic acid—To 1.0 g of Bismuth Subgallate add 20 mL of ethanol (95), shake for 1 minute, and filter. ‌556 ‌Evaporate the filtrate on a water bath to dryness: the mass of the residue is not more than 5.0 mg. Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, 3 hours).
Weigh accurately about 0.5 g of Bismuth Subgallate, previously dried, ignite at about 5009C for 30 minutes, and cool. Dissolve the residue in 5 mL of diluted nitric acid (2 in 5) by warming, and add water to make exactly 100 mL. Measure exactly 30 mL of this solution, add 200 mL of water, and titrate <2.50> with 0.02 mol/L disodium dihydro-gen ethylenediamine tetraacetate VS until the color of the so-lution changes from red-purple to yellow (indicator: 2 to 3 drops of xylenol orange TS). Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.180 mg of Bi
Bismuth Subnitrate, when dried, contains not less than 71.
Bismuth Subnitrate occurs as a white powder. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It readily dissolves in hydrochloric acid and in nitric acid without effervescence. It is slightly hygroscopic, and changes moistened blue litmus paper to red.
Bismuth Subnitrate responds to Qualitative Tests <1.09> for bismuth salt and nitrate.
(1) Chloride <1.03>—Dissolve 0.7 g of Bismuth Subnitrate in 2 mL of water and 2 mL of nitric acid, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 2 mL of nitric acid on a water bath to dryness, add 0.70 mL of 0.01 mol/L hydro-chloric acid VS, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.035z). Sulfate—Dissolve 3.0 g of Bismuth Subnitrate in 3.0 mL of warmed nitric acid, pour this solution into 100 mL of water, shake, and filter. Concentrate the filtrate on a water bath to 30 mL, filter, and use this filtrate as the sample solu-tion. To 5 mL of the sample solution add 2 to 3 drops of barium nitrate TS: no turbidity is produced. Ammonium—Boil 0.10 g of bismuth Subnitrate with 5 mL of sodium hydroxide TS: the gas evolved does not change moistened red litmus paper to blue. Copper—To 5 mL of the sample solution obtained in (2) add 2 mL of ammonia TS, and filter: no blue color de-velops. Lead—To 1.0 g of Bismuth Subnitrate add 5 mL of a solution of sodium hydroxide (1 in 6), boil carefully for 2 minutes, cool and centrifuge. Transfer the supernatant liq-uid to a test tube, add 10 drops of potassium chromate TS, and add dropwise acetic acid (31) to render the solution acid: no turbidity or yellow precipitate is produced. Silver—To 5 mL of the sample solution obtained in (2) add 0.5 mL of nitric acid and 2 to 3 drops of dilute hy-drochloric acid: no turbidity is produced. Alkaline earth metals and alkali metals—Boil 2.0 g of Bismuth Subnitrate with 40 mL of diluted acetic acid (31) (1 in 2) for 2 minutes, cool, add water to make 40 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute hydrochlo-ric acid, boil, immediately pass hydrogen sulfide thoroughly through the solution, filter, and wash the residue with water. Combine the filtrate and the washings, add 5 drops of sulfu-ric acid, evaporate to dryness, and ignite as directed under Residue on Ignition <2.44>: the residue is not exceed 5.0 mg Arsenic <1.11>—To 0.20 g of Bismuth Subnitrate add mL of sulfuric acid, heat until white fumes evolve, dilute cautiously with water to 5 mL, use this solution as the test solution, and perform the test (not more than 10 ppm). Loss on drying <2.41> Not more than 3.0z (2 g, 1059C, 2 hours).
Weigh accurately about 0.4 g of Bismuth Subnitrate, previously dried, dissolve in 5 mL of diluted nitric acid (2 in 5) by warming, and add water to make exactly 100 mL. Pipet 25 mL of the solution, add 200 mL of water and titrate <2.50> with 0.02 mol/L disodium dihydrogen ethylene- diamine tetraacetate VS until the color of the solution changes from red-purple to yellow (indicator: 5 drops of xylenol orange TS) Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.180 mg of Bi
Bisoprolol Fumarate, when dried, contains not less than 98.
Bisoprolol Fumarate occurs as white crystals or a white crystalline powder. It is very soluble in water and in methanol, and freely soluble in ethanol (99.5) and in acetic acid (100). A solution of Bisoprolol Fumarate (1 in 10) shows no opti-cal rotation.
(1) Determine the absorption spectrum of a solution of Bisoprolol Fumarate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the ‌JP XVIII Official Monographs / Bisoprolol Fumarate Tablets 557 same wavelengths. Determine the infrared absorption spectrum of Bi-soprolol Fumarate as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Bisoprolol Fumarate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Bisoprolol Fumarate in 100 mL of a mixture of water and acetonitrile (4:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and acetonitrile (4:1) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly Fumarate, previously dried, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). The endpoint of titration is when the purple color of the solution turns blue and then blue-green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.35 mg of (C18H31NO4)2.C4H4O4
Bleomycin Hydrochloride is the hydrochloride of a mixture of substances having antitumor activity pro-duced by the growth of Streptomyces verticillus. It contains not less than 1400 mg (potency) and not more than 2000 mg (potency) per mg, calculated on the dried basis.
Bleomycin Hydrochloride occurs as a white to yellow-white powder. It is freely soluble in water, and slightly soluble in ethanol (95). It is hygroscopic.
(1) To 4 mg of Bleomycin Hydrochloride add 5 mL of copper (II) sulfate TS, and dissolve in water to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Bleomycin Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Bleomycin Hydrochloride (1 in 100) re-sponds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Bleomycin Hydrochloride in 20 mL of water is be-tween 4.5 and 6.0. Content ratio of the active principle Dissolve 10 mg of Bleomycin Hydrochloride in 20 mL of water, and use this so-lution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions, and deter-mine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak of bleomycin A2 (the first principal peak) is between 55z and 70z, that of bleomycin B2 (the second principal peak) is between 25z and 32z, the total amount of the peak of bleomycin A2 and bleomycin B2 is not less than 85z, the amount of the peak of demethylbleomy-cin A2 (a peak having the relative retention time of 1.5 – 2.5 to bleomycin A2) is not more than 5.5z, and the total amount of the rest peaks is not more than 9.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase stock solution: Dissolve 0.96 g of sodium 1-pentanesulfonate and 1.86 g of disodium dihydrogen ethyl-enediamine tetraacetate dihydrate in 1000 mL of water and 5 mL of acetic acid (100), and adjust the pH to 4.3 with ammonia TS. Mobile phase A: A mixture of the mobile phase stock so-lution and methanol (9:1). Mobile phase B: A mixture of the mobile phase stock solu-tion and methanol (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 60 100 → 0 0 → 100 60 – 75 Flow rate: About 1.2 mL per minute. Time span of measurement: 20 minutes after elution of the peak of demethylbreomycin A2, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, bleomycin A2 and bleomycin B2 are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of bleomycin A2 is not more than 2.0z.
(1) Clarity and color of solution—A solution ob-tained by dissolving 80 mg of Bleomycin Hydrochloride in 4 mL of water is clear and colorless. Copper—Dissolve exactly 75 mg of Bleomycin Hy-drochloride in exactly 10 mL of diluted nitric acid (1 in 100), and use this solution as the sample solution. Separately, to exactly 15 mL of Standard Copper Solution add diluted nitric acid (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions: the absorbance of the sample solu-tion is not more than that of the standard solution (not more than 200 ppm). Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Copper hollow-cathode lamp. Wavelength: 324.8 nm. Loss on drying <2.41> Not more than 5.0z (60 mg, in vacuum, phosphorus (V) oxide, 609C, 3 hours. Take the sample to be tested while avoiding moisture absorption).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Mycobacterium smegmatis ATCC 607 Agar medium for seed, base layer and transferring the test organism Mix all the components, and sterilize. Adjust the pH after sterilization to 6.9 – 7.1 with sodium hydroxide TS. Liquid media for suspending the test organism Glycerin 10.0 g Peptone 10.0 g Meat extract 10.0 g Sodium chloride 3.0 g Water 1000 mL Mix all the components, and sterilize. Adjust the pH after sterilization to 6.9 – 7.1 with sodium hydroxide TS. Preparation of seeded agar layer—Cultivate the test organism on the slant of the agar medium for transferring the test organism at 279C for 40 to 48 hours, then inoculate the test organism thus obtained in 100 mL of the liquid me-dia for suspending the test organism, cultivate with shaking at between 259C and 279C for 5 days, and use this as the sus-pension of test organism. Store the suspension of test organ-ism at a temperature not exceeding 59C, and use within 14 days. Add 0.5 mL of the suspension of test organism in 100 mL of the agar medium for seed previously kept at 489C, mix thoroughly, and use as the seeded agar layer. Preparation of cylinder-agar plate—Proceed as di-rected in 1.7. Preparation of cylinder-agar plates under the Microbial Assay for Antibiotics, dispensing 5.0 mL of agar medium for base layer and 8.0 mL of the agar medium for seed into the Petri dish. Standard solutions—Weigh accurately an amount of Bleomycin A2 Hydrochloride RS, previously dried under reduced pressure not exceeding 0.67 kPa at an ordinary tem-perature for 3 hours, equivalent to about 15 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below, and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 6.8) to make solutions so that each mL contains 30 mg (potency) and 15 mg (po-tency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Bleomycin Hydrochloride, equivalent to about 15 mg (po-tency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer so-lution (pH 6.8) to make solutions so that each mL contains 30 mg (potency) and 15 mg (potency), and use these solutions as the high concentration sample solution and low concen-tration sample solution, respectively.
Bleomycin Sulfate is the sulfate of a mixture of sub-stances having antitumor activity produced by the growth of Streptomyces verticillus. It contains not less than 1400 mg (potency) and not more than 2000 mg (potency) per mg, calculated on the dried basis.
Bleomycin Sulfate occurs as a white to yellow-white powder. It is freely soluble in water, and slightly soluble in ethanol (95). It is hygroscopic.
(1) To 4 mg of Bleomycin Sulfate add 5 mL of copper (II) sulfate TS, and dissolve in water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bleomycin Sulfate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Bleomycin Sulfate (1 in 200) responds to Qualitative Tests <1.09> (1) and (2) for sulfate. pH <2.54> The pH of a solution obtained by dissolving 10 mg of Bleomycin Sulfate in 20 mL of water is between 4.5 and 6.0. Content ratio of the active principle Dissolve 10 mg of Bleomycin Sulfate in 20 mL of water, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak of bleomycin A2 (the first principal peak) is between 55z and 70z, that of bleomycin B2 (the second principal peak) is between 25z and 32z, the total amount of the peak of bleomycin A2 and bleomycin B2 is not less than 85z, the amount of the peak of demethylbleomy-cin A2 (a peak having the relative retention time of 1.5 – 2.5 to bleomycin A2) is not more than 5.5z, and the total amount of the rest peaks is not more than 9.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase stock solution: Dissolve 0.96 g of sodium 1-pentanesulfonate and 1.86 g of disodium dihydrogen ethyl-enediamine tetraacetate dihydrate in 1000 mL of water and 5 mL of acetic acid (100), and adjust the pH to 4.3 with am-monia TS. Mobile phase A: A mixture of the mobile phase stock so-lution and methanol (9:1). Mobile phase B: A mixture of the mobile phase stock solu-tion and methanol (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 60 100 → 0 0 → 100 60 – 75 Flow rate: About 1.2 mL per minute. ‌Time span of measurement: Twenty minutes after elution of the peak of demethylbleomycin A2, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, bleomycin A2 and bleomycin B2 are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of bleomycin A2 is not more than 2.0z.
(1) Clarity and color of solution—A solution ob-tained by dissolving 80 mg of Bleomycin Sulfate in 4 mL of water is clear and colorless. Copper—Dissolve exactly 75 mg of Bleomycin Sul-fate in 10 mL of diluted nitric acid (1 in 100), and use this so-lution as the sample solution. Separately, to exactly 15 mL of Standard Copper Solution add diluted nitric acid (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions: the absorbance of the sample solution is not more than that of the standard solution (not more than 200 ppm). Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Copper hollow-cathode lamp. Wavelength: 324.8 nm. Loss on drying <2.41> Not more than 3.0z (60 mg, in vacuum, phosphorus (V) oxide, 609C, 3 hours. Take the sample to be tested while avoiding moisture absorption).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Mycrobacterium smegmatis ATCC 607 Agar medium for seed, base layer and transferring the test organism Glycerin 10.0 g Peptone 10.0 g Meat extract 10.0 g Sodium chloride 3.0 g Agar 15.0 g Water 1000 mL Mix all the components and sterilize. Adjust the pH after sterilization to 6.9 – 7.1 with sodium hydroxide TS. Liquid media for suspending the test organism Glycerin 10.0 g Peptone 10.0 g Meat extract 10.0 g Sodium chloride 3.0 g Water 1000 mL Mix all the components and sterilize. Adjust the pH after sterilization to 6.9 – 7.1 with sodium hydroxide TS. Preparation of seeded agar layer—Cultivate the test organism on the slant of the agar medium for transferring the test organism at 279C for 40 to 48 hours, then inoculate the test organism thus obtained in 100 mL of the liquid me-dia for suspending the test organism, cultivate with shaking at between 259C and 279C for 5 days, and use this as the sus-pension of test organism. Store the suspension of test organ-ism at a temperature not exceeding 59C, and use within 14 days. Add 0.5 mL of the suspension of test organism in 100 mL of the agar medium for seed previously kept at 489C, mix thoroughly, and use as the seeded agar layer. Preparation of cylinder-agar plate—Proceed as di-rected in 1.7. Preparation of cylinder-agar plates under the Microbial Assay for Antibiotics, dispensing 5.0 mL of agar medium for base layer and 8.0 mL of the agar medium for seed into the Petri dish. Standard solutions—Weigh accurately an amount of Bleomycin A2 Hydrochloride RS, previously dried under reduced pressure not exceeding 0.67 kPa at an ordinary tem-perature for 3 hours, equivalent to about 15 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below, and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 6.8) to make solutions so that each mL contains 30 mg (potency) and 15 mg (po-tency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Bleomycin Sulfate, equivalent to about 15 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 6.8) to make solutions so that each mL contains 30 mg (potency) and 15 mg (potency), and use these solutions as the high con-centration sample solution and low concentration sample so-lution, respectively.
Bofutsushosan Extract contains not less than 9 mg and not more than 36 mg of paeoniflorin (C23H28O11: 480.
Bofutsushosan Extract is a yellow-brown to brown powder or black-brown viscous extract. It has a slightly odor and a sweet and slightly bitter taste. Identiˆcation (1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, add 10 mL of sodium hydroxide TS, shake, centrifuge, and use the diethy ether layer as the sam-ple solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of butyl acetate and hexane (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main Bofutsushosan Extract / Crude Drugs and Related Drugs wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Japanese Angelica Root; Cnidium Rhizome). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 1 minute: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple to purple spot from the standard solution (Peony Root). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, and heat the plate at 1059C for 1 minute: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple to purple spot from the standard solution (Gardenia Fruit). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 1.0 g of pulverized forsythia fruit add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution as bands on the original line on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and am-monia solution (28) (10:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple spot (Rf value: about 0.4) from the standard solution (Forsythia Fruit). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of diluted phosphoric acid (1 in 30), shake, then add 15 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the sample solution. Separately, shake 0.2 g of pulverized mentha herb with 10 mL of diluted phosphoric acid (1 in 30), add 15 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, ethyl acetate, water, and acetic acid (100) (10:10:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzo-quinone monoimine TS on the plate, heat the plate at 1059C for 5 minutes: one of the spot among the several spots ob-tained from the sample solution has the same color tone and Rf value with the red-brown spot (Rf value: around 0.4) from the standard solution (Mentha Herb). Perform the test according to the following (i) or (ii) (Ginger). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution. To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of [6]-shogaol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution. To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of 0.1 mol/L hydrochloric acid TS, shake, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use the solution as the sample solution. Sepa-rately, dissolve 1 mg of rosmarinic acid for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (60:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the green-ish brown spot from the standard solution (Schizonepeta Spike; Mentha Herb). JP XVIII Crude Drugs and Related Drugs / Bofutsushosan Extract 1963 For preparation prescribed Saposhnikovia Root and Rhizome—To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of 4?-O-glycosyl-5-O-methylvisamminol for thin-layer chromatog-raphy in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 2 minutes, then exa-mine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluores-cent spot from the standard solution (Saposhnikovia Root and Rhizome). For preparation prescribed Glehnia Root and Rhi- zome—To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 5 mL of ethyl acetate, and heat under a reflux con-denser for 30 minutes. After cooling, filter, and use the fil-trate as the sample solution. Separately, dissolve 1 mg of scopoletin for thin-layer chromatography in 10 mL of meth-anol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution (Glehnia Root and Rhizome). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of sodium hydroxide TS, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 15 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (4:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot is observed at about 0.5 of Rf value (Ephedra Herb). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, dissolve 1 mg of rhein for thin-layer chromatography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the orange fluo-rescent spot from the standard solution (Rhubarb). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, dis-solve 1 mg of atractylenolide III for thin-layer chromatogra-phy in 2 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air- dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the red to red-purple spot from the standard solution (Atractylodes Rhi-zome). To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of sodium carbonate TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 2.0 g of pulver-ized platycodon root add 10 mL of sodium carbonate TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate and water (4:4:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly 1,3-naphthalenediol TS on the plate, heat the plate at 1059C for 5 minutes: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot (Rf value: about 0.4) from the standard solution (Platycodon Root). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, centrifuge, then add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown to grayish brown spot from the standard solution (Scutellaria Root). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand- Bofutsushosan Extract / Crude Drugs and Related Drugs ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C H O ) color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). ＝ MS × AT/AS × 5/8 23 28 11 Place 2.0 g of the dry extract (or 6.0 g of the viscous extract) in a porcelain crucible, ignite to incinerate at 5509C, then to the residue add 60 mL of water, shake, centrifuge, and use the supernatant as the sample solution. Add ammo-nium oxalate TS to the sample solution: a white precipitate is formed. The precipitate does not dissolve in diluted acetic acid, but dissolve on the addition of diluted hydrochloric acid (Gypsum). Place 2.0 g of the dry extract (or 6.0 g of the viscous extract) in a porcelain crucible, ignite to incinerate at 5509C. To the residue add 60 mL of water, shake well, centrifuge, and use the supernatant as the sample solution. The sample solution responds to the Qualitative Tests <1.09> (1) for sulfate (Gypsum; Sodium Sulfate or Anhydrous Sodium Sul-fate). Place 2.0 g of the dry extract (or 6.0 g of the viscous extract) in a crucible, and ignite at 5509C for 5 hours to in-cinerate. To the residue add 3 mL of diluted sulfuric acid (1 in 3), and heat until white fumes are evolved. After cooling, add 20 mL of water, shake, and filter. To 5 mL of the fil-trate add ammonia TS until a white gelatinous precipitate is formed, centrifuge, and remove the supernatant liquid. To the residue add 5 mL of water, shake, centrifuge, and re-move the supernatant liquid. Then, to the residue add 5 mL of water, shake, centrifuge, and remove the supernatant liq-uid. To the obtained residue add 5 drops of alizarin red S TS, and shake occasionally in lukewarm water: the residue shows red to red-brown in color (Kasseki).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not less than 10.0z and more than 22.0z, calculated on the dried basis.
(1) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, elute through a column packed with 2 g of polyamide for column chromatography using 20 mL of water, then add 1 mL of acetic acid (100), add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Total alkaloids (ephedrine and pseudoephedrine)—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as above, and remove the diethyl ether layer. To the aqueous layer add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. In addition, repeat twice in the same manner for the aqueous layer using 1.0 mL of ammonia TS and 20 mL of diethyl ether. Combine all the extracts, evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in diluted methanol (1 in 2) to make exactly 50 mL, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 10 mg of ephedrine hydrochloride for assay of crude drug, previously dried at 1059C for 3 hours, dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas, ATE and ATP, of ephedrine and pseudoephedrine ob-tained with the sample solution, and the peak area, AS, of ephedrine obtained with the standard solution. Amount (mg) of total alkaloids (ephedrine and pseudoe-phedrine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 JP XVIII Crude Drugs and Related Drugs / Bofutsushosan Extract 1965 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drug taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, then add 650 mL of water and 1 mL of phosphoric acid. Flow rate: 1.0 mL per minute (the retention time of ephe-drine is about 27 minutes). System suitability— System performance: Dissolve 1 mg each of ephedrine hy-drochloride for assay of crude drug and pseudoephedrine hy-drochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephe-drine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 1.5z. ‌Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Boiogito Extract contains not less than 4 mg and not more than 16 mg of sinomenine, and not less than 10 mg and not more than 30 mg of glycyrrhizic acid rately, dissolve 1.
Boiogito Extract is a light yellow-brown to reddish brown powder or black-brown viscous extract. It has a slightly odor, and a sweet taste at first and then a slight hot and bitter taste later.
(1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 15 mL of sodium hydroxide TS, shake, centrifuge, and separate the supernatant liquid. To this liquid add 10 mL of 1-butanol, shake, centrifuge, and separate 1-butanol layer. To this liquid add 10 mL of water, shake, centrifuge, separate the 1-butanol layer, then evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in 1 mL of methanol, and use the solution as the sample solution. Separately, dissolve 1 mg of sinomenine for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red to red-brown spot from the standard solution (Sinomenium Stem and Rhi-zome). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 15 mL of sodium hydroxide TS, shake, centrifuge, and separate the supernatant liquid. To this liquid add 10 mL of 1-butanol, shake, centrifuge, and separate 1-butanol layer. To the aqueous layer add 10 mL of 1-butanol, and proceed in the same manner as above. Combine the 1-butanol layers, add 10 mL of water, shake, centrifuge, sepa-rate the 1-butanol layer, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue in exactly 1 mL of methanol, and use this solution as the sample solution. Sepa- heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-brown spot obtained from the standard solution, and the spot is larger and more intense than the spot from the standard solution (Astragalus Root). For preparation prescribed Atractylodes Rhizome—To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of Atrac-tylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red to red-purple spot from the standard solu-tion (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), then add 0.5 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying on the plate, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhizome). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air- JP XVIII Crude Drugs and Related Drugs / Boiogito Extract 1967 dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 11.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not less than 8.0z, calculated on the dried basis. However, for the dry extract prepared by adding Light Anhydrous Silicic Acid, between 9.0z and 18.0z.
(1) Sinomenine—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 254 nm). Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 309C. Mobile phase: To 3 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, then add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of sinomenine is about 18 minutes). System suitability— System performance: When the procedure is run with 10 mL each of the sample solution, the sinomenine standard so-lution and the glycyrrhizic acid standard solution obtained in Assay (2) under the above operating conditions, peaks of sinomenine and glycyrrhizic acid are observed in the sample solution, glycyrrhizic acid and sinomenine are eluted in this order with the resolution between these peaks being not less than 4.5. Furthermore, except for the peak of glycyrrhizic acid, distinct peaks are observed before and after the peak of sinomenine, and the resolutions between sinomenine and these peaks are respectively not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sinomenine is not more than 1.5z. (2) Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. diethyl ether, shake, then add 5.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes, centrifuge, and remove the diethyl ether layer. To the aqueous layer add Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) 20 mL of diethyl ether, and proceed in the same manner as described above. To the aqueous layer add 5.0 mL of diluted sodium hydroxide TS (1 in 10) and 10 mL of methanol, shake for 15 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 15 minutes, centrifuge, and take the super-natant liquid. Combine all the supernatant liquids, add dilut-ed methanol (1 in 2) to make exactly 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 5 mg of sinomenine for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of sinomenine in each solution. Amount (mg) of sinomenine ＝ MS × AT/AS × 1/2 MS: Amount (mg) of sinomenine for assay taken MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu ‌1968‌ Brown Rice / Crude Drugs and Related Drugs tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Boric Acid, when dried, contains not less than 99.
Boric Acid occurs as colorless or white, crys-tals or crystalline powder. It is odorless, and has a slight, characteristic taste. It is freely soluble in warm water, in hot ethanol (95) and in glycerin, soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. The pH of a solution of 1.0 g of Boric Acid in 20 mL of water is between 3.5 and 4.1.
A solution of Boric Acid (1 in 20) responds to Qualitative Tests <1.09> for borate.
(1) Clarity and color of solution—Dissolve 1.0 g of Boric Acid in 25 mL of water or in 10 mL of hot ethanol (95): the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Boric Acid according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution ‌JP XVIII Official Monographs / Bromfenac Sodium Hydrate 563‌‌ (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Boric Acid according to Method 1, and perform the test (not more than 5 ppm). Loss on drying <2.41> Not more than 0.5z (2 g, silica gel, 5 hours).
Weigh accurately about 1.5 g of Boric Acid, previ-ously dried, add 15 g of D-sorbitol and 50 mL of water, and dissolve by warming. After cooling, titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 drops of phenol-phthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 61.83 mg of H3BO3
Bromazepam, when dried, contains not less than 99.
Bromazepam occurs as white to light yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in methanol, in ethanol (99.5) and in acetone, and practically insoluble in water. Melting point: about 2459C (with decomposition).
(1) Determine the absorption spectrum of a solution of Bromazepam in ethanol (99.5) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bromazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bromazepam in a platinum crucible according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Bromazepam in 5 mL of a mixture of acetone and methanol (3:2), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mixture of acetone and methanol (3:2) to make exactly 50 mL. Pipet 5 mL of this solution, add the mixture of acetone and methanol (3:2) to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ammonia so-lution (28) and ethanol (99.5) (38:1:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution and the spot of the starting point are not more than 2, and not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Bromazepam, pre-viously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.62 mg of C14H10BrN3O
Bromhexine Hydrochloride, when dried, contains not less than 98.
Bromhexine Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in formic acid, sparingly soluble in methanol, and slightly soluble in water and in ethanol (95). The pH of its saturated solution is between 3.0 and 5.0. Melting point: about 2399C (with decomposition).
(1) Dissolve 3 mg of Bromhexine Hydro-chloride in 0.01 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bromhexine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. Add 20 mL of water to 1 g of Bromhexine Hydro-chloride. After thorough shaking, add 3 mL of sodium hy-droxide TS, and extract with four 20-mL portions of diethyl ether. Neutralize the water layer with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Bromhexine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 50 ‌566 mg of Bromhexine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: each peak area other than bromhexine obtained from the sample solution is not larger than the peak area of bromhexine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column about 5 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of potassium dihydrogen phosphate in 900 mL of water, adjust the pH to 7.0 with 0.5 mol/L sodium hydroxide TS, and add water to make 1000 mL. To 200 mL of this solution add 800 mL of acetonitrile. Flow rate: Adjust so that the retention time of bromhexine is about 6 minutes. Selection of column: To 0.05 g of bamethan sulfate add 0.5 mL of the sample solution, and add the mobile phase to make 10 mL. Proceed with 5 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of bamethan and bromhexine in this order with the resolution between these peaks being not less than 7. Detection sensitivity: Adjust the detection sensitivity so that the peak height of bromhexine from 5 mL of the stand-ard solution is between 5 mm and 15 mm. Time span of measurement: About 2 times as long as the retention time of bromhexine, beginning after the solvent peak. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Bromhexine Hydro-chloride, previously dried, dissolve in 2 mL of formic acid, add 60 mL of acetic anhydride, and warm in a water bath at 509C for 15 minutes. After cooling, titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-green to yellow-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.26 mg of C14H20Br2N2.HCl
Bromocriptine Mesilate contains not less than 98.
Bromocriptine Mesilate occurs as a white to pale yellowish white or pale brownish white crystalline powder. It is odorless, or has a faint characteristic odor. It is very soluble in acetic acid (100), freely soluble in methanol, sparingly soluble in ethanol (95), very slightly soluble in acetic anhydride, in dichloromethane and in chlo-roform, and practically insoluble in water and in diethyl ether. It is gradually colored by light.
(1) Dissolve 2 mg of Bromocriptine Mesi-late in 1 mL of methanol, add 2 mL of 4-dimethylaminoben-zaldehyde-ferric chloride TS, and shake: a purplish blue color develops. Determine the absorption spectrum of a solution of Bromocriptine Mesilate in methanol (3 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bromocriptine Mesilate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Bromocriptine Mesilate as di-rected under Flame Coloration Test <1.04> (2): a green color appears. D Optical rotation <2.49> [a]20: ＋95 – ＋1059[0.1 g, calcu-lated on the dried basis, a mixture of methanol and dichloro-methane (1:1), 10 mL, 100 mm].
(1) Clarity and color of solution—Dissolve 0.10 g of Bromocriptine Mesilate in 10 mL of methanol: the solu-tion is clear, and has no more color than the following con-trol solution. Control solution: To 2.5 mL of Cobalt (II) Chloride CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II) Sulfate CS add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. Heavy metals <1.07>—Proceed with 1.0 g of Bromocriptine Mesilate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). ‌JP XVIII Official Monographs / Bromovalerylurea 567 Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Bromocriptine Mesilate in 10 mL of a mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and chloroform (1:1) to make exactly 200 mL, and use this solution as the standard solution (1). Pipet 10 mL of the standard solution (1), add a mixture of methanol and chloroform (1:1) to make exactly 20 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2), as a band with 1 cm in width, on a plate of silica gel for thin-layer chromatography. Develop the plate immediately with a mixture of dichloro-methane, 1,4-dioxane, ethanol (95) and ammonia solution (28) (1800:150:50:1) to a distance of about 10 cm, and dry the plate under reduced pressure for 30 minutes. Spray evenly Dragendorff's TS for spraying on the plate, then spray evenly hydrogen peroxide TS, cover the plate with a glass plate, and examine: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and the spot other than the principal spot, which is more intense than the spot from the standard solution (2), is not more than one. Loss on drying <2.41> Not more than 3.0z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, 809C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Bromocriptine Mesilate, dissolve in 80 mL of a mixture of acetic anhydride and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 75.07 mg of C32H40BrN5O5.CH4O3S
Containers—Tight containers. Storage—Light-resistant, and not exceeding －189C.
Bromovalerylurea, when dried, contains not less than 98.
Bromovalerylurea occurs as colorless or white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is soluble in ethanol (95), sparingly soluble in diethyl ether, and very slightly soluble in water. It dissolves in sulfuric acid, in nitric acid and in hydro-chloric acid, and precipitates are produced on the addition of water. It dissolves in sodium hydroxide TS.
(1) Boil 0.2 g of Bromovalerylurea with 5 mL of a solution of sodium hydroxide (1 in 10): the gas evolved changes moistened red litmus paper to blue. Boil this solution with an excess of dilute sulfuric acid: the odor of valeric acid is perceptible. (2) To 0.1 g of Bromovalerylurea add 0.5 g of anhydrous sodium carbonate, and decompose thoroughly by gentle heating. Dissolve the residue in 5 mL of hot water, cool, acidify with acetic acid (31), and filter: the filtrate responds to Qualitative Tests <1.09> (2) for bromide.
(1) Acidity or alkalinity—To 1.5 g of Bromovalerylurea add 30 mL of water, shake for 5 minutes, and filter: the filtrate is neutral. Chloride <1.03>—Perform the test with a 10-mL por- tion of the filtrate obtained in (1). Prepare the control solu-tion with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.028z). Sulfate <1.14>—Perform the test with 10 mL of the filtrate obtained in (1). Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Bromovalerylurea according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Dissolve 0.5 g of Bromovalerylurea in 5 mL of sodium hydroxide TS, use this solution as the test solution, and perform the test (not more than 4 ppm). Readily carbonizable substances <1.15>—Perform the test with 0.5 g of Bromovalerylurea: the solution is not more colored than Matching Fluid A. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Bromovalerylurea, previously dried, in a 300-mL conical flask, add 40 mL of sodium hydroxide TS, and boil gently for 20 minutes under a reflux condenser. Cool, wash the lower part of the reflux condenser and the mouth of the flask with 30 mL of water, and combine the washings with the solution in the conical flask. Add 5 mL of nitric acid and exactly 30 mL of 0.1 mol/L silver nitrate VS, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indica-tor: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 22.31 mg of C6H11BrN2O2
Brotizolam, when dried, contains not less than 98.
Brotizolam occurs as a white or pale yellow crystalline powder. It is sparingly soluble in methanol, slightly soluble in acetonitrile and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Brotizolam in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Brotizolam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Brotizolam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Brotizolam in 50 mL of acetonitrile, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add aceto-nitrile to make exactly 100 mL. Pipet 1 mL of this solution, add acetonitrile to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: each peak area other than brotizolam obtained from the sample solution is not larger than 1/2 times the peak area of brotizolam from the stand-ard solution, and the total area of the peaks other than brotizolam from the sample solution is not larger than the peak area of brotizolam from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Flow rate: About 2 mL per minute. Time span of measurement: About 2 times as long as the retention time of brotizolam, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add acetonitrile to make exactly 20 mL. Con-firm that the peak area of brotizolam obtained with 5 mL of this solution is equivalent to 18 to 32z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of brotizolam are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of brotizolam is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of Brotizolam, previ-ously dried, dissolve in 75 mL of a mixture of acetic anhy-dride and acetic acid (100) (2:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.68 mg of C15H10BrClN4S
Bucillamine, when dried, contains not less than 98.
Bucillamine occurs as white, crystals or crys-talline powder. It is freely soluble in methanol and in ethanol (95), and slightly soluble in water.
(1) To 5 mL of a solution of Bucillamine (1 in 250) add 2 mL of sodium hydroxide TS and 2 drops of so-dium pentacyanonitrosylferrate (III) TS: the solution reveals a red-purple color. (2) Determine the infrared absorption spectrum of Bucil-lamine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋33.0 – ＋36.59(after dry-ing, 2 g, ethanol (95), 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bucillamine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Bucillamine according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 60 mg of Bucillamine in 20 mL of a mixture of water and methanol (1:1), and use this solution as the sample solution. Pipet 3 mL of the sam-ple solution, add the mixture of water and methanol (1:1) to make exactly 200 mL, and use this solution as the standard solution. Immediately perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak areas of related sub-stances, having the relative retention time of about 2.3 and about 3.1 to bucillamine, obtained from the sample solution are not larger than 8/15 times and 2/5 times the peak area of bucillamine from the standard solution, respectively, and the area of the peak other than the bucillamine and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of bucillamine from the standard so-lution. The total area of the peaks other than bucillamine from the sample solution is not larger than the peak area of bucillamine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.01 mol/L citric acid TS and methanol (1:1). Flow rate: Adjust so that the retention time of bucillamine is about 5 minutes. Time span of measurement: About 7 times as long as the retention time of bucillamine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of bucillamine obtained with 20 mL of this solution is equiva-lent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 0.10 g of bucillamine and 10 mg of 4-fluorobenzoic acid in 100 mL of methanol. To 10 mL of this solution add water to make exactly 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, bucillamine and 4-fluorobenzoic acid are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bucillamine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Bucillamine, dis-solve in 35 mL of methanol, add 15 mL of water, and titrate <2.50> with 0.05 mol/L iodine VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L iodine VS ＝ 11.17 mg of C7H13NO3S2
Bucumolol Hydrochloride, when dried, contains not less than 99.
Bucumolol Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water, sparingly soluble in methanol and in ethanol (95), slightly soluble in acetic acid (100), and practically insoluble in diethyl ether. Melting point: about 2289C (with decomposition).
(1) Dissolve 0.01 g of Bucumolol Hydro-chloride in 10 mL of diluted ethanol (95) (1 in 2), and ob-serve under ultraviolet light (main wavelength: 365 nm): the solution shows a yellow-green fluorescence. Render this so-lution alkaline by adding sodium hydroxide TS: the fluores-cence disappears. Acidify the solution by adding dilute hy-drochloric acid: the fluorescence reappears. Dissolve 0.1 g of Bucumolol Hydrochloride in 5 mL of water, and add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Bucumolol Hydrochloride (1 in 60,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bucumolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of solution, add methanol to make exactly 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol and ammonia-ammonium chloride buffer solu-tion (pH 10.7) (30:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Bucumolol Hydro-chloride, previously dried, add 45 mL of acetic acid (100), dissolve by warming at 609C, and cool. Add 105 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.18 mg of C17H23NO4.HCl
Bufetolol Hydrochloride, when dried, contains not less than 98.
(1) To 5 mL of a solution of Bufetolol Hydrochloride (1 in 100) add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Bufetolol Hydrochloride (1 in 20,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bufetolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho- JP XVIII Official Monographs / Buformin Hydrochloride 573 tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. ‌A solution of Bufetolol Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Bucumolol Hydrochloride in 20 mL of water: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 1.0 g of Bucumolol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Bucumolol Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Bucumolol Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this ble in diethyl ether. A solution of Bufetolol Hydrochloride (1 in 10) shows no optical rotation. Identification (1) To 5 mL of a solution of Bufetolol Hydrochloride (1 in 100) add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Bufetolol Hydrochloride (1 in 20,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bufetolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho- JP XVIII Official Monographs / Buformin Hydrochloride 573 tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. ‌A solution of Bufetolol Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride. Melting point <2.60> 153 – 1579C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Bufetolol Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.5 g of Bufetolol Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Bufetolol Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of Bufetolol Hy-drochloride in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, acetone, ethanol (95) and ammonia solution (28) (40:20:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Bufetolol Hydro-chloride, previously dried, dissolve in 10 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.99 mg of C18H29NO4.HCl
A round disk with slightly dented bottom and protuberant surface, about 8 cm in diameter, about 1.5 cm in thickness, the mass of one disk being about 80 to 90 g; or a round disk with almost ‰attened surfaces on both sides, about 3 cm in diameter, and about 0.5 cm in thickness, the mass of one disk being about 8 g; externally red-brown to black-brown, somewhat lustrous, approximately uniform and horny, hard in texture, and diŠicult to break; fractured surface nearly ‰at, and edges of broken pieces red-brown and translucent. Odorless.
To 0.3 g of pulverized Toad Cake add 3 mL of acetone, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of resibufogenin for thin-layer chromatography in 2 mL of ace-tone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of cyclohexane and acetone (3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard so-lution. Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of pulverized Toad Amount (mg) of resibufogenin ＝ MSR × QTR/QSR MSB: Amount (mg) of bufalin for assay taken MSC: Amount (mg) of cinobufagin for assay taken MSR: Amount (mg) of resibufogenin for assay taken Internal standard solution—A solution of indometacin in methanol (1 in 4000). Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 300 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (11:9). Flow rate: Adjust so that the retention time of the internal standard is 16 to 19 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, bufalin, cinobufagin, resibufogenin and the internal standard are eluted in this order with the resolution among these peaks being not less than 1.5.
Buformin Hydrochloride, when dried, contains not less than 98.
Buformin Hydrochloride occurs as a white crystalline powder. It is freely soluble in water and in ethanol (99.5).
(1) To 5 mL of a solution of Buformin Hydrochloride (1 in 2000) add 1 mL of dilute sodium penta-cyanonitrosylferrate (III)-potassium hexacyanoferrate (III) TS: a red-brown color develops. Determine the absorption spectrum of a solution of Buformin Hydrochloride (1 in 125,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Buformin Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Buformin Hydrochloride (1 in 20) re-sponds to the Qualitative Tests <1.09> for chlorides.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Buformin Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Buformin Hydrochloride according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Buformin Hydrochloride in 200 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the area of the peak other than buformin obtained from the sample solution is not larger than 1/5 times the peak area of buformin from the standard solution. Further-more, the total areas of all peaks other than the buformin peak from the sample solution is not larger than 1/2 times the peak area of buformin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of a solution of sodium perchlorate monohydrate in diluted phosphoric acid (1 in 1000) (7 in 250) and acetonitrile (7:1). Flow rate: Adjust so that the retention time of buformin is about 6 minutes. Time span of measurement: About 2 times as long as the retention time of buformin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of buformin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of buformin are not less than 5000 and ‌574 not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of buformin is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of Buformin Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and immediately titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 9.684 mg of C6H15N5.HCl
Bumetanide, when dried, contains not less than 98.
Bumetanide occurs as white, crystals or crys-talline powder. It is freely soluble in pyridine, soluble in methanol and in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in potassium hydroxide TS. It is gradually colored by light.
(1) Dissolve 0.01 g of Bumetanide in 1 mL of pyridine, add 2 drops of copper (II) sulfate TS, shake, add 3 mL of water and 5 mL of chloroform, shake, and allow to stand: a light blue color develops in the chloroform layer. Dissolve 0.04 g of Bumetanide in 100 mL of phos-phate buffer solution (pH 7.0) and dilute 10 mL of the solu-tion with water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bumetanide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 50 mg of Bumetanide in 2 mL of a solution of potassium hydroxide (1 in 30) and 8 mL of water: the solution is clear, and is not more colored than the following control solution. Control solution: Pipet 0.5 mL each of Cobalt (II) Chlo-ride CS, Iron (III) Chloride CS and Copper (II) Sulfate CS, washing, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Heavy metals <1.07>—Proceed with 2.0 g of Bumetanide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Bumetanide according to Method 3, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Bumetanide in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetic acid (100), cyclohexane and methanol (32:4:4:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Bumetanide, previ-ously dried, dissolve in 50 mL of ethanol (95), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 36.44 mg of C17H20N2O5S
Bunazosin Hydrochloride occurs as a white crystalline powder. It is very soluble in formic acid, slightly soluble in water and in methanol, very slightly soluble in ethanol (99.5), and practically insoluble in diethyl ether. Melting point: about 2739C (with decomposition).
(1) Dissolve 0.1 g of Bunazosin Hydrochlo-ride in 10 mL of 0.2 mol/L hydrochloric acid TS, and boil for 3 minutes over a flame: butylic acid like odor is percepti-ble. Determine the infrared absorption spectrum of Bunazosin Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Bunazosin Hydrochloride (1 in 100) re-sponds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bunazosin Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.05 g of Bunazosin Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. To exactly 1 mL of the sam-ple solution add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography 500 mL of acetonitrile and water to make 1000 mL. Flow rate: Adjust so that the retention time of bunazosin is about 5 minutes. Selection of column: Proceed with 20 mL of a mixture of the standard solution and a solution of procaine hydrochlo-ride in the mobile phase (1 in 20,000) (1:1) under the above operating conditions, and calculate the resolution. Use a column giving elution of procaine and bunazosin in this order with the resolution between these peaks being not less than 3.0. Detection sensitivity: Adjust the detection sensitivity so that the peak height of bunazosin obtained from 20 mL of the standard solution is 20 to 60z of the full-scale. Time span of measurement: About 6 times of the retention time of bunazosin. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Bunazosin Hydro-chloride, previously dried, dissolve in 6 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid, and heat for 20 minutes on a water bath. After cooling, add 20 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.99 mg of C19H27N5O3.HCl
Bupivacaine Hydrochloride Hydrate contains not less than 98.
Bupivacaine Hydrochloride Hydrate occurs as a white crystalline powder. It is freely soluble in acetic acid (100), and soluble in water, in methanol and in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. A solution of 0.5 g of Bupivacaine Hydrochloride Hydrate in 50 mL of a mixture of ethanol (99.5), water and 5 mol/L sodium hydroxide TS (34:15:1) shows no optical rotation. Melting point: about 2529C (with decomposition).
(1) Determine the absorption spectrum of a solution of Bupivacaine Hydrochloride Hydrate in 0.01 mol/L hydrochloric acid TS (1 in 2000) as directed under Ul- ‌578 traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bupivacaine Hydrochloride Hydrate as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A solution of Bupivacaine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Bupivacaine Hydrochloride Hydrate in 100 mL of freshly boiled and cooled water is between 4.5 to 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Bupivacaine Hydrochloride Hydrate in 50 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Bupivacaine Hydrochloride Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). 2,6-Dimethylaniline—Dissolve exactly 0.50 g of Bupivacaine Hydrochloride Hydrate in 10 mL of methanol. To 2 mL of this solution add 1 mL of a freshly prepared solution of 4-dimethylaminobenzaldehyde in methanol (1 in 100) and 2 mL of acetic acid (100), and allow to stand for 10 minutes: the color of the solution is not more colored than the following control solution. Control solution: Prepare by proceeding in the same man-ner as above, using 2 mL of a solution of 2,6-dimethylaniline in methanol (1 in 200,000). Related substances—Dissolve 50 mg of Bupivacaine Hydrochloride Hydrate in 2.5 mL of water, add 2.5 mL of 2 mol/L sodium hydroxide TS and 5 mL of the internal stand-ard solution, shake, collect the lower layer, filter, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, and add the internal standard solution to make ex-actly 100 mL. Pipet 1 mL of this solution, add the internal standard solution to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the ratio of the area of the peak other than bupivacaine to the peak area of the internal standard obtained from the sample solution is not larger than the ratio of the peak area of bupivacaine to that of the internal standard from the standard solution. Internal standard solution—A solution of methyl behenate in dichloromethane (1 in 20,000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A quartz tube 0.32 mm in inside diameter and 30 m in length, coated the inside surface with 5z diphenyl-95z dimethylpolysiloxane for gas chromatography 0.25 mm in thickness. Column temperature: Rise the temperature from 1809C to 2309C at the rate of 59C per minute, and maintain at 2309C for 5 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of bupivacaine is about 10 minutes. Split ratio: 1:12. Time span of measurement: About 1.5 times as long as the retention time of bupivacaine. System suitability— System performance: To 1 mL of the sample solution add the internal standard solution to make 100 mL, and use this solution as the solution for system suitability test. When the procedure is run with 1 mL of the solution for system suita-bility test under the above operating conditions, bupivacaine and the internal standard are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 6 times with 1 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the ratio of the peak area of bupivacaine to that of the in-ternal standard is not more than 2.0z. Water <2.48> 4.0 – 6.0z (0.25 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Bupivacaine Hydro-chloride Hydrate, dissolve in 20 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.49 mg of C18H28N2O.HCl
Bupranolol Hydrochloride, when dried, contains not less than 98.
Bupranolol Hydrochloride occurs as a white crystalline powder. It is sparingly soluble in methanol, slightly soluble in water, in ethanol (95) and in acetic acid (100), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Bupranolol Hydrochlo-ride in 1000 mL of water is between 5.2 and 6.2.
(1) Take 0.01 g of Bupranolol Hydrochlo-ride in a test tube, mix with 25 mg of potassium iodide and 25 mg of oxalic acid dihydrate, cover the mouth of the test tube with filter paper moistened with a solution of 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol (95) (1 in 100), and heat gently for several minutes. Expose the filter paper to ammonia gas: the filter paper acquires a blue color. Determine the absorption spectrum of a solution of ‌JP XVIII Official Monographs / Buprenorphine Hydrochloride 579 Bupranolol Hydrochloride in 0.1 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bupranolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Bupranolol Hydrochloride (1 in 200) responds to Qualitative Tests <1.09> for chloride. 1 cm Absorbance <2.24> E 1z (275 nm): 57 – 60 (after drying, 50 mg, 0.1 mol/L hydrochloric acid TS, 500 mL).
(1) Clarity and color of solution—Dissolve 0.10 g of Bupranolol Hydrochloride in 15 mL of water: the solu-tion is clear and colorless. Acidity—Dissolve 0.10 g of Bupranolol Hydrochlo-ride in 15 mL of freshly boiled and cooled water, and add 1 drop of methyl red TS: a light red color develops. To this so-lution add 0.05 mL of 0.01 mol/L sodium hydroxide VS: the color changes to yellow. Sulfate <1.14>—Perform the test with 0.10 g of Bupranolol Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.168z). Heavy metals <1.07>—Proceed with 1.0 g of Bupranolol Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Bupranolol Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.30 g of Bupranolol Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of polyamide with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol, ammonia solution (28) and water (16:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.18 g of Bupranolol Hy-drochloride, previously dried, dissolve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (2:1) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.82 mg of C14H22ClNO2.HCl
Buprenorphine Hydrochloride occurs as white to yellowish white, crystals or a crystalline powder. It is freely soluble in methanol and in acetic acid (100), and sparingly soluble in water and in ethanol (99.5). Melting point: about 2689C (with decomposition).
(1) Determine the absorption spectrum of a solution of Buprenorphine Hydrochloride (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Buprenorphine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Buprenorphine Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: －92 – －989(after drying, 0.4 g, methanol, 20 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Buprenorphine Hydrochloride in 200 mL of water is between 4.0 and 6.0.
(1) Clarity and color of solution—A solution ob-tained by dissolving 0.1 g of Buprenorphine Hydrochloride in 10 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Buprenorphine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Buprenor-phine Hydrochloride in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than buprenorphine ob-tained from the sample solution is not larger than 1/4 times the peak area of buprenorphine obtained from the standard solution. Furthermore, the total area of the peaks other than ‌580‌ buprenorphine from the sample solution is not larger than 13/20 times the peak area of buprenorphine from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 288 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, ammonium acetate solution (1 in 100), and acetic acid (100) (6000:1000:1). Flow rate: Adjust so that the retention time of buprenor-phine is about 17 minutes. Time span of measurement: About 2.5 times as long as the retention time of buprenorphine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of buprenorphine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of buprenorphine are not less than 6500 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of buprenorphine is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1159C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Buprenorphine Hy-drochloride, previously dried, dissolve in 5 mL of acetic acid Identification (1) To 0.1 g of Busulfan add 10 mL of water and 5 mL of sodium hydroxide TS, dissolve by heat-ing, and use this solution as the sample solution. To 7 mL of the sample solution add 1 drop of potas-sium permanganate TS: the red-purple color of potassium permanganate TS changes from blue-purple through blue to green. Acidify 7 mL of the sample solution with dilute sul-furic acid, and add 1 drop of potassium permanganate TS: the color of potassium permanganate TS remains. (2) Determine the infrared absorption spectrum of Busulfan, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 115 – 1189C Purity (1) Sulfate <1.14>—To 1.0 g of Busulfan add 40 mL of water, and dissolve by heating. Cool in ice for 15 minutes, and filter. Wash the residue with 5 mL of water, combine the washings with the filtrate, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). (2) Heavy metals <1.07>—Proceed with 1.0 g of Busulfan according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Busulfan, add 40 mL of water, and boil gently under a reflux condenser for 30 minutes. Cool, and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (indicator: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS (100), add 50 mL of acetic anhydride, and titrate <2.50> with mol/L perchloric acid VS (potentiometric titration). Per- ＝ 12.32 mg of C6H 14O6S2 form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 50.41 mg of C29H41NO4.HCl
Dried Aluminum Potassium Sulfate occurs as white masses or white powder. It is odorless. It has a slightly sweet, astringent taste. It is freely soluble in hot water and practically insoluble in ethanol (95). It dissolves slowly in water.
A solution of Dried Aluminum Potassium Sulfate (1 in 20) responds to Qualitative Tests <1.09> for alu-minum salt, to Qualitative Tests <1.09> (1), (3) and (4) for potassium salt, and to Qualitative Tests <1.09> (1) and (3) for sulfate.
(1) Water-insoluble substances—To 2.0 g of Dried Aluminum Potassium Sulfate add 40 mL of water, shake frequently, and allow to stand for 48 hours. Collect the insoluble residue on a glass filter (G4), wash with 50 mL of water, and dry at 1059C for 2 hours: the mass of the residue is not more than 50 mg. Heavy metals <1.07>—Dissolve 0.5 g of Dried Alumi- num Potassium Sulfate in 45 mL of water, and filter, if necessary. Add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 40 ppm). Iron <1.10>—Prepare the test solution with 0.54 g of Dried Aluminum Potassium Sulfate according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 37 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Dried Aluminum Potassium Sulfate, according to Method 1, and perform the test (not more than 5 ppm). Loss on drying <2.41> Not more than 15.0z (2 g, 2009C, 4 hours).
Weigh accurately about 1.2 g of Dried Aluminum Potassium Sulfate, previously dried, add 80 mL of water, and heat on a water bath with occasional shaking for 20 minutes. Cool, add water to make exactly 100 mL, and filter, if necessary. Discard the first 30 mL of the filtrate, take exactly the subsequent 20 mL of the filtrate, and add exactly 30 mL of 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS and 20 mL of acetic acid-ammo-nium acetate buffer solution (pH 4.8), boil for 5 minutes, and cool. Add 55 mL of ethanol (95), and titrate <2.50> with 2 3 0.05 mol/L zinc acetate VS (indicator: 2 mL of dithizone TS), until the color of the solution changes from light dark ‌442‌ green to light red. Perform a blank determination in the same manner. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 12.91 mg of AlK (SO4)2
Butenafine Hydrochloride, when dried, contains not less than 99.
Busulfan occurs as a white crystalline powder. It is slightly soluble in diethyl ether, very slightly soluble in ethanol (95), and practically insoluble in water. C23H27N.HCl: 353.93 N-[4-(1,1-Dimethylethyl)benzyl]-N-methyl-1-(naphthalen-1-yl)methylamine monohydrochloride [101827-46-7] Butenafine Hydrochloride, when dried, contains not less than 99.0z and not more than 101.0z of butenafine hydrochloride (C23H27N.HCl). Description Butenafine Hydrochloride occurs as white, crystals or crystalline powder. JP XVIII Official Monographs / Butenafine Hydrochloride Cream 581 ‌It is very soluble in formic acid, freely soluble in methanol and in ethanol (99.5), and slightly soluble in water. The pH of a solution dissolved 0.20 g of Butenafine Hy-drochloride in 100 mL of water by warming and cooled is 3.0 to 4.0. Melting point: about 2149C (with decomposition). 0 – 10 60 → 20 40 → 80 10 – 60 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz)
(1) Determine the absorption spectrum of a solution of Butenafine Hydrochloride in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Butenafine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Butenafine Hydrochloride in dilute ethanol (1 in 200) responds to Qualitative Tests <1.09> (1) for chloride.
(1) Heavy metals <1.07>—Dissolve 2.0 g of Butenafine Hydrochloride in 20 mL of ethanol (99.5), add 2 mL of dilute acetic acid and ethanol (99.5) to make 50 mL, and perform the test using this solution as the test solution. The control solution: To 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid, and add ethanol (99.5) to make 50 mL (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Butenafine Hydrochloride in 50 mL of a mixture of water and aceto-nitrile for liquid chromatography (3:2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water and acetonitrile for liquid chroma-tography (3:2) to make exactly 50 mL. Pipet 1 mL of this solution, add a mixture of water and acetonitrile for liquid chromatography (3:2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.16 to butenafine, obtained from the sample solution is not larger than 3/10 times the peak area of butenafine from the standard solution, and the area of the peak other than butenafine and the peak mentioned above from the sample solution is not larger than the peak area of butenafine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 217 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Diluted 0.5 mol/L ammonium acetate TS (1 in 1000). Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: 0.4 mL per minute. Time span of measurement: For 60 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of water and acetonitrile for liq-uid chromatography (3:2) to make exactly 10 mL. Confirm that the peak area of butenafine obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of butenafine are not less than 20,000 and 0.9 to 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of butenafine is not more than 2.0z. Loss on drying <2.41> Not more than 0.1z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Butenafine Hydro-chloride, previously dried, dissolve in 5 mL of formic acid, add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.39 mg of C23H27N.HCl
Butropium Bromide, when dried, contains not less than 98.
Butropium Bromide occurs as white, crystals or crystalline powder. It is very soluble in formic acid, freely soluble in metha-nol, soluble in ethanol (95), slightly soluble in water, and practically insoluble in diethyl ether and in acetic anhydride.
(1) To 1 mg of Butropium Bromide add 3 drops of fuming nitric acid, and evaporate on a water bath to dryness. Dissolve the residue in 1 mL of N,N-dimethylfor-mamide, and add 5 to 6 drops of tetraethylammonium hy-droxide TS: a red-purple color develops. Determine the absorption spectrum of a solution of Butropium Bromide in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, determine the absorption spectrum of a solution of Butropium Bromide in methanol (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Butropium Bromide in methanol (1 in 20) responds to Qualitative Tests <1.09> (1) for bromide. D Optical rotation <2.49> [a]20: －14.0 – －17.09(after dry-ing, 0.5 g, methanol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Dissolve 1.0 g of Butro-pium Bromide in 40 mL of ethanol (95), add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test, using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Butropium ‌584 Bromide in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the peak area, having the relative retention time about 0.5 to butropium obtained from the sample solution is not larger than 1/4 times the peak area from the standard solution, and the total area of all peaks other than the peak eluted first, the peak, having the relative retention time to butropium about 0.5 and butropium peak from the sample solution is not larger than the peak area from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column about 5 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.15 g of sodium lauryl sulfate in 1000 mL of a mixture of acetonitrile and 0.005 mol/L sulfu-ric acid (3:2). Flow rate: Adjust so that the retention time of butropium is about 5 minutes. Selection of column: Dissolve 0.50 g of Butropium Bromide in 9 mL of ethanol (99.5) and 1 mL of 0.1 mol/L potassium hydroxide-ethanol TS, and heat at 709C for 15 minutes. After cooling, to 1 mL of this solution add the mo-bile phase to make 100 mL. Proceed with 5 mL of this solu-tion under the above operating conditions, and calculate the resolution. Use a column giving elution of the peak of butro-pium and the peak having a ratio of the retention time about 0.7 to butropium with the resolution between these peaks being not less than 2.5. Detection sensitivity: Adjust the detection sensitivity so that the peak height of the butropium obtained from 5 mL of the standard solution is between 10 mm and 30 mm. Time span of measurement: About twice as long as the retention time of butropium. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.8 g of Butropium Bromide, previously dried, dissolve in 5 mL of formic acid, add 100 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid-dioxane VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid-dioxane VS ＝ 53.25 mg of C28H38BrNO4
Butyl Parahydroxybenzoate contains not less than 98.
Butyl Parahydroxybenzoate occurs as color-less crystals or white crystalline powder. It is very soluble in methanol, freely soluble in ethanol (95) and in acetone, and practically insoluble in water.◆
Determine the infrared absorption spectrum of Butyl Parahydroxybenzoate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Butyl Parahydroxybenzoate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Butyl Parahydroxybenzoate in ethanol (95) to make 10 mL: the solution is clear and not more intensely colored than the following control solution. Control solution: To 5.0 mL of Cobalt (II) Chloride CS, 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity—To 2 mL of the solution of Butyl Parahy-droxybenzoate obtained in (1) add 3 mL of ethanol (95), add 5 mL of freshly boiled and cooled water and 0.1 mL of bromocresol green-sodium hydroxide-ethanol TS, then add 0.1 mol/L sodium hydroxide VS until the solution shows a blue color: the volume of 0.1 mol/L sodium hydroxide VS used does not exceed 0.1 mL. (3) Heavy metals <1.07>—Dissolve 1.0 g of Butyl Par- ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm).◆ Related substances—Dissolve 50.0 mg of Butyl Par-ahydroxybenzoate in 2.5 mL of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL of this so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly ‌JP XVIII Official Monographs / Cabergoline 585‌ 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak area of parahydroxybenzoic acid having a relative retention time of about 0.1 to butyl parahy-droxybenzoate obtained from the sample solution is not larger than the peak area of butyl parahydroxybenzoate from the standard solution (0.5z). For the area of the peak of parahydroxybenzoic acid multiply the correction factor, 1.4. Furthermore, the area of the peak other than butyl par-ahydroxybenzoate and parahydroxybenzoic acid from the sample solution is not larger than the peak area of butyl par-ahydroxybenzoate from the standard solution (0.5z), and the total area of the peaks other than butyl parahydroxyben-zoate is not larger than 2 times the peak area of butyl par-ahydroxybenzoate from the standard solution (1.0z). For this calculation the peak area not larger than 1/5 times the peak area of butyl parahydroxybenzoate from the standard solution is excluded (0.1z). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of butyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of butyl parahydroxybenzo-ate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of butyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50.0 mg each of Butyl Par-ahydroxybenzoate and Butyl Parahydroxybenzoate RS, dis-solve separately in 2.5 mL each of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of butyl parahydroxybenzoate in each solution. Mobile phase: A mixture of methanol and potassium dihy-drogen phosphate solution (17 in 2500) (1:1). Flow rate: 1.3 mL per minute. System suitability— System performance: Dissolve 5 mg each of Butyl Parahy-droxybenzoate, propyl parahydroxybenzoate and parahy-droxybenzoic acid in the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the solution for system suitability test (1). Separately, dissolve 5 mg of isobutyl parahydroxybenzoate in the mobile phase to make exactly 100 mL. Pipet 0.5 mL of this solution, add the stand-ard solution to make exactly 50 mL, and use this solution as the solution for system suitability test (2). When the proce-dure is run with 10 mL each of the solution for system suita-bility test (1) and (2) under the above operating conditions, parahydroxybenzoic acid, propyl parahydroxybenzoate, isobutyl parahydroxybenzoate and butyl parahydroxybenzo-ate are eluted in this order, the relative retention times of parahydroxybenzoic acid, propyl parahydroxybenzoate and isobutyl parahydoxybenzoate to butyl parahydroxybenzoate are about 0.1, about 0.5 and about 0.9, respectively, the resolution between the peaks of propyl parahydroxybenzoate and butyl parahydroxybenzoate is not less than 5.0, and the resolution between the peaks of isobutyl parahydroxybenzo-ate and butyl parahydroxybenzoate is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of butyl parahydroxybenzoate is not more than 0.85z.
Byakkokaninjinto Extract contains not less than 9 mg and not more than 36 mg of mangiferin, not less than 13 mg and not more than 39 mg of glycyrrhizic acid (C42H62O16: 822.
Byakkokaninjinto Extract occurs as a very pale yellow-brown to light brown powder or blackish brown viscous extract. It has a slight odor, and has a slightly sweet and slightly bitter taste.
(1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 1 g of pulverized Anemarrhena Rhizome add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centri-fuge, and use the 1-butanol layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solu-tion and 1 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 2 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellowish red to dark red spot (at an Rf value of about 0.3) from the standard solution (Anemarrhena Rhizome). Place 2.0 g of the dry extract (or 6.0 g of the viscous extract) in a porcelain crucible, and ignite to incinerate at 500 – 5509C. To the residue add 60 mL of water, shake, cen-trifuge, and use the supernatant as the sample solution. Add ammonium oxalate TS to the sample solution: a white pre-cipitate is formed. The precipitate does not dissolve in diluted acetic acid, but dissolves on the addition of diluted hydrochloric acid (Gypsum). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). To 5.0 g of the dry extract (or 15 g of the viscous ex-tract) add 15 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of cycloartenyl ferulate for thin-layer chromatography in 1 mL of ethyl ace- tate, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 30 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (50:20:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly a mixture of sulfuric acid and ethanol (99.5) (1:1), heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the light yellow-white to yellow fluorescent spot from the standard solution. (Brown Rice). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rg1 RS or ginsenoside Rg1 for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple to dark purple spot from the standard solution (Ginseng).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 20.0z, calculated on the dried basis.
(1) Mangiferin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, cen-trifuge, and use the supernatant as the sample solution. Separately, weigh accurately about 10 mg of mangiferin for assay, dissolve in diluted methanol (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of mangiferin in each so-lution. Amount (mg) of mangiferin ＝ MS × AT/AS × 1/4 MS: Amount (mg) of mangiferin for assay taken, calcu-lated on the basis of the content obtained by qNMR JP XVIII Crude Drugs and Related Drugs / Byakkokaninjinto Extract 1971 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 367 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (1780:220:1). Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mangiferin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mangiferin is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly to about 1 g of the dried substance), add 25 mL of diluted methanol (3 in 5), shake for 30 minutes, then allow to stand, and separate the supernatant liquid. To the residue add 8 mL of water, shake for 15 minutes, then add 12 mL of metha-nol, shake for 15 minutes, centrifuge, and separate the su-pernatant liquid. Combine all the supernatant liquids, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of sodium hydroxide TS, allow to stand for 30 minutes, then add 3 mL of 1 mol/L hy-drochloric acid TS, and add water to make exactly 20 mL. Apply exactly 10 mL of this solution to a column [about 10 mm in inside diameter, packed with 0.36 g of octadecyl-silanized silica gel for pre-treatment (55 – 105 mm in particle size), and washed just before using with methanol and then diluted methanol (3 in 10)], and wash the column in sequence with 2 mL of diluted methanol (3 in 10), 1 mL of sodium carbonate TS and 10 mL of diluted methanol (3 in 10). Fi-nally, elute with methanol to collect exactly 5 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of Ginsenoside Rb1 RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog- Amount (mg) of ginsenoside Rb1 ＝ MS × AT/AS × 1/10 (C54 H92O 23) raphy <2.01> according to the following conditions, and de- termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute. System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Ginsenoside Rb1—Weigh accurately about 1 g of the dry extract (or an amount of the viscous extract, equivalent MS: Amount (mg) of Ginsenoside Rb1 RS taken, calcu-lated on the anhydrous basis For the prescription 1) Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with carbamoyl group bound silica gel for liquid chromatography (3.5 mm in particle di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (1700:300:1). Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ginsenoside Rb1 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z. For the prescription 2) Operating conditions— Detector, column temperature, and flow rate: Proceed as directed in the operating conditions in the prescription 1). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with carbamoyl group bound silica gel for liquid chromatography (5 mm in particle diame-ter). Mobile phase: A mixture of acetonitrile, water and phos- ‌1972‌‌‌‌ Cacao Butter / Crude Drugs and Related Drugs phoric acid (400:100:1). System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the prescription 1).
Cabergoline occurs as a white crystalline pow-der. It is very soluble in methanol, freely soluble in ethanol (95), and very slightly soluble in water. It is gradually colored to yellow by light. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Cabergoline in ethanol (95) (1 in 30,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cabergoline RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. lution. Amount (mg) of cabergoline (C 26H37N5O2) ＝ M S × AT/AS (2) Determine the infrared absorption spectrum of Cabergoline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cabergoline RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. If any differ-ence appears between the spectra, dissolve Cabergoline and Cabergoline RS in ethanol (95), respectively, then evaporate the ethanol, dry the residues, and repeat the test on the residues. D Optical rotation<<2.49> [a]20: －77 – －839(0.1 g calculated on the anhydrous basis, ethanol (95), 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cabergoline according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Perform the test with 20 mL of the sample solution obtained in the Assay as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area by the automatic integra-tion method, and calculate the amounts of them by the area percentage method: the amount of related substances A and B, having the relative retention times of about 0.8 and about 2.8 to cabergoline are not more than 0.5z, respectively, and the amount of the peak other than cabergoline and the peaks mentioned above is not more than 0.1z. Furthermore, the total amount of the peaks other than cabergoline is not more than 1.5z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of cabergoline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Use the diluted sample so-lution (1 in 500) as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of cabergoline obtained with 20 mL of this so-lution is equivalent to 18 to 32z of that with 20 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cabergoline is not more than 2.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration).
Conduct this procedure using light-resistant vessels. Weigh accurately about 30 mg each of Cabergoline and Cabergoline RS (separately determine the water <2.48> in the same manner as Cabergoline), dissolve each in the mobile phase to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of cabergoline in each so- MS: Amount (mg) of Cabergoline RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.0 with phos-phoric acid, and add water to make 1000 mL. To this solu-tion add 0.2 mL of triethylamine. To 840 mL of this solution add 160 mL of acetonitrile. Flow rate: Adjust so that the retention time of cabergoline is about 12 minutes. System suitability— System performance: Suspend 50 mg of Cabergoline in 10 mL of 0.1 mol/L sodium hydroxide TS, and stir for 15 minutes. To 1 mL of this solution add 1 mL of 0.1 mol/L hydrochloric acid TS, and add the mobile phase to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution be-tween the peaks of related substance A having the relative retention time of about 0.8 to cabergoline and cabergoline is not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cabergoline is not more than 1.0z.
Cadralazine, when dried, contains not less than 98.
Cadralazine occurs as a pale yellow to light yellow crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in ethanol (99.5), and slightly soluble in water. It dissolves in 0.05 mol/L sulfuric acid TS. A solution of Cadralazine in methanol (1 in 40) shows no optical rotation. Melting point: about 1659C (with decomposition).
(1) Determine the absorption spectrum of a solution of Cadralazine in 0.05 mol/L sulfuric acid TS (1 in 125,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cadralazine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.40 g of Cadrala-zine in 15 mL of methanol, add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS by adding 15 mL of methanol, 6 mL of dilute nitric acid, and water to make 50 mL (not more than 0.036z). Heavy metals <1.07>—Proceed with 1.0 g of Cadrala- zine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Cadralazine in 20 mL of 0.05 mol/L sulfuric acid TS, add water to 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak, having the relative retention time of about 2.1 to cadralazine, obtained from the sample solution is not larger than the peak area of cadralazine from the standard solution, and the area of the peak other than cadralazine and the peak mentioned above is not larger than 2/5 times the peak area of cadralazine from the standard solution. Furthermore, the total area of the peaks other than cadralazine from the sample solution is not larger than 2 times the peak area of cadralazine from the standard solu-tion. For the areas of the peaks, having the relative retention time of about 0.49 and about 2.1 to cadralazine, multiply their correction factors, 0.65 and 1.25, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 13.6 g of sodium acetate trihydrate in 800 mL of water, adjust the pH to 5.8 with dilute acetic acid, and add water to make 1000 mL. To 860 mL of this so-lution add 140 mL of acetonitrile. Flow rate: Adjust so that the retention time of cadralazine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of cadralazine. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add water to make exactly 25 mL. Confirm that the peak area of cadralazine obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cadralazine are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cadralazine is not more than 4.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Cadralazine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.33 mg of C12H21N5O3
Caffeine Hydrate, when dried, contains not less than 98.
Caffeine Hydrate occurs as white, soft crystals or powder. It is odorless, and has a slightly bitter taste. It is freely soluble in chloroform, sparingly soluble in water, in acetic acid (100) and in acetic anhydride, slightly soluble in ethanol (95), and very slightly soluble in diethyl ether. The pH of a solution of 1.0 g of Caffeine Hydrate in 100 mL of water is between 5.5 and 6.5. It effloresces in dry air.
(1) To 2 mL of a solution of Caffeine Hy-drate (1 in 500) add tannic acid TS dropwise: a white precipi-tate, which dissolves upon the dropwise addition of tannic acid TS, is produced. To 0.01 g of Caffeine Hydrate add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, and evaporate to dryness on a water bath: the residue acquires a yellow-red color. Invert the residue over a vessel containing 2 to 3 drops of ammonia TS: the color turns red-purple, and disappears upon the addition of 2 to 3 drops of sodium hy-droxide TS. Dissolve 0.01 g of Caffeine Hydrate in water to make 50 mL. To 5 mL of this solution add 3 mL of diluted acetic acid (31) (3 in 100) and 5 mL of a solution of pyridine (1 in 10), mix, add 2 mL of diluted sodium hypochlorite TS (1 in 5), and allow to stand for 1 minute. Add 2 mL of sodium thiosulfate TS and 5 mL of sodium hydroxide TS to the solu-tion: a yellow color develops.
(1) Chloride <1.03>—Dissolve 2.0 g of Caffeine Hydrate in 80 mL of hot water, cool rapidly to 209C, add water to make 100 mL, and use this solution as the sample solution. To 40 mL of the sample solution add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Sulfate <1.14>—To 40 mL of the sample solution ob- tained in (1) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Caffeine Hydrate according to Method 2, and perform the test. Pre- ‌590 pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Caffeine Hy-drate in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 100 mL. Pipet 1 mL of this solu-tion, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of chloroform and ethanol (95) (9:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Readily carbonizable substances <1.15>—Perform the test using 0.5 g of Caffeine Hydrate: the solution is not more colored than Matching Fluid D. Loss on drying <2.41> 0.5 – 8.5z (1 g, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.4 g of Caffeine Hydrate, previously dried, dissolve in 70 mL of a mixture of acetic an-hydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the solution changes from purple through green to yellow (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.42 mg of C8H10N4O2
Calcitonin Salmon is synthetic salmon calcitonin, and is a peptide consisting of 32 amino acid residues. It contains not less than 4000 Units of calcitonin salmon per 1 mg of peptide.
Calcitonin Salmon occurs as a white powder. It is freely soluble in water. It dissolves in dilute acetic acid. Dissolve 20 mg of Calcitonin Salmon in 2 mL of water: the pH of the solution is between 5.0 and 7.0. It is hygroscopic.
Dissolve 1 mg of Calcitonin Salmon in 1 mL of dilute acetic acid. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. ture of about 1309C. Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B, C, D and E according to the following table. 1 cm Absorbance <2.24> E1z (275 nm): 3.3 – 4.0 (1 mg, dilute acetic acid, 1 mL). Mobile phase A Mobile phase B Mobile phase C Mobile phase D Mobile phase E Citric acid monohydrate 19.80 g 22.00 g 12.80 g 6.10 g — Trisodium citrate 6.19 g dihydrate 7.74 g 13.31 g 26.67 g — Sodium hydroxide — — — — 8.00 g Sodium chlo- ride 5.66 g 7.07 g 3.74 g 54.35 g — Ethanol (99.5) 130.0 mL 20.0 mL 4.0 mL — 100.0 mL Benzyl alcohol — — — 5.0 mL — Thiodiglycol 5.0 mL 5.0 mL 5.0 mL — — Peptide content (z) ＝ 3431.85 × 5/M × A/11 × 100 A: Total (mmol/mL) of the amino acid analysis values of valine, leucine, glycine and proline M: Amount (mg) of Calcitonin Salmon taken 11: Total of the theoretical residue numbers of valine, leu-cine, glycine and proline per one mole of calcitonin salmon
(1) Acetic acid—Weigh accurately about 10 mg of Calcitonin Salmon, dissolve in water to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 1 g of acetic acid (100), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solu- Lauromacrogol solution (1 in 4) 4.0 mL 4.0 mL 4.0 mL 4.0 mL 4.0 mL tion, add water to make exactly 200 mL, and use this solu-tion as the standard solution. Perform the test with exactly Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL Water a sufficient a sufficient a sufficient a sufficient a sufficient amount amount amount amount amount Total volume 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B, C, D and E as directed in the follow-ing table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) Mobile phase D (volz) Mobile phase E (volz) 0 – 1.5 1.5 – 4 4 – 12 12 – 26 26 – 30 Reaction reagent: Mix 407 g of lithium acetate dihydrate, 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for 10 minutes while passing nitrogen, and use this solution as solution A. Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for 30 minutes while passing nitrogen, and use this solution as solu- 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of acetic acid in each solution, and calculate the amount of acetic acid by the following equation: the amount of acetic acid is not more than 7.0z. Amount (z) of acetic acid (CH3COOH) ＝ MS/MT × AT/AS × 1/10 MS: Amount (mg) of acetic acid (100) taken MT: Amount (mg) of Calcitonin Salmon taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: To 0.7 mL of phosphoric acid add 900 mL of water, adjust the pH to 3.0 with 8 mol/L sodium hy-droxide TS, and add water to make 1000 mL. Mobile phase B: Methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. tion B. Mix solution A and solution B before use. Flow rate of mobile phase: About 0.4 mL per minute. Flow rate of reaction reagent: About 0.35 mL per minute. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, aspartic acid, threonine, serine, glutamic acid, pro-line, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and argi-nine are eluted in this order with the resolutions between the peaks of threonine and serine, glycine and alanine, and isoleucine and leucine being not less than 1.2, 1.0 and 1.2, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak areas of aspartic acid, proline, valine and arginine are not more than 2.0z, respectively. Peptide content Calculate the peptide content in Calcitonin Salmon by the following equation using amino acid analysis values (mmol/mL) obtained in the Constituent amino acids: it is not less than 80.0z. 0 – 5 95 5 5 – 10 95 → 50 5 → 50 10 – 20 50 50 20 – 22 50 → 95 50 → 5 22 – 30 95 5 Flow rate: Adjust so that the retention time of acetic acid is about 4 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of acetic acid are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of acetic acid is not more than 2.0z. (2) Related substances—Dissolve 2 mg of Calcitonin Salmon in 2 mL of dilute acetic acid, and use this solution as the sample solution. Perform the test with 20 mL of the sam-ple solution as directed under Liquid Chromatography JP XVIII Official Monographs / Calcitonin Salmon 593 <2.01> according to the following conditions. Determine each peak area from the sample solution by the automatic integra-tion method, and calculate the amount of them by the area percentage method: the total amount of the peaks other than calcitonin salmon is not more than 3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 1z trimethylamine-phosphate buffer solution (pH 3.0) and acetonitrile (27:13). Flow rate: Adjust so that the retention time of calcitonin salmon is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of calcitonin salmon, beginning after the sol-vent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak at room temperature for about 30 minutes, and centrifuge at 3000 revolutions per minute for 10 minutes to separate serum. Serum calcium determination: Pipet 0.1 mL of the serum, add exactly 6.9 mL of strontium TS, mix well, and use this solution as the sample solution for calcium determi-nation. Separately, pipet a suitable volume of Standard Cal-cium Solution for Atomic Absorption Spectrophotometry, dissolve in strontium TS to make a solution so that each mL contains 0.2 to 3 mg of calcium (Ca: 40.08), and use this solution as the standard solution for calcium determination. Perform the test as directed under Atomic Absorption Spec-trometry <2.23> according to the following conditions, and calculate the calcium content of the sample solution for calcium determination from the calibration curve obtained from the absorbance of the standard solution for calcium determination. Amount (mg) of Calcium (Ca) in 100 mL of the serum ＝ Calcium content (ppm) in the sample solution for calcium determination × 7 Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Calcium hollow-cathode lamp. Wavelength: 422.7 nm. Calculation: Amounts of calcium in the serum ob- area of calcitonin salmon obtained with 20 mL of this solu- tion is equivalent to 5 to 15z of that with 20 mL of the solu- tained with SH, SL, TH and TL are symbolized as y1, y2, y3 tion for system suitability test. System performance: Dissolve 5 mg of methyl parahy-droxybenzoate and 7 mg of ethyl parahydroxybenzoate in 100 mL of acetonitrile. When the procedure is run with 20 and y4, respectively. Sum up y1, y2, y3 and y4 on each set to obtain Y1, Y2, Y3 and Y4, respectively. Units per mg of peptide ＝ antilog M × b/a × 1/c × 5 M ＝ 0.3010 × (Y /Y ) mL of this solution under the above operating conditions a b methyl parahydroxybenzoate and ethyl parahydroxybenzo-ate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak areas of calcitonin salmon is not more than 2.0z. Water <2.48> Not more than 10.0z (5 mg, coulometric titration).
(i) Test animals: Select healthy albino rats weigh-ing between 55 and 180 g, fasted for 24 hours before the test but allowed to drink water ad libitum. Standard solutions: Dissolve a quantity of Calcitonin Salmon RS in acetic acid buffer solution containing 0.1z bovine serum albumin, and prepare a high-dose standard solution SH and a low-dose standard solution SL containing exactly 0.050 and 0.025 Units per mL, respectively. Sample solutions: According to the labeled units, Ya ＝ －Y1 － Y2 ＋ Y3 ＋ Y4 Yb ＝ Y1 － Y2 ＋ Y3 － Y4 a: Amount (mg) of Calcitonin Salmon taken b: Total volume (mL) of the high-dose sample solution prepared by dissolving Calcitonin Salmon in acetic acid buffer solution containing 0.1z bovine serum albumin. c: Peptide content (z) F? computed by the following equation should be smaller than F1 shown in the table against n with which s2 is calcu-lated. Calculate L (P ＝ 0.95) by use of the following equa-tion: L should be not more than 0.20. If F? exceeds F1, or if L exceeds 0.20, repeat the test, increasing the number of animals or arranging the assay conditions so that F? is not more than F1 and L is not more than 0.20. F? ＝ (－Y1 ＋ Y2 ＋ Y3 － Y4)2/4fs2 f: Number of the test animals of each group. s 2 ＝ {Sy2 － (Y/f )}/n Sy2: The sum of squares of y1, y2, y3 and y4 in each group. Y ＝ Y 2 ＋ Y2 ＋ Y 2 ＋ Y 2 1 2 3 4 weigh accurately a suitable amount of Calcitonin Salmon, and dissolve in acetic acid buffer solution containing 0.1z bovine serum albumin, and prepare a high-dose sample solu- n ＝ 4( f － 1) (C － 1)(CM 2 ＋ 0.09062) L ＝ 2 C ＝ Y 2/(Y 2 － 4fs2t2) b b tion TH and the low-dose sample solution TL having Units equal to the standard solutions in equal volumes, respec-tively. Dose for injection: Inject 0.3 mL per animal. Procedure: Divide the test animals at random into 4 groups, A, B, C and D, with not less than 8 animals and equal numbers in each group. Inject SH, SL, TH and TL into the tail vein or subcutaneously into the neck of each animal of the respective groups. At 1 hour after the injection, collect blood from the abdominal aorta in a way that minimizes the suffering of the animals, allow the blood samples to stand t2: Value shown in the following table against n used to calculate s2. ‌594‌ n t2 ＝ F1 n t2 ＝ F1 n t2 ＝ F1 as directed under Flame Coloration Test <1.04> (1): no green color appears. 161.45 4.667 4.242 (4) Magnesium and alkali metals—Dissolve 1.0 g of 18.51 4.600 4.225 Precipitated Calcium Carbonate in a mixture of 20 mL of 10.129 4.543 4.210 water and 10 mL of dilute hydrochloric acid, boil, neutralize 7.709 4.494 4.196 with ammonia TS, and add ammonium oxalate TS until 6.608 4.451 4.183 precipitation of calcium oxalate is completed. Heat the mix- 5.987 4.414 4.171 ture on a water bath for 1 hour, cool, dilute with water to 5.591 4.381 4.085 100 mL, shake well, and filter. To 50 mL of the filtrate add 5.318 4.351 4.001 0.5 mL of sulfuric acid, evaporate to dryness, and ignite at 5.117 4.325 3.920 6009C to constant mass: the mass of the residue is not more 4.965 4.301 ∞ 3.841 than 5.0 mg. 11 4.844 12 4.747 23 4.279 24 4.260 (5) Arsenic <1.11>—Moisten 0.40 g of Precipitated Cal-cium Carbonate with 1 mL of water, then dissolve in 4 mL of dilute hydrochloric acid, use this solution as the test solu-
Camostat Mesilate, when dried, contains not less than 98.
Camostat Mesilate occurs as white, crystals or QT and QS, of the peak area of camostat to that of the inter-nal standard. Amount (mg) of camostat mesilate (C20H22N4O5.CH4O3S) ＝ MS × QT/QS MS: Amount (mg) of Camostat Mesilate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in ethanol (95) (1 in 1500). ‌JP XVIII Official Monographs / dl-Camphor 611 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, a solution of so-dium 1-heptane sulfonate (1 in 500), a solution of sodium lauryl sulfate (1 in 1000) and acetic acid (100) (200:100:50:1). Flow rate: Adjust so that the retention time of camostat is about 10 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, camostat and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of camostat to that of the internal standard is not more than 1.0z.
(1) To 4 mL of a solution of Camostat Mesilate (1 in 2000) add 2 mL of 1-naphthol TS and 1 mL of diacetyl TS, and allow to stand for 10 minutes: a red color develops. Determine the absorption spectrum of a solution of Camostat Mesilate (1 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Camostat Mesilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. A 0.1 g portion of Camostat Mesilate responds to Qualitative Tests <1.09> (1) for mesilate.
(1) Heavy metals <1.07>—Dissolve 1.0 g of Camostat Mesilate in 40 mL of water by warming, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid (not more than 20 ppm). Arsenic <1.11>—Dissolve 2.0 g of Camostat Mesilate in 20 mL of 2 mol/L hydrochloric acid TS by heating in a water bath, and continue to heat for 20 minutes. After cool-ing, centrifuge, take 10 mL of the supernatant liquid, and use this solution as the test solution. Perform the test (not more than 2 ppm). Related substances—Dissolve 30 mg of Camostat Mesilate in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand overnight in iodine vapor: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, silica gel, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 50 mg each of Camostat Mesilate and Camostat Mesilate RS, previously dried, and dissolve each in water to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the test with 2 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac- cording to the following conditions, and calculate the ratios, Dimethylcarbamoylmethyl 4-(4-guanidinobenzoyloxy)phenylacetate monomethanesulfonate [59721-29-8] Camostat Mesilate, when dried, contains not less than 98.5z of camostat mesilate (C20H22N4O5. CH4O3S). Description Camostat Mesilate occurs as white, crystals or QT and QS, of the peak area of camostat to that of the inter-nal standard. Amount (mg) of camostat mesilate (C20H22N4O5.CH4O3S) ＝ MS × QT/QS MS: Amount (mg) of Camostat Mesilate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in ethanol (95) (1 in 1500). ‌JP XVIII Official Monographs / dl-Camphor 611 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, a solution of so-dium 1-heptane sulfonate (1 in 500), a solution of sodium lauryl sulfate (1 in 1000) and acetic acid (100) (200:100:50:1). Flow rate: Adjust so that the retention time of camostat is about 10 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, camostat and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of camostat to that of the internal standard is not more than 1.0z.
Candesartan Cilexetil contains not less than 99.
Candesartan Cilexetil occurs as white, crystals or crystalline powder. It is soluble in acetic acid (100), sparingly soluble in meth-anol, slightly soluble in ethanol (99.5), and practically in-soluble in water. A solution of Candesartan Cilexetil in methanol (1 in 100) shows no optical rotation. Candesartan Cilexetil shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Candesartan Cilexetil in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Can-desartan Cilexetil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the RS according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals.
(1) Water—Shake 1.0 g of dl-Camphor with 10 mL of carbon disulfide: the solution is clear. Chlorinated compounds—Mix 0.20 g of finely pow-dered dl-Camphor with 0.4 g of sodium peroxide in a dried porcelain crucible. Heat the crucible gently by the open flame until the incineration is complete. Dissolve the residue in 20 mL of warm water, acidify with 12 mL of dilute nitric acid, and filter the solution into a Nessler tube. Wash the filter paper with three 5-mL portions of hot water, adding the washings to the filtrate. After cooling, add water to make 50 mL, then add 1 mL of silver nitrate TS, mix well, and allow to stand for 5 minutes: the turbidity of the solu-tion does not exceed that of the following control solution. Control solution: Prepare in the same manner as described above, using 0.20 mL of 0.01 mol/L hydrochloric acid VS. Non-volatile residue—Heat 2.0 g of dl-Camphor on a water bath until sublimation is complete, then dry the residue at 1059C for 3 hours: the mass of the residue does not exceed 1.0 mg.
Weigh accurately about 0.1 g each of dl-Camphor and dl-Camphor RS, add exactly 5 mL each of the internal standard solution, dissolve in ethanol (99.5) to make 100 mL, and use these solutions as the sample solution and standard solution, respectively. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and calculate the ratios, QT and QS, of the peak area of dl-camphor to that of the internal standard. Amount (mg) of dl-camphor (C10H16O) ＝ MS × QT/QS MS: Amount (mg) of dl-Camphor RS taken Internal standard solution—A solution of methyl salicylate in ethanol (99.5) (1 in 25). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 3 m in length, which is packed with 10z of polyethylene glycol 20 M for gas chromatography supported on 180 to 250 mm mesh silanized siliceous earth for gas chromatography. Column temperature: A constant temperature of about 1609C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of dl-camphor is about 6 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, dl-camphor and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. C33H34N6O6: 610.66 (1RS)-1-(Cyclohexyloxycarbonyloxy)ethyl-2-ethoxy-1-{[2?-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl}- 1H-benzimidazole-7-carboxylate [145040-37-5] Candesartan Cilexetil contains not less than 99.0z and not more than 101.0z of candesartan cilexetil (C33H34N6O6), calculated on the anhydrous basis. Description Candesartan Cilexetil occurs as white, crystals or crystalline powder. It is soluble in acetic acid (100), sparingly soluble in meth-anol, slightly soluble in ethanol (99.5), and practically in-soluble in water. A solution of Candesartan Cilexetil in methanol (1 in 100) shows no optical rotation. Candesartan Cilexetil shows crystal polymorphism. Identification (1) Determine the absorption spectrum of a solution of Candesartan Cilexetil in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Can-desartan Cilexetil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the RS according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Candesartan Cilexetil according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Candesartan Cilexetil in 50 mL of a mixture of acetonitrile and water (3:2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- ‌JP XVIII Official Monographs / Candesartan Cilexetil Tablets 613 lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peaks, having the relative retention time of about 0.4 and about 2.0 to can-desartan cilexetil, obtained from the sample solution is not larger than 1/5 times the peak area of candesartan cilexetil from the standard solution, the area of the peak, having the relative retention time of about 0.5 to candesartan cilexetil, from the sample solution is not larger than 3/10 times the peak area of candesartan cilexetil from the standard solu-tion, the area of the peak other than candesartan cilexetil and the peaks mentioned above from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the standard solution, and the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 3/5 times the peak area of candesartan cilex-etil from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Mobile phase B: A mixture of acetonitrile, water and acetic acid (100) (90:10:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Each mL of 0.1 mol/L perchloric acid VS ＝ 61.07 mg of C33H34N6O6
Capsicum and Salicylic Acid Spirit is a light brown-yellow liquid. Specific gravity d 20: about 0.84
(1) Shake 10 mL of Capsicum and Salicylic Acid Spirit with 15 mL of sodium hydrogen carbonate TS and 10 mL of diethyl ether, and separate the aqueous layer. To 1 mL of the solution add hydrochloric acid-potassium chloride buffer solution (pH 2.0) to make 200 mL, and to 5 mL of this solution add 5 mL of a solution of iron (III) ni-trate enneahydrate (1 in 200): a red-purple color is produced (salicylic acid). To 0.5 mL of Capsicum and Salicylic Acid Spirit add 20 mL of water and 5 mL of dilute hydrochloric acid, extract with 20 mL of diethyl ether, wash the diethyl ether extract with two 5-mL portions of sodium hydrogen carbonate TS, and then extract with 20 mL of dilute sodium hydroxide TS. To 1 mL of the extract add 1 mL of sodium nitrite TS and 1 mL of dilute hydrochloric acid, shake, and allow to stand for 10 minutes. Add 3 mL of sodium hydroxide TS: a yellow color is produced (phenol). To 0.2 mL of Capsicum and Salicylic Acid Spirit add 5 mL of dilute hydrochloric acid, extract with 5 mL of chlo-roform, and use the extract as the sample solution. Dissolve 0.01 g of salicylic acid and 0.02 g of phenol in 5 mL and 25 mL of chloroform, respectively, and use both solutions as the standard solution (1) and the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solu-tion and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots of the several spots obtained from the sample solution exhibit the same Rf values as those from standard solution (1) and standard solution (2). Spray evenly iron (III) chloride TS upon the plate: one of the sever-al spots from the sample solution has the same color tone and Rf value with the spot from the standard solution (1). Alcohol number <1.01> Not less than 8.1 (Method 2). Prepare the sample solution as follows: Pipet 5 mL of Capsicum and Salicylic Acid Spirit at 15 ± 29C into a glass- stoppered, conical flask containing exactly 45 mL of water while shaking vigorously, allow to stand, and filter the lower layer. Discard the first 15 mL of the filtrate. Pipet 25 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, and add water to make exactly 100 mL.
Capsicum Tincture contains not less than 0.
Capsicum Tincture is a yellow-red liquid. It has a burning, pungent taste. Specific gravity d 20: about 0.82
Proceed as directed in the Identification under Capsicum, using Capsicum Tincture as the sample solution. Spot 20 mL each of the sample solution and stand-ard solution. Alcohol number <1.01> Not less than 9.7 (Method 2).
Pipet 2 mL of Capsicum Tincture, add methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of (E)-capsaicin for assay, previously dried in a desiccator (in vacu-um, phosphorus (V) oxide, 409C) for 5 hours, dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 25 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, ATC and ATD, of (E)-capsaicin and dihydrocapsaicin (the relative retention time to (E)-capsaicin is about 1.3) obtained with the sample solution, and the peak area, AS, of (E)-capsaicin obtained with the standard solution. Amount (mg) of total capsaicins ＝ MS × (ATC ＋ ATD)/AS × 0.032 MS: Amount (mg) of (E)-capsaicin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 281 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of (E)-capsai-cin is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of (E)-capsaicin for assay and 4-hydroxy-3-methoxybenzyl nonylic acid amide in methanol to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-hydroxy-3-methoxybenzyl nonylic acid amide and (E)-capsaicin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times ‌JP XVIII Crude Drugs and Related Drugs / Carnauba Wax 1975‌‌ with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of (E)-capsaicin is not more than 1.5z.
Captopril occurs as white, crystals or crystal-line powder. It is very soluble in methanol, freely soluble in ethanol (99.5), and soluble in water.
Determine the infrared absorption spectrum of Captopril as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: －125 – －1349(after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Captopril according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So- ‌JP XVIII Official Monographs / Carbamazepine 623 lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Captopril according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Captopril in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 15 mg of 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-diproline in methanol to make exactly 250 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop with a mixture of toluene and acetic acid (100) (13:7) to a distance of about 15 cm, and air-dry the plate. Place the plate in a chamber filled with iodine vapor, and allow to stand for 30 minutes: the number of the spots other than the spot cor-responding to that obtained from the standard solution and the principal spot from the sample solution is not more than two, and they are not more intense than the spot from the standard solution. 1,1?-[3,3?-Dithiobis(2-methyl-1-oxopropyl)]-L-diproline—Dissolve 0.10 g of Captopril in methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-diproline in methanol to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of 1,1?-[3,3?-dithiobis(2-methyl-1- oxopropyl)]-L-diproline in each solution: AT is not larger than AS. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, methanol and phos-phoric acid (1000:1000:1). Flow rate: Adjust so that the retention time of 1,1?-[3,3?- dithiobis(2-methyl-1-oxopropyl)]-L-diproline is about 10 minutes. System suitability— System performance: Dissolve 25 mg each of Captopril and 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-diproline in 200 mL of methanol. When the procedure is run with 20 mL of this solution under the above operating conditions, captopril and 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-diproline are eluted in this order with the resolution be-tween these peaks being not less than 3. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of 1,1?-[3,3?-dithiobis(2-methyl-1-oxopropyl)]-L-dipro-line is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Captopril, dissolve in 100 mL of water, add 20 mL of dilute sulfuric acid and 1 g of potassium iodide, and shake. Titrate <2.50> with 1/60 mol/L potassium iodate VS (indicator: 2 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 1/60 mol/L potassium iodate VS ＝ 21.73 mg of C9H15NO3S
Carbamazepine, when dried, contains not less than 97.
Carbamazepine occurs as a white to slightly yellow-white powder. It is odorless and tasteless at first, and leaves a slightly bitter aftertaste. It is freely soluble in chloroform, sparingly soluble in ethanol (95) and in acetone, and very slightly soluble in water and in diethyl ether.
(1) To 0.1 g of Carbamazepine add 2 mL of nitric acid, and heat on a water bath for 3 minutes: an orange-red color is produced. To 0.1 g of Carbamazepine add 2 mL of sulfuric acid, and heat on a water bath for 3 minutes: a yellow color is produced with a green fluorescence. Examine Carbamazepine under ultraviolet light: the solution shows an intense blue fluorescence. Determine the absorption spectrum of the solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 1.0 g of Carbamazepine in 10 mL of chloroform: the solution is clear and colorless to pale yellow. Acidity—To 2.0 g of Carbamazepine add exactly 40 mL of water, stir well for 15 minutes, and filter through a glass filter (G3). To 10 mL of this filtrate add 1 drop of phenolphthalein TS and 0.50 mL of 0.01 mol/L sodium hy-droxide VS: a red color is produced. Alkalinity—To 10 mL of the filtrate obtained in (2) add 1 drop of methyl red TS and 0.50 mL of 0.01 mol/L hydrochloric acid VS: a red color is produced. Chloride <1.03>—Dissolve 0.25 g of Carbamazepine in 30 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.20 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.028z). Heavy metals <1.07>—Proceed with 2.0 g of Car- bamazepine according to Method 2, and perform the test. ‌624 Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.25 g of Carbamaze-pine in 10 mL of chloroform, and use this solution as the sample solution. Separately, dissolve 5.0 mg of iminodiben-zyl in chloroform to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of toluene and methanol (19:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly potassium dichromate-sulfuric acid TS on the plate: the spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 50 mg of Carbamazepine, previously dried and accurately weighed, in ethanol (95) to make ex-actly 250 mL. Pipet 5 mL of this solution and add ethanol (95) to make exactly 100 mL. Perform the test as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbance A of this solution at the wavelength of maximum absorption at about 285 nm. Amount (mg) of carbamazepine (C15H12N2O) ＝ A/490 × 50,000
Carbidopa Hydrate contains not less than 98.
Carbidopa Hydrate occurs as a white to yel-lowish white powder. It is sparingly soluble in methanol, slightly soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 1979C (with decomposition).
(1) Dissolve 0.01 g of Carbidopa Hydrate in 250 mL of a solution of hydrochloric acid in methanol (9 in 1000). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Carbidopa RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Car-bidopa Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Carbidopa RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －21.0 – －23.59(1 g, alumi- num (III) chloride TS, 100 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Carbidopa Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Carbidopa Hydrate in 70 mL of the mobile phase, by warming and using ultrasonication, if necessary. After cooling, add the mobile phase to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area from both so-lutions by the automatic integration method: the total area of the peaks other than carbidopa obtained from the sample solution is not larger than the peak area of carbidopa from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of carbidopa. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of carbidopa obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. Loss on drying <2.41> 6.9 – 7.9z (1 g, in vacuum not exceeding 0.67 kPa, 1009C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Carbidopa Hydrate and Carbidopa RS (separately determine the loss on drying <2.41> under the same conditions as Carbidopa Hy-drate), and dissolve each in 70 mL of the mobile phase, by warming and using ultrasonication if necessary. After cool-ing, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of carbidopa in each solution. Amount (mg) of carbidopa hydrate (C10H14N2O4.H2O) ＝ MS × AT/AS × 1.080 MS: Amount (mg) of Carbidopa RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 950 mL of 0.05 mol/L sodium dihydro-gen phosphate TS add 50 mL of ethanol (95), and adjust the pH to 2.7 with phosphoric acid. ‌626 Flow rate: Adjust so that the retention time of carbidopa is about 6 minutes. System suitability— ‌System performance: Dissolve 50 mg each of Carbidopa Hydrate and methyldopa in 100 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, methyldopa and carbidopa are eluted in this order with the resolution between these peaks being not less than 0.9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carbidopa is not more than 1.0z.
Carbon Dioxide contains not less than 99.
Carbon Dioxide is a colorless gas at room tem-perature and under atmospheric pressure. It is odorless. A 1 mL volume of Carbon Dioxide dissolves in 1 mL of water, and the solution is slightly acid. 1000 mL of Carbon Dioxide at 09C and under a pressure of 101.3 kPa weighs 1.978 g.
(1) Pass 100 mL of Carbon Dioxide through a carbon dioxide measuring detector tube: the detec-tor tube is changed to a stipulated color tone by each detec-tor tube, provided that the detector tube with a upper limit of measurement of not less than 10z is used. (2) Pass Carbon Dioxide into calcium hydroxide TS: a white precipitate is produced. Collect the precipitate, and add acetic acid (31): it dissolves with effervescence.
(1) Acidity—Place 50 mL of freshly boiled and cooled water in a Nessler tube, and pass 1000 mL of Carbon Dioxide into it for 15 minutes through an introducing tube about 1 mm in diameter extending to 2 mm from the bottom of the Nessler tube, then add 0.10 mL of methyl orange TS: the solution is not more colored than the following control solution. Control solution: To 50 mL of freshly boiled and cooled water in a Nessler tube add 0.10 mL of methyl orange TS and 1.0 mL of 0.01 mol/L hydrochloric acid VS. Hydrogen phosphide, hydrogen sulfide or reducing ＝ MS × QT/QS × V/4 5 9 4 organic substances—Place 25 mL of silver nitrate-ammonia TS and 3 mL of ammonia TS in each of two Nessler tubes MS: Amount of L-carbocisteine for assay taken Internal standard solution—A solution of nicotinic acid (9 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave- A and B, and designate the solution in each tube as solution A and solution B, respectively. Pass 1000 mL of Carbon Di-oxide into solution A in the same manner as directed in (1): the turbidity and color of this solution are the same as that of solution B. ‌628 Carbon monoxide—Pass a specified amount of Car-bon Dioxide through a carbon monoxide measuring detector tube: the concentration of carbon monoxide is less than 15 ppm, provided that the passing amount (mL) of Carbon Dioxide is stipulated according to each detector tube.
Place 125 mL of a solution of potassium hydroxide (1 in 2) in a gas pipet of suitable capacity. Measure exactly about 100 mL of Carbon Dioxide in a 100-mL gas buret filled with water. Force the entire volume of gas into the gas pipet, and shake for 5 minutes. Draw some of the unab-sorbed gas into the gas buret, measure the volume, force the residual back upon the surface of the liquid in the gas pipet, and repeat this procedure until a constant volume of the residual reading is obtained. Determine the volume V (mL) of the residual gas. Calculate the volume of the sample and V on the basis of the gas volume at 209C and at 101.3 kPa. Volume (mL) of carbon dioxide (CO2) ＝ volume (mL) of the sample － V (mL)
Containers—Cylinders. Storage—Not exceeding 409C.
Carboplatin occurs as white, crystals or crys-talline powder. It is sparingly soluble in water, and very slightly soluble in ethanol (99.5). Melting point: about 2009C (with decomposition).
(1) To 2 mL of a solution of Carboplatin (1 in 100) add 2 to 3 drops of diluted tin (II) chloride TS (1 in 15), and allow to stand for 30 minutes: a yellowish brown precipitate is formed. (2) Determine the infrared absorption spectrum of Carboplatin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Carboplatin RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. pH <2.54> Dissolve 0.10 g of Carboplatin in 10 mL of water: the pH of this solution is 5.0 to 7.0.
(1) 1,1-Cyclobutanedicarboxylic acid—Weigh ac-curately about 40 mg of Carboplatin, dissolve in the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of 1,1-cyclobutanedicarboxylic acid, and dissolve in the mobile phase to make exactly 100 mL. Pipet 4 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard are eluded in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 1,1-cyclobutanedicarboxylic acid is not more than 2.0z. (2) Related substances—Dissolve 25 mg of Carboplatin in 25 mL of water, and use this solution as the sample solu-tion. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of the peaks by the area percentage method: the amount of the peak, having the relative retention time of about 0.8 to car-boplatin, is not more than 0.25z, the amount of the peak other than carboplatin and the peak mentioned above is not more than 0.1z, and the total amount of the peaks other than carboplatin is not more than 0.5z. Operating conditions— Detector, column, column temperature, mobile phases A and B, and flow rate: Proceed as directed in the operating conditions in the Assay. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. ‌JP XVIII Official Monographs / Carboplatin Injection 629 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 100 0 15 – 35 100 → 0 0 → 100 35 – 50 0 100 Time span of measurement: About 2.5 times as long as the retention time of carboplatin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the sample solution add water to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add water to make exactly 20 mL. Confirm that the peak area of carboplatin obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of carboplatin is not more than 2.0z. Loss on drying <2.41> Not more than 0.1z (0.5 g, 1059C, 4 hours).
Weigh accurately about 25 mg each of Carboplatin and Carboplatin RS (separately determine the loss on drying <2.41> under the same conditions as Carboplatin), dissolve separately in water to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of carboplatin in each solution. Amount (mg) of carboplatin (C6H12N2O4Pt) ＝ MS × AT/AS MS: Amount (mg) of Carboplatin RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylhexylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 279C. Mobile phase A: Dissolve 8.5 g of tetrabutylammonium hydrogensulfate in 80 mL of water, add 3.4 mL of phos-phoric acid, and adjust to pH 7.5 with a solution of sodium hydroxide (43 in 100). To 20 mL of this solution add water to make 1000 mL. Mobile phase B: Dissolve 8.5 g of tetrabutylammonium hydrogensulfate in 80 mL of water, add 3.4 mL of phos-phoric acid, and adjust to pH 7.5 with a solution of sodium hydroxide (43 in 100). To 20 mL of this solution add water to make 800 mL, and add 200 mL of acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 0 – 15 100 0 15 – 35 100 → 0 0 → 100 Flow rate: 0.5 mL per minute. System suitability— System performance: To 9 mL of the standard solution add 1 mL of diluted hydrogen peroxide TS (1 in 60), and allow to stand at room temperature for not less than 1 hour. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak of carboplatin and the peak having the relative reten-tion time about 0.93 to carboplatin is not less than 1.2. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carboplatin is not more than 1.0z.
Carmellose occurs as a white powder. It is practically insoluble in ethanol (95). It swells with water to form suspension. It becomes viscid in sodium hydroxide TS. It is hygroscopic.◆
(1) Determine the infrared absorption spec-trum of Carmellose as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) The pH <2.54> of a suspension, obtained by shaking 1 g of Carmellose with 100 mL of water, is between 3.5 and 5.0.
(1) Chloride—Shake well 0.8 g of Carmellose with 50 mL of water, add 10 mL of sodium hydroxide TS to dis- JP XVIII Official Monographs / Carmellose Calcium 631 ‌solve, and add water to make 100 mL. Heat 20 mL of this solution with 10 mL of dilute nitric acid in a water bath until a flocculent precipitate is produced, cool, centrifuge, and take out the supernatant liquid. Wash the precipitate with three 10-mL portions of water by centrifuge each time, com-bine the supernatant liquid and the washings, and add water to make 100 mL. Take 25 mL of this solution in a Nessler tube, add 6 mL of dilute nitric acid and water to make 50 mL, and use this solution as the test solution. Separately, to 0.40 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL, and use this solu-tion as the control solution. To the test solution and the con-trol solution add 1 mL each of silver nitrate TS, ◆mix,◆ and allow to stand protected from light for 5 minutes. Compare the opalescence developed in both solutions ◆against a black background by viewing downward or transversely◆. The opalescence in the test solution is not more intense than that in the control solution (not more than 0.36z). Sulfate—Shake well 0.40 g of Carmellose with 25 mL of water, add 5 mL of sodium hydroxide TS to dissolve, and add 20 mL of water. Heat this solution with 2.5 mL of hydrochloric acid in a water bath until a flocculent precipi-tate is produced, cool, centrifuge, and take out the superna-tant liquid. Wash the precipitate with three 10-mL portions of water by centrifuge each time, combine the supernatant liquid and the washings, and add water to make 100 mL. Filter this solution, discard the first 5 mL of the filtrate, take 25 mL of the subsequent filtrate in a Nessler tube, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and use this solution as the test solution. Separately, to 1.5 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydro-chloric acid and water to make 50 mL, and use this solution as the control solution. To the test solution and the control solution add 2 mL each of barium chloride TS, mix, and allow to stand for 10 minutes. Compare the opalescence de-veloped in both solutions ◆against a black background by viewing downward or transversely◆. The opalescence in the test solution is not more intense than that in the control solu-tion (not more than 0.72z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Car- mellose according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm).◆ Loss on drying <2.41> Not more than 8.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 1.5z (after dry-ing, 1 g). ◆Containers and storage Containers—Tight containers.◆ Carmellose Calcium Carboxymethylcellulose Calcium
Carmofur, when dried, contains not less than 98.
Carmofur occurs as a white crystalline pow-der. It is very soluble in N,N-dimethylformamide, freely solu-ble in acetic acid (100), soluble in diethyl ether, sparingly soluble in methanol and in ethanol (99.5), and practically in-soluble in water. Melting point: about 1119C (with decomposition).
(1) Proceed with 5 mg of Carmofur as di-rected under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid, and prepare the test solution: the test solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Carmofur in a mixture of methanol and phosphoric acid-acetic acid-boric acid buffer solution (pH 2.0) (9:1) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Car-mofur, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Carmofur according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Dissolve 0.20 g of Carmofur in 10 mL of a mixture of methanol and acetic acid (100) (99:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and acetic acid (100) (99:1) to make exactly 500 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 15 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of toluene and acetone (5:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. After exposure of the plate to bromine vapor for 30 second, spray evenly a solution of fluorescein in ethanol (95) (1 in 2500): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 509C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Carmofur, previ-ously dried, dissolve in 20 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hy-droxide-methanol VS until the color of the solution changes from yellow through blue-green to blue (indicator: 3 drops of thymol blue-dimethylformamide TS). Each mL of 0.1 mol/L tetramethylammonium hydroxide-methanol VS ＝ 25.73 mg of C11H16FN3O3
Carteolol Hydrochloride occurs as white, crys tals or crystalline powder. It is soluble in water, sparingly soluble in methanol, very slightly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Carteolol Hydrochloride in 100 mL of water is between 5.0 and 6.0. The solution of Carteolol Hydrochloride (1 in 20) shows no optical rotation. Melting point: about 2779C (with decomposition).
(1) Dissolve 0.1 g of Carteolol Hydrochlo-ride in 5 mL of water, and add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Carteolol Hydrochloride (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Car-teolol Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Carteolol Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Carteolol Hydrochloride in 30 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Car- teolol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Carteolol Hydrochloride according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Carteolol Hy-drochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia solution (28) (50:20:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Carteolol Hydro-chloride, previously dried, add 30 mL of acetic acid (100), dissolve by heating on a water bath, and cool. After adding 70 mL of acetic anhydride, titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.88 mg of C16H24N2O3.HCl
Carvedilol, when dried, contains not less than 99.
Carvedilol occurs as white to pale yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. A solution of Carvedilol in methanol (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Carvedilol in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Car-vedilol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Wrap 2.0 g of Car-vedilol with a filter paper for quantitative analysis, then proceed according to Method 4, and perform the test. Pre-pare the control solution as follows: Put a filter paper for quantitative analysis in a crucible, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), then proceed as directed for the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 10 ppm). (2) Related substances—Dissolve 65 mg of Carvedilol in 100 mL of the mobile phase. To 1 mL of this solution add the mobile phase to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak other than carvedilol obtained from the sample solution is not larger than 3/20 times the peak area of carvedilol from the stand-ard solution, and the total area of the peaks other than car-vedilol from the sample solution is not larger than 1/2 times the peak area of carvedilol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). ‌638 Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 559C. Mobile phase: Dissolve 2.72 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.0 with phos-phoric acid, and add water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of carvedilol is about 4 minutes. Time span of measurement: About 9 times as long as the retention time of carvedilol, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, add the mobile phase to make exactly 20 mL. Con-firm that the peak area of carvedilol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of carvedilol are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carvedilol is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Carvedilol, previ-ously dried, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform the blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.65 mg of C24H26N2O4
Cefaclor contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calcu-lated on the anhydrous basis.
Cefaclor occurs as a white to yellow-white crystalline powder. It is slightly soluble in water and in methanol, and practi-cally insoluble in N,N-dimethylformamide and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Cefaclor (1 in 50,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefaclor as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 40 mg of Cefaclor in 0.5 mL of heavy water for nuclear magnetic resonance spectroscopy and 1 drop of deuterated hydrochloric acid for nuclear magnetic resonance spectroscopy, and determine the 1H spectrum of this solution as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropanesulfonate for nuclear magnetic resonance spectroscopy as an internal ref-erence compound: it exhibits an AB type quartet signal A at around d 3.7 ppm, and a singlet signal or a sharp multiplet signal B at around d 7.6 ppm. The ratio of the integrated in-tensity of each signal, A:B, is about 2:5. Perform the test with Cefaclor as directed under Flame Coloration Test <1.04> (2): a green color appears. D Optical rotation <2.49> [a]20: ＋105 – ＋1209(0.1 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cefaclor according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution by suspend- ing 1.0 g of Cefaclor in 10 mL of N,N-dimethylformamide, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Cefaclor in 10 mL of sodium dihydrogen phosphate TS (pH 2.5), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peaks other than cefaclor obtained from the sample solution are not larger than 1/2 times the peak area of cefaclor from the standard solution, and the total area of the peaks other than cefaclor from the sample solution is not larger than 2 times of the peak area of cefaclor from the standard solution. If necessary, proceed with 20 mL of sodium dihydrogen phosphate TS (pH 2.5) in the same manner as above to compensate the base line. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 7.8 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust the pH to 4.0 with phosphoric acid. Mobile phase B: To 550 mL of the mobile phase A add 450 mL of acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 30 95 → 75 5 → 25 30 – 45 75 → 0 25 → 100 45 – 55 Flow rate: 1.0 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of cefaclor, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add sodium dihydrogen phos-phate TS, pH 2.5 to make exactly 20 mL. Confirm that the peak area of cefaclor obtained with 20 mL of this solution is equivalent to 4 to 6z of that with 20 mL of the standard so-lution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefaclor are not less than 40,000 and 0.8 to 1.3, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak areas and the retention times of cefaclor are not more than 2.0z, respectively. Water <2.48> Not more than 6.5z (0.2 g, volumetric titra-tion, back titration).
Weigh accurately an amount of Cefaclor and Cefaclor RS, equivalent to about 50 mg (potency), and dis-solve each in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution, add 0.1 JP XVIII Official Monographs / Cefaclor Capsules 641 ‌mol/L phosphate buffer solution (pH 4.5) to make 50 mL, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of cefaclor to that of the internal standard. Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 3.4 with diluted phosphoric acid (3 in 500). To 940 mL of this solution add 60 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefaclor is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefaclor and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefaclor to that of the internal standard is not more than 1.0z.
Cefadroxil contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Cefadroxil occurs as a white to light yellow-white powder. It is sparingly soluble in water, slightly soluble in metha-nol, and very slightly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Cefadroxil (1 in 50,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefadroxil RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefadroxil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefadroxil RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefadroxil in a mixture of heavy water for nuclear magnetic resonance spectroscopy and deuterated hydrochloric acid (3:1) (1 in 10), using sodium 3-(trimethylsilyl)propionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits a singlet signal A at around d 2.1 ppm, a doublet signal B at around d 7.0 ppm, and a doublet signal C at around d 7.5 ppm. The ratio of integrated intensity of each signal, A:B:C, is about 3:2:2. D Optical rotation <2.49> [a]25: ＋164 – ＋1829(0.6 g calcu-lated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefadroxil in 200 mL of water: pH of the solution is between 4.0 and 6.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of ‌646 Cefadroxil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.1 g of Cefadroxil in 4 mL of a mixture of ethanol (99.5), water and diluted hydro-chloric acid (1 in 5) (75:22:3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of ethanol (99.5), water and diluted hydrochloric acid (1 in 5) (75:22:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop with a mixture of ethyl acetate, water, ethanol (99.5) and formic acid (14:5:5:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly ninhydrin-citric acid-acetic acid TS on the plate, and heat the plate at 1009C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> Not less than 4.2z and not more than 6.0z (0.5 g, volumetric titration, direct titration).
Weigh accurately an amount of Cefadroxil and Cefadroxil RS, equivalent to about 50 mg (potency), dissolve each in water to make exactly 500 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefadroxil in each solution. Amount [ mg (potency)] of cefadroxil (C16H17N3O5S) ＝ MS × AT/AS × 1000 MS: amount [mg (potency)] of Cefadroxil RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 262 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of potassium dihy-drogenphosphate (17 in 12,500) and methanol (17:3). Flow rate: Adjust so that the retention time of cefadroxil is about 5 minutes. System suitability— System performance: Dissolve about 5 mg (potency) of Cefadroxil and about 10 mg (potency) of propylene glycol cefatrizine in 50 mL of water. When the procedure is run with 10 mL of this solution under the above operating condi-tions, cefadroxil and cefatrizine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefadroxil is not more than 1.0z.
Cefalexin contains not less than 950 mg (potency) and not more than 1030 mg (potency) per mg, calcu-lated on the anhydrous basis.
Cefalexin occurs as a white to light yellow-white, crystals or crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, and practically insoluble in ethanol (95) and in N,N-dimethylformamide. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Cefalexin (3 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefalexin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefalexin in heavy water for nuclear magnetic resonance spectroscopy (1 in 200) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropane-sulfonate for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a singlet signal A at around d 1.8 ppm, and a singlet or a sharp multiplet signal B at around d 7.5 ppm. The ratio of integrated intensity of these signals, A:B, is about 3:5. D Optical rotation <2.49> [a]20: ＋144 – ＋1589(0.125 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefalexin according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefalexin by suspending in 10 mL of N,N-dimethylfor-mamide, and perform the test (not more than 2 ppm). Related substances—Dissolve about 25 mg of Cefalexin in a solution of potassium dihydrogenphosphate (9 in 500) to make 5 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a solution of potassium dihydrogenphosphate (9 in 500) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the areas of each peak by the automatic integra-tion method. If necessary, correct the change of the base-line due to the potassium dihydrogenphosphate solution by proceeding in the same manner with 20 mL of a solution of potassium dihydrogenphosphate (9 in 500): each peak area other than cefalexin from the sample solution is not larger equivalent to 1.8 to 2.2z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefalexin are not less than 150,000 and between 0.8 and 1.3, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the reten-tion time and the peak areas of cefalexin are not more than 2.0z, respectively. Water <2.48> Not more than 8.0z (0.2 g, volumetric titra-tion, back titration).
Weigh accurately an amount of Cefalexin and Cefalexin RS, equivalent to about 25 mg (potency), dissolve each in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 25 mL. Pipet 10 mL of these solutions, add exactly 5 mL of the internal standard solution, then add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefalexin to that of the internal standard. than the peak area of cefalexin from the standard solution, and the total area of the peaks other than cefalexin from the sample solution which are larger than 1/50 times the peak Amount [ mg (potency)] of cefalexin (C ＝ MS × QT/QS × 1000 16H17N3O4S) area of cefalexin from the standard solution is not larger than 5 times of the peak area of cefalexin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 1.0 g of sodium 1-pentanesul-fonate in 1000 mL of water, add 15 mL of triethylamine, and adjust to pH 2.5 with phosphoric acid. Mobile phase B: Dissolve 1.0 g of sodium 1-pentanesul-fonate in 300 mL of water, add 15 mL of triethylamine, and adjust to pH 2.5 with phosphoric acid. To this solution add 350 mL of acetonitrile and 350 mL of methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxy-acetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 1500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen-phosphate in 1000 mL of water, adjust to pH 3.0 with diluted phosphoric acid (3 in 500). To 800 mL of this solu-tion add 200 mL of methanol. Flow rate: Adjust so that the retention time of cefalexin is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) ditions, cefalexin and the internal standard are eluted in this order with the resolution between these peaks being not less 0 – 1 100 0 1 – 34.5 100 → 0 0 → 100 34.5 – 35.5 0 100 Flow rate: 1.0 mL per minute. Time span of measurement: About 2 times as long as the retention time of cefalexin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, add a solution of potassium dihydrogenphosphate (9 in 500) to make exactly 100 mL. Confirm that the peak area of cefalexin obtained with 20 mL of this solution is than 6. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefalexin to that of the internal standard is not more than 1.0z.
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Cefatrizine Propylene Glycolate contains not less than 816 mg (potency) and not more than 876 mg (potency) per mg, calculated on the anhydrous basis.
Cefatrizine Propylene Glycolate occurs as a white to yellowish white powder. It is sparingly soluble in water, and practically insoluble in methanol and in ethanol (95).
(1) Determine the absorption spectrum of a solution of Cefatrizine Propylene Glycolate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefatrizine Propylene Glyco-late RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefatrizine Propylene Glycolate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Cefatrizine Propylene Glycolate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefatri-zine Propylene Glycolate in a mixture of heavy water for nuclear magnetic resonance spectroscopy and deuterated hy-drochloric acid for nuclear magnetic resonance spectroscopy (3:1) (1 in 10), using sodium 3-(trimethylsilyl)propionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits a doublet signal A at around d 1.2 ppm, a doublet signal B at around d 7.0 ppm, a doublet signal C at around d 7.5 ppm and a singlet signal D at around d 8.3 ppm. The ratio of integrated inten-sity of these signals, A:B:C:D, is about 3:2:2:1. D Optical rotation <2.49> [a]20: ＋52 – ＋589(2.5 g calculated on the anhydrous bases, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cefatrizine Propylene Glycolate according to Method 2, and ‌JP XVIII Official Monographs / Cefatrizine Propylene Glycolate for Syrup 655 perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefatrizine Propylene Glycolate according to Method 3, and perform the test (not more than 2 ppm). Use a solution of magnesium nitrate hexahydrate in ethanol (1 in 25). Related substances—Dissolve 25 mg of Cefatrizine Propylene Glycolate in 5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly ninhydrin-citric acid-acetic acid TS on the plate, and heat the plate at 1009C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Cefatrizine Propy-lene Glycolate and Cefatrizine Propylene Glycolate RS, equivalent to about 0.1 g (potency), dissolve each in water to make exactly 500 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefatrizine in each solution. Cefatrizine Propylene Glycolate for Syrup
セフカペン ピボキシル塩酸塩水和物
Cefcapene Pivoxil Hydrochloride Hydrate contains not less than 722 mg (potency) and not more than 764 mg (potency) per mg, calculated on the anhydrous basis.
Cefcapene Pivoxil Hydrochloride Hydrate oc-curs as a white to pale yellow-white, crystalline powder or mass. It has slightly a characteristic odor. It is freely soluble in N,N-dimethylformamide and in methanol, soluble in ethanol (99.5), slightly soluble in water, and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Cefcapene Pivoxil Hydrochloride Hydrate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cef-capene Pivoxil Hydrochloride RS prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectra of Cefca- JP XVIII Official Monographs / Cefcapene Pivoxil Hydrochloride Hydrate 661 pene Pivoxil Hydrochloride Hydrate and Cefcapene Pivoxil Hydrochloride RS as directed in the paste method under In-frared Spectrophotometry <2.25>, and compare these spec-tra: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefca-pene Pivoxil Hydrochloride Hydrate in deuterated methanol for nuclear magnetic resonance spectroscopy (1 in 50) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic reso- nance spectroscopy as an internal reference compound: it exhibits a triplet signal A at around d 6.3 ppm, and a single signal B at around d 6.7 ppm, and the ratio of integrated intensity of each signal, A:B, is about 1:1. Dissolve 10 mg of Cefcapene Pivoxil Hydrochloride Hydrate in 2 mL of a mixture of water and methanol (1:1), and add 1 drop of silver nitrate TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋51 – ＋549(0.1 g calculated on the anhydrous basis, methanol, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefcapene Pivoxil Hydrochloride Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substance I—Dissolve an amount of Cefca-pene Pivoxil Hydrochloride Hydrate, equivalent to about 10 mg (potency), in 2 mL of methanol, add a mixture of water and methanol (1:1) to make 50 mL, and use this solution as the sample solution. Perform the test with 30 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. If nec-essary, compensate the base-line by performing in the same manner as the test with 30 mL of a mixture of water and methanol (1:1). Measure the amount of the peak other than cefcapene pivoxil by the area percentage method: the amounts of the peaks, having the relative retention times of about 1.5 and about 1.7 to cefcapene pivoxil, are not more than 0.2z, respectively. The amount of the peaks other than the peaks mentioned above is not more than 0.1z, and the total of them is not more than 1.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase A: Dissolve 5.99 g of potassium dihydrogen phosphate in water to make 1100 mL. To this solution add a solution prepared by dissolving 1.89 g of tetra-n-pentylam-monium bromide in methanol to make 1000 mL. Mobile phase B: A mixture of methanol and water (22:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 20 20 – 40 98 → 50 2 → 50 40 – 50 Flow rate: 0.8 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of cefcapene pivoxil. System suitability— Test for required detectability: To exactly 1 mL of the sample solution add a mixture of water and methanol (1:1) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of cefcapene pivoxil obtained with 30 mL of this solution is equivalent to 7 to 13z of that with 30 mL of the solution for system suitability test. System performance: Dissolve 10 mg of Cefcapene Pivoxil Hydrochloride Hydrate and 10 mg of propyl parahydroxy-benzoate in 25 mL of methanol, and add water to make 50 mL. To 5 mL of this solution add the mixture of water and methanol (1:1) to make 50 mL. When the procedure is run with 30 mL of this solution under the above operating condi-tions, cefcapene pivoxil and propyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 3 times with 30 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cefcapene pivoxil is not more than 4.0z. Related substance II—Dissolve an amount of Cefca-pene Pivoxil Hydrochloride Hydrate, equivalent to about 2 mg (potency), in N,N-dimethylformamide for liquid chro-matography to make 20 mL, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks which appear earlier than cefcapene pivoxil is not more than 1.7z of the total area of the peaks other than the solvent. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase: A solution of lithium bromide in N,N-dimethylformamide for liquid chromatography (13 in 5000). Flow rate: Adjust so that the retention time of cefcapene pivoxil is about 22 minutes. Time span of measurement: About 1.8 times as long as the retention time of cefcapene pivoxil. System suitability— Test for required detectability: To exactly 1 mL of the sample solution add N,N-dimethylformamide for liquid chromatography to make 100 mL, and use this solution as the solution for system suitability test. Pipet 3 mL of the so-lution for system suitability test, and add N,N-dimethylfor-mamide for liquid chromatography to make exactly 10 mL. Conform that the peak area of cefcapene pivoxil obtained with 20 mL of this solution is equivalent to 20 to 40z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, the number of theoretical plates of the peak of cefca-pene pivoxil is not less than 12,000. System repeatability: When the test is repeated 6 times ‌662 with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak areas of cefcapene pivoxil is not more than 4.0z. Water <2.48> Not less than 2.8z and not more than 3.7z (0.5 g, volumetric titration, back titration).
Weigh accurately an amount of Cefcapene Pivoxil Hydrochloride Hydrate and Cefcapene Pivoxil Hydrochlo-ride RS, equivalent to about 20 mg (potency), and dissolve each in a mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution and the mixture of water and methanol (1:1) to them to make 50 mL, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of cefcapene pivoxil to that of the internal standard. Cefcapene Pivoxil Hydrochloride Fine Granules
Cefdinir contains not less than 930 mg (potency) and not more than 1020 mg (potency) per mg.
Cefdinir occurs as a white to light yellow crys-talline powder. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in 0.1 mol/L phosphate buffer solution (pH 7.0).
(1) Determine the absorption spectra of so-lutions of Cefdinir and Cefdinir RS in 0.1 mol/L phosphate buffer solution (pH 7.0) (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare these spectra: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectra of Cef-dinir and Cefdinir RS as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare these spec-tra: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefdinir in a mixture of deuterated dimethyl sulfoxide for nuclear magnetic resonance spectroscopy and heavy water for nuclear magnetic resonance spectroscopy (4:1) (1 in 10), using tetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits multiplet signals, A at around d 5.0 – 6.1 ppm and B at around d 6.4 – 7.5 ppm. The ratio of integrated intensity of each signal, A:B is about 2:1. D Optical rotation <2.49> [a]20: －58 – －669 (0.25 g, 0.1 mol/L phosphate buffer solution (pH 7.0), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefdinir according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Dissolve about 0.1 g of Cefdinir in 10 mL of 0.1 mol/L phosphate buffer solution (pH 7.0). To 3 mL of this solution add tetramethylammonium hydrox-ide TS (pH 5.5) to make 20 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of their peaks by the area percentage method: the amount of the peaks, having the relative retention time of about 0.7, about 1.2 and about 1.5 to cefdinir, are not more than 0.7z, not more than 0.3z and not more than 0.8z, respectively, the total amount of the peaks, having the rela-tive retention time of about 0.85, about 0.93, about 1.11 and about 1.14, is not more than 0.4z, and the amount of the peak other than cefdinir and the peaks mentioned above is not more than 0.2z. And the total amount of the peaks other than cefdinir is not more than 3.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: To 1000 mL of tetramethylammonium hydroxide TS (pH 5.5) add 0.4 mL of 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate TS. ‌JP XVIII Official Monographs / Cefdinir Capsules 665 Mobile phase B: To 500 mL of tetramethylammonium hy-droxide TS (pH 5.5) add 300 mL of acetonitrile for liquid chromatography and 200 mL of methanol, and add 0.4 mL of 0.1 mol/L disodium dihydrogen ethylenediamine tetraace-tate TS. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase: To 1000 mL of tetramethylammonium hydroxide TS (pH 5.5) add 0.4 mL of 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate TS. To 900 mL of 0 – 2 95 5 2 – 22 95 → 75 5 → 25 22 – 32 75 → 50 25 → 50 32 – 37 50 50 Flow rate: 1.0 mL per minute (the retention time of cefdinir is about 22 minutes). Time span of measurement: For 37 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add tetramethylammonium hydroxide TS (pH 5.5) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, add tetramethylammonium hydroxide TS (pH 5.5) to make exactly 10 mL. Confirm that the peak area of cefdinir obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: Dissolve 30 mg of Cefdinir RS and 2 mg of cefdinir lactam ring-cleavage lactones in 3 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), add tetramethy-lammonium hydroxide TS (pH 5.5) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, peak 1 and peak 2 of cefdinir lactam ring-cleavage lactones separated into 4 peaks, cefdinir, peak 3 and peak 4 of remaining cefdinir lactam ring-cleavage lactones are eluted in this order. Relative reten-tion time of peak 3 of cefdinir lactam ring-cleavage lactone to cefdinir is about 1.11. The number of theoretical plates and the symmetry factor of the peak of cefdinir are not less than 7000 and not more than 3.0, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak areas of cefdinir is not more than 2.0z. Water <2.48> Not more than 2.0z (1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination).
Weigh accurately an amount of Cefdinir and Cef-dinir RS equivalent to about 20 mg (potency), dissolve each in 0.1 mol/L phosphate buffer solution (pH 7.0) to make ex-actly 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 5 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefdinir in each solution. Amount [ mg (potency)] of cefdinir (C14H13N5O5S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefdinir RS taken this solution add 60 mL of acetonitrile for liquid chromatog-raphy and 40 mL of methanol. Flow rate: Adjust so that the retention time of cefdinir is about 8 minutes. System suitability— System performance: Dissolve 2 mg of Cefdinir RS and 5 mg of cefdinir lactam ring-cleavage lactones in 10 mL of 0.1 mol/L phosphate buffer solution, pH 7.0. When the proce-dure is run with 5 mL of this solution under the above operat-ing conditions, peak 1 and peak 2 of cefdinir lactam ring-cleavage lactones separated into 4 peaks, cefdinir, peak 3 and peak 4 of remaining cefdinir lactam ring-cleavage lactones are eluted in this order. The resolution between the peak 2 of cefdinir lactam ring-cleavage lactone and that of cefdinir is not less than 1.2. The number of theoretical plates and the symmetry factor of the peak of cefdinir are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas of cefdinir is not more than 1.0z.
Cefditoren Pivoxil contains not less than 770 mg (potency) and not more than 820 mg (potency) per mg, Optical rotation <2.
Cefditoren Pivoxil occurs as a light yellow-white to light yellow crystalline powder. It is sparingly soluble in methanol, slightly soluble in acetonitrile and in ethanol (95), very slightly soluble in diethyleter and practically insoluble in water. It dissolves in dilute hydrochloric acid.
(1) Dissolve 5 mg of Cefditoren Pivoxil in 3 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 5 minutes, add 1 mL of acidic ammonium iron (III) sulfate TS and shake: a red-brown color develops. Dissolve 1 mg of Cefditoren Pivoxil in 1 mL of dilute hydrochloric acid and 4 mL of water, add 3 drops of sodium nitrite TS under ice-cooling, shake, and allow to stand for 2 minutes. Then add 1 mL of ammonium amidosulfate TS, shake well, and allow to stand for 1 minute, and add 1 mL of N,N-diethyl-N?-1-naphthylethylenediamine oxalate TS: a purple color develops. Determine the absorption spectrum of a solution of Cefditoren Pivoxil in methanol (1 in 50,000) as directed MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken Internal standard solution—A solution of propyl p-hydrox-ybenzoate in acetonitrile (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.58 g of ammonium formate in 900 mL of water, adjust to pH 6.0 with diluted formic acid (1 in 250), and add water to make 1000 mL. To 450 mL of this solution add 275 mL of acetonitrile and 275 mL of methanol. Flow rate: Adjust so that the retention time of cefditoren pivoxil is about 15 minutes. ‌668‌ System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefditoren pivoxil are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefditoren pivoxil to that of the internal standard is not more than 1.0z.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefditoren Pivoxil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 1.5z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition Being specified separately when the drug is granted approval based on the Law.
Conduct this procedure using light-resistant vessels. Weigh accurately an amount of Cefditoren Pivoxil and Cef-ditoren Pivoxil RS, equivalent to about 40 mg (potency), dis-solve in 40 mL of acetonitrile, add exactly 10 mL each of the internal standard solution, and add acetonitrile to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of cefditoren pivoxil to that of the internal standard. calculated on the anhydrous basis. The potency of Cefditoren Pivoxil is expressed as mass (potency) of cefditoren (C H N O S : 506.58). Amount [ mg (potency)] of cefditoren (C ＝ MS × QT/QS × 1000 19H18N6O5S3) 19 18 6 5 3 Description Cefditoren Pivoxil occurs as a light yellow-white to light yellow crystalline powder. It is sparingly soluble in methanol, slightly soluble in acetonitrile and in ethanol (95), very slightly soluble in diethyleter and practically insoluble in water. It dissolves in dilute hydrochloric acid. Identification (1) Dissolve 5 mg of Cefditoren Pivoxil in 3 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 5 minutes, add 1 mL of acidic ammonium iron (III) sulfate TS and shake: a red-brown color develops. Dissolve 1 mg of Cefditoren Pivoxil in 1 mL of dilute hydrochloric acid and 4 mL of water, add 3 drops of sodium nitrite TS under ice-cooling, shake, and allow to stand for 2 minutes. Then add 1 mL of ammonium amidosulfate TS, shake well, and allow to stand for 1 minute, and add 1 mL of N,N-diethyl-N?-1-naphthylethylenediamine oxalate TS: a purple color develops. Determine the absorption spectrum of a solution of Cefditoren Pivoxil in methanol (1 in 50,000) as directed MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken Internal standard solution—A solution of propyl p-hydrox-ybenzoate in acetonitrile (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.58 g of ammonium formate in 900 mL of water, adjust to pH 6.0 with diluted formic acid (1 in 250), and add water to make 1000 mL. To 450 mL of this solution add 275 mL of acetonitrile and 275 mL of methanol. Flow rate: Adjust so that the retention time of cefditoren pivoxil is about 15 minutes. ‌668‌ System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefditoren pivoxil are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefditoren pivoxil to that of the internal standard is not more than 1.0z.
Cefepime Dihydrochloride Hydrate contains not less than 835 mg (potency) and not more than 886 mg (potency) per mg, calculated on the anhydrous basis.
Cefepime Dihydrochloride Hydrate occurs as a white to yellowish white, crystals or crystalline powder. It is freely soluble in water and in methanol, and slightly soluble in ethanol (95), and practically in soluble in diethyl ether.
(1) Dissolve 0.02 g of Cefepime Dihydro-chloride Hydrate in 2 mL of water, add 1 mL of a solution of hydroxylammonium chloride (1 in 10) and 2 mL of so-dium hydroxide TS, allow to stand for 5 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS and 3 drops of iron (III) chloride TS: a red-brown color develops. Determine the absorption spectra of solutions (1 in 20,000) of Cefepime Dihydrochloride Hydrate and Cefepime Dihydrochloride RS as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare these spectra: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectra of Cefepime Dihydrochloride Hydrate and Cefepime Dihydro-chloride RS as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare these spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefepime Dihydrochloride Hydrate in heavy water for nuclear mag-netic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits single signals, A and B, at around d 3.1 ppm and at around d 7.2 ppm, respectively, and the ratio of integrated intensity of each signal, A:B, is about 3:1. Amount [mg (potency)] of cefditoren (C ＝ MS × QT/QS × 25 19H18N6O5S3) Dissolve 15 mg of Cefepime Dihydrochloride Hy-drate in 5 mL of water, and add 2 drops of silver nitrate TS: a white turbidity is produced. MS: Amount [mg (potency)] of Cefditoren Pivoxil RS 1 cm taken Absorbance <2.24> E 1z (259 nm): 310 – 340 (50 mg calcu- lated on the anhydrous basis, water, 1000 mL). Internal standard solution—A solution of propyl parahy- D Optical rotation <2.49> [a]20: ＋39 – ＋479(60 mg calcu-lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 0.1 g of Cefepime Dihydrochloride Hy-drate in 10 mL of water: the pH of this solution is between 1.6 and 2.1.
(1) Clarity and color of solution—Dissolve 0.5 g of Cefepime Dihydrochloride Hydrate in 5 mL of a solution of L-arginine (3 in 50): the solution is clear and has no more color than Matching Fluid H. Heavy metals <1.07>—Proceed with 1.0 g of Cefepime Dihydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). N-Methylpyrrolidine—Weigh accurately an amount of Cefepime Dihydrochloride Hydrate equivalent to about 80 mg (potency), dissolve in diluted nitric acid (2 in 3125) to make exactly 10 mL, and use this solution as the sample so-lution. Separately, put 30 mL of water in a 100-mL volumet-ric flask, weigh accurately the mass of flask, then add about 0.125 g of N-methylpyrrolidine, weigh accurately the mass of the flask again, and add water to make exactly 100 mL. Cefepime Dihydrochloride Hydrate in the mobile phase A to make 50 mL, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the area of each peak by the automatic integration method. Calculate the total amount of the peaks other than cefepime by the area percen-tage method: not more than 0.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 0.57 g of ammonium dihydro-genphosphate in 1000 mL of water. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Pipet 4 mL of this solution, add diluted nitric acid (2 in 3125) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each Time after injection of the sample (min) Mobile phase A (volz) Mobile phase B (volz) of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of N-methylpyrrolidine by the automatic integration method. Calculate the amount of N-methylpyrrolidine per 1 mg (potency) of Cefepime Dihydrochloride Hydrate by the following equation: not more than 0.5z. The sample solu-tion must be tested within 20 minutes after preparation. Amount (z) of N-methylpyrrolidine ＝ (MS × f )/MT × AT/AS × 1/250 MS: Amount (mg) of N-methylpyrrolidine taken MT: Amount [mg (potency)] of Cefepime Dihydrochloride Hydrate taken f: Purity (z) of N-methylpyrrolidine Operating conditions— Detector: An electric conductivity detector. Column: A plastic tube 4.6 mm in inside diameter and 5 cm in length, packed with hydrophilic silica gel for liquid chromatography carrying sulfonic acid groups having the exchange capacity of about 0.3 meq per g (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 990 mL of diluted nitric acid (2 in 3125) add 10 mL of acetonitrile. Flow rate: 1.0 mL per minute. System suitability— System performance: To 20 mL of a solution of sodium chloride (3 in 1000) add 0.125 g of N-methylpyrrolidine, and add water to make 100 mL. To 4 mL of this solution add diluted nitric acid (2 in 3125) to make 100 mL. When the procedure is run with 100 mL of this solution under the above operating conditions, sodium and N-methylpyrroli-dine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 5 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of N-methylpyrrolidine is not more than 4.0z. Related substances—Dissolve about 0.1 g of 0 – 25 100 → 75 0 → 25 Flow rate: Adjust so that the retention time of cefepime is about 9.5 minutes. Time span of measurement: About 2.5 times as long as the retention time of cefepime. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase A to make 10 mL, and use this solution as the solution for system suitability test. To 1 mL of the solution for system suitability test add the mobile phase A to make 10 mL, and use this solution as the solution for test for required detectability. Pipet 1 mL of the solution for test for required detectability, add the mobile phase A to make exactly 10 mL. Conform that the peak area of cefepime obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for test for re-quired detectability. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates of the peak of cefepime is not less than 6000. System repeatability: When the test is repeated 3 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak areas of cefepime is not more than 2.0z. Water <2.48> Not less than 3.0z and not more than 4.5z (Weigh accurately about 50 mg of Cefepime Dihydrochlo-ride Hydrate, dissolve in exactly 2 mL of methanol for water determination and perform the test with exactly 0.5 mL of this solution; coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Bacterial endotoxins <4.01> Less than 0.04 EU/mg (po-tency).
Weigh accurately an amount of Cefepime Dihydro-chloride Hydrate and Cefepime Dihydrochloride RS, equiva-lent to about 60 mg (potency), dissolve in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the JP XVIII Official Monographs / Cefepime Dihydrochloride for Injection 671 ‌test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefepime in each solution. Amount [ mg (potency)] of cefepime (C19H24N6O5S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefepime Dihydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust a solution of sodium 1-pentanesul-fonate (261 in 100,000) to pH 3.4 with acetic acid (100), then adjust this solution to pH 4.0 with a solution of potassium hydroxide (13 in 20). To 950 mL of this solution add 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefepime is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates of the peak of cefepime is not less than 1500. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefepime is not more than 2.0z.
Cefixime Hydrate occurs as a white to light yellow crystalline powder. It is freely soluble in methanol and in dimethylsulfoxide, sparingly soluble in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Cefixime Hydrate in 0.1 mol/L phosphate buffer solution (pH 7.0) (1 in 62,500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefixime RS prepared in the same manner as the sample so-lution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefixime Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefixime RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 50 mg of Cefixime Hydrate in 0.5 mL of a mixture of deuterated dimethylsulfoxide for nuclear mag-netic resonance spectroscopy and heavy water for nuclear magnetic resonance spectroscopy (4:1). Determine the 1H spectrum of this solution, as directed under Nuclear Mag-netic Resonance Spectroscopy <2.21>, using tetramethylsi-lane for nuclear magnetic resonance spectroscopy as an in-ternal reference compound: it exhibits a singlet signal A at around d 4.7 ppm, and a multiplet signal B between d 6.5 ppm and d 7.4 ppm. The ratio of integrated intensity of these signals, A:B, is about 1:1. Optical rotation <2.49> [a]20: －75 – －889(0.45 g calcu-lated on the anhydrous bases, a solution of sodium hydrogen carbonate (1 in 50), 50 mL, 100 mm). D
Dissolve 0.1 g of Cefixime Hydrate in 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), and use this solu-tion as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than cefixime is not more than 1.0z, and the total amount of the peaks other than cefixime is not more than 2.5z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cefixime, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Confirm that the peak area of cefixime obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of cefixime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak areas of cefixime is not more than 2.0z. Water <2.48> Not less than 9.0 and not more than 12.0z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Cefixime Hydrate, equivalent to about 0.1 g (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL. Pipet 10 mL of this solution, add 0.1 mol/L phos-phate buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefixime RS, equivalent to about 20 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 20 mL. Pipet 10 mL of this solution, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefixime in each solution. Amount [mg (potency)] of cefixime (C16H15N5O7S2) ＝ MS × AT/AS × 5000 MS: Amount [mg (potency)] of Cefixime RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌JP XVIII Official Monographs / Cefixime Capsules 673 Column: A stainless steel column 4 mm in inside diameter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 25 mL of a solution of tetrabutylammo-nium hydroxide TS (10 in 13) add water to make 1000 mL, and adjust to pH 6.5 with diluted phosphoric acid (1 in 10). To 300 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefixime is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefixime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of peak area of cefixime is not more than 2.0z.
Cefmenoxime Hydrochloride contains not less than 890 mg (potency) and not more than 975 mg (potency) per mg, calculated on the anhydrous basis.
Cefmenoxime Hydrochloride occurs as white to light orange-yellow, crystals or crystalline powder. It is freely soluble in formamide and in dimethylsulfoxide, slightly soluble in methanol, very slightly soluble in water, and practically insoluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Cefmenoxime Hydrochloride in 0.1 mol/L phos-phate buffer solution (pH 6.8) (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefmenoxime Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cef-menoxime Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefmenoxime Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cef- Amount [mg (potency)] of ceˆxime (C ＝ MS × AT/AS × 5 16H15N5O7S2) menoxime Hydrochloride in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy MS: Amount [mg (potency)] of Ceˆxime RS taken
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Cefmenoxime Hydrochloride in 10 mL of diluted sodium carbonate TS (1 in 4) is clear and colorless to light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Cef- menoxime Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefmenoxime Hydrochloride according to Method 4 and adding 10 mL of dilute hydrochloric acid to the residue after cooling, and perform the test (not more than 2 ppm). Related substances—Weigh accurately about 0.1 g of Cefmenoxime Hydrochloride, dissolve in 20 mL of 0.1 mol/L phosphate buffer solution (pH 6.8) and add the mobile phase to make exactly 100 mL. Pipet 4 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of 1-methyl-1H-tetrazol-5-thiol, and dissolve in the mobile phase to make exactly 100 mL. Pipet 4 mL of this solution, add the mobile phase to make exactly 250 mL, and use this solution as the standard solution (1). Weigh accurately about 0.1 g of Cefmenoxime Hydrochlo-ride RS, dissolve in 20 mL of 0.1 mol/L phosphate buffer solution (pH 6.8) and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 250 mL, and use this solution as the standard solution (2). Perform the test immediately after preparation of these solutions with exactly 10 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following the standard solution (1), add the mobile phase to make exactly 100 mL. Confirm that the peak area of 1-methyl-1H-tetrazol-5-thiol obtained with 10 mL of this solution is equivalent to 4.5 to 5.5z of that with 10 mL of the standard solution (1). Then, measure exactly 2 mL of the standard so-lution (2), add the mobile phase to make exactly 100 mL. Confirm that the peak area of cefmenoxime obtained with 10 mL of this solution is equivalent to 1.5 to 2.5z of that with 10 mL of the standard solution (2). System repeatability: When the test is repeated 6 times with 10 mL of the standard solution (1) under the above operating conditions, the relative standard deviation of the peak area of 1-methyl-1H-tetrazol-5-thiol is not more than 1.0z. Water <2.48> Not more than 1.5z (1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1)).
Weigh accurately an amount of Cefmenoxime Hy-drochloride and Cefmenoxime Hydrochloride RS, equiva-lent to about 50 mg (potency), dissolve each in 10 mL of 0.1 mol/L phosphate buffer solution (pH 6.8) and add the mobile phase to make exactly 50 mL. Pipet 4 mL each of these solutions, add exactly 20 mL of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of cefmenoxime to that of the internal standard. conditions. Determine each peak area of these solutions by the automatic integration method, and calculate the amounts of 1-methyl-1H-tetrazol-5-thiol and the total related sub- Amount [ mg (potency)] of cefmenoxime (C ＝ MS × QT/QS × 1000 16H17N9O5S3) stance by the following formula: the amount of 1-methyl-1H-tetrazol-5-thiol is not more than 1.0z, and the total related substance is not more than 3.0z. Amount (z) of 1-methyl-1H-tetrazol-5-thiol ＝ MSa/MT × ATa/ASa × 20 Amount (z) of total related substances MS: Amount [mg (potency)] of Cefmenoxime Hydrochlo-ride RS taken Internal standard solution—A solution of phthalimide in methanol (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave- ＝ MSa/MT × ATa/ASa × 20 length: 254 nm). Column: A stainless steel column 4 mm in inside diameter ＋ MSb/MT × ST/ASb × 5 MSa: Amount (g) of 1-methyl-1H-tetrazol-5-thiol taken MSb: Amount (g) of Cefmenoxime Hydrochloride RS taken MT: Amount (g) of Cefmenoxime Hydrochloride taken ASa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the standard solution (1) ASb: Peak area of cefmenoxime from the standard solu-tion (2) ATa: Peak area of 1-methyl-1H-tetrazol-5-thiol from the sample solution ST: Total area of the peaks other than 1-methyl-1H-tetrazol-5-thiol and other than cefmenoxime from the sample solution Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of cefmenoxime. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 5 mL of and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (50:10:1). Flow rate: Adjust so that the retention time of cef-menoxime is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefmenoxime and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak areas of cefmenoxime to that of the internal standard is not more than 1.0z.
Cefotetan contains not less than 960 mg (potency) and not more than 1010 mg (potency) per mg, calcu-lated on the anhydrous basis.
Cefotetan occurs as white to light yellow-white powder. It is sparingly soluble in methanol, and slightly soluble in water and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Cefotetan in phosphate buffer solution for anti-biotics, pH 6.5 (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefotetan RS prepared in the same manner as the sample so- than 0.4z. 1-Methyl-1H-tetrazole-5-thiol (z) lution: both spectra exhibit similar intensities of absorption at the same wavelengths. ＝ MSa/MT × QTa/QSa × 1/100 Determine the infrared absorption spectrum of Cefotetan as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefotetan RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Dissolve 50 mg of Cefotetan in 0.5 mL of a solution of sodium hydrogen carbonate in heavy water for nuclear Cefotetan lactone (z) ＝ MSb/MT × QTb/QSb × 1/100 D2-Cefotetan (z) ＝ MSb/MT × QTc/QSb × 1/100 Isothiazole substance (z) ＝ MSb/MT × QTd/QSb × 1/100 Each of other related substances (z) magnetic resonance spectroscopy (1 in 25). Determine the 1H spectrum of this solution as directed under Nuclear Magnetic ＝ MSb/MT × QTe/QSb × 1/100 Resonance Spectroscopy <2.21>, using sodium 3-trimethyl- silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits singlet signals, A, B, C and D, at around d 3.6 ppm, at around d 4.0 ppm, at around d 5.1 ppm and at around d 5.2 ppm, respectively. The ratio of the integrated intensity of each signal, A:B:C:D, is about 3:3:1:1. D Optical rotation <2.49> [a]20: ＋112 – ＋1249(0.5 g calcu-lated on the anhydrous basis, a solution of sodium hydrogen carbonate (1 in 200), 50 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Cefotetan in 10 mL of a solution of sodium hydrogen car-bonate (1 in 30): the solution is clear, and colorless or light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Cefote- tan according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Weigh accurately about 0.1 g of Cefotetan, dissolve in a suitable amount of methanol, add exactly 2 mL of the internal standard solution and methanol to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 3 mg of 1-methyl-1H-tetrazole-5-thiol for liquid chromatography, previously dried in a desiccator (in vacuum, silica gel) for 2 hours, and about 2 mg of Cefotetan RS, calculated on the anhydrous basis, dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solu-tion and methanol to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QTa, QTb, QTc, QTd, QTe and QTf, of the peak areas of 1-methyl-1H-tetrazole-5-thiol, cefotetan lactone having the relative retention time of about 0.5 to cefotetan, D2-cefotetan having the relative retention time of about 1.2, isothiazole substance having the relative retention time of about 1.3, each of other related substances and the total of other related substances, to the peak area of the internal standard, respectively, obtained from the sample solution, and the ratios, QSa and QSb, of the peak areas of 1-methyl-1H-tetrazole-5-thiol and cefotetan, to the peak area of the internal standard, respectively, from the standard so-lution. Calculate the amount of 1-methyl-1H-tetrazole-5-thiol, cefotetan lactone, D2-cefotetan, isothiazole substance, each of other related substances and the total of other related substances from the following equations: the amount of 1-methyl-1H-tetrazole-5-thiol is not more than 0.3z, cefotetan lactone is not more than 0.3z, D2-cefotetan is not more than 0.5z, isothiazole substance is not more than 0.5z, each of other related substances is not more than 0.2z and the total of other related substances is not more Total of other related substances (z) ＝ MSb/MT × QTf/QSb × 1/100 MSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiol taken MSb: Amount (mg) of Cefotetan RS, calculated on the an-hydrous basis taken MT: Amount (g) of Cefotetan taken Internal standard solution—A solution of anhydrous caffeine in methanol (3 in 10,000). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3.5 times as long as the retention time of cefotetan. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 15 mL of the standard solution, and add methanol to make exactly 100 mL. Confirm that the peak area of cefotetan obtained with 5 mL of this solution is equivalent to 12 to 18z of that with 5 mL of the standard solution. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cefotetan to that of the internal standard is not more than 2.0z. Water <2.48> Not more than 2.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 10 mg of Cefotetan in 20 mL of methanol, and use this solution as the sample solution. Per-form the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method. Calculate the amount of the adjacent two peaks appeared at around the retention time of 40 minutes, one having shorter retention time is l-substance and another having longer retention time is d-substance, by the area percentage method: the amount of l-substance is not less than 35z and not more than 45z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.1 mol/L phosphate buffer ‌JP XVIII Official Monographs / Cefotiam Hexetil Hydrochloride 687 solution (pH 7.0), water and a solution of tetrabutylammo-nium hydrogensulfate in acetonitrile (1 in 150) (9:9:2). Flow rate: Adjust so that the retention time of l-substance is about 40 minutes. System suitability— System performance: When the procedure is run with 5 mL of the sample solution under the above operating conditions, l-substance and d-substance are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: To exactly 1 mL of the sample solu-tion add methanol to make exactly 10 mL. When the test is repeated 6 times with 5 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of l-substance is not more than 5.0z.
Weigh accurately an amount of Cefotetan and Cefotetan RS, equivalent to about 50 mg (potency), and dis-solve each in phosphate buffer solution for antibiotics, pH 6.5 to make exactly 50 mL. Pipet 15 mL each of these solu-tions, add exactly 10 mL of the internal standard solution and phosphate buffer solution for antibiotics, pH 6.5 to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefotetan to that of the internal standard. Amount [ mg (potency)] of cefotetan (C17H17N7O8S4) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefotetan RS taken Internal standard solution—A solution of anhydrous caffeine (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 11.53 g of phosphoric acid in 1000 mL of water. To 850 mL of this solution add 50 mL of acetonitrile, 50 mL of acetic acid (100) and 50 mL of metha-nol. Flow rate: Adjust so that the retention time of cefotetan is about 17 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and cefotetan are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 5 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cefotetan to that of the internal standard is not more than 1.0z.
Cefotiam Hexetil Hydrochloride contains not less than 615 mg (potency) and not more than 690 mg (po-tency) per mg, calculated on the anhydrous basis.
Cefotiam Hexetil Hydrochloride occurs as a white to light yellow powder. It is very soluble in water, in methanol and in ethanol (95), freely soluble in dimethylsulfoxide, and slightly soluble in acetonitrile. It dissolves in 0.1 mol/L hydrochloric acid TS. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Cefotiam Hexetil Hydrochloride in 0.1 mol/L hydrochloric acid TS (3 in 125,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefotiam Hexetil Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the 1H spectrum of a solution of Cefotiam Hexetil Hydrochloride in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 20) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits two singlet signals, A and B, at around d 2.8 ppm and at around d 6.6 ppm, and a multiplet signal, C, at around d 6.9 ppm. The ratio of the integrated intensity of each signal, A:B:C, is about 6:1:1. To a solution of Cefotiam Hexetil Hydrochloride (1 in 200) add 2 mL of dilute nitric acid and 1 mL of silver nitrate TS, and mix: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋52 – ＋609(0.1 g calculated on the anhydrous basis, 0.1 mol/L hydrochloric acid TS, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefotiam Hexetil Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Cefotiam Hexetil Hydrochloride according to Method 3, and perform the test, using a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 5) (not more than 1 ppm). Related substance 1—Weigh accurately about 50 mg of Cefotiam Hexetil Hydrochloride, and dissolve in a mix-ture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL. Pipet 10 mL of this solution, add a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make exactly 25 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg of Cefotiam Hexetil Hydrochloride RS, and dissolve in a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL. Pipet 1 mL of this solution, add a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of the related substances by the following equa-tion: the amount of the related substance having the relative retention time of about 1.2 to one of the peaks of cefotiam hexetil, which has the larger retention time, is not more than 2.0z, and each amount of the other related substances is not more than 0.5z. For the peak area, having the relative retention time of about 1.2 to one of the peaks of cefotiam hexetil, which has the larger retention time, multiply the cor-rection factor, 0.78. Amount (z) of related substance ＝ MS/MT × AT/AS × 5 System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the resolution between the two peaks of cefotiam hexetil is not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the total of the two peak areas of cefotiam hexetil is not more than 2.0z. Related substance 2—Weigh accurately about 20 mg of Cefotiam Hexetil Hydrochloride, dissolve in 2 mL of methanol, add a mixture of a solution of diammonium hydrogen phosphate (79 in 20,000) and acetic acid (100) (200:3) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Cefotiam Hydrochloride RS, and dissolve in the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of the related substances by the following equation: the amounts of the related substances having the relative reten-tion time of about 0.1 and about 0.9 are not more than 1.0z, respectively, and each amount of the related sub-stances other than the related substances having the relative retention time of about 0.1 and about 0.9 is not more than 0.5z. For the peak area, having the relative retention time of about 0.9 to cefotiam, multiply the correction factor, MS: Amount (g) of Cefotiam Hexetil Hydrochloride RS taken MT: Amount (g) of Cefotiam Hexetil Hydrochloride taken AS: Total of two peak areas of cefotiam hexetil from the 0.76. Amount (z) of related substance ＝ MS/MT × AT/AS × 4 standard solution AT: Each peak area of related substance from the sample solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of diluted 0.2 mol/L potas-sium dihydrogen phosphate TS (1 in 2), acetonitrile and acetic acid (100) (72:28:1). Mobile phase B: A mixture of acetonitrile, diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in 2) and acetic acid (100) (60:40:1). Flowing of mobile phase: Adjust so that the mixing rate of the mobile phase A and the mobile phase B is changed lineal-ly from 1:0 to 0:1 for 30 minutes. Flow rate: 0.7 mL per minute. Time span of measurement: As long as about 3 times of the retention time of one of the cefotiam hexetil peaks, which appears first, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add a mixture of diluted phos-phoric acid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL. Confirm that each area of the two peaks of cefotiam hexetil obtained with 10 mL of this solution is equivalent to 1.6 to 2.4z of that with 10 mL of the standard solution. MS: Amount (g) of Cefotiam Hydrochloride RS taken MT: Amount (g) of Cefotiam Hexetil Hydrochloride taken AS: Peak area of cefotiam from the standard solution AT: Each peak area from the sample solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of diammonium hydrogen phosphate (79 in 20,000), methanol and acetic acid (100) (200:10:3). Flow rate: Adjust so that the retention time of cefotiam is about 15 minutes. Time span of measurement: As long as about 2 times of the retention time of cefotiam, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of cefotiam obtained with 10 mL of this solution is equivalent to 1.6 to 2.4z of that with 10 mL of the standard solution. System performance: To 1 mL of a solution of acetamino-phen in the mobile phase (1 in 50,000) add 3 mL of the standard solution, and mix well. When the procedure is run with 10 mL of this solution under the above operating condi-tions, acetaminophen and cefotiam are eluted in this order ‌JP XVIII Official Monographs / Cefotiam Hydrochloride 689 with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefotiam is not more than 2.0z. Total amount of related substances—The total of the amount of related substances obtained in the Related sub-stance 1 and the Related substance 2 is not more than 6.5z. Water <2.48> Not more than 3.5z (0.1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Proceed the test with 20 mL of the sample so-lution obtained in the Assay as directed under Liquid Chro-matography <2.01> according to the conditions directed in the Assay, and determine the areas of the two peaks, Aa for the faster peak and Ab for the later peak, closely appeared each other at the retention time of around 10 minutes: Aa/ (Aa ＋ Ab) is not less than 0.45 and not more than 0.55.
Weigh accurately an amount of Cefotiam Hexetil Hydrochloride and Cefotiam Hexetil Hydrochloride RS, equivalent to about 30 mg (potency), and dissolve each in a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make exactly 50 mL. Measure exactly 5 mL each of these solutions, add exactly 5 mL of the internal standard solution and a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefotiam hexetil to that of the internal standard. For this calculation, the total of the areas of the two peaks appeared closely each other at the retention time of around 10 minutes is used as the peak area of cefotiam hexetil. ing conditions, the relative standard deviation of the ratios of the peak area of cefotiam hexetil to that of the internal standard is not more than 1.0z.
Cefotiam Hydrochloride contains not less than 810 mg (potency) and not more than 890 mg (potency) per mg, calculated on the anhydrous basis.
Cefotiam Hydrochloride occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in water, in methanol and in for-mamide, slightly soluble in ethanol (95), and practically in-soluble in acetonitrile. Amount [ mg (potency)] of cefotiam (C ＝ MS × QT/QS × 1000 18H23N9O4S3)
(1) Determine the absorption spectrum of a solution of Cefotiam Hydrochloride (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and MS: Amount [mg (potency)] of Cefotiam Hexetil Hydro-chloride RS taken Internal standard solution—A solution of benzoic acid in a mixture of diluted phosphoric acid (1 in 100) and acetonitrile (4:1) (7 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in 2), acetonitrile and acetic acid (100) (72:28:1). Flow rate: Adjust so that the retention time of the faster peak of cefotiam hexetil is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and cefotiam hexetil are eluted in this order with the resolution between the two peaks of cefotiam hexetil being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefotiam Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefotiam Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefotiam Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefotiam Hydrochloride in heavy water for nuclear magnetic reso-nance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropanesulfonate for nuclear magnetic reso-nance spectroscopy as an internal reference compound: it exhibits singlet signals, A and B, at around d 3.1 ppm and at around d 6.7 ppm, respectively. The ratio of integrated in-tensity of each signal, A:B, is about 6:1. Dissolve 0.1 g of Cefotiam Hydrochloride in 5 mL of dilute nitric acid, and immediately add 1 mL of silver nitrate TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋60 – ＋729(1 g calculated on the anhydrous bases, water, 100 mL, 100 mm). ‌690 pH <2.54> Dissolve 1.0 g of Cefotiam Hydrochloride in 10 mL of water: the pH of the solution is between 1.2 and 1.7.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefotiam Hydrochloride in 10 mL of water: the solution is clear, and colorless to yellow. Heavy metals <1.07>—To 1.0 g of Cefotiam Hydro- chloride add 1 mL of sulfuric acid, and heat gently to car-bonize. After cooling, add 10 mL of a solution of magne-sium nitrate hexahydrate in ethanol (95) (1 in 10), fire the ethanol to burn, then heat gradually to incinerate. If a car-bonized residue still retains, moisten the residue with a little amount of sulfuric acid, and ignite again to incinerate. After cooling, add 2 mL of hydrochloric acid to the residue, heat on a water bath to dissolve, then heat to dryness. Add 10 mL of water, and heat to dissolve. After cooling, add ammonia TS dropwise to adjust to pH 3 – 4, if necessary, filter, wash the residue on the filter with 10 mL of water, transfer the fil-trate and washings into a Nessler tube, add water to make 50 mL, and use this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution in the same manner as for preparation of the test solution (not more than 20 ppm). Arsenic <1.11>—Incinerate 1.0 g of Cefotiam Hydro- chloride according to Method 4. After cooling, add 10 mL of dilute hydrochloric acid to the residue, heat to dissolve on the water bath, and use this solution as the test solution. Per-form the test (not more than 2 ppm). Water <2.48> Not more than 7.0z (0.25 g, volumetric titration, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination).
Weigh accurately an amount of Cefotiam Hydro-chloride and Cefotiam Hydrochloride RS, equivalent to about 0.1 g (potency), and dissolve each in the mobile phase to make exactly 100 mL, and use these solutions as the sam-ple solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefotiam in each solution. Amount [ mg (potency)] of cefotiam (C18H23N9O4S3) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefotiam Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: To 800 mL of 0.05 mol/L disodium hydrogenphosphate TS add 0.05 mol/L potassium dihydro-genphosphate TS to adjust the pH to 7.7. To 440 mL of this solution add 60 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefotiam is about 14 minutes. System suitability— System performance: Dissolve 0.04 g of orcine in 10 mL of the standard solution. When the procedure is run with 10 mL of the standard solution under the above operating condi-tions, orcine and cefotiam are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefotiam is not more than 1.0z.
Cefozopran Hydrochloride contains not less than 860 mg (potency) and not more than 960 mg (potency) per mg, calculated on the anhydrous basis.
Cefozopran Hydrochloride occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in dimethylsulfoxide and in formamide, slightly soluble in water, in methanol and in ethanol (95), and practically insoluble in acetonitrile and diethyleter.
(1) Dissolve 0.02 g of Cefozopran Hydro-chloride in 10 mL of water, add 1 mL of a solution of hy-droxylammonium chloride (1 in 10) and 2 mL of sodium hy-droxide TS, allow to stand for 5 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS and 3 drops of iron (III) chlo-ride TS, and mix: a red-purple color develops. Determine the absorption spectra of solutions of Cefozopran Hydrochloride and Cefozopran Hydrochloride RS in a mixture of sodium chloride TS and methanol (3:2) (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare these spectra: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the 1H spectrum of a solution of Cefozopran Hydrochloride in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 20) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic reso- nance spectroscopy as an internal reference compound: it exhibits a singlet signal A at around d 3.9 ppm, a doublet signal B at around d 5.2 ppm, and a quartet signal C at around d 8.0 ppm, and the ratio of integrated intensity of each signal, A:B:C, is about 3:1:1. Dissolve 0.01 g of Cefozopran Hydrochloride in 1 mL of water and 2 mL of acetic acid (100), add 2 drops of silver nitrate TS, and mix: a white turbidity is formed. 1 cm Absorbance <2.24> E 1z (238 nm): 455 – 485 (50 mg calcu-lated on the anhydrous basis, a mixture of sodium chloride TS and methanol (3:2), 5000 mL). D Optical rotation <2.49> [a]20: －73 – －789(0.1 g calculated on the anhydrous basis, a mixture of sodium chloride TS and methanol (3:2), 10 mL, 100 mm).
(1) Clarity and color of solution—Being specified separately when the drug is granted approval based on the Law. Heavy metals <1.07>—Proceed with 2.0 g of Cefozopran Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic—Being specified separately when the drug is granted approval based on the Law. Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Residue on ignition Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po-tency).
Weigh accurately an amount of Cefozopran Hydro-chloride and Cefozopran Hydrochloride RS, equivalent to about 50 mg (potency), and dissolve each in the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution and the mobile phase to make 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefozopran to that of the internal standard. Amount [ mg (potency)] of cefozopran (C19H17N9O5S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefozopran Hydrochlo-ride RS taken Internal standard solution—A solution of 2,4-dihydroxyben-zoic acid in the mobile phase (1 in 1250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Mix 0.366 g of diethylamine with water to make 1000 mL, and add 60 mL of acetonitrile and 5 mL of acetic acid (100). Flow rate: Adjust so that the retention time of cefozopran ‌692‌ is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefozopran and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefozopran to that of the internal stand-ard is not more than 1.0z.
Cefpirome Sulfate contains not less than 760 mg (potency) per mg, calculated on the anhydrous basis.
Cefpirome Sulfate occurs as a white to pale yellow-white crystalline powder, and has a slight, character-istic odor. It is soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic.
(1) Dissolve 10 mg of Cefpirome Sulfate in 2 mL of water, add 3 mL of hydroxylammonium hydrochlo-ride-ethanol TS, allow to stand for 5 minutes, add 1 mL of acidic ammonium iron (III) sulfate TS, and shake: a red-brown color develops. Dissolve 1 mg of Cefpirome Sulfate in 4 mL of water, add 1 mL of dilute hydrochloric acid while cooling in ice, add 1 mL of a freshly prepared solution of sodium nitrite (1 in 100), and allow to stand for 2 minutes. Add 1 mL of am-monium amidosulfuric acid TS while cooling in ice bath, allow to stand for 1 minute, and add 1 mL of a solution of lated on the anhydrous basis, 0.01 mol/L hydrochloric acid TS, 2500 mL). D Optical rotation <2.49> [a]20: －27 – －339(50 mg calcu-lated on the anhydrous basis, a solution prepared by addi-tion of water to 25 mL of acetonitrile to make 50 mL, 20 mL, 100 mm). pH <2.54> Dissolve 0.1 g of Cefpirome Sulfate in 10 mL of water: the pH of the solution is between 1.6 and 2.6.
(1) Clarity and color of solution—Being specified separately when the drug is granted approval based on the Law. Heavy metals <1.07>—Proceed with 1.0 g of Cefpi- rome Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic—Being specified separately when the drug is granted approval based on the Law. Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency).
Weigh accurately an amount of Cefpirome Sulfate and Cefpirome Sulfate RS, equivalent to about 50 mg (po-tency), dissolve each in water to make exactly 100 mL. Pipet 5 mL of these solutions, add each in water to make exactly 20 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefpirome in each solution. N-1-naphthylethylene dihydrochloride (1 in 1000): a purple color develops. Take 5 mg of Cefpirome Sulfate, dissolve in 1 mL of Amount [ mg (potency)] of cefpirome (C ＝ MS × AT/AS × 1000 22H22N6O5S2) ethanol (95) and 1 mL of water, add 100 mg of 1-chloro-2,4-dinitrobenzene, and heat on a water bath for 5 minutes. After cooling, add 2 or 3 drops of a solution of sodium hy-droxide (1 in 10) and 3 mL of ethanol (95): a red-brown color develops. MS: Amount [mg (potency)] of Cefpirome Sulfate RS taken ‌JP XVIII Official Monographs / Cefpodoxime Proxetil 695 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.45 g of ammonium dihydrogen-phosphate in 1000 mL of water, and adjust the pH to 3.3 with phosphoric acid. To 800 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefpirome is about 7.5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates of the peak of cefpi-rome is not less than 3600. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefpirome is not more than 1.0z.
Containers—Hermetic containers. Storage—At a temperature between 29C and 89C.
Cefpodoxime Proxetil contains not less than 706 mg (potency) and not more than 774 mg (potency) per mg, calculated on the anhydrous basis.
Cefpodoxime Proxetil occurs as a white to light brown-white powder. It is very soluble in acetonitrile, in methanol and in chlo-roform, freely soluble in ethanol (99.5), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Cefpodoxime Proxetil in acetonitrile (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Cefpodoxime Proxetil RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cef- podoxime Proxetil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefpodoxime Proxetil RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cef-podoxime Proxetil in deuterochloroform for nuclear mag-netic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spectros-copy as an internal reference compound: it exhibits doublet signals, A and B, at around d 1.3 ppm and at around d 1.6 ppm, and singlet signals, C and D, at around d 3.3 ppm and at around d 4.0 ppm. The ratio of the integrated intensity of these signals, A:B:C:D, is about 2:1:1:1. D Optical rotation <2.49> [a]20: ＋24.0 – ＋31.49(0.1 g calcu-lated on the anhydrous basis, acetonitrile, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cefpodoxime Proxetil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Cefpodoxime Proxetil in 50 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), and use this solution as the sam-ple solution. Perform the test with 20 mL of the sample solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the amount of the peak, having the relative retention time of about 0.8 to the isomer B of cefpodoxime proxetil, is not more than 2.0z, the amount of the peak other than cefpodoxime proxetil is not more than 1.0z, and the total amount of the peaks other than cefpodoxime proxetil is not more than 6.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 229C. Mobile phase A: A mixture of water, methanol and a solution of formic acid (1 in 50) (11:8:1). Mobile phase B: A mixture of methanol and a solution of formic acid (1 in 50) (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) 0 – 65 95 5 65 – 145 95 → 15 5 → 85 145 – 155 15 85 Flow rate: 0.7 mL per minute (the retention time of the isomer B of cefpodoxime proxetil is about 60 minutes). Time span of measurement: For 155 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 5 mL of the sample so-lution add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 200 mL, and use this solution as the ‌solution for system suitability test. Pipet 2 mL of the solu-tion for system suitability test, and add the mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make exactly 100 mL. Confirm that the peak areas of the isomer A and the isomer B of cefpodoxime proxetil obtained with 20 mL of this solution are equivalent to 1.4 to 2.6z of them with 20 mL of the solution for system suitability test, respectively. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the isomer A and the isomer B of cef-podoxime proxetil are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 5 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of the isomer A and the isomer B of cefpodoxime proxetil is not more than 2.0z. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g). Isomer ratio Perform the test with 5 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas of the two isomers of cefpodoxime proxetil, Aa, for the isomer having the smaller retention time, and Ab, for the isomer having the larger retention time, by the automatic integration method: Ab/ (Aa ＋ Ab) is between 0.50 and 0.60. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol (11:9). Flow rate: Adjust so that the retention time of the internal standard is about 11 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard, the isomer A and the isomer B are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 5 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of the isomer B of cefpodoxime proxetil to that of the internal standard is not more than 1.0z.
Cefroxadine Hydrate contains not less than 930 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Cefroxadine Hydrate occurs as pale yellow-white to light yellow, crystalline particles or powder. It is very soluble in formic acid, slightly soluble in water and in methanol, and very slightly soluble in acetonitrile and in ethanol (95). It dissolves in 0.001 mol/L hydrochloric acid TS and in dilute acetic acid.
(1) Determine the absorption spectrum of a solution of Cefroxadine Hydrate in 0.001 mol/L hydrochlo-ric acid TS (1 in 50,000) as directed under Ultraviolet-visible JP XVIII Official Monographs / Cefroxadine Hydrate 699 Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefroxadine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Cefroxadine Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefroxadine RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefroxa-dine Hydrate in deuterated formic acid for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethyl-silane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits three sharp singlet signals, A, B and C, at around d 2.8 ppm, at around d 4.1 ppm and at around d 6.3 ppm. The ratio of the integrated intensity of each signal, A:B:C, is about 4:3:1. D Optical rotation <2.49> [a]20: ＋95 – ＋1089(0.1 g calcu-lated on the anhydrous basis, diluted acetic acid (100) (3 in 25), 100 mL, 100 mm).
(1) Heavy metals <1.07>—Weigh 1.0 g of Cefroxa-dine Hydrate in a porcelain crucible, add 10 mL of a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), mix, burn the ethanol, and carbonize by gently heating. After cooling, add 2 mL of nitric acid, heat carefully, and incinerate by ignition at 500 – 6009C. If a carbonized sub-stance still remains, moisten it with a small amount of nitric acid, and incinerate again by ignition. After cooling, add 6 mL of hydrochloric acid, and evaporate on a water bath to dryness. Moisten the residue with 3 drops of hydrochloric acid, and add 10 mL of hot water to dissolve the residue by heating on a water bath. After cooling, adjust the pH be-tween 3 and 4 with ammonia TS, add 2 mL of dilute acetic acid, filter if necessary, transfer to a Nessler tube, wash the crucible with 10 mL of water, and add the washing and water to the tube to make 50 mL. Perform the test with this solution. Prepare the control solution as follows: Put 2.0 mL of Standard Lead Solution and 10 mL of a solution of mag- Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.4 g of sodium perchlorate in 1000 mL of a mixture of water and acetonitrile (489:11). Flow rate: Adjust so that the retention time of cefroxadine is about 20 minutes. Time span of measurement: About 2 times as long as the retention time of cefroxadine. System suitability— Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mobile phase to make ex-actly 20 mL. Confirm that the peak area of cefroxadine ob-tained with 40 mL of this solution is equivalent to 7 to 13z of that with 40 mL of the standard solution. System performance: Dissolve 3 mg of Cefroxadine Hy-drate and 15 mg of orcin in 100 mL of the mobile phase. When the procedure is run with 40 mL of this solution under the above operating conditions, orcin and cefroxadine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefroxadine is not more than 2.0z. Water <2.48> Not less than 8.5z and not more than 12.0z (0.1 g, volumetric titration, direct titration).
Weigh accurately an amount of Cefroxadine Hy-drate and Cefroxadine RS, equivalent to about 50 mg (po-tency), dissolve each in a suitable amount of a mixture of dilute acetic acid and phosphoric acid (500:1), add exactly 5 mL of the internal standard solution and a mixture of dilute acetic acid and phosphoric acid (500:1) to make 200 mL, and use these solutions as the sample solution and standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of cefroxadine to that of the internal standard. nesium nitrate hexahydrate in ethanol (95) (1 in 10) in a por-celain crucible, and proceed as directed for the preparation of the test solution (not more than 20 ppm). Amount [ mg (potency)] of cefroxadine (C ＝ MS × QT/QS × 1000 16H19N3O5S) (2) Related substances—Dissolve 10 mg of Cefroxadine Hydrate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 40 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the areas of the peaks, having the relative retention times of about 0.07, about 0.6 and about 0.8 to cefroxadine obtained from the sample solution are not larger than 2 times, 4 times and 1 time the peak area of cefroxadine from the standard solu-tion, respectively, and any peak area other than cefroxadine and other than the peaks mentioned above is not larger than 1/2 times the peak area of cefroxadine from the standard so-lution, and the total area of the peaks other than cefroxadine is not larger than 6 times the peak area of cefroxadine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). MS: Amount [mg (potency)] of Cefroxadine RS taken Internal standard solution—Dissolve 1.6 g of vanillin in 5 mL of methanol, and add a mixture of dilute acetic acid and phosphoric acid (500:1) to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 50) and acetonitrile (97:3). Flow rate: Adjust so that the retention time of cefroxadine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefroxadine and the internal standard are eluted in this order with the resolution between these peaks being not ‌700 less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak areas of cefroxadine to that of the internal stand-ard is not more than 1.0z.
Ceftazidime Hydrate contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Ceftazidime Hydrate occurs as a white to light yellow-white crystalline powder. It is slightly soluble in water, and very slightly soluble in acetonitrile and in ethanol (95).
(1) Determine the absorption spectrum of a solution of Ceftazidime Hydrate in phosphate buffer solu-tion (pH 6.0) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ceftazidime RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Ceftazidime Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Ceftazidime RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. To 0.05 g of Ceftazidime Hydrate add 5 mg of dried sodium carbonate, and add 0.5 mL of heavy water for nuclear magnetic resonance spectroscopy to dissolve. Deter-mine the 1H spectrum of this solution as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using so-dium 3-trimethylsilylpropanesulfonate for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits singlet signals, A and B, at around d 1.5 ppm and at around d 6.9 ppm, and a multiplet signal C between d 7.9 ppm and d 9.2 ppm. The ratio of integrated intensity of each signal, A:B:C, is about 6:1:5. D Optical rotation <2.49> [a]20: －28 – －349(0.5 g calculated on the anhydrous basis, phosphate buffer solution (pH 6.0), 100 mL, 100 mm). pH <2.54> Dissolve 0.5 g of Ceftazidime Hydrate in 100 mL of water: the pH of the solution is between 3.0 and 4.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Ceftazidime Hydrate in 10 mL of a solution obtained by dissolving 5 g of anhydrous disodium hydrogen phosphate and 1 g of potassium dihydrogen phosphate in water to make 100 mL: the solution is clear, and its absorbance at 420 nm, determined as directed under Ultraviolet-visible Spectropho-tometry <2.24>, is not more than 0.20. JP XVIII Official Monographs / Ceftazidime Hydrate 703 Heavy metals <1.07>—Proceed with 1.0 g of Ceftazi-dime Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances (i) Trityl-t-butyl substance and t-butyl substance—Dissolve 0.10 g of Ceftazidime Hydrate in 2 mL of diluted disodium hydrogen phosphate TS (1 in 3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted disodium hydrogen-phosphate TS (1 in 3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop with a mixture of acetic acid (100), n-butyl acetate, acetate buffer solution (pH 4.5) and 1-butanol (16:16:13:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots which appear upper in posi-tion than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. (ii) Other related substances—Dissolve 20 mg of Ceftazi-dime Hydrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ceftazidime obtained from the sample solution is not larger than that of ceftazidime from the standard solution, and the total of peak areas other than ceftazidime from the sample solution is not larger than 5 times the peak area of ceftazidime from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.0 g of ammonium dihydrogen-phosphate in 750 mL of water, adjust to pH 3.5 with phos-phoric acid, and add water to make 870 mL. To this solution add 130 mL of acetonitrile. Flow rate: Adjust so that the retention time of ceftazidime is about 4 minutes. Time span of measurement: About 3 times as long as the retention time of ceftazidime, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add the mobile phase to make exactly 5 mL, and confirm that the peak area of ceftazidime obtained with 5 mL of this solution is equivalent to 15 to 25z of that with 5 mL of the standard solution. System performance: Dissolve about 10 mg each of Ceftazidime Hydrate and acetanilide in 20 mL of the mobile phase. When the procedure is run with 5 mL of this solution under the above operating conditions, ceftazidime and acetanilide are eluted in this order with the resolution be-tween these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ceftazidime is not more than 2.0z. Free pyridine—Weigh accurately about 50 mg of Ceftazidime Hydrate, dissolve in the mobile phase to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of pyridine, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak heights, HT and HS, of pyridine in each solution: the amount of free pyridine is not more than 0.3z. Amount (mg) of free pyridine ＝ MS × HT/HS × 1/1000 MS: Amount (mg) of pyridine taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.88 g of ammonium dihydrogen-phosphate in 500 mL of water, add 300 mL of acetonitrile and water to make 1000 mL, and adjust to pH 7.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of pyridine is about 4 minutes. System suitability— System performance: Dissolve 5 mg of Ceftazidime Hy-drate in 100 mL of a solution of pyridine in the mobile phase (1 in 20,000). When the procedure is run with 10 mL of this solution under the above operating conditions, ceftazidime and pyridine are eluted in this order with the resolution be-tween these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak height of pyridine is not more than 5.0z. Water <2.48> 13.0 – 15.0z (0.1 g, volumetric titration, direct titration).
Weigh accurately an amount of Ceftazidime Hy-drate, equivalent to about 0.1 g (potency), and dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to make ex-actly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, then add 0.05 mol/L phos-phate buffer solution (pH 7.0) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Ceftazidime RS, equivalent to about 20 mg (potency), dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 20 mL. Pipet 10 mL of this solu-tion, add exactly 5 mL of the internal standard solution, then add 0.05 mol/L phosphate buffer solution (pH 7.0) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ceftazidime to that of the internal standard. ‌704‌ Amount [mg (potency)] of ceftazidime (C22H22N6O7S2) ＝ MS × QT/QS × 5000 MS: Amount [mg (potency)] of Ceftazidime RS taken Internal standard solution—A solution of dimedon in 0.05 mol/L phosphate buffer solution (pH 7.0) (11 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with hexasilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.26 g of anhydrous disodium hydrogen phosphate and 2.72 g of potassium dihydrogen phosphate in 980 mL of water, and add 20 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of ceftazidime is about 4 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and ceftazidime are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of ceftazidime to that of the internal standard is not more than 1.0z.
Cefteram Pivoxil contains not less than 743 mg (po-tency) and not more than 824 mg (potency) per mg, JP XVIII Official Monographs / Cefteram Pivoxil 705 calculated on the anhydrous basis.
Cefteram Pivoxil occurs as a white to pale yellow-white powder. It is very soluble in acetonitrile, freely soluble in metha-nol, in ethanol (95) and in chloroform, and practically in-soluble in water.
(1) Determine the absorption spectrum of a solution of Cefteram Pivoxil in 0.05 mol/L hydrochloric acid-methanol TS (1 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefteram Pivoxil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefteram Pivoxil in deuterated chloroform for nuclear magnetic reso-nance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethy-lsilane for nuclear magnetic resonance spectroscopy as an mL. Confirm that the peak area of cefteram pivoxil obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefteram pivoxil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefteram pivoxil is not more than 3.0z. Water <2.48> Not more than 3.0z (0.3 g, coulometric titration).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cefteram Pivoxil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Cefteram Pivoxil in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the each area of the peaks, having the relative retention time of about 0.2 and about 0.9 to cefteram pivoxil, obtained from the sample so-lution is not larger than 1/2 times and 1.25 times the peak area of cefteram pivoxil from the standard solution, respec-tively, the area of the peak other than cefteram pivoxil and the peaks mentioned above is not larger than 1/4 times the peak area of cefteram pivoxil from the standard solution, and the total area of the peaks other than cefteram pivoxil is not larger than 2.75 times the peak area of cefteram pivoxil from the standard solution. For the area of the peak, having the relative retention time of about 0.1 to cefteram pivoxil, multiply the correction factor, 0.74. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of cefteram pivoxil. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 10 MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 100 mL of acetic acid-sodium acetate buffer solution (pH 5.0) add 375 mL of acetonitrile and water to make 1000 mL. Flow rate: Adjust so that the retention time of cefteram pivoxil is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefteram pivoxil are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefteram pivoxil to that of the internal standard is not more than 1.0z.
Weigh accurately an amount of Cefteram Pivoxil and Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to about 40 mg (potency), dissolve each in 20 mL of diluted acetonitrile (1 in 2), add exactly 5 mL of the internal stand-ard solution and diluted acetonitrile (1 in 2) to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefteram pivoxil to that of the internal standard. internal reference compound: it exhibits singlet signals A, B Amount [ mg (potency)] of cefteram (C H N O S ) and C, at around d 1.2 ppm, at around d 2.5 ppm and at around d 4.0 ppm, respectively. The ratio of the integrated ＝ MS × QT/QS × 1000 16 17 9 5 2 intensity of these signals, A:B:C, is about 3:1:1. D Optical rotation <2.49> [a]20: ＋35 – ＋439(0.4 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Cefteram Pivoxil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Cefteram Pivoxil in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the each area of the peaks, having the relative retention time of about 0.2 and about 0.9 to cefteram pivoxil, obtained from the sample so-lution is not larger than 1/2 times and 1.25 times the peak area of cefteram pivoxil from the standard solution, respec-tively, the area of the peak other than cefteram pivoxil and the peaks mentioned above is not larger than 1/4 times the peak area of cefteram pivoxil from the standard solution, and the total area of the peaks other than cefteram pivoxil is not larger than 2.75 times the peak area of cefteram pivoxil from the standard solution. For the area of the peak, having the relative retention time of about 0.1 to cefteram pivoxil, multiply the correction factor, 0.74. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of cefteram pivoxil. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 10 MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 100 mL of acetic acid-sodium acetate buffer solution (pH 5.0) add 375 mL of acetonitrile and water to make 1000 mL. Flow rate: Adjust so that the retention time of cefteram pivoxil is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefteram pivoxil are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefteram pivoxil to that of the internal standard is not more than 1.0z.
Ceftibuten Hydrate contains not less than 900 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Ceftibuten Hydrate occurs as a white to pale yellow-white crystalline powder. It is freely soluble in N,N-dimethylformamide and in dimethyl sulfoxide, and practically insoluble in water, in ethanol (95) and in diethyl ether.
(1) Determine the absorption spectrum of a solution of Ceftibuten Hydrate in 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ceftibuten Hydrate as directed in the paste method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Ceftibut-en Hydrate in deuterated dimethyl sulfoxide for nuclear magnetic resonance spectroscopy (1 in 30), using tetra-methylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits doublet signals A and B, at around d 3.2 ppm and at around d 5.1 ppm, a quartet signal C, at around d 5.8 ppm, and a singlet signal D, at around d 6.3 ppm. The ratio of integrated inten-sity of each signal except the signal at around d 3.2 ppm, B:C:D is about 1:1:1. D Optical rotation <2.49> [a]20: ＋135 – ＋1559(0.3 g calcu-lated on the anhydrous basis, 0.1 mol/L phosphate buffer solution for antibiotics, pH 8.0, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ceftibuten Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—(i) Keep the sample solution and the standard solution at not exceeding 59C and use within 2 hours after preparation. Dissolve 25 mg of Cefti-buten Hydrate in 20 mL of 0.1 mol/L phosphate buffer solu-tion for antibiotics (pH 8.0). To 4 mL of this solution add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make 20 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ceftibuten obtained from the sample solution is not larger than 1/5 times the peak area of ceftibuten from the standard solution, and the total area of the peaks other than ceftibuten from the sample solution is not larger than the peak area of ceftibuten from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.7 times as long as the retention time of ceftibuten, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make exactly 20 mL. Confirm that the peak area of ceftibuten obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard so-lution. System performance: Dissolve 5 mg of Ceftibuten Hydrate in 20 mL of 0.1 mol/L hydrochloric acid TS, and allow to stand at 409C for 1 hour. To 4 mL of this solution add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make 25 mL. When the procedure is run with 5 mL of this so-lution under the above operating conditions, trans-isomer of ceftibuten and ceftibuten are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 5 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ceftibuten is not more than 2.0z. (ii) Keep the sample solution at not exceeding 59C, and use within 24 hours after preparation. To 5 mg of Ceftibuten Hydrate add 20 mL of the mobile phase, sonicate, if neces-sary, then shake to dissolve, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Perform the test with 10 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late their amounts by the area percentage method: the total amount of the peaks that are eluted faster than ceftibuten is not more than 5.0z. For the areas of these peaks, multiply the correction factor, 1.63, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with glycol etherified silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.05 g of disodium hydrogen phos-phate dodecahydrate and 0.58 g of potassium dihydrogen phosphate in water to make 1000 mL. Flow rate: Adjust so that the retention time of ceftibuten is about 20 minutes. Time span of measurement: About 1.6 times as long as the retention time of ceftibuten. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 20 mL, and use this so-lution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of ceftibuten obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of ceftibuten are not less than 10,000 and 0.8 – 1.2, respectively. System repeatability: When the test is repeated 5 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ceftibuten is not more than 1.7z. Water <2.48> Not less than 8.0z and not more than 13.0z (0.2 g, volumetric titration, direct titration. Use a mixture of pyridine for water determination and ethylene glycol for water determination (5:1) instead of methanol for water de-termination). Residue on ignition <2.44> Not more than 0.1z (1 g).
Keep the sample solution and the standard solution at not exceeding 59C and use within 2 hours after prepara-tion. Weigh accurately an amount of Ceftibuten Hydrate and Ceftibuten Hydrochloride RS, equivalent to about 10 mg (potency), dissolve each in about 36 mL of 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0), add ex-actly 4 mL each of the internal standard solution, shake, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of ceftibuten to that of the internal standard. JP XVIII Official Monographs / Ceftizoxime Sodium 709 Amount [mg (potency)] of ceftibuten (C15H14N4O6S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Ceftibuten Hydrochloride RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile (3 in 4000). Operating conditions— ‌Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A stainless steel column 4 mm in inside diameter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.005 mol/L n-decyl trimethy-lammonium bromide TS and acetonitrile (4:1). Flow rate: Adjust so that the retention time of ceftibuten is about 10 minutes. System suitability— System performance: Dissolve 5 mg of Ceftibuten Hydrate in 20 mL of 0.1 mol/L hydrochloric acid TS, and allow to stand at 409C for 1 hour. To 4 mL of this solution add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make 25 mL. When the procedure is run with 5 mL of this so-lution under the above operating conditions, trans-isomer of ceftibuten and ceftibuten are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ceftibuten to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage—Light-resistant, and not exceeding 59C.
Cefuroxime Axetil contains not less than 800 mg (potency) and not more than 850 mg (potency) per mg, calculated on the anhydrous and acetone-free basis.
Cefuroxime Axetil occurs as white to yellow-white non-crystalline powder. It is freely soluble in dimethylsulfoxide, soluble in metha-nol, sparingly soluble in ethanol (95), and very slightly solu-ble in water.
(1) Determine the absorption spectrum of a solution of Cefuroxime Axetil in methanol (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefuroxime Axetil RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Cefuroxime Axetil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefuroxime Axetil RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefuroxime Axetil in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 20) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a doublet signal or a pair of doublet signals A at around d 1.5 ppm, a pair of singlet signals B at around d 2.1 ppm, and a singlet signal C at around d 3.9 ppm. The ratio of the inte-grated intensity of each signal, A:B:C, is about 1:1:1. D Optical rotation <2.49> [a]20: ＋41 – ＋479(0.5 g, metha-nol, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefuroxime Axetil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 25 mg of Cefuroxime Axetil in 4 mL of methanol, add a solution of ammonium dihydrogenphosphate (23 in 1000) to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add 40 mL of methanol and a solution of am-monium dihydrogenphosphate (23 in 1000) to make exactly JP XVIII Official Monographs / Cefuroxime Axetil 713 100 mL, and use this solution as the standard solution. Per-form the test with exactly 2 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the area of the peak other than cefuroxime axetil obtained from the sample solution is not larger than 1.5 times the total area of the two peaks of cefuroxime axetil from the standard solution, and the total area of the peaks other than cefuroxime axetil from the sample solution is not larger than 4 times the total area of the two peaks of cefuroxime axetil from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of the peak having the larger retention time of the two peaks of cefuroxime axetil, beginning after the sol-vent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 4 mL of methanol and a solution of am-monium dihydrogenphosphate (23 in 1000) to make exactly 10 mL. Confirm that the total area of the two peaks of cefuroxime axetil obtained with 2 mL of this solution is equivalent to 7 to 13z of that with 2 mL of the standard so-lution. System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the resolution between the two peaks of cefuroxime axetil is not less than 1.5. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the total area of the two peaks of cefuroxime axetil is not more than 2.0z. Acetone—Weigh accurately about 1 g of Cefuroxime Axetil, add exactly 0.2 mL of the internal standard solution and dimethylsulfoxide to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.5 g of acetone, and add dimethylsulfoxide to make exactly 100 mL. Pipet 0.2 mL of this solution, add exactly 0.2 mL of the internal standard solution and dimethylsulfoxide to make 10 mL, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of acetone to that of Temperature of injection port: A constant temperature of about 1159C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of the internal standard is about 4 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, acetone and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of acetone to that of the internal standard is not more than 5.0z. Water <2.48> Not more than 2.0z (0.4 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (0.5 g). Isomer ratio Perform the test with 10 mL of the sample so-lution obtained in the Assay as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the area, Aa, of the peak having the smaller retention time and the area, Ab, of the peak having the big-ger retention time of the two peaks of cefuroxime axetil: Ab/(Aa ＋ Ab) is between 0.48 and 0.55. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay.
Weigh accurately an amount of Cefuroxime Axetil and Cefuroxime Axetil RS, equivalent to about 50 mg (po-tency), and dissolve each in methanol to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 5 mL of the internal standard solution, 5 mL of methanol and a solution of ammonium dihydrogen phosphate (23 in 1000) to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the sum area of the two peaks of cefuroxime axetil to the peak area of the internal standard. the internal standard: the amount of acetone is not more Amount [ mg (potency)] of cefuroxime (C H N O S) than 1.3z. ＝ MS × QT/QS × 1000 16 16 4 8 Amount (z) of acetone ＝ MS/MT × QT/QS × 1/5 MS: Amount (g) of acetone taken MT: Amount (g) of Cefuroxime Axetil taken Internal standard solution—A solution of 1-propanol in dimethylsulfoxide (1 in 200). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 2 m in length, packed with siliceous earth for gas chromatogra-phy coated with a mixture of polyethylene glycol 600 for gas chromatography and polyethylene glycol 1500 for gas chro-matography (1:1) in the ratio of 20z (125 – 150 mm in parti-cle diameter). Column temperature: A constant temperature of about 909C. MS: Amount [mg (potency)] of Cefuroxime Axetil RS taken Internal standard solution—A solution of acetanilide in methanol (27 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with trimethylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of ammonium di-hydrogen phosphate (23 in 1000) and methanol (5:3). Flow rate: Adjust so that the retention time of the peak ‌714 having the smaller retention time of the two peaks of cefuroxime axetil is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefuroxime axetil are eluted in this order with the resolution between the two peaks of cefuroxime axetil being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the sum area of the two peaks of cefuroxime axetil to the peak area of the internal standard is not more than 1.0z.
Celecoxib occurs as a white, powder or crystal-line powder. It is freely soluble in methanol, soluble in ethanol (99.5), and practically insoluble in water. Melting point: 161 – 1649C Celecoxib shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Celecoxib in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and Pipet 1 mL of this solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area, AT, in the sample solution and the peak area of celecoxib, AS, in the standard solution by the automatic integration method, and calculate the amount of the related substances by the follow-ing equation: the amount of related substance A having the relative retention time of about 0.94 to celecoxib is not more than 0.4z, and each amount of the related substances other than related substance A is not more than 0.10z. Further-more, the total amount of the related substances is not more than 0.5z. Amount (z) of related substance ＝ MS/MT × AT/AS MS: Amount (mg) of Celecoxib RS taken MT: Amount (mg) of Celecoxib taken Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of celecoxib, beginning after the solvent peak. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. Confirm that the peak area of celecox-ib obtained with 25 mL of this solution is equivalent to 3.5 to 6.5z of that with 25 mL of the standard solution. Water <2.48> Not more than 0.5z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1.0 g, platinum crucible).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Celecoxib according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Separately, weigh accurately about 50 mg of Celecoxib RS, dissolve in a mix-ture of methanol and water (3:1) to make exactly 100 mL. MS: Amount (mg) of Celecoxib RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase: Adjust 0.02 mol/L potassium dihydrogen phosphate TS to pH 3.0 with phosphoric acid. To 600 mL of this solution add 300 mL of methanol for liquid chromatog-raphy and 100 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of celecoxib is about 22 minutes. ‌JP XVIII Official Monographs / Cellacefate 715 System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of celecoxib are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of celecoxib is not more than 1.0z.
Weigh accurately about 50 mg each of Celecoxib and Celecoxib RS, and dissolve each in a mixture of metha-nol and water (3:1) to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of celecoxib in each solution. compare the spectrum with the Reference Spectrum or the spectrum of a solution of Celecoxib RS prepared in the same manner as the sample solution: both spectra exhibit similar Amount (mg) of celecoxib (C ＝ MS × AT/AS 17H14F3N3O2S) intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Celecoxib, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Celecoxib RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Celecoxib according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Separately, weigh accurately about 50 mg of Celecoxib RS, dissolve in a mix-ture of methanol and water (3:1) to make exactly 100 mL. MS: Amount (mg) of Celecoxib RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase: Adjust 0.02 mol/L potassium dihydrogen phosphate TS to pH 3.0 with phosphoric acid. To 600 mL of this solution add 300 mL of methanol for liquid chromatog-raphy and 100 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of celecoxib is about 22 minutes. ‌JP XVIII Official Monographs / Cellacefate 715 System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of celecoxib are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of celecoxib is not more than 1.0z.
Cellacefate occurs as a white, powder or grain. It is freely soluble in acetone, and practically insoluble in water and in ethanol (99.5).◆
Determine the infrared absorption spectrum of Cellacefate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cellacefate for Identification RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Viscosity <2.53> Weigh accurately a quantity of Cellace-fate, equivalent to 15 g calculated on the anhydrous basis, dissolve in 85 g of a mixture of acetone and water (249: 1 in mass), and use this solution as the sample solution. Perform the test with the sample solution at 25 ± 0.29C as directed in Method 1 to obtain the kinematic viscosity n. Separately, de-termine the density, r, of the sample solution as directed under Determination of Specific Gravity and Density <2.56>, and calculate the viscosity of the sample solution, h, as h ＝ rn: not less than 45 mPa･s and not more than 90 mPa･s.
(1) ◆Heavy metals <1.07>—Proceed with 2.0 g of Cellacefate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ (2) Free acids—Weigh accurately about 3 g of Cellace-fate, put in a glass-stoppered conical flask, add 100 mL of diluted methanol (1 in 2), stopper tightly, and filter after shaking for 2 hours. Wash both the flask and residue with two 10-mL portions each of diluted methanol (1 in 2), com-bine the washes to the filtrate, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 2-3 drops of phenol-phthalein TS). Perform the blank determination with 120 mL of diluted methanol (1 in 2), and make any necessary correction. Amount (z) of free acids ＝ 0.8306A/M A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed M: Amount (g) of Cellacefate taken, calculated on the an-hydrous basis The amount of free acids is not more than 3.0z, calcu-lated as phthalic acid (C8H6O4: 166.13). Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration, using a mixture of ethanol (99.5) and dichloromethane (3:2) instead of methanol for water deter-mination). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Carboxybenzoyl group—Weigh accurately about 1 g of Cellacefate, dissolve in 50 mL of a mixture of ethanol (95) and acetone (3:2), and titrate <2.50> with 0.1 mol/L so- dium hydroxide VS (indicator: 2 – 3 drops of phenolphthal-ein TS). Perform a blank determination in the same manner, and make any necessary correction. Content (z) of carboxybenzoyl group (C8H5O3) 1.491 × A－(1.795 × B) M ＝ 100 － B ×100 A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed B: Amount (z) of free acids obtained in the Purity (2) M: Amount (g) of Cellacefate taken, calculated on the an-hydrous basis (2) Acetyl group—Weigh accurately about 0.1 g of Cel-lacefate, put in a glass-stoppered conical flask, add exactly 25 mL of 0.1 mol/L sodium hydroxide VS, and boil for 30 minutes under a reflux condenser. After cooling, add 2 – 3 drops of phenolphthalein TS, and titrate <2.50> the excess of sodium hydroxide with 0.1 mol/L hydrochloric acid VS. Perform a blank determination in the same manner. Content (z) of free acids and bound acetyl group (C2H3O) ＝ 0.4305A/M A: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed, corrected by the blank determination M: Amount (g) of Cellacefate taken, calculated on the an-hydrous basis ‌716 Content (z) of acetyl group (C2H3O) ＝100 × (P － 0.5182B)/(100 － B) － 0.5772C B: Amount (z) of free acids obtained in the Purity (2) C: Content (z) of carboxybenzoyl group P: Content (z) of free acids and bound acetyl group (C2H3O) ◆Containers and storage Containers—Tight containers.◆ Microcrystalline Cellulose
Celmoleukin (Genetical Recombination) is a recom-binant human interleukin-2, and is a protein consisting of 133 amino acid residues. It is a solution. It contains not less than 0.
Celmoleukin (Genetical Recombination) oc-curs as a clear and colorless liquid.
(1) Add 100 mL of protein digestive enzyme TS to 100 mL of Celmoleukin (Genetical Recombination), shake, leave standing at 379C for 18 to 24 hours, and then add 2 mL of 2-mercaptoethanol. Leave at 379C for a further 30 minutes, and add 5 mL of trifluoroacetic acid solution (1 in 10). Use this solution as the sample solution. Separately, process with celmoleukin for liquid chromatography by using the same method. Use this solution as the standard so-lution. Perform the test with 50 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and compare the chromatograms obtained from the sample solu-tion and standard solution: the similar peaks are observed at the same retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (particle size: 5 mm). Column temperature: A constant temperature of about 259C. Mobile phase A: A solution of trifluoroacetic acid (1 in 1000). Mobile phase B: A solution of trifluoroacetic acid in a mixture of acetonitrile and water (17:3) (1 in 1000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. and phenol (100:10:1). Replace the vial interior with inert gas or reduce the pressure, and heat at about 1159C for 24 hours. After drying under vacuum, dissolve in 0.5 mL of 0.02 mol/L hydrochloric acid TS, and use this solution as 0 – 5 5 – 45 100 → 60 0 → 40 45 – 75 60 → 0 40 → 100 75 – 85 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) the sample solution (1). To the other hydrolysis tube, add 100 mL of ice cold performic acid, oxidize for 1.5 hours on ice, add 50 mL of hydrobromic acid, and dry under vacuum. Add 200 mL of water, repeat the dry under vacuum proce-dure two more times, place the hydrolysis tube in a vial, and humidify the inside of the vial with 200 mL of diluted hydro-chloric acid (59 in 125). Replace the vial interior with inert gas or reduce the pressure, and heat at about 1159C for 24 Flow rate: Adjust so that the retention time of celmoleu-kin is about 70 minutes. System suitability— System performance: Add 2 mL of 2-mercaptoethanol to 100 mL of celmoleukin for liquid chromatography, leave at 379C for 2 hours, and then run this solution under the above operating conditions. Celmoleukin and its reduced form are eluted in this order with the resolution between these peaks being not less than 1.5. (2) Accurately measure an appropriate amount of Cel-moleukin (Genetical Recombination), dilute by adding cul-ture medium for celmoleukin, and prepare a sample solution containing 800 units per mL. Add 25 mL of the sample solu-tion to 2 wells (A and B) of a flat-bottomed microtest plate for tissue culture, and then add 25 mL of reference anti-inter-leukin-2 antiserum solution diluted with culture medium for celmoleukin to well A and 25 mL of culture medium for cel-moleukin to well B. Add 50 mL of culture medium for cel-moleukin to another well (well C). After shaking the microtest plate, warm in air containing 5z carbon dioxide at 379C for 30 minutes to 2 hours. Next, add to each well 50 mL of culture medium for celmoleukin containing the interleu-kin-2 dependent mouse natural killer cells NKC3 and culture at 379C for 16 to 24 hours. Add 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide TS, culture at 379C for 4 to 6 hours, and add sodium lauryl sulfate TS and leave for 24 to 48 hours. When the absorbance at 590 nm of the solution in each well is measured, the difference in absor-bance between the solutions from wells A and C is not more than 3z of the difference in absorbance between the solu-tions from wells B and C. Constituent amino acid When hydrolyze Celmoleukin (Genetical Recombination) according to Method 1 and Method 4 described in ``1. Hydrolysis of Protein and Pep-tide'', and perform the test according to Method 1 described in ``2. Methodologies of Amino Acid Analysis'' under Ami-no Acid Analysis of Proteins <2.04>, the molar ratios of the respective amino acids are as follows: glutamic acid (or glutamine) is 17 or 18, threonine is 11 to 13, aspartic acid (or asparagine) is 11 or 12, lysine is 11, isoleucine is 7 or 8, ser- ine is 6 to 9, phenylalanine is 6, alanine is 5, proline is 5 or 6, arginine is 4, methionine is 4, cysteine is 3 or 4, valine is 3 or 4, tyrosine is 3, histidine is 3, glycine is 2, and tryptophan is Procedure Hydrolysis Based on the results of the Assay (1), place an amount of Celmoleukin (Genetical Recombination), equivalent to about 50 mg as the total protein in two hydroly-sis tubes, and evaporate to dryness under vacuum. To one of the hydrolysis tubes add 100 mL of a mixture of diluted hy-drochloric acid (59 in 125), mercapto acetic acid and phenol (100:10:1), and shake. Place this hydrolysis tube in a vial and humidify the inside of the vial with 200 mL of the mixture of diluted hydrochloric acid (59 in 125), mercapto acetic acid hours. After drying under vacuum, dissolve in 0.5 mL of 0.02 mol/L hydrochloric acid TS, and use this solution as the sample solution (2). Separately, weigh exactly 60 mg of L-aspartic acid, 100 mg of L-glutamic acid, 17 mg of L-ala- nine, 23 mg of L-methionine, 21 mg of L-tyrosine, 24 mg of L-histidine hydrochloride monohydrate, 58 mg of L-threo-nine, 22 mg of L-proline, 14 mg of L-cystine, 45 mg of L-isoleucine, 37 mg of L-phenylalanine, 32 mg of L-arginine hydrochloride, 32 mg of L-serine, 6 mg of glycine, 18 mg of L-valine, 109 mg of L-leucine, 76 mg of L-lysine hydrochlo-ride, and 8 mg of L-tryptophan, dissolve with 0.1 mol/L hy-drochloric acid TS to make exactly 500 mL, and use this so-lution as the standard solution. Transfer 40 mL each of the standard solution to two hydrolysis tubes, evaporate to dry-ness under vacuum, and proceed in the same way for each respective sample solution to make the standard solutions (1) and (2). Amino acid analysis Perform the test with exactly 250 mL each of the sample solutions (1) and (2) and standard solutions (1) and (2) as directed under Liquid Chromatogra-phy <2.01> according to the following conditions, and from the peak areas for each amino acid obtained from the sample solutions (1) and (2) and standard solutions (1) and (2) calcu-late the molar number of the amino acids contained in 1 mL of the sample solutions (1) and (2). Furthermore, calculate the number of amino acids assuming there are 22 leucine residues in one mole of celmoleukin. Operating conditions— Detector: A visible absorption photometer [wavelength: 440 nm (proline) and 570 nm (amino acids other than pro-line)]. Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with strongly acidic ionex-change resin for liquid chromatography (Na type) (sulfonic acid group bound divinylbenzenepolystyrene) (5 mm in parti-cle diameter). Column temperature: Maintaining a constant temperature of about 489C for 28 minutes after sample injection, then a constant temperature of about 629C until 121 minutes after the injection. Reaction temperature: A constant temperature of about 1359C. Color developing time: About 1 minute. Mobile phases A, B, C and D: Prepare according to the following table. JP XVIII Official Monographs / Celmoleukin (Genetical Recombination) 721 Mobile phase A B C D Citric acid monohydrate 17.70 g 10.50 g 6.10 g — Trisodium citrate dihydrate 7.74 g 15.70 g 26.67 g — Sodium chloride 7.07 g 2.92 g 54.35 g — Sodium hydroxide — — 2.30 g 8.00 g Methanol (99.5) 40 mL — — — Benzyl alcohol — 10 mL 5 mL — Thiodiglycol 5 mL 5 mL 5 mL — Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL
◆(1) Heavy metals <1.07>—Proceed with 2.0 g of Powdered Cellulose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Water-soluble substances—Shake 6.0 g of Powdered Cellulose with 90 mL of recently boiled and cooled water, and allow to stand for 10 minutes with occasional shaking. Filter, with the aid of vacuum through a filter paper, discard the first 10 mL of the filtrate, and pass the subsequent fil-trate through the same filter, if necessary, to obtain a clear filtrate. Evaporate a 15.0-mL portion of the filtrate in a tared evaporating dish to dryness without charring, dry at 1059C for 1 hour, and weigh after allowing to cool in a desiccator: the difference between the mass of the residue and the mass obtained from a blank determination does not exceed 15.0 mg (1.5z). Diethyl ether-soluble substances—Place 10.0 g of Powdered Cellulose in a column having an internal diameter of about 20 mm, and pass 50 mL of peroxide-free diethyl [94218-72-1] Celmoleukin (Genetical Recombination) is a recom-binant human interleukin-2, and is a protein consisting of 133 amino acid residues. It is a solution. It contains not less than 0.5 and not more than 1.5 mg of protein per mL, and 1 mg of this protein con-tains potency not less than 8.0 × 106 units. Description Celmoleukin (Genetical Recombination) oc-curs as a clear and colorless liquid. Identification (1) Add 100 mL of protein digestive enzyme TS to 100 mL of Celmoleukin (Genetical Recombination), shake, leave standing at 379C for 18 to 24 hours, and then add 2 mL of 2-mercaptoethanol. Leave at 379C for a further 30 minutes, and add 5 mL of trifluoroacetic acid solution (1 in 10). Use this solution as the sample solution. Separately, process with celmoleukin for liquid chromatography by using the same method. Use this solution as the standard so-lution. Perform the test with 50 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and compare the chromatograms obtained from the sample solu-tion and standard solution: the similar peaks are observed at the same retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (particle size: 5 mm). Column temperature: A constant temperature of about 259C. Mobile phase A: A solution of trifluoroacetic acid (1 in 1000). Mobile phase B: A solution of trifluoroacetic acid in a mixture of acetonitrile and water (17:3) (1 in 1000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. and phenol (100:10:1). Replace the vial interior with inert gas or reduce the pressure, and heat at about 1159C for 24 hours. After drying under vacuum, dissolve in 0.5 mL of 0.02 mol/L hydrochloric acid TS, and use this solution as 0 – 5 5 – 45 100 → 60 0 → 40 45 – 75 60 → 0 40 → 100 75 – 85 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) the sample solution (1). To the other hydrolysis tube, add 100 mL of ice cold performic acid, oxidize for 1.5 hours on ice, add 50 mL of hydrobromic acid, and dry under vacuum. Add 200 mL of water, repeat the dry under vacuum proce-dure two more times, place the hydrolysis tube in a vial, and humidify the inside of the vial with 200 mL of diluted hydro-chloric acid (59 in 125). Replace the vial interior with inert gas or reduce the pressure, and heat at about 1159C for 24 Flow rate: Adjust so that the retention time of celmoleu-kin is about 70 minutes. System suitability— System performance: Add 2 mL of 2-mercaptoethanol to 100 mL of celmoleukin for liquid chromatography, leave at 379C for 2 hours, and then run this solution under the above operating conditions. Celmoleukin and its reduced form are eluted in this order with the resolution between these peaks being not less than 1.5. (2) Accurately measure an appropriate amount of Cel-moleukin (Genetical Recombination), dilute by adding cul-ture medium for celmoleukin, and prepare a sample solution containing 800 units per mL. Add 25 mL of the sample solu-tion to 2 wells (A and B) of a flat-bottomed microtest plate for tissue culture, and then add 25 mL of reference anti-inter-leukin-2 antiserum solution diluted with culture medium for celmoleukin to well A and 25 mL of culture medium for cel-moleukin to well B. Add 50 mL of culture medium for cel-moleukin to another well (well C). After shaking the microtest plate, warm in air containing 5z carbon dioxide at 379C for 30 minutes to 2 hours. Next, add to each well 50 mL of culture medium for celmoleukin containing the interleu-kin-2 dependent mouse natural killer cells NKC3 and culture at 379C for 16 to 24 hours. Add 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide TS, culture at 379C for 4 to 6 hours, and add sodium lauryl sulfate TS and leave for 24 to 48 hours. When the absorbance at 590 nm of the solution in each well is measured, the difference in absor-bance between the solutions from wells A and C is not more than 3z of the difference in absorbance between the solu-tions from wells B and C. Constituent amino acid When hydrolyze Celmoleukin (Genetical Recombination) according to Method 1 and Method 4 described in ``1. Hydrolysis of Protein and Pep-tide'', and perform the test according to Method 1 described in ``2. Methodologies of Amino Acid Analysis'' under Ami-no Acid Analysis of Proteins <2.04>, the molar ratios of the respective amino acids are as follows: glutamic acid (or glutamine) is 17 or 18, threonine is 11 to 13, aspartic acid (or asparagine) is 11 or 12, lysine is 11, isoleucine is 7 or 8, ser- ine is 6 to 9, phenylalanine is 6, alanine is 5, proline is 5 or 6, arginine is 4, methionine is 4, cysteine is 3 or 4, valine is 3 or 4, tyrosine is 3, histidine is 3, glycine is 2, and tryptophan is Procedure Hydrolysis Based on the results of the Assay (1), place an amount of Celmoleukin (Genetical Recombination), equivalent to about 50 mg as the total protein in two hydroly-sis tubes, and evaporate to dryness under vacuum. To one of the hydrolysis tubes add 100 mL of a mixture of diluted hy-drochloric acid (59 in 125), mercapto acetic acid and phenol (100:10:1), and shake. Place this hydrolysis tube in a vial and humidify the inside of the vial with 200 mL of the mixture of diluted hydrochloric acid (59 in 125), mercapto acetic acid hours. After drying under vacuum, dissolve in 0.5 mL of 0.02 mol/L hydrochloric acid TS, and use this solution as the sample solution (2). Separately, weigh exactly 60 mg of L-aspartic acid, 100 mg of L-glutamic acid, 17 mg of L-ala- nine, 23 mg of L-methionine, 21 mg of L-tyrosine, 24 mg of L-histidine hydrochloride monohydrate, 58 mg of L-threo-nine, 22 mg of L-proline, 14 mg of L-cystine, 45 mg of L-isoleucine, 37 mg of L-phenylalanine, 32 mg of L-arginine hydrochloride, 32 mg of L-serine, 6 mg of glycine, 18 mg of L-valine, 109 mg of L-leucine, 76 mg of L-lysine hydrochlo-ride, and 8 mg of L-tryptophan, dissolve with 0.1 mol/L hy-drochloric acid TS to make exactly 500 mL, and use this so-lution as the standard solution. Transfer 40 mL each of the standard solution to two hydrolysis tubes, evaporate to dry-ness under vacuum, and proceed in the same way for each respective sample solution to make the standard solutions (1) and (2). Amino acid analysis Perform the test with exactly 250 mL each of the sample solutions (1) and (2) and standard solutions (1) and (2) as directed under Liquid Chromatogra-phy <2.01> according to the following conditions, and from the peak areas for each amino acid obtained from the sample solutions (1) and (2) and standard solutions (1) and (2) calcu-late the molar number of the amino acids contained in 1 mL of the sample solutions (1) and (2). Furthermore, calculate the number of amino acids assuming there are 22 leucine residues in one mole of celmoleukin. Operating conditions— Detector: A visible absorption photometer [wavelength: 440 nm (proline) and 570 nm (amino acids other than pro-line)]. Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with strongly acidic ionex-change resin for liquid chromatography (Na type) (sulfonic acid group bound divinylbenzenepolystyrene) (5 mm in parti-cle diameter). Column temperature: Maintaining a constant temperature of about 489C for 28 minutes after sample injection, then a constant temperature of about 629C until 121 minutes after the injection. Reaction temperature: A constant temperature of about 1359C. Color developing time: About 1 minute. Mobile phases A, B, C and D: Prepare according to the following table. JP XVIII Official Monographs / Celmoleukin (Genetical Recombination) 721 Mobile phase A B C D Citric acid monohydrate 17.70 g 10.50 g 6.10 g — Trisodium citrate dihydrate 7.74 g 15.70 g 26.67 g — Sodium chloride 7.07 g 2.92 g 54.35 g — Sodium hydroxide — — 2.30 g 8.00 g Methanol (99.5) 40 mL — — — Benzyl alcohol — 10 mL 5 mL — Thiodiglycol 5 mL 5 mL 5 mL — Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL Purity (1) Related substances—Perform the test with 10 mL each of Celmoleukin (Genetical Recombination) and 0.01 mol/L acetic acid buffer solution (pH 5.0) as directed under Liquid Chromatography <2.01> under the following condi-tions, and measure the area of each peak by an automatic in-tegration method. When the amounts of related substances other than celmoleukin are calculated by the area percentage method, the total amount is not more than 5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: Stainless steel tube with an inside diameter of 4 Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL Water a sufficient quantity a sufficient quantity a sufficient quantity a sufficient quantity Total 1000 mL 1000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B, C and D as directed in the following table. mm and a length of 30 cm packed with octadecylsilanized silica gel for liquid chromatography (particle size: 5 mm). Column temperature: A constant temperature of about 259C. Mobile phase A: A solution of trifluoroacetic acid in a mixture of acetic acid and water (3:2) (1 in 1000). Mobile phase B: A solution of trifluoroacetic acid in a mixture of acetic acid and water (13:7) (1 in 1000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) Mobile phase D (volz) 0 – 35 35 – 60 60 – 111 111 – 121 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 60 70 → 10 30 → 90 Reaction reagent: Mix 407 g of lithium acetate dihydrate, 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for 10 minutes while passing a current of nitrogen, and assign as solution A. Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for 30 minutes while passing a current of nitrogen, and assign as solution B. Mix solutions A and B before use. Flow rate of mobile phase: Adjust so that the retention times of serine and leucine are about 30 minutes and about 73 minutes, respectively (about 0.21 mL per minute). Flow rate of reaction reagent: About 0.25 mL per minute. System suitability— System performance: To 2 mL of the standard solution add 0.02 mol/L hydrochloric acid TS to make 25 mL. When the procedure is run with 250 mL of this solution under the above operating conditions, the resolution between the peaks of threonine and serine is not less than 1.2. System repeatability: To 2 mL of the standard solution add 0.02 mol/L hydrochloric acid TS to make 25 mL. When the test is repeated 3 times with 250 mL of this solution under the above operating conditions, the relative standard devia-tion of the peak area of aspartic acid, serine, arginine and proline is not more than 2.4z. Molecular mass Based on the results of the Assay (1), add buffer for celmoleukin and dilute to prepare a sample solu-tion so that there is about 0.5 mg of protein per mL. To ver-tical uncontinuous buffer SDS-polyacrylamide gel prepared from resolving gel for celmoleukin and stacking gel for cel-moleukin add 20 mL of the sample solution or 20 mL of mar-ker protein for celmoleukin molecular mass determination to each stacking gel well, and perform the electrophoresis. The molecular mass of the main electrophoretic band is between 12,500 and 13,800 when the band is stained by immersion in Coomassie staining TS. pH <2.54> 4.5 – 5.5 Flow rate: Adjust so that the retention time of celmoleu-kin is about 50 minutes. Time span of measurement: About 1.3 times as long as the retention time of celmoleukin, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 0.5 mL of Celmoleukin (Genetical Recombination), and add 0.01 mol/L acetic acid buffer solution (pH 5.0) to make exactly 50 mL. Confirm that the peak area of celmoleukin obtained with 10 mL of this solution is equivalent to 0.9 to 1.1z of the peak area with 10 mL of Celmoleukin (Genetical Recombina-tion). System performance: Add 2 mL of 2-mercaptoethanol to 100 mL of Celmoleukin (Genetical Recombination), leave at 379C for 2 hours, and then run this solution under the above conditions. Celmoleukin and its reduced form are eluted in this order with the resolution between these peaks being not less than 3.0. Multimers—Dilute (at least 4 steps) the sample solu-tion prepared in the Molecular mass with buffer solution for celmoleukin so that the protein content is within the range of 2 to 32 mg per mL to prepare a series of standard solutions. Pipet 20 mL each of the sample solution and the standard solutions into the stacking gel well, and perform vertical uncoupled buffer SDS-polyacrylamide gel electrophoresis followed by immersion in Coomassie staining TS. Each elec-trophoretic band is stained blue. Next, determine the peak area of the electrophoretic bands obtained from each stand-ard solution using a densitometer and calculate the protein content using the calibration curve mentioned above. When determining the polymer proteins derived from celmoleukin, other than celmoleukin monomer, the amount is not more than 2z in relation to the total protein. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 100 EU/mL. ‌Ammonium acetate Measure exactly 0.1 mL of Celmoleu-kin (Genetical Recombination), add water to make exactly 10 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.1 g of ammonium chloride, and add water to make exactly 100 mL. Measure exactly 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard stock solution. Measure ex-actly 3 mL of the standard stock solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. When determining the area of the ammonium ion peak AT and AS, Celmoleukin (Genetical Recombination) contains not less than 0.28 mg and not more than 0.49 mg of ammonium ace-tate per mL. Amount (mg) of ammonium acetate (CH3COONH4) per mL ＝ AT/AS × MS × 0.003 × 1.441 MS: Amount (mg) of ammonium chloride taken 0.003: Dilution correction factor 1.441: Molecular mass conversion coefficient for convert-ing ammonium chloride to ammonium acetate Operating conditions— Detector: An electric conductivity detector. Column: Resin column 5 mm in inside diameter and 25 cm in length, packed with weakly acidic ion exchange resin for liquid chromatography (particle size: 5.5 mm). Column temperature: A constant temperature of about 409C. Mobile phase: Diluted 0.1 mol/L methanesulfonic acid TS (3 in 10). Flow rate: Adjust so that the retention time of ammonium is about 8 minutes. System suitability— System performance: Measure exactly 1 mL of Standard Sodium Stock Solution and 0.2 mL of Standard Potassium Stock Solution, and then add water to make exactly 100 mL. Measure exactly 5 mL of this solution and 3 mL of Standard Ammonium Solution, and then add water to make exactly 50 mL. When 25 mL of this solution is run under the above con-ditions, sodium, ammonium and potassium are eluted in this order with the resolution between the peaks of sodium and ammonium being not less than 3.0. System repeatability: When the test is repeated 5 times with 25 mL of the standard solution under the above condi-tions, the relative standard deviation of the ammonium peak area is not more than 10z.
(1) Total protein content—Measure accurately 1 mL of Celmoleukin (Genetical Recombination) and add water to make exactly 10 mL. Use this solution as the sample solution. Separately, weigh accurately about 50 mg of bovine serum albumin for assay in water to prepare standard dilu-tion solutions of 50, 100, and 150 mg/mL. Measure exactly 1 mL of the sample solution and each standard dilution solu-tion, add exactly 2.5 mL of alkaline copper TS for protein content determination, shake, and leave for 15 minutes. Next, add exactly 2.5 mL of water and 0.5 mL of dilute Folin's TS, and leave at 379C for 30 minutes. Measure the absorbances of these solutions at 750 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using 1 mL of water processed in the same way as control. Using the calibration curve prepared from the absorbance of the stand-ard dilution solution, calculate the protein content of Cel- moleukin (Genetical Recombination). (2) Specific activity—Measure exactly 0.1 mL of Cel-moleukin (Genetical Recombination) and add exactly 0.9 mL of culture medium for celmoleukin to make the sample solution. Separately, take one Interleukin-2 RS and add exactly 1 mL of water to dissolve. This is the standard solu-tion. Dilute exactly the sample and standard solutions in serially two-fold steps with culture medium for celmoleukin, and add equal volumes of interleukin-2 dependent mouse natural killer NKC3 cells to the serially diluted solutions. The control solution is a mixture of equal volumes of inter-leukin-2 dependent mouse natural killer NKC3 and culture medium for celmoleukin. Incubate these solutions at 379C for 16 to 24 hours. Following this, add a volume of 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-2H-tetrazolium bromide TS that is 1/5 that of the volume of culture medium for cel-moleukin, incubate at 379C for 4 to 6 hours, add a volume of sodium lauryl sulfate TS equivalent to the volume of the culture medium for celmoleukin, and leave for 24 to 48 hours. After eluting the blue-colored pigment generated, perform the test on these solutions as directed under Ultravi-olet-visible Spectrophotometry <2.24>, and measure the ab-sorbance at 590 nm. Taking the absorbance obtained when 1000 to 2000 units of celmoleukin per mL are added as 100z and the absorbance of the control solution as 0z, determine the dilution factor (A) of the Interleukin-2 RS that shows an absorbance of 50z and dilution factor of Celmoleukin (Genetical Recombination) (B). Multiply the B/A value by the unit number of the Interleukin-2 RS to calculate the bio-logical activity of 1 mL of Celmoleukin (Genetical Recombi-nation). Calculate the ratio of biological activity in relation to protein content determined in the total protein content test.
Containers—Tight containers. Storage—At －209C or lower.
White Beeswax occurs as white to yellowish white masses. It has a characteristic odor. It is comparatively brittle when cooled, and the fractured surface is granular, and non-crystalline. It is slightly soluble in diethyl ether, and practically insolu-ble in water and in ethanol (99.5). Acid value <1.13> 5 – 9 or 17 – 22 Weigh accurately about 6 g of White Beeswax, place in a glass-stoppered 250-mL flask, and add 50 mL of ethanol (99.5). Warm the mixture to dissolve the wax, add 1 mL of phenolphthalein TS, and proceed as directed in the Acid value. Perform a blank deter-mination using solvent which is not previously neutralized, and make any necessary correction. Saponification value <1.13> 80 – 100 Weigh accurately about 3 g of White Beeswax, place in a glass-stoppered 250-mL flask, and add exactly 25 mL of 0.5 mol/L potas-sium hydroxide-ethanol VS and 50 mL of ethanol (95), heat under a reflux condenser for 4 hours, and proceed as di-rected in the Saponification value.
Paraffin, fat, Japan wax or resin—Melt White Beeswax at the lowest possible temperature, drip the liquid into a vessel containing ethanol (95) to form granules, and allow them to stand in air for 24 hours. Drop the granules into two mixtures of ethanol (95) and water, one adjusted so as to have a specific gravity of 0.95 and the other 0.97: the granules sink or are suspended in the mixture with the spe-cific gravity of 0.95, and float or are suspended in the other mixture.
Carnauba Wax occurs as light yellow to light brown, hard and brittle masses or white to light yellow pow-der. It has a slight, characteristic odor. It is tasteless. It is practically insoluble in water, in ethanol (95), in diethyl ether and in xylene. Specific gravity d 20: 0.990 – 1.002 Melting point: 80 – 869C Acid value <1.13> Not more than 10.0. Use a mixture of xylene and ethanol (95) (2:1) as solvent. Saponification value <1.13> 78 – 95 Weigh accurately about 3 g of Carnauba Wax in a 300-mL flask, add 25 mL of xylene, and dissolve by warming. To this solution add 50 mL of ethanol (95) and exactly 25 mL of 0.5 mol/L potassium hydroxide-ethanol VS, and proceed as directed in the Saponification value. The time of heating should be 2 hours ‌1976‌‌ Cassia Seed / Crude Drugs and Related Drugs and the titration should be done by warming. Iodine value <1.13> 5 – 14 (Dissolve the sample by shaking a glass-stoppered flask in warm water.) ‌Containers and storage Containers—Well-closed contain-ers. Cassia Seed Cassiae Semen
Yellow Beeswax occurs as light yellow to brownish yellow masses. It has a characteristic odor, which is not rancid. It is comparatively brittle when cooled, and the fractured surface is granular, and non-crystalline. Acid value <1.13> 5 – 9 or 17 – 22 Weigh accurately about 6 g of Yellow Beeswax, place in a glass-stoppered 250-mL flask, and add 50 mL of ethanol (99.5). Warm the mixture to dissolve the wax, add 1 mL of phenolphthalein TS, and proceed as directed in the Acid value. Perform a blank deter-mination using solvent which is not previously neutralized, and make any necessary correction. ‌JP XVIII Crude Drugs and Related Drugs / Belladonna Root 1957 Saponification value <1.13> 80 – 100 Weigh accurately about 3 g of Yellow Beeswax, place in a 250-mL glass-stoppered flask, and add 25 mL of 0.5 mol/L potassium hy-droxide-ethanol and 50 mL of ethanol (95), heat under a reflux condenser for 4 hours, and proceed as directed in the Saponification value.
Paraffin, fat, Japan wax or resin—Melt Yellow Beeswax at the lowest possible temperature, drip the liquid into a glass vessel containing ethanol (95) to form granules, and allow them to stand in air for 24 hours. Drop the gran-ules into two mixtures of ethanol (95) and water, one adjusted so as to have a specific gravity of 0.95 and the other 0.97: the granules sink or are suspended in the mixture with the specific gravity of 0.95, and float or are suspended in the other mixture.
Cetanol occurs as unctuous, white, flakes, granules, or masses. It has a faint, characteristic odor. It is tasteless. It is very soluble in pyridine, freely soluble in ethanol (95), in ethanol (99.5) and in diethyl ether, very slightly soluble in acetic anhydride, and practically insoluble in water.
(1) Clarity of solution—Dissolve 3.0 g of Cetanol in 25 mL of ethanol (99.5) by warming: the solution is clear. (2) Alkalinity—To the solution obtained in (1) add 2 drops of phenolphthalein TS: no red color develops. Residue on ignition <2.44> Not more than 0.05z (2 g).
Cetirizine Hydrochloride, when dried, contains not less than 99.
Cetirizine Hydrochloride occurs as a white crystalline powder. It is very soluble in water, and slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. A solution of Cetirizine Hydrochloride (1 in 10) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Cetirizine Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cetirizine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Cetirizine Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cetirizine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Cetirizine Hy-drochloride in 50 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 2 mL of the sample solu-tion, add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area of each solution by the automatic in-tegration method: the area of each peak other than cetirizine obtained from the sample solution is not larger than the peak area of cetirizine from the standard solution. And the total area of the peaks other than cetirizine from the sample solu-tion is not larger than 2.5 times the peak area of cetirizine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and diluted 0.5 mol/L sulfuric acid TS (2 in 25) (47:3). Flow rate: Adjust so that the retention time of cetirizine is about 9 minutes. Time span of measurement: About 3 times as long as the retention time of cetirizine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cetirizine obtained with 10 mL of this solution is equivalent to 35 to 65z of that with 10 mL of the standard solution. System performance: Dissolve 20 mg of Cetirizine Hydro-chloride in the mobile phase to make 100 mL. To 5 mL of this solution, add 3 mL of a solution of aminopyrine in the mobile phase (1 in 2500), and add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, cetirizine and aminopyrine are eluted in this order with the resolution be-tween these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cetirizine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.1 g of Cetirizine Hydro-chloride, previously dried, dissolve in 70 mL of a mixture of acetone and water (7:3), and titrate <2.50> to the second equivalence point with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 15.39 mg of C21H25ClN2O3.2HCl
Cetotiamine Hydrochloride Hydrate contains not less than 98.
Cetotiamine Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is odorless or has a faint characteristic odor. It is freely soluble in water and in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. Melting point: about 1329C (with decomposition).
(1) Determine the absorption spectrum of a solution of Cetotiamine Hydrochloride Hydrate in 0.01 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cetotiamine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cetotiamine Hydrochloride Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cetotiamine Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Cetotiamine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Cetotiamine Hydrochloride Hy-drate in 10 mL of water is clear and has no more color than the following control solution. Control solution: Mix exactly 1.5 mL of Cobalt (II) Chlo-ride CS, exactly 36 mL of Iron (III) Chloride CS and exactly 12.5 mL of diluted dilute hydrochloric acid (1 in 10). Pipet 1 mL of this mixture, and add diluted dilute hydrochloric acid (1 in 10) to make exactly 100 mL. Heavy metals <1.07>—Proceed with 1.0 g of Cetotia- mine Hydrochloride Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Cetotiamine Hydrochloride Hydrate in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than cetotiamine obtained from the sample solution is not larger than the peak area of cetotia-mine from the standard solution, and the total area of the peaks other than cetotiamine from the sample solution is not larger than 2 times the peak area of cetotiamine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cetotiamine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of cetotiamine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cetotiamine are not less than 3000 and 0.7 – 1.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cetotiamine is not more than 2.0z. Water <2.48> 3.0 – 5.0z (40 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 30 mg each of Cetotiamine Hydrochloride Hydrate and Cetotiamine Hydrochloride RS (separately determine the water <2.48> in the same manner as Cetotiamine Hydrochloride Hydrate), add exactly 10 mL each of the internal standard solution, then add a mixture of water and methanol (1:1) to make 50 mL. To 2 mL each of these solutions add a mixture of water and methanol (1:1) to make 10 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cetotiamine to that of the internal standard. Amount (mg) of cetotiamine hydrochloride (C18H26N4O6S.HCl) ＝ MS × QT/QS MS: Amount (mg) of Cetotiamine Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌726 Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.0 g of sodium 1-heptanesul-fonate in diluted acetic acid (100) (1 in 100) to make 1000 mL. To 1 volume of this solution add 1 volume of methanol. Flow rate: Adjust so that the retention time of cetotiamine is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cetotiamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cetotiamine to that of the internal standard is not more than 1.0z.
Cetraxate Hydrochloride occurs as white, crys-tals or crystalline powder. It is soluble in methanol, sparingly soluble in water and in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 2369C (with decomposition).
(1) Determine the absorption spectrum of a solution of Cetraxate Hydrochloride in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Dissolve 0.5 g of Cetraxate Hydrochloride in 5 mL of a mixture of water and 2-propanol (1:1) by warming, cool to below 259C. Filter, dry the formed crystals in vacuum for 4 hours, and further dry at 1059C for 1 hour. Determine the infrared absorption spectrum of the dried matter as directed in the potassium chloride disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Cetraxate Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cetraxate Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cetraxate Hydrochloride according to Method 3, and per-form the test with a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 5) (not more than 2 ppm). cis Isomer—Dissolve 0.10 g of Cetraxate Hydrochlo-ride in 10 mL of water, and use this solution as the sample solution. To exactly 5 mL of the sample solution add water to make exactly 100 mL. To exactly 2 mL of this solution add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of the peak, having the relative retention time 1.3 to 1.6 to cetraxate from the sample solution is not larger than the peak area of cetraxate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust the pH of a mixture of water, meth-anol and 0.5 mol/L ammonium acetate TS (15:10:4) to 6.0 with acetic acid (31). Flow rate: Adjust so that the retention time of cetraxate is about 10 minutes. System suitability— System performance: Dissolve 0.02 g of Cetraxate Hydro-chloride and 0.01 g of phenol in 100 mL of water. To 2 mL of this solution add water to make 20 mL. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, cetraxate and phenol are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cetraxate is not more than 2.0z. 3-( p-Hydroxyphenyl)propionic acid—To 0.10 g of Cetraxate Hydrochloride add exactly 2 mL of the internal standard solution and methanol to make 10 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of 3-( p-hydroxyphenyl)propionic acid in methanol to make exactly 100 mL. To exactly 2 mL of this solution add exactly 2 mL of the internal standard solution and methanol to make 10 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of 3-( p-hydroxy-phenyl)propionic acid to that of the internal standard: QT is not larger than QS. Internal standard solution—A solution of caffeine in metha-nol (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust the pH of a mixture of water, meth-anol and 0.5 mol/L ammonium acetate TS (15:5:2) to 5.5 JP XVIII Official Monographs / Chenodeoxycholic Acid 727 ‌with acetic acid (31). Flow rate: Adjust so that the retention time of 3-( p-hydroxyphenyl)propionic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, 3-(p-hydroxyphenyl)propionic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of 3-( p-hydroxyphenyl)propionic acid to that of the internal standard is not more than 1.0z. Related substances—Dissolve 0.10 g of Cetraxate Hy-drochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and acetic acid (100) (20:4:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 909C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Cetraxate Hydro-chloride, previously dried, dissolve in 100 mL of water, and adjust the pH of this solution to between 7.0 and 7.5 with dilute sodium hydroxide TS. To this solution add 10 mL of formaldehyde solution, stir for about 5 minutes, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS by taking over about 20 minutes (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 34.18 mg of C17H23NO4.HCl
Chenodeoxycholic Acid, when dried, contains not less than 98.
Chenodeoxycholic Acid occurs as white, crys- (2), and the spot corresponding to the spot from the stand- ‌728‌ ard solution (3) is not more intense than the spot from the standard solution (3). As compared to the spots with the standard solutions A, B, C, D and E, the spots other than the principal spot and the spots mentioned above from the sample solution are not more intense than the spot from the standard solution E, and the total amount of them is not more than 1.5z. Loss on drying <2.41> Not more than 1.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Determine the infrared absorption spectrum of Chenodeoxycholic Acid, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋11.0 – ＋13.09(after dry-ing, 0.4 g, ethanol (99.5), 20 mL, 100 mm).
(1) Chloride <1.03>—Dissolve 0.36 g of Chenodeoxycholic Acid in 30 mL of methanol, add 10 mL of dilute nitric acid and water to make 50 mL, and perform the test with this solution. Prepare the control solution as follows: To 1.0 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of methanol, 10 mL of dilute nitric acid and water to make 50 mL (not more than 0.1z). Heavy metals <1.07>—Proceed with 1.0 g of Chenodeoxycholic Acid according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Barium—To 2.0 g of Chenodeoxycholic Acid add 100 mL of water, and boil for 2 minutes. To this solution add 2 mL of hydrochloric acid, boil for 2 minutes, filter after cooling, and wash the filter with water until to get 100 mL of the filtrate. To 10 mL of the filtrate add 1 mL of dilute sulfuric acid: no turbidity appears. Related substances—Dissolve 0.20 g of Chenodeoxy-cholic Acid in a mixture of acetone and water (9:1) to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of lithocholic acid for thin-layer chromatography in the mixture of acetone and water (9:1) to make exactly 10 mL. Pipet 2 mL of this solution, add the mixture of acetone and water (9:1) to make exactly 100 mL, and use this solution as the standard solution (1). Separately, dissolve 10 mg of ursodeoxycholic acid in the mixture of ace-tone and water (9:1) to make exactly 100 mL, and use this solution as the standard solution (2). Separately, dissolve 10 mg of cholic acid for thin-layer chromatography in the mix-ture of acetone and water (9:1) to make exactly 100 mL, and use this solution as the standard solution (3). Pipet 1 mL of the sample solution, and add the mixture of acetone and water (9:1) to make exactly 20 mL. Pipet 0.5 mL, 1 mL, 2 mL, 3 mL and 5 mL of this solution, add the mixture of ace-tone and water (9:1) to each of them to make exactly 50 mL, and designate these solutions as standard solution A, stand-ard solution B, standard solution C, standard solution D and standard solution E, respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution, standard solu- tions (1), (2), (3) and standard solutions A, B, C, D and E on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 4-methyl-2-pentanone, toluene and formic acid (16:6:1) to a distance of about 15 cm, air-dry the plate, and further dry at 1209C for 30 minutes. Im-mediately, spray evenly a solution of phosphomolybdic acid n-hydrate in ethanol (95) (1 in 5) on the plate, and heat the plate at 1209C for 2 to 3 minutes: the spot corresponding to the spot obtained from the standard solution (1) is not more intense than the spot from the standard solution (1), the spot chenodeoxycholic acid (C 24H40 O4). corresponding to the spot from the standard solution (2) is not more intense than the spot from the standard solution Description Chenodeoxycholic Acid occurs as white, crys- (2), and the spot corresponding to the spot from the stand- ‌728‌ ard solution (3) is not more intense than the spot from the standard solution (3). As compared to the spots with the standard solutions A, B, C, D and E, the spots other than the principal spot and the spots mentioned above from the sample solution are not more intense than the spot from the standard solution E, and the total amount of them is not more than 1.5z. Loss on drying <2.41> Not more than 1.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Chenodeoxycholic Acid, previously dried, dissolve in 40 mL of ethanol (95) and 20 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 39.26 mg of C24H40O4
Chloral Hydrate contains not less than 99.
Chloral Hydrate occurs as colorless crystals. It has a pungent odor and an acrid, slightly bitter taste. It is very soluble in water, and freely soluble in ethanol (95) and in diethyl ether. It slowly volatilizes in air.
(1) Dissolve 0.2 g of Chloral Hydrate in 2 mL of water, and add 2 mL of sodium hydroxide TS: the turbidity is produced, and it separates into two clear layers by warming. (2) Heat 0.2 g of Chloral Hydrate with 3 drops of aniline and 3 drops of sodium hydroxide TS: the disagreeable odor of phenylisocyanide (poisonous) is perceptible.
(1) Clarity and color of solution—Dissolve 1.0 g of Chloral Hydrate in 2 mL of water: the solution is clear and colorless. Acidity—Dissolve 0.20 g of Chloral Hydrate in 2 mL of water, and add 1 drop of methyl orange TS: a yellow color develops. Chloride <1.03>—Perform the test with 1.0 g of Chlo- ral Hydrate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Chloral alcoholate—Warm 1.0 g of Chloral Hydrate with 10 mL of sodium hydroxide TS, filter the upper layer, add iodine TS to the filtrate until a yellow color develops, and allow the solution to stand for 1 hour: no yellow precipi-tate is produced. Benzene—Warm the solution obtained in (1) with 3 mL of water: no odor of benzene is perceptible. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 4 g of Chloral Hydrate in a glass-stoppered flask, add 10 mL of water and exactly 40 mL of 1 mol/L sodium hydroxide VS, and allow the mixture to stand for exactly 2 minutes. Titrate <2.50> the excess sodium hydroxide immediately with 0.5 mol/L sulfuric acid VS (in-dicator: 2 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 1 mol/L sodium hydroxide VS ＝ 165.4 mg of C2H3Cl3O2
クロラムフェニコールパルミチン酸エステル
Chloramphenicol Palmitate contains not less than 558 mg (potency) and not more than 587 mg (potency) per mg, calculated on the dried basis.
Chloramphenicol Palmitate occurs as a white to grayish white, crystalline powder. It is freely soluble in acetone, sparingly soluble in metha-nol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Chloramphenicol Palmitate in ethanol (99.5) (1 in 33,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Chloram-phenicol Palmitate RS prepared in the same manner as the exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. The test should be performed within 30 minutes after the sample solution and standard solution are prepared. Determine each peak area by the automatic integration method: the total area of the peaks other than the peak of chloramphenicol palmitate obtained from the sample solution is not larger than 3.5 times the peak area of chloramphenicol palmitate from the standard solution. For the peak areas for chloramphenicol, having the relative retention time of about 0.5 to chloramphenicol palmitate, and for chloramphenicol dipalmitate, having the relative retention time of about 5.0, multiply their correction factors, 0.5 and 1.4, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Methanol. Flow rate: Adjust so that the retention time of chloram-phenicol palmitate is about 5 minutes. Time span of measurement: About 6 times as long as the retention time of chloramphenicol palmitate. System suitability— Test for required detectability: Dissolve 50 mg of Chlo-ramphenicol Palmitate in 50 mL of methanol. To 1 mL of this solution, add methanol to make 100 mL, and use this so-lution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add methanol to make exactly 50 mL. Confirm that the peak area of chlo-ramphenicol palmitate obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution ‌730 for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates of the peak of chloramphenicol palmitate is not less than 5000. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of chloramphenicol palmitate is not more than 1.0z. Loss on drying <2.41> Not more than 1.0z (1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours).
Weigh accurately an amount of Chloramphenicol Palmitate and Chloramphenicol Palmitate RS, equivalent to about 37 mg (potency), dissolve each in 40 mL of methanol and 1 mL of acetic acid (100), and add methanol to make ex-actly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of chloramphenicol palmitate in each solution. Amount [ mg (potency)] of chloramphenicol (C11H12Cl2N2O5) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Chloramphenicol Palmitate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water and acetic acid (100) (172:27:1). Flow rate: Adjust so that the retention time of chloram-phenicol palmitate is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates of the peak of chlo-ramphenicol palmitate is not less than 2400. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of chloramphenicol palmitate is not more than 1.0z.
Chloramphenicol contains not less than 980 mg (po-tency) and not more than 1020 mg (potency) per mg, calculated on the dried basis.
Chloramphenicol occurs as white to yellow-white, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (99.5), and slightly soluble in water.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Chloramphenicol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Chloramphenicol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Chloramphenicol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋18.5 – ＋21.59 (1.25 g, ethanol (99.5), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Chloramphenicol according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 25 ppm). (2) Related substances—Dissolve 0.10 g of Chloram-phenicol in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 10 mL of the standard solu-tion (1), add methanol to make exactly 20 mL, and use this ‌JP XVIII Official Monographs / Chloramphenicol Palmitate 729 solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, methanol and acetic acid (100) (10:1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution (1), and the total amount of these spots from the sample so-lution is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Chloramphenicol and Chloramphenicol RS, equivalent to about 50 mg (po-tency), dissolve each in 10 mL of methanol, and add water to make exactly 50 mL. Pipet 20 mL each of these solutions, and add water to make exactly 100 mL. Pipet 10 mL each of these solutions, add water to make exactly 100 mL, and use these solutions as the sample solution and standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 278 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve 5 mg each of Chloramphenicol Palmitate and Chloramphenicol Palmitate RS in 1 mL of acetone, and use these solutions as the sample solution and standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of acetone and cyclohexane (1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. D Optical rotation <2.49> [a]25: ＋21 – ＋259(1 g calculated on the dried basis, ethanol (99.5), 20 mL, 100 mm). Melting point <2.60> 91 – 969C Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Chloramphenicol Palmitate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Chloramphenicol Palmitate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Chloram-phenicol Palmitate in 50 mL of methanol, and use this solu- Amount [mg (potency)] of chloramphenicol (C ＝ MS × AT/AS × 1000 11H12Cl2N2O5) tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with MS: Amount [mg (potency)] of Chloramphenicol RS taken
Chlordiazepoxide, when dried, contains not less diazepoxide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths.
Chlordiazepoxide occurs as white to light yel- low, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in ethanol (95), very slightly soluble in diethyl ether, and practi-cally insoluble in water. It dissolves in dilute hydrochloric acid. It is gradually affected by light. Melting point: about 2409C (with decomposition).
(1) Determine the absorption spectrum of a solution of Chlordiazepoxide in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Chlor- Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.98 mg of C16H14ClN3O
(1) Heavy metals <1.07>—Proceed with 1.0 g of Chlordiazepoxide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.20 g of Chlordiazepoxide in exactly 10 mL of a mixture of methanol and ammonia TS (97:3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and ammonia TS (97:3) to make ex-actly 200 mL, and use this solution as the standard solution (1). Separately, dissolve 10 mg of 2-amino-5-chloroben-zophenone for thin-layer chromatography in methanol to make exactly 200 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL of the sample solution and 5 mL each of the standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and ethanol (99.5) (19:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1). Spray evenly a solution of sodium nitrite in 1 mol/L hydrochloric acid TS (1 in 100) on the plate, allow to stand for 1 minute, and spray evenly N-(1-naphthyl)-N?-diethylethylenediamine oxalate-acetone TS on the plate: the spots from the sample solution are not more intense than the spots from the standard solution (2). Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Chlordiazepoxide, previously dried, and dissolve in 50 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS until the than 98.5z of chlordiazepoxide (C 16H14 ClN3O). color of the supernatant liquid changes from purple through blue-purple to blue (indicator: 3 drops of crystal violet TS). Description Chlordiazepoxide occurs as white to light yel- low, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in ethanol (95), very slightly soluble in diethyl ether, and practi-cally insoluble in water. It dissolves in dilute hydrochloric acid. It is gradually affected by light. Melting point: about 2409C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Chlordiazepoxide in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Chlor- Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.98 mg of C16H14ClN3O
Chlorhexidine Hydrochloride, when dried, contains not less than 98.
Chlorhexidine Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is soluble in formic acid, slightly soluble in methanol and in warm methanol, and practically insoluble in water, in ethanol (95) and in diethyl ether. It is gradually colored by light.
(1) Dissolve 0.01 g of Chlorhexidine Hy-drochloride in 5 mL of methanol by warming, and add 1 mL of bromine TS and 1 mL of 8 mol/L sodium hydroxide TS: a deep red color is produced. Dissolve 0.3 g of Chlorhexidine Hydrochloride in 10 ‌736‌ mL of 6 mol/L hydrochloric acid TS, cool in ice, and add 10 mL of 8 mol/L sodium hydroxide TS dropwise with stirring: a white precipitate is produced. Collect the precipitate, wash with water, recrystallize from diluted ethanol (95) (7 in 10), and dry at 1059C for 30 minutes: the crystals so obtained melt <2.60> between 1309C and 1349C. Dissolve 0.1 g of Chlorhexidine Hydrochloride in 50 mL of dilute nitric acid: the solution responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Chlorhexidine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—To 1.0 g of Chlorhexidine Hydro- chloride in a crucible add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), fire the ethanol (95) to burn, and heat gradually to incinerate. If a car-bonized substance remains, moisten with a small amount of nitric acid, and ignite to incinerate. Cool, add 10 mL of dilute hydrochloric acid to the residue, dissolve by warming on a water bath, use this solution as the test solution, and perform the test (not more than 2 ppm). p-Chloroaniline—Dissolve 0.10 g of Chlorhexidine Hydrochloride in 2 mL of formic acid, and add 15 mL of 1 mol/L hydrochloric acid TS and 20 mL of water immedi-ately. Add 0.3 mL of sodium nitrite TS, shake, and allow to stand for 2 minutes. Add 4 mL of ammonium amidosulfate TS, and then allow to stand for 1 minute. Add 5 mL of N-(1-naphthyl)-N?-diethylethylenediamine oxalate-acetone TS, allow to stand for 10 minutes, and add 1 mL of ethanol (95) and water to make 50 mL: the solution has no more color than the following control solution. Control solution: Dissolve 20 mg of 4-chloroaniline in 10 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of the solution, and add water to make exactly 100 mL. To 2.0 mL of the solution add 2 mL of formic acid, 15 mL of 1 mol/L hydrochloric acid TS and 20 mL of water, and proceed in the same manner. Loss on drying <2.41> Not more than 2.0z (1 g, 1309C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Chlorhexidine Hy-drochloride, previously dried, dissolve in 2.0 mL of formic acid, add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 14.46 mg of C22H30Cl2N10.2HCl
Chlorinated Lime contains not less than 30.
Chlorinated Lime occurs as a white powder. It has a chlorine-like odor. It dissolves partially in water. The solution changes red litmus paper to blue, then gradually decolorizes.
(1) To Chlorinated Lime add dilute hydro-chloric acid: a gas, which has the odor of chlorine, evolves, and the gas changes moistened starch-potassium iodide paper to blue. (2) Shake 1 g of Chlorinated Lime with 10 mL of water, and filter: the filtrate responds to Qualitative Tests <1.09> (2) and (3) for calcium salt.
Weigh accurately about 5 g of Chlorinated Lime, transfer to a mortar, and triturate thoroughly with 50 mL of water. Transfer to a 500-mL volumetric flask with the aid of water, and add water to make 500 mL. Mix well, imme-diately take exactly 50 mL of the mixture in an iodine flask, add 10 mL of potassium iodide TS and 10 mL of dilute hy-drochloric acid, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 3.545 mg of Cl
Chlormadinone Acetate occurs as white to light yellow, crystals or crystalline powder. It is odorless. It is freely soluble in chloroform, soluble in acetonitrile, slightly soluble in ethanol (95) and in diethyl ether, and prac-tically insoluble in water.
(1) Dissolve 2 mg of Chlormadinone Acetate in 1 mL of ethanol (95), and add 1 mL of 1,3-dinitrobenzene TS and 1 mL of a solution of potassium hy-droxide (1 in 5): a red-purple color develops. To 0.05 g of Chlormadinone Acetate add 2 mL of po-tassium hydroxide-ethanol TS, and boil on a water bath for 5 minutes. After cooling, add 2 mL of diluted sulfuric acid (2 in 7), and boil gently for 1 minute: the odor of ethyl ace-tate is perceptible. Determine the infrared absorption spectrum of Chlormadinone Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of previously dried Chlor-madinone Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Chlormadinone Acetate as di-rected under Flame Coloration Test <1.04> (2): a green color ‌JP XVIII Official Monographs / Chlorobutanol 737 appears. D Optical rotation <2.49> [a]20: －10.0 – －14.09(after dry-ing, 0.2 g, acetonitrile, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Chlormadinone Acetate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Chlormadinone Acetate according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 20 mg of Chlormadi-none Acetate in 10 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of peaks other than the peak of chlormadinone acetate obtained from the sample solution is not larger than the peak area of chlor- visible Spectrophotometry <2.24>, and determine the absor-bances, AT and AS, at 285 nm. Amount (mg) of chlormadinone acetate (C23H29ClO4) ＝ MS × AT/AS MS: Amount (mg) of Chlormadinone Acetate RS taken
Chlorobutanol contains not less than 98.
Chlorobutanol occurs as colorless or white crystals. It has a camphoraceous odor. It is very soluble in methanol, in ethanol (95) and in diethyl ether, and slightly soluble in water. It slowly volatilizes in air. Melting point: not lower than about 769C.
(1) To 5 mL of a solution of Chlo-robutanol (1 in 200) add 1 mL of sodium hydroxide TS, then slowly add 3 mL of iodine TS: a yellow precipitate is pro-duced and the odor of iodoform is perceptible. (2) To 0.1 g of Chlorobutanol add 5 mL of sodium hy-droxide TS, shake well the mixture, add 3 to 4 drops of ani-line, and warm gently: the disagreeable odor of phenyl isoc-yanide (poisonous) is perceptible.
(1) Acidity—Shake thoroughly 0.10 g of the pow-der of Chlorobutanol with 5 mL of water: the solution is neutral. (2) Chloride <1.03>—Dissolve 0.5 g of Chlorobutanol in 25 mL of dilute ethanol, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS by adding 25 mL of dilute ethanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.071z). Water <2.48> Not more than 6.0z (0.2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Chlormadi-none Acetate and Chlormadinone Acetate RS, previously dried, and dissolve in ethanol (95) to make exactly 100 mL. Pipet 5 mL each of these solutions, to each add ethanol (95) to make exactly 100 mL, and use these solutions as the sam-ple solution and the standard solution, respectively. Perform the test with these solutions as directed under Ultraviolet- Description Chlorobutanol occurs as colorless or white crystals. It has a camphoraceous odor. It is very soluble in methanol, in ethanol (95) and in diethyl ether, and slightly soluble in water. It slowly volatilizes in air. Melting point: not lower than about 769C. Identification (1) To 5 mL of a solution of Chlo-robutanol (1 in 200) add 1 mL of sodium hydroxide TS, then slowly add 3 mL of iodine TS: a yellow precipitate is pro-duced and the odor of iodoform is perceptible. (2) To 0.1 g of Chlorobutanol add 5 mL of sodium hy-droxide TS, shake well the mixture, add 3 to 4 drops of ani-line, and warm gently: the disagreeable odor of phenyl isoc-yanide (poisonous) is perceptible. Purity (1) Acidity—Shake thoroughly 0.10 g of the pow-der of Chlorobutanol with 5 mL of water: the solution is neutral. (2) Chloride <1.03>—Dissolve 0.5 g of Chlorobutanol in 25 mL of dilute ethanol, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS by adding 25 mL of dilute ethanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.071z). Water <2.48> Not more than 6.0z (0.2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Transfer about 0.1 g of Chlorobutanol, accurately weighed, to a 200-mL conical flask, and dissolve in 10 mL of ethanol (95). Add 10 mL of sodium hydroxide TS, boil under a reflux condenser for 10 minutes, cool, add 40 mL of dilute nitric acid and exactly 25 mL of 0.1 mol/L silver ni-trate VS, and shake well. Add 3 mL of nitrobenzene, and shake vigorously until the precipitate is coagulated. Titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. ‌738 Each mL of 0.1 mol/L silver nitrate VS ＝ 5.915 mg of C4H7Cl3O
クロルフェネシンカルバミン酸エステル
Chlorphenesin Carbamate, when dried, contains not less than 98.
Chlorphenesin Carbamate occurs as white, crystals or a crystalline powder. It is freely soluble in methanol, in ethanol (95) and in pyri-dine, and slightly soluble in water. A solution of Chlorphenesin Carbamate in ethanol (95) (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Chlorphenesin Carbamate in ethanol (95) (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Chlorphenesin Carbamate, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Chlorphenesin Carbamate as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Dissolve 2.0 g of Chlor-phenesin Carbamate in 20 mL of ethanol (95), and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution as follows: to 2.0 mL of Standard Lead Solu-tion add 20 mL of ethanol (95), 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Chlorphenesin Carbamate according to Method 3, and perform the test (not more than 2 ppm). Chlorphenesin-2-carbamate—Dissolve 0.10 g of Chlorphenesin Carbamate in 20 mL of a mixture of hexane for liquid chromatography and 2-propanol (7:3), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine the peak area, Aa, of chlorphenesin carbamate and the peak area, Ab, of chlorphenesin-2-carbamate by the au-tomatic integration method: the ratio, Ab/(Aa ＋ Ab), is not more than 0.007. Operating conditions— Detector: An ultraviolet absorption photometer (wave- System performance: Dissolve 0.1 g of Chlorphenesin Carbamate in 50 mL of methanol. To 25 mL of this solution add 25 mL of dilute sodium hydroxide TS, and warm at 609C for 20 minutes. To 20 mL of this solution add 5 mL of 1 mol/L hydrochloric acid TS, shake well with 20 mL of ethyl acetate, and allow to stand to separate the upper layer. When the procedure is run with 10 mL of this layer under the above operating conditions, chlorphenesin, chlorphenesin carbamate and chlorphenesin-2-carbamate are eluted in this order, with the relative retention times of chlorphenesin and chlorphenesin-2-carbamate to chlorphenesin carbamate being about 0.7 and about 1.2, respectively, and with the resolution between the peaks of chlorphenesin and chlor-phenesin carbamate being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak areas of chlorphenesin carbamate is not more than 2.0z. Related substances—Dissolve 0.10 g of Chlorphene-sin Carbamate in 10 mL of ethanol (95), and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add ethanol (95) to make exactly 20 mL. Pipet 2 mL of this solution, add ethanol (95) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 50 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, methanol and ammonia solution (28) (17:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 20 minutes: the spot other than the principal spot obtained from the sample solution is not more than one, and it is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Chlorphenesin Car-bamate, previously dried, dissolve in 20 mL of pyridine, add exactly 50 mL of 0.1 mol/L potassium hydroxide-ethanol TS, and warm at 709C for 40 minutes. After cooling, add 100 mL of ethanol (95), and titrate <2.50> the excess potas-sium hydroxide with 0.1 mol/L hydrochloric acid VS until ‌JP XVIII Official Monographs / Chlorphenesin Carbamate Tablets 739 the color of the solution changes from blue through blue-green to yellow (indicator: 1 mL of thymol blue TS). Per-form a blank determination in the same manner. Each mL of 0.1 mol/L potassium hydroxide-ethanol TS ＝ 24.57 mg of C10H12ClNO4
クロルフェニラミンマレイン酸塩
Chlorpheniramine Maleate, when dried, contains not less than 98.
Chlorpheniramine Maleate occurs as white, fine crystals. It is very soluble in acetic acid (100), freely soluble in water and in methanol, and soluble in ethanol (99.5). It dissolves in dilute hydrochloric acid. A solution of Chlorpheniramine Maleate (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Chlorpheniramine Maleate in 0.1 mol/L hydro-chloric acid TS (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Chlorpheniramine Maleate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Chlorpheniramine Maleate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Chlorpheniramine Maleate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.10 g of Chlorpheniramine Maleate in 5 mL of methanol, and use this solution as the sample solution. Separately, dissolve 56 mg of maleic acid in 10 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, methanol, acetic acid (100) and water (70:20:7:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot among two of the spots obtained with the sample solution shows the same intense and Rf value with the spot obtained with the standard solution. pH <2.54> Dissolve 1.0 g of Chlorpheniramine Maleate in 100 mL of freshly boiled and cooled water: the pH of this so-lution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Chlorpheniramine Maleate in 50 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Chlor- pheniramine Maleate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Chlor-pheniramine Maleate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 3 mL of the sam-ple solution, and add the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak other than maleic acid and chlorpheniramine obtained from the sample solu-tion is not larger than 2/3 times the peak area of chlor-pheniramine from the standard solution, and the total area of the peaks other than maleic acid and chlorpheniramine from the sample solution is not larger than the peak area of chlorpheniramine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 8.57 g of ammonium dihydrogen-phosphate and 1 mL of phosphoric acid in water to make 1000 mL. To 800 mL of this solution add 200 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of chlor-pheniramine is about 11 minutes. Time span of measurement: About 4 times as long as the retention time of chlorpheniramine, beginning after the sol-vent peak. System suitability— Test for required detectability: To exactly 2.5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of chlorpheniramine ob-tained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of chlorpheniramine are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of chlorpheniramine is not more than 4.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 0.4 g of Chlorpheniramine Maleate, previously dried and accurately weighed, in 20 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-green to green (indicator: 2 drops of crystal violet TS). ‌JP XVIII Official Monographs / Chlorpheniramine Maleate Powder 741‌ Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.54 mg of C H ClN .C H O lution. Determine the absorbances AT and AS of the sample solution and standard solution at a wavelength of the maxi-mum absorbance at about 265 nm as directed under Ultravi-olet-visible Spectrophotometry <2.24>. 16 19 2 4 4 4
Chlorpromazine Hydrochloride occurs as a white to pale yellow crystalline powder. It is odorless, or has a faint, characteristic odor. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), sparingly soluble in acetic anhy-dride, and practically insoluble in diethyl ether. It is gradually colored by light.
(1) To 5 mL of a solution of Chlorproma-zine Hydrochloride (1 in 1000) add 1 drop of iron (III) chlo-ride TS: a red color develops. Dissolve 0.1 g of Chlorpromazine Hydrochloride in JP XVIII Official Monographs / Chlorpromazine Hydrochloride Tablets 745 ‌20 mL of water and 3 drops of dilute hydrochloric acid, add 10 mL of 2,4,6-trinitrophenol TS, and allow to stand for 5 hours. Collect the resulting precipitate, wash with water, recrystallize from a small portion of acetone, and dry at 1059C for 1 hour: the crystals so obtained melt <2.60> be-tween 1759C and 1799C.‌ Dissolve 0.5 g of Chlorpromazine Hydrochloride in 5 mL of water, add 2 mL of ammonia TS, and heat on a water bath for 5 minutes. Cool, filter, and render the filtrate acidic with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—A solution of 1.0 g of Chlorpromazine Hydrochloride in 20 mL of water, when observed within 10 minutes, is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Chlor- promazine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Transfer an exactly measured volume of Chlor-promazine Hydrochloride Injection, equivalent to about pH <2.54> Dissolve 1.0 g of Chlorpromazine Hydrochlo- ride in 20 mL of freshly boiled and cooled water, and meas- 0.15 g of chlorpromazine hydrochloride (C 17H 19ClN2S.HCl) ure within 10 minutes: the pH of this solution is between 4.0 and 5.0. Purity (1) Clarity and color of solution—A solution of 1.0 g of Chlorpromazine Hydrochloride in 20 mL of water, when observed within 10 minutes, is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Chlor- promazine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.7 g of Chlorpromazine Hydrochloride, previously dried, dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.53 mg of C17H19ClN2S.HCl
Chlorpropamide, when dried, contains not less than 98.
Chlorpropamide occurs as white, crystals or crystalline powder. It is freely soluble in methanol and in acetone, soluble in ethanol (95), and slightly soluble in diethyl ether, and practi-cally insoluble in water.
(1) Dissolve 0.08 g of Chlorpropamide in 50 mL of methanol. To 1 mL of the solution add 0.01 mol/L hydrochloric acid TS to make 200 mL. Determine the ab-sorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Chlorpropamide, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. Perform the test with Chlorpropamide as directed under Flame Coloration Test <1.04> (2): a green color ap-pears.
(1) Acidity—To 3.0 g Chlorpropamide add 150 mL of water, and warm at 709C for 5 minutes. Allow to stand in ice water for 1 hour, and filter. To 25 mL of the filtrate add 2 drops of methyl red TS and 0.30 mL of 0.1 mol/L sodium hydroxide VS: a yellow color develops. Chloride <1.03>—To 40 mL of the filtrate obtained in (1) add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.25 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.011z). Sulfate <1.14>—To 40 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.021z). Heavy metals <1.07>—Proceed with 2.0 g of Chlor- propamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.6 g of Chlor-propamide in acetone to make exactly 10 mL, and use this Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Chlorpropamide, previously dried, dissolve in 30 mL of neutralized ethanol, and add 20 mL of water. Titrate <2.50> with 0.1 mol/L so-dium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 27.67 mg of C10H13ClN2O3S
Cholesterol occurs as white to pale yellow, crystals or grains. It is odorless, or has a slight odor. It is tasteless. It is freely soluble in chloroform and in diethyl ether, spar-ingly soluble in ethanol (99.5) and in acetone, and practically insoluble in water. It gradually changes to a yellow to light yellow-brown color by light.
(1) Dissolve 0.01 g of Cholesterol in 1 mL of chloroform, add 1 mL of sulfuric acid, and shake: a red color develops in the chloroform layer, and the sulfuric acid layer shows a green fluorescence. (2) Dissolve 5 mg of Cholesterol in 2 mL of chloroform, add 1 mL of acetic anhydride and 1 drop of sulfuric acid, and shake: a red color is produced, and it changes to green through blue. D Optical rotation <2.49> [a]25: －29 – －369(after drying, 0.2 g, acetone, 10 mL, 100 mm).
(1) Clarity of solution—Place 0.5 g of Cholesterol in a glass-stoppered flask, dissolve in 50 mL of warm ethanol (95), and allow to stand at room temperature for 2 hours: no turbidity or deposit is produced. (2) Acidity—Place 1.0 g of Cholesterol in a flask, dis-solve in 10 mL of diethyl ether, add 10.0 mL of 0.1 mol/L sodium hydroxide VS, and shake for 1 minute. Expel the diethyl ether, and boil for 5 minutes. Cool, add 10 mL of water, and titrate <2.50> with 0.05 mol/L sulfuric acid VS (indicator: 2 drops of phenolphthalein TS). Perform a blank determination in the same manner. The volume of 0.1 mol/L sodium hydroxide VS consumed is not more than 0.30 mL. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Chotosan Extract contains not less than 24 mg and not more than 72 mg of hesperidin, not less than 6 mg and not more than 18 mg of glycyrrhizic acid (C42H62O16: 822.
Chotosan Extract is a light brown to yellow-brown powder or black-brown viscous extract. It has a slight odor, and has a pungent and slightly sweet first, then bitter taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 20 mL of water and 2 mL of am-monia TS, add 20 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pres-sure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg each of rhyncophylline for thin-layer chromatography and hirsutine for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with one of the two dark purple spots Chotosan Extract / Crude Drugs and Related Drugs from the standard solution (Uncaria Hook). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of hesperidin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, acetone, water and acetic acid (100) (10:6:3:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and allow to stand in an ammonia gas: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (Citrus Unshiu Peel). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 5 mL of 1-butanol, shake, centrifuge, remove the 1-butanol layer, and use the aqueous layer as the sample solution. Separately, heat 3.0 g of pulver-ized ophiopogon root in 50 mL of water under a reflux con-denser for 1 hour. After cooling, shake 20 mL of the extract with 5 mL of 1-butanol, centrifuge, remove the 1-butanol layer, and use the aqueous layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL of the sample solu-tion and 5 mL of the standard solution as bands on the origi-nal line on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethanol (99.5), water and acetic acid (100) (120:80:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzal-dehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark blue-green spot (around Rf value 0.3) from the standard solution (Ophiopogon Root). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of 4?-O-glycosyl-5-O-methylvisamminol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (Saposhnikovia Root and Rhizome). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 20 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of luteolin for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 3 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, hexane and formic acid (5:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yel-low-brown spot from the standard solution (Chrysanthe-mum Flower). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple spot from the standard solution (Ginger). Shake 1.0 g of a dry extract (or 3.0 g of the viscous extract) with 30 mL of methanol, centrifuge, and separate the supernatant liquid. Shake the residue with 30 mL of water, centrifuge, and separate the supernatant liquid. Add ammonium oxalate TS to this solution: a white precipitate is formed, and it does not dissolve by addition of dilute acetic acid, but it dissolve by addition of dilute hydrochloric acid. (Gypsum) JP XVIII Crude Drugs and Related Drugs / Chotosan Extract 1979
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 7.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Com-bine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Total ash <5.01> Not more than 15.0z, calculated on the dried basis. Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16)
(1) Hesperidin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.1 g of dried substance), add exactly 50 mL of diluted tetrahydrofuran (1 in 4), shake for 30 minutes, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 10 mg of hesperidin for assay, previously dried in a desiccator (silica gel) for 24 hours, dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add diluted tetrahydrofuran (1 in 4) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of hesperidin in each solution. MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr- Amount (mg) of hesperidin ＝ MS × AT/AS × 1/20 rhizic acid is about 15 minutes). System suitability— MS: Amount (mg) of hesperidin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (82:18:1). Flow rate: 1.0 mL per minute (the retention time of hesperidin is about 15 minutes). System suitability— System performance: Dissolve 1 mg each of hesperidin for assay and naringin for thin-layer chromatography in diluted methanol (1 in 2) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, naringin and hesperidin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hesperidin is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as de-scribed above, and remove the ethyl acetate layer. To the resultant aqueous layer add 10 mL of methanol, shake for 30 System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Total alkaloid (rhyncophylline and hirsutine)—Weigh accurately about 1 g of the dry extract (or an amount of the viscous extract, equivalent to about 1 g of dried sub-stance), add 20 mL of diethyl ether, shake, add 3 mL of 1 mol/L hydrochloric acid TS and 7 mL of water, shake for 10 minutes, centrifuge, and remove the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, and repeat the above process. To the aqueous layer add 10 mL of sodium hydroxide TS and 20 mL of diethyl ether, shake for 10 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 20 mL of diethyl ether, proceed in the same manner, and repeat the procedure twice. Combine all the ex-tracts, evaporate the solvent under low pressure (in vacuo) at not more than 409C, and dissolve the residue in the mobile phase to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 5 mg of rhyncophylline for assay and about 5 mg of hirsutine for assay, and dissolve in a mixture of methanol and dilute acetic acid (7:3) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of methanol and dilute acetic acid (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each Chrysanthemum Flower / Crude Drugs and Related Drugs ‌of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas of rhyn-cophylline and hirsutine, ATR and ATH, and ASR and ASH, in each solution.‌ Amount (mg) of the total alkaloid (rhyncophylline and hirsutine) ＝ MSR × ATR/ASR × 1/50 ＋ MSH × ATH/ASH × 1/50 Odor, characteristic; taste, slightly bitter. Identification To 1 g of pulverized Chrysanthemum Flower add 20 mL of methanol, shake for 10 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 1 mL of methanol, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of luteolin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu- MSR : Amount (mg) of rhyncophylline for assay taken tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu- MSH: Amount (mg) of hirsutine for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 5 g of sodium lauryl sulfate in 1150 mL of acetonitrile and 1350 mL of water, mix with 1 mL of phosphoric acid. Flow rate: 1.0 mL per minute (the retention time of rhyn-cophylline is about 12 minutes and that of hirsutine is about 27 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rhyncophylline and hirsutine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of rhyncophylline and hirsutine are not more than 1.5z, respectively.
Cibenzoline Succinate, when dried, contains not less than 98.
Cibenzoline Succinate occurs as a white crys-talline powder. It is freely soluble in methanol and in acetic acid (100), and sparingly soluble in water and in ethanol (99.5). A solution of Cibenzoline Succinate in methanol (1 in 10) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Cibenzoline Succinate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cibenzoline Succinate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Shake 0.4 g of Cibenzoline Succinate with 2.5 mL of sodium hydroxide TS and 5 mL of ethyl acetate, allow to stand, and to 1 mL of the water layer so obtained add 0.5 mL of 1 mol/L hydrochloric acid TS and 0.5 mL of iron (III) chloride TS: a blown precipitate is formed.
(1) Clarity and color of solution—A solution ob-tained by dissolving 0.20 g of Cibenzoline Succinate in 10 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Cibenzo- line Succinate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cibenzoline Succinate according to Method 3, using a so-lution of magnesium nitrate hexahydrate in ethanol (95) (1 in 25), and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Cibenzoline Succinate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 5 mL and 2 mL of this solution, add methanol to make them exactly 10 mL, and use these solutions as the standard solution (1) and the standard solution (2). Perform the test with these solutions and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (20:3:2) to a distance of about 10 cm, air-dry the plate, and dry more at 809C for 30 minutes. After cooling, examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solu-tion is not more intense than the spot from the standard so-lution (1). Allow the plate to stand for 30 minutes in iodine vapor: the spot other than the principal spot from the sample solution is not more intense than the spot from the standard solution (1), and the spot, which is more intense than the spot from the standard solution (2), is not more than two. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Cibenzoline Suc-cinate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from violet to blue-green through blue (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.04 mg of C18H18N2.C4H6O4
Ciclacillin contains not less than 920 mg (potency) and not more than 1010 mg (potency) per mg, calcu-lated on the anhydrous basis.
Ciclacillin occurs as white to light yellow-white crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, and practically insoluble in acetonitrile and in ethanol (99.5).
Determine the infrared absorption spectrum of Ciclacillin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ciclacillin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋300 – ＋3159(2 g, water, Amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4) ＝ MS × QT/QS MS: Amount (mg) of cibenzoline succinate for assay taken Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl parahydroxybenzoate in methanol to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.67 g of sodium di-2-ethylhexyl 100 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ciclacillin according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of Ciclacillin according to Method 3, and perform the test (not more than 2 ppm). Water <2.48> Not more than 2.0z (1 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Ciclacillin and Ciclacillin RS, equivalent to about 50 mg (potency), dissolve each in a suitable amount of the mobile phase, add exactly 5 ‌mL of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ciclacillin to that of the internal standard. culated on the dried basis. Description Ciclosporin occurs as a white powder. It is very soluble in acetonitrile, in methanol and in ethanol (95), freely soluble in diethyl ether, and practically insoluble in water. Identification Determine the infrared absorption spectrum Amount [ mg (potency)] of ciclacillin (C ＝ MS × QT/QS × 1000 15H23N3O4S) of Ciclosporin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec- MS: Amount [mg (potency)] of Ciclacillin RS taken Internal standard solution—A solution of orcin in the mo-bile phase (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.771 g of ammonium acetate in about 900 mL of water, adjust the pH to 4.0 with acetic acid (100), and add water to make 1000 mL. To 850 mL of this solution add 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of ciclacillin is about 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ciclacillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak areas of ciclacillin to that of the internal stand-ard is not more than 1.0z.
Ciclosporin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 2 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard soluton as directed un-der Liquid Chromatography <2.01> according to the follow-ing conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than the ciclosporin obtained from the sample solution is not larger than 7/10 times the peak area of ciclosporin from the standard solution, and the total area of all peaks other than the ciclosporin from the sample solution is not larger than 1.5 times the peak area of ciclosporin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of ciclosporin, beginning after the solvent peak. System suitability— C62H N11O 12: 1202.61 System performance: Proceed as directed in the system suitability in the Assay. cyclo{-[(2S,3R,4R,6E )-3-Hydroxy-4-methyl-2- methylaminooct-6-enoyl]-L-2-aminobutanoyl- N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl- L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl-} [59865-13-3] Ciclosporin contains not less than 98.
(1) Clarity and color of solution—Dissolve 1.0 g of Ciclosporin in 10 mL of ethanol (95): the solution is clear, and has no more color than the following control solutions (1), (2) or (3). Control solution (1): To exactly 3.0 mL of Iron (III) Chlo-ride CS and exactly 0.8 mL of Cobalt (II) Chloride CS add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. Control solution (2): To exactly 3.0 mL of Iron (III) Chlo-ride CS, exactly 1.3 mL of Cobalt (II) Chloride CS and ex-actly 0.5 mL of Copper (II) Sulfate CS add diluted hydro- chloric acid (1 in 40) to make exactly 100 mL. Control solution (3): To exactly 0.5 mL of Iron (III) Chlo-ride CS and exactly 1.0 mL of Cobalt (II) Chloride CS add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. Heavy metals <1.07>—Proceed with 1.0 g of Ciclosporin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 2 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard soluton as directed un-der Liquid Chromatography <2.01> according to the follow-ing conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak other than the ciclosporin obtained from the sample solution is not larger than 7/10 times the peak area of ciclosporin from the standard solution, and the total area of all peaks other than the ciclosporin from the sample solution is not larger than 1.5 times the peak area of ciclosporin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of ciclosporin, beginning after the solvent peak. System suitability— C62H N11O 12: 1202.61 System performance: Proceed as directed in the system suitability in the Assay. cyclo{-[(2S,3R,4R,6E )-3-Hydroxy-4-methyl-2- methylaminooct-6-enoyl]-L-2-aminobutanoyl- N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl- L-leucyl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl-} [59865-13-3] Ciclosporin contains not less than 98.5z and not Test for required detectability: To exactly 2 mL of the standard solution add a mixture of water and acetonitrile (1:1) to make exactly 20 mL. Confirm that the peak area of ciclosporin obtained from 20 mL of this solution is equiva-lent to 7 to 13z of that of ciclosporin obtained from 20 mL of the standard solution. System repeatability: When the test is repeated 6 times more than 101.5z of ciclosporin (C 62H111 N11 O12 ), cal- with 20 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak ‌JP XVIII Official Monographs / Cilastatin Sodium 753 area of ciclosporin is not more than 3.0z. Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 30 mg each of Ciclosporin and Ciclosporin RS (separately determine the loss on drying <2.41> under the same conditions as Ciclosporin), and dis- solve each in a mixture of water and acetonitrile (1:1) to make exactly 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ciclosporin in each solution. Amount (mg) of ciclosporin (C62H111N11O12) ＝ MS × AT/AS MS: Amount (mg) of Ciclosporin RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Con-nect the sample injection port and the column with a stain-less steel tube 0.3 mm in inside diameter and 1 m in length. Column temperature: A constant temperature of about 809C (including the sample injection port and the connecting tube). Mobile phase: A mixture of water, acetonitrile, tert-butyl methyl ether and phosphoric acid (520:430:50:1). Flow rate: Adjust so that the retention time of ciclosporin is about 27 minutes. System suitability— System performance: Dissolve 3 mg of Ciclosporin U in 2.5 mL of a mixture of water and acetonitrile (1:1), and add 2.5 mL of the standard solution. When the procedure is run with 20 mL of this solution under the above operating condi-tions, ciclosporin U and ciclosporin are eluted in this order with the resolution between these peaks being not less than 1.2. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ciclosporin is not more than 1.0z.
Cilazapril Hydrate contains not less than 98.
Cilazapril Hydrate occurs as white to yellowish white, crystals or crystalline powder. It is very soluble in methanol, freely soluble in ethanol (99.5) and in acetic acid (100), and slightly soluble in water. It gradually turns yellow on exposure to light. Melting point: about 1019C (with decomposition). with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, acetic acid (100), hexane, and water (62:15:10:10:3) to a dis-tance of about 15 cm, and air-dry the plate. Leave the plate in iodine vapor for 2 hours, and examine the plate under ul-traviolet light (main wavelength: 254 nm): of the spots other than the principal spot with an Rf value close to 0.40 ob-tained from the sample solution, the spot in the vicinity of Rf value 0.17 is not more intense than the spot from the standard solution (1), and the spot in the vicinity of Rf value 0.44 is not more intense than the spot from the standard so-lution (2). The number of all other spot does not exceed 3, and of these spots, no more than one is more intense than the spot from the standard solution (3) and none are more intense than the spot from the standard solution (2). Water <2.48> 3.5 – 5.0z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
(1) To 4 mL of a solution of Cilazapril Hy-drate (1 in 1000) add 2 mL of Dragendorff's TS: an orange precipitate is produced. (2) Determine the infrared absorption spectrum of Cilazapril Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －53 – －589(0.2 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm).
Weigh accurately about 0.2 g of Cilazapril Hydrate, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.02 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L perchloric acid VS ＝ 8.350 mg of C H N O 22 31 3 5 Identification (1) To 4 mL of a solution of Cilazapril Hy-drate (1 in 1000) add 2 mL of Dragendorff's TS: an orange precipitate is produced. (2) Determine the infrared absorption spectrum of Cilazapril Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －53 – －589(0.2 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm).
Cilnidipine, when dried, contains not less than 98.
Cilnidipine occurs as a faint yellow crystalline powder. It is freely soluble in acetonitrile, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. A solution of Cilnidipine in acetonitrile (1 in 100) shows no optical rotation. It is gradually colored to reddish yellow and decomposed by light.
(1) Determine the absorption spectrum of a solution of Cilnidipine in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cilnidipine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of previ-ously dried Cilnidipine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Cilnidipine RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cilnidipine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Cilnidipine in 20 mL of acetonitrile, add the mobile phase to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.5 to cilnidipine, obtained from the sample solution is not larger than 2/5 times the peak area of cilnidipine from the standard solution, the area of the peaks other than cil-nidipine and the above mentioned peak from the sample so-lution is not larger than 1/5 times the peak area of cilnidi-pine from the standard solution, and the total area of the peaks other than cilnidipine from the sample solution is not larger than the peak area of cilnidipine from the standard so-lution. For the area of the peak, having the relative retention time of about 1.15, about 1.6, and about 1.7 to cilnidipine, multiply the correction factor, 1.5, 1.4, and 1.6, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cilnidipine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 50 mL. Con-firm that the peak area of cilnidipine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilnidipine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1.0 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 50 mg each of Cilnidipine and Cil-nidipine RS, both previously dried, dissolve in 20 mL of acetonitrile, and add the mobile phase to make exactly 100 mL, respectively. Pipet 5 mL each of these solutions, add ex-actly 5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilnidipine to that of the in-ternal standard. Amount (mg) of cilnidipine (C27H28N2O7) ＝ MS × QT/QS MS: Amount (mg) of Cilnidipine RS taken Internal standard solution—A solution of butyl parahy- ‌758 droxybenzoate in acetonitrile (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with perfluorohexylpropyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate in water to make 1000 mL, and adjust to pH 5.5 with diluted acetic acid (100) (1 in 100). To 400 mL of this solution add 600 mL of methanol. Flow rate: Adjust so that the retention time of cilnidipine is about 20 minutes. System suitability— System performance: After exposing Cilnidipine to a fluo-rescent light (15,000 lx･h), take 10 mg, dissolve in 4 mL of acetonitrile, and add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak of cilnidipine and the peak having the relative retention time of about 1.07 to cilnidipine is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cilnidipine to that of the internal standard is not more than 1.0z.
Cilostazol, when dried, contains not less than 98.
Cilostazol occurs as white to pale yellow-white, crystals or crystalline powder. It is slightly soluble in methanol, in ethanol (99.5) and in acetonitrile, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Cilostazol in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cilostazol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cilostazol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the Amount (mg) of cilnidipine (C 27H28N2O7) ＝ M S × QT/QS spectrum with the Reference Spectrum or the spectrum of Cilostazol RS: both spectra exhibit similar intensities of ab- MS: Amount (mg) of Cilnidipine RS taken sorption at the same wave numbers.
Weigh accurately about 50 mg each of Cilostazol and Cilostazol RS, previously dried, dissolve each in a suita-ble amount of methanol, add exactly 5 mL of the internal standard solution and methanol to make 50 mL. To 1 mL each of these solutions add methanol to make 10 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilostazol to that of the internal standard. Amount (mg) of cilostazol (C20H27N5O2) ＝ MS × QT/QS MS: Amount (mg) of Cilostazol RS taken Internal standard solution—A solution of benzophenone in methanol (1 in 250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and metha-nol (10:7:3). Flow rate: Adjust so that the retention time of cilostazol is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cilostazol and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cilostazol to that of the internal standard is not more than 1.0z.
Cimetidine, when dried, contains not less than 99.
Cimetidine occurs as a white crystalline pow-der. It is odorless, and has a bitter taste. It is freely soluble in methanol and in acetic acid (100), sparingly soluble in ethanol (95), slightly soluble in water, and practically insoluble in diethyl ether. It dissolves in dilute hydrochloric acid. It is gradually colored by light.
(1) To 0.1 mL of a solution of Cimetidine in ethanol (95) (1 in 100) add 5 mL of citric acid-acetic anhy-dride TS, and heat in a water bath for 15 minutes: a red-purple color develops. (2) Determine the infrared absorption spectrum of Cimetidine, previously dried, as directed in the potassium bromide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 0.5 g of Cimetidine in 50 mL of freshly boiled and cooled water, shake for 5 minutes and filter: the pH of the filtrate is between 9.0 and 10.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Cimetidine in 10 mL of methanol: the solution is clear and colorless to pale yellow in color. Heavy metals <1.07>—Proceed with 2.0 g of Cimeti- dine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Amount (mg) of cilostazol (C 20H27N5O2) ＝ M S × QT/QS Arsenic <1.11>—Dissolve 1.0 g of Cimetidine in 5 mL of dilute hydrochloric acid, and perform the test with this so- ‌762 lution (not more than 2 ppm). ‌Related substances—Dissolve 0.5 g of Cimetidine in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add meth-anol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 4 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and am-monia solution (28) (21:2:2) to a distance of about 15 cm, air-dry the plate, and then dry at 809C for 30 minutes. Allow the plate to stand in iodine vapor for 45 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.24 g of Cimetidine, previ-ously dried, dissolve in 75 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 25.23 mg of C10H16N6S
Dibucaine Hydrochloride occurs as white, crystals or crystalline powder. It is very soluble in water, in ethanol (95) and in acetic acid (100), freely soluble in acetic anhydride, and practically in-soluble in diethyl ether. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Dibucaine Hydrochloride in 1 mol/L hydrochlo-ric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dibucaine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Dibucaine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Dibucaine Hydrochloride in 50 mL of water: the pH of this solution is between 5.0 and 6.0. nation, and dry in vacuum over phosphorus (V) oxide at 809C for 5 hours. Seal immediately the open end of the tube, and determine the melting point.
(1) Clarity and color of solution—Dissolve 1.0 g of Dibucaine Hydrochloride in 20 mL of water: the solution is clear and colorless. Determine the absorbance of this solu-tion at 430 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>, using water as the blank: it is not more than 0.03. Sulfate <1.14>—Perform the test with 0.30 g of Dibucaine Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.056z). Heavy metals <1.07>—Proceed with 1.0 g of Dibucaine Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Dibucaine Hydrochloride in 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 20 mL, then pipet 2 mL of this solution, add ethanol (95) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let (main wavelength: 254 nm): the spots other than the prin-cipal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, phosphorus (V) oxide, 809C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Dibucaine Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.00 mg of C20H29N3O2.HCl
Cinoxacin, when dried, contains not less than 98.
Cinoxacin occurs as a white to pale yellow crystalline powder. It is odorless or has a slight, characteris-tic odor. It has a bitter taste. It is slightly soluble in N,N-dimethylformamide and in acetone, very slightly soluble in ethanol (99.5), and practi-cally insoluble in water. It dissolves in sodium hydroxide TS. Melting point: about 2659C (with decomposition).
(1) Dissolve 30 mg of Cinoxacin in 10 mL of dilute sodium hydroxide TS, and add water to make 100 mL. To 1 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref- erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cinoxacin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—Dissolve 0.20 g of Cinoxacin in 10 mL of dilute sodium hydroxide TS, add 20 mL of 0.1 mol/L hydrochloric acid TS, shake, filter, and add water to the filtrate to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.20 mL of 0.005 mol/L sulfuric acid VS by adding 10 mL of dilute sodium hydroxide TS, 20 mL of 0.1 mol/L hydrochlo-ric acid TS, and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Cinoxa- cin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 10 mg of Cinoxacin in 10 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of acetonitrile, water and ammonia solution (28) (14:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.4 g of Cinoxacin, previ-ously dried, add 60 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and dissolve by warming. After cool-ing, titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.22 mg of C12H10N2O5
シプロフロキサシン塩酸塩水和物
Ciprofloxacin Hydrochloride Hydrate contains not less than 98.
Ciprofloxacin Hydrochloride Hydrate occurs as a white to pale yellow crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, and very slightly soluble in ethanol (99.5). It is gradually colored to a slightly brownish light yellow by light.
(1) Determine the infrared absorption spec-trum of Ciprofloxacin Hydrochloride Hydrate, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Ciprofloxacin Hydrochloride Hydrate in 5 mL of water, and use this solution as the sample solution. Separately, dissolve 45 mg of Ciprofloxacin RS in 5 mL of ammonia TS, and use this solution as the standard solution. Perform the test with these solutions, as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. After allowing to stand the plate in the vapor of ammonia for 15 minutes, develop the plate with a mixture of metha-nol, dichloromethane, ammonia solution (28) and aceto-nitrile (4:4:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot obtained from the standard solution show the same Rf value. A solution of Ciprofloxacin Hydrochloride Hydrate (1 in 500) responds to Qualitative Tests <1.09> for chloride.
(1) Sulfate <1.14>—Perform the test with 0.5 g of Ciprofloxacin Hydrochloride Hydrate. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Ciprofloxacin Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 1.0 ml of Standard Lead Solution (not more than 10 ppm). Fluoroquinolonic acid—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Ciprofloxacin Hydrochloride Hydrate in water to make exactly 5 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of fluoroquinolonic acid for thin-layer chromatogra- phy in 0.1 mL of ammonia TS and water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. After allowing to stand the plate in the vapor of ammonia for 15 minutes, develop the plate with a mixture of methanol, dichloromethane, ammonia solution (28) and acetonitrile (4:4:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution, corresponding to the spot from the standard solution, is not more intense than that from the standard solution. Related substances—Conduct this procedure using light-resistant vessels. Dissolve 25 mg of Ciprofloxacin Hy-drochloride Hydrate in 50 mL of mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample so-lution, add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peaks other than ciprofloxacin obtained from the sample solution is not larger than the peak area of ciprofloxacin from the standard solution, and the total area of the peaks other than ciprofloxacin from the sample solution is not larger than 2.5 times the peak area of ciprofloxacin from the standard solution. For the area of the peaks, having the rela-tive retention times of about 0.4, about 0.5, and about 1.2 to ciprofloxacin, multiply the correction factors, 6.7, 1.3, and 1.4, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of ciprofloxacin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of ciprofloxacin obtained with 50 mL of this solution is equivalent to 20 to 30z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ciprofloxacin are not less than 3500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ciprofloxacin is not more than 2.0z. Water <2.48> 4.7 – 6.7z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 25 mg of Ciprofloxacin Hydrochlo-ride Hydrate, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 22.5 mg of Ciprofloxacin RS, previ- ‌766 ously dried at 1209C in vacuum for 6 hours, add 2 mL of a mixture of water and phosphoric acid (13:1), then add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing operating conditions, and determine the peak areas, AT and AS, of ciprofloxacin in each solution. Amount (mg) of ciprofloxacin hydrochloride (C H FN O .HCl)＝ M × A /A × 1.110 the spectrum with the Reference Spectrum or the spectrum of a solution of Cisplatin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cisplatin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cisplatin RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. 17 18 3 3 S T S A solution of Cisplatin (1 in 2000) responds to MS: Amount (mg) of Ciprofloxacin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 2.88 g of phosphoric acid add water to make 1000 mL, and adjust to pH 3.0 with triethylamine. To 870 mL of this solution add 130 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of ciprofloxa-cin is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ciprofloxacin are not less than 3500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ciprofloxacin is not more than 1.0z.
Ciprofloxacin, when dried, contains not less than 98.
Ciprofloxacin occurs as a white to light yellow-white, crystalline powder. It is practically insoluble in water and in ethanol (99.5). It dissolves in ammonia TS. It is gradually colored to yellow tint by light. Melting point: about 2709C (with decomposition).
(1) Determine the infrared absorption spec-trum of Ciprofloxacin, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of the Ciprofloxacin RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. (2) Conduct this procedure using light-resistant vessels. Dissolve 50 mg each of Ciprofloxacin and Ciprofloxacin RS in 5 mL of ammonia TS, and use these solutions as the sam-ple solution and the standard solution, respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu- tion and the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. After allowing to stand this plate in the vapor of ammonia for 15 minutes, develop the plate with a mixture of methanol, dichloromethane, ammonia solution (28) and acetonitrile (4:4:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value.
(1) Chloride <1.03>—To 1.5 g of Ciprofloxacin add 75 mL of water, and boil for 5 minutes. After cooling, add water to make 75 mL, and filter. To 25 mL of the fil-trate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute sulfuric acid and water to make 50 mL (not more than 0.021z). Heavy metals <1.07>—Proceed with 2.0 g of Ciprofloxacin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Fluoroquinolonic acid—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Ciprofloxacin in ammonia TS to make exactly 5 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of fluoro-quinolonic acid for thin-layer chromatography in 0.1 mL of ammonia TS and water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. After allowing to Time span of measurement: About 2.3 times as long as the retention time of ciprofloxacin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of ciprofloxacin obtained with 50 mL of this solution is equivalent to 20 to 30z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ciprofloxacin are not less than 3500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ciprofloxacin is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (2 g, in vacu-um, 1209C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (2 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 25 mg each of Ciprofloxacin and Ciprofloxacin RS, both dried previously, add 2 mL of a mix-ture of water and phosphoric acid (13:1), add the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of ciprofloxacin in each solution. stand this plate in the vapor of ammonia for 15 minutes, de-velop the plate with a mixture of methanol, dichlorometh-ane, ammonia solution (28) and acetonitrile (4:4:2:1) to a Amount (mg) of ciprofloxacin (C ＝ MS × AT/AS 17H18FN3O3) distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution, corresponding to the spot obtained from the standard solution, is not more in-tense than that obtained from the standard solution. Related substances—Conduct this procedure using light-resistant vessels. To 25 mg of Ciprofloxacin add 2 mL of a mixture of water and phosphoric acid (13:1), then add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 20 mL. Pipet 1 mL of this so-lution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than ciprofloxacin obtained from the sample so-lution is not larger than the peak area of ciprofloxacin from the standard solution, and the total area of the peaks other than ciprofloxacin from the sample solution is not larger than 2.5 times the peak area of ciprofloxacin from the stand-ard solution. For the area of peak, having the relative reten-tion time of about 0.4, about 0.5, and about 1.2 to ciprofloxacin, multiply the correction factor, 6.7, 1.3, and 1.4, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. MS: Amount (mg) of Ciprofloxacin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 2.88 g of phosphoric acid add water to make 1000 mL, and adjust to pH 3.0 with triethylamine. To 870 mL of this solution add 130 mL of acetonitrile. Flow rate: Adjust so that the retention time of ciprofloxa-cin is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ciprofloxacin are not less than 3500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ciprofloxacin is not more than 1.0z.
Cisplatin, when dried, contains not less than 98.
Cisplatin occurs as a yellow crystalline pow-der. It is sparingly soluble in N,N-dimethylformamide, slightly soluble in water, and practically insoluble in ethanol (99.5).
(1) To 5 mL of a solution of Cisplatin (1 in 2000) add 2 to 3 drops of a solution of tin (II) chloride dihy-drate (1 in 100): a brown precipitate is formed. (2) Determine the absorption spectrum of a solution of Cisplatin in a solution of sodium chloride in 0.01 mol/L hy- Qualitative Tests <1.09> (1) for chloride.
Ammonium amminetrichloroplatinate—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Cisplatin in a solution of sodium chloride (9 in 1000) to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of ammonium am-minetrichloroplatinate for liquid chromatography, previ-ously dried at 809C for 3 hours, in the solution of sodium chloride (9 in 1000) to make exactly 200 mL. Pipet 2 mL of this solution, add the solution of sodium chloride (9 in 1000) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak area of ammonium am-minetrichloroplatinate by the automatic integration method: the peak area obtained from the sample solution is not larger than that from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 209 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel for liquid chromatography having quaternary ammonium groups (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A solution of ammonium sulfate (1 in 800). Flow rate: Adjust so that the retention time of ammonium amminetrichloroplatinate is about 8 minutes. System suitability— System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ammonium amminetrichloroplatinate are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ammonium amminetrichloroplatinate is not more than 3.0z. Loss on drying <2.41> Not more than 0.1z (1 g, 1059C, 4 hours).
Conduct this procedure using light-resistant vessels. Weigh accurately about 25 mg each of Cisplatin and Cisplatin RS, previously dried, dissolve in N,N-dimethylfor-mamide to make exactly 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cisplatin in each solution. drochloric acid TS (9 in 1000) (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare Amount (mg) of cisplatin (Cl2H6N2Pt) ＝ M S × AT/AS ‌JP XVIII Official Monographs / Citicoline 767 MS: Amount (mg) of Cisplatin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 310 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with aminopropylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of ethyl acetate, methanol, water and N,N-dimethylformamide (25:16:5:5). Flow rate: Adjust so that the retention time of cisplatin is about 4 minutes. System suitability— System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cisplatin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cisplatin is not more than 1.0z.
Citicoline occurs as a white crystalline powder. It is very soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS.
(1) Determine the absorption spectrum of a solution of Citicoline in 0.01 mol/L hydrochloric acid TS (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Citicoline RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Citicoline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Citicoline RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Amount (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 10.32 M: Amount (mg) of Citicoline taken, calculated on the dried basis Related substances—Dissolve 0.10 g of Citicoline in water to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peaks other than citicoline obtained from the sample solution is not larger than 3/5 times the peak area of citicolins from the standard solution, and the total area of the peaks other than citicoline from the sample solution is not larger than the peak area of citicoline from the standard solution. For the area of the peaks, hav-ing the relative retention times of about 0.62, about 0.64 and about 1.3 to citicoline, multiply the correction factors, 1.2, 0.7 and 0.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of citicoline. System suitability— Test for required detectability: Pipet 4 mL of the standard solution, and add water to make exactly 50 mL. Confirm that the peak area of citicoline obtained with 10 mL of this solution is equivalent to 5.6 to 10.4z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- ‌768 ditions, the number of theoretical plates and the symmetry factor of the peak of citicoline are not less than 2000 and 0.9 to 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of citicoline is not more than 2.0z. Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, phosphorus (V) oxide, 1009C, 4 hours).
(1) Clarity and color of solution—Dissolve 1.0 g of Citicoline in 8 mL of water: the solution is clear and col-orless. Heavy metals <1.07>—Proceed with 2.0 g of Citico- line according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Citicoline according to Method 4, and perform the test (not more than 2 ppm). Free phosphoric acid—Weigh accurately about 0.1 g of Citicoline, dissolve in water to make exactly 10 mL, and use this solution as the sample solution. Separately, pipet 4 mL of Standard Phosphoric Acid Solution, add water to make exactly 10 mL, and use this solution as the standard solution. To each of the sample solution and the standard solution, add exactly 1 mL of hexaammonium heptamolyb-date-sulfuric acid TS and exactly 0.5 mL of 1-amino-2-naphthol-4-sulfonic acid TS, and after shaking, allow to stand for 30 minutes at 20 ± 19C. To exactly 2 mL each of these solutions add water to make exactly 10 mL, and deter-mine the absorbances, AT and AS, of the solutions obtained from the sample solution and the standard solution at 730 nm as directed under Ultraviolet-visible Spectrometry <2.24>, using the solution, obtained by proceeding with 10 mL of water in the same manner as the sample solution, as the blank. The amount of free phosphoric acid is not more than 0.1z. C14H26N4O11P2: 488.32 P?-[2-(Trimethylammonio)ethyl] cytidine 5?-(monohydrogen diphosphate) [987-78-0] Citicoline contains not less than 98.0z and not more than 102.0z of citicoline (C14H26N4O11P2), cal-culated on the dried basis. Description Citicoline occurs as a white crystalline powder. It is very soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. Identification (1) Determine the absorption spectrum of a solution of Citicoline in 0.01 mol/L hydrochloric acid TS (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Citicoline RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Citicoline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Citicoline RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Amount (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 10.32 M: Amount (mg) of Citicoline taken, calculated on the dried basis Related substances—Dissolve 0.10 g of Citicoline in water to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peaks other than citicoline obtained from the sample solution is not larger than 3/5 times the peak area of citicolins from the standard solution, and the total area of the peaks other than citicoline from the sample solution is not larger than the peak area of citicoline from the standard solution. For the area of the peaks, hav-ing the relative retention times of about 0.62, about 0.64 and about 1.3 to citicoline, multiply the correction factors, 1.2, 0.7 and 0.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of citicoline. System suitability— Test for required detectability: Pipet 4 mL of the standard solution, and add water to make exactly 50 mL. Confirm that the peak area of citicoline obtained with 10 mL of this solution is equivalent to 5.6 to 10.4z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- ‌768 ditions, the number of theoretical plates and the symmetry factor of the peak of citicoline are not less than 2000 and 0.9 to 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of citicoline is not more than 2.0z. Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, phosphorus (V) oxide, 1009C, 4 hours).
Weigh accurately about 0.1 g of Citicoline, and dis-solve in water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 25 mg of Citicoline RS (separately determine the loss on drying <2.41> under the same conditions as Citicoline), and dissolve in water to make exactly 25 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of citicoline in each solution. Amount (mg) of citicoline (C14H26N4O11P2) ＝ MS × AT/AS × 4 MS: Amount (mg) of Citicoline RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: Combine 2 stainless steel columns (4 mm in in-side diameter and 25 cm in length) packed with strongly bas-ic ion exchange resin for liquid chromatography (10 mm in particle diameter) in series. Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 8.17 g of potassium dihydrogen phosphate in water to make 1000 mL. Adjust the pH of this solution to 3.5 with phosphoric acid. Flow rate: Adjust so that the retention time of citicoline is about 26 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of citicoline are not less than 2000 and 0.9 to 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of citicoline is not more than 1.0z.
Irregular pieces of pericarp, about 2 mm in thickness; externally yellow-red to dark yellow-brown, with numerous small dents associated with oil sacs; internally white to light grayish yellow-brown; light and brittle in texture. Odor, characteristic aroma; taste, bitter and slightly pun-gent.
To 0.5 g of pulverized Citrus Unshiu Peel add 10 mL of methanol, warm on a water bath for 2 minutes, and filter. To 5 mL of the filtrate add 0.1 g of mag-nesium in ribbon-form and 1 mL of hydrochloric acid, and allow to stand: a red-purple color develops.
Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Citrus Unshiu Peel provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.2 mL.
Weigh accurately about 0.1 g of pulverized Citrus Unshiu Peel, add 30 mL of methanol, heat under a reflux condenser for 15 minutes, centrifuge after cooling, and sepa-rate the supernatant liquid. To the residue add 20 mL of methanol, and proceed in the same manner. Combine the ex-tracts, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of hesperidin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, and dis-solve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of hesperidin in each solution. Amount (mg) of hesperidin ＝ MS × AT/AS × 1/2 MS: Amount (mg) of hesperidin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (82:18:1). Flow rate: 1.0 mL per minute (the retention time of hesperidin is about 15 minutes). System suitability— System performance: Dissolve 1 mg each of hesperidin for assay and naringin for thin-layer chromatography in 10 mL of methanol, and add water to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, naringin and hesperidin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hesperidin is not more than 1.5z.
Citric Acid Hydrate contains not less than 99.
Citric Acid Hydrate occurs as colorless crys-tals, white granules or crystalline powder. It is very soluble in water, and freely soluble in ethanol (99.5). It is efflorescent in dry air.◆
Determine the infrared absorption spectrum of Citric Acid Hydrate, previously dried at 1059C for 2 hours, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 2.0 g of Citric Acid Hydrate in water to make 10 mL: the solution is clear and colorless or has no more color than the following control solutions (1), (2) or (3). Control solution (1): To 1.5 mL of Cobalt (II) Chloride CS and 6.0 mL of Iron (III) Chloride CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Control solution (2): To 2.5 mL of Cobalt (II) Chloride CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Control solution (3): To 0.15 mL of Cobalt (II) Chloride CS, 7.2 mL of Iron (III) Chloride CS and 0.15 mL of Cop-per (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Sulfates—Dissolve 2.0 g of Citric Acid Hydrate in water to make 30 mL, and use this solution as the sample so-lution. Separately, dissolve 0.181 g of potassium sulfate in diluted ethanol (3 in 10) to make exactly 500 mL. Pipet 5 mL of this solution, and add diluted ethanol (3 in 10) to make exactly 100 mL. To 4.5 mL of this solution add 3 mL of a so-lution of barium chloride dihydrate (1 in 4), shake, and allow to stand for 1 minute. To 2.5 mL of this solution add 15 mL of the sample solution and 0.5 mL of acetic acid (31), and allow to stand for 5 minutes: the solution has no more turbidity than the following control solution. (not more than 150 ppm). Control solution: Dissolve 0.181 g of potassium sulfate in water to make exactly 500 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and proceed in the same manner as above using this solution instead of the sample so-lution. Oxalic acid—Dissolve 0.80 g of Citric Acid Hydrate in 4 mL of water, add 3 mL of hydrochloric acid and 1 g of zinc, and boil for 1 minute. After allowing to stand for 2 minutes, take the supernatant liquid, add 0.25 mL of a solu-tion of phenylhydrazinium chloride (1 in 100), heat to boil, and then cool quickly. To this solution add the equal volume of hydrochloric acid and 0.25 mL of a solution of potassium hexacyanoferrate (III) (1 in 20), mix, and allow to stand for 30 minutes: the solution has no more color than the follow-ing control solution prepared at the same time (not more than 360 ppm expressed as oxalic anhydride). Control solution: To 4 mL of a solution of oxalic acid di-hydrate (1 in 10,000) add 3 mL of hydrochloric acid and 1 g of zinc, and proceed in the same manner as the test solution. (4) Heavy metals <1.07>—Proceed with 2.0 g of Citric Acid Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Readily carbonizable substances—Place 1.0 g of Citric Acid Hydrate in a Nessler tube, add 10 mL of sulfuric acid, immediately heat in a 90 ± 19C water bath for 60 minutes, and cool quickly. Compare the color of 2.0 mL each of this solution and Matching Fluid K, using test tubes 12 mm in outside diameter, from a side against white back-ground: the solution is not more colored than the matching fluid. ‌770 Water <2.48> Not less than 7.5z and not more than 9.0z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.55 g of Citric Acid Hy-drate, dissolve in 50 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 1 drop of phenol-phthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 64.04 mg of C6H8O7 ◆Containers and storage Containers—Tight containers.◆ Clarithromycin
Clarithromycin is a derivative of erythromycin. It contains not less than 950 mg (potency) and not more than 1050 mg (potency) per mg, calculated on the anhydrous basis.
Clarithromycin occurs as a white crystalline powder and has a bitter taste. It is soluble in acetone and in chloroform, slightly soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) To 5 mg of Clarithromycin add 2 mL of sulfuric acid, and shake gently: a red-brown color develops. Dissolve 3 mg of Clarithromycin in 2 mL of acetone, and add 2 mL of hydrochloric acid: an orange color develops and changes immediately to red to deep purple. Determine the infrared absorption spectra of Clarithromycin and Clarithromycin RS as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare these spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －96 – －1069(0.25 g calcu-lated on the anhydrous basis, acetone, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Clarithromycin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Weigh accurately about 0.1 g of Clarithromycin, dissolve in the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 10 mg of Clarithromycin RS, dissolve in the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the each peak area by the automatic integration method: the amount of each related substance calculated on the anhydrous basis is not more than 2.0z, and the total amount of them is not more than 5.0z. For these calculations, exclude any peak with an area of less than 0.05z. Amount (z) of each related substance calculated on the anhydrous basis ＝ MS/MT × AT/AS × 100 Total amount (z) of the related substances calculated on the anhydrous basis ＝ MS/MT × SAT/AS × 100 MS: Amount (mg) of Clarithromycin RS taken MT: Amount (mg) of Clarithromycin taken, calculated on the anhydrous basis AS: Peak area of clarithromycin obtained with the stand-ard solution AT: Peak area of each related substance obtained with the sample solution SAT: Total area of the peaks other than clarithromycin obtained with the sample solution Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of the main peak, beginning from 2 minutes after injection of the sample solution. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL, and use this solution as the solution for system suitabil-ity test. Pipet 1 mL of the solution for system suitability test, add the mobile phase to make exactly 10 mL. Pipet 2.5 mL of this solution, add the mobile phase to make exactly 10 mL. Confirm that the peak area of clarithromycin obtained with 10 mL of this solution is equivalent to 0.25 – 0.75z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of clarithromycin are not less than 2500 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of clarithromycin is not more than 3.0z. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (2 g).
Weigh accurately an amount of Clarithromycin and Clarithromycin RS, equivalent to about 50 mg (potency), JP XVIII Official Monographs / Clarithromycin for Syrup 771 ‌and dissolve each in the mobile phase to make exactly 10 mL. Pipet 2 mL each of these solutions, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak methanol, ethyl acetate and acetic acid (100) (90:10:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: the principal spot obtained from the sample solu-tion and the spot from the standard solution show a black-purple color and the same Rf value. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration). area of clarithromycin to that of the internal standard. Amount [mg (potency)] of clarithromycin (C ＝ MS × QT/QS × 1000 38H69NO13) Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Clarithromycin for Syrup in single-dose packages meet the requirement of the Content MS: Amount [mg (potency)] of Clarithromycin RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in the mobile phase (1 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in 3) and acetonitrile (13:7). Flow rate: Adjust so that the retention time of clarithromycin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, clarithromycin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of clarithromycin to that of the internal standard is not more than 2.0z.
Clebopride Malate, when dried, contains not less than 98.
Clebopride Malate occurs as a white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in water, and slightly soluble in ethanol (99.5). A solution of Clebopride Malate in methanol (1 in 25) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Clebopride Malate in methanol (1 in 80000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, Amount [mg (potency)] of clarithromycin (C38 ＝ MS × QT/QS × 1/5 H69NO13) and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. MS: Amount [mg (potency)] of Clarithromycin RS taken Internal standard solution (1)—A solution of butyl parahy-droxybenzoate in the mobile phase (1 in 1000). Internal standard solution (2)—To exactly 1 mL of the inter-nal standard solution (1) add the mobile phase to make exactly 20 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted 0.2 mol/L potassium Determine the infrared absorption spectrum of Cle-bopride Malate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. Perform the test with Clebopride Malate under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—Dissolve 1.0 g of Clebopride Malate in 20 mL of acetic acid (100), add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS by adding 20 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.009z). ‌774 Heavy metals <1.07>—Proceed with 2.0 g of Cle-bopride Malate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Clebopride Malate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 0.2 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of the peaks other than clebopride obtained from the sample solution is not larger than the peak area of clebopride from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.85 g of ammonium acetate in water to make 500 mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. To 400 mL of the fil-trate add 600 mL of methanol. Flow rate: Adjust so that the retention time of clebopride is about 15 minutes. Time span of measurement: About 2 times as long as the retention time of clebopride. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add water to make exactly 100 mL. Con-firm that the peak area of clebopride obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 30 mg Clebopride Malate and 5 mg of propyl parahydroxybenzoate in the mobile phase to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, propyl parahydroxybenzoate and clebopride are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clebopride is not more than 2.5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Clebopride Malate, previously dried, dissolve in 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 50.80 mg of C20H24ClN3O2.C4H6O5
Clemastine Fumarate, when dried, contains not less than 98.
Clemastine Fumarate occurs as a white, crys-talline powder. It is odorless. It is sparingly soluble in methanol and in acetic acid (100), slightly soluble in ethanol (95), very slightly soluble in diethyl ether, and practically insoluble in water.
(1) To 5 mg of Clemastine Fumarate add 5 mL of sulfuric acid, and shake to dissolve: a yellow color de-velops. Slowly drop this solution into 10 mL of water: the yellow color immediately disappears. To 0.01 g of Clemastine Fumarate add 1 mL of fum-ing nitric acid, and evaporate on a water bath to dryness. Then add 2 mL of diluted hydrochloric acid (1 in 2) and 0.2 g of zinc powder, heat for 10 minutes on a water bath, cool, and filter. Add 20 mL of water to the filtrate. The solu-tion responds to Qualitative Tests <1.09> for primary aro-matic amines. To 5 mL of a solution of Clemastine Fumarate (1 in 50,000), add 5 mL of 4-dimethylaminobenzaldehyde TS, and warm for 10 minutes: a red-purple color develops. Perform the test with Clemastine Fumarate as di-rected under Flame Coloration Test <1.04> (2): a green color appears. Dissolve 0.04 g of Clemastine Fumarate and 0.01 g of fumaric acid for thin-layer chromatography in 2 mL each of a mixture of ethanol (95) and water (4:1) by gentle warming, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of isopropyl ether, formic acid and water (90:7:3) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spot with larger Rf value from the sample solution has the same Rf value as the spot from the standard solution. D Optical rotation <2.49> [a]20: ＋16 – ＋189(after drying, 0.1 g, methanol, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Clemastine Fumarate in 10 mL of methanol by warming: the solution is clear and colorless. Heavy metals <1.07>—Perform the test with 1.0 g of Clemastine Fumarate according to Method 2. Prepare the control solution with 2.0 mL of Standard Lead Solution (not ‌JP XVIII Official Monographs / Clindamycin Hydrochloride 775 more than 20 ppm). Arsenic <1.11>—Take 1.0 g of Clemastine Fumarate, prepare the test solution according to Method 3, and per-form the test (not more than 2 ppm). Related Substances—Dissolve 0.10 g of Clemastine Fumarate in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 250 mL, and use this solution as the standard solution (1). Pipet 5 mL of the standard solu-tion (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, methanol and ammonia solution (28) (90:10:1) to a distance of about 15 cm, and air-dry the plate. After spraying evenly Dragendorff 's TS on the plate, immediately spray evenly hydrogen peroxide TS: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and not more than 2 spots from the sample solution are more intense than the spot from the standard solution (2). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.4 g of Clemastine Fumarate, previously dried, dissolved in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS soluble in ethanol (99.5). Identification (1) Determine the infrared absorption spec-trum of Clindamycin Hydrochloride as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Clindamycin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of Clindamycin Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]25: ＋135 – ＋1509(0.5 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Clindamycin Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of clindamycin B, having the relative retention time of about 0.7 to clindamycin, and that of 7-epiclindamy-cin, having the relative retention time of about 0.8, obtained from the sample solution are not larger than 2 times the peak area of clindamycin from the standard solution, the area of the peak other than clindamycin and the peaks mentioned above from the sample solution is not larger than the peak area of clindamycin from the standard solution, and the ＝ 46.00 mg of C21H26 ClNO.C4H4O4 total area of the peaks other than clindamycin from the sam-ple solution is not larger than 4 times the peak area of clin-
Clindamycin Hydrochloride is the hydrochloride of a derivative of lincomycin. It contains not less than 838 mg (potency) and not more than 940 mg (potency) per mg, calculated on the anhydrous basis.
Clindamycin Hydrochloride occurs as white to grayish white, crystals or crystalline powder. It is freely soluble in water and in methanol, and slightly damycin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of clindamycin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of clindamycin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clindamycin are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clindamycin is not more than 2.0z. Water <2.48> Not more than 6.0z (0.3 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Clindamycin Hydro-chloride and Clindamycin Hydrochloride RS, equivalent to about 20 mg (potency), dissolve each in the mobile phase to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the ‌776 test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of clindamycin in each solution. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Clindamycin Hydrochloride Capsules add Amount [mg (potency)] of clindamycin (C ＝ MS × AT/AS × 1000 18H33ClN2O5S) a suitable amount of the mobile phase, shake for 30 minutes, and add the mobile phase to make exactly V mL so that each mL contains 0.75 mg (potency) of Clindamycin Hydrochlo- MS: Amount [mg (potency)] of Clindamycin Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 0.05 mol/L potassium dihydrogen phos-phate TS add 8 mol/L potassium hydroxide TS to adjust the pH to 7.5. To 550 mL of this solution add 450 mL of aceto-nitrile for liquid chromatography. Flow rate: Adjust so that the retention time of clindamy-cin is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clindamycin are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clindamycin is not more than 1.0z.
クリンダマイシンリン酸エステル
Clindamycin Phosphate is a derivative of clindamy- ＝ MS × AT/AS 18 33 2 5 cin. It contains not less than 800 mg (potency) and not MS: Amount [mg (potency)] of Clindamycin Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm).
Clindamycin Phosphate occurs as a white to pale yellow-white crystalline powder. It is freely soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (95).
Determine the infrared absorption spectrum of Clindamycin Phosphate, previously dried at 1009C for 2 hours, as directed in the paste method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Clindamycin Phos-phate RS previously dried at 1009C for 2 hours: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋115 – ＋1309(0.25 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Clindamycin Phosphate in 10 mL of water. The pH of the solution is between 3.5 and 4.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Clindamycin Phosphate in 10 mL of freshly boiled and cooled water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Clin- damycin Phosphate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Clindamycin Phosphate according to Method 4, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.1 g of Clindamycin Phosphate in 100 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak area of clindamycin, having the relative retention time of about 1.8 to clindamycin phosphate, obtained from the sam-ple solution is not larger than 1/2 times the peak area of clin-damycin phosphate from the standard solution, and the total ‌778 area of the peaks other than clindamycin phosphate from the sample solution is not larger than 4 times the peak area of clindamycin phosphate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of clindamycin phosphate, beginning after the solvent peak. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make ex-actly 10 mL. Confirm that the peak area of clindamycin phosphate obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. Water <2.48> Not more than 6.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Clindamycin Phos-phate and Clindamycin Phosphate RS, equivalent to about 20 mg (potency), add exactly 25 mL of the internal standard solution and the mobile phase to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clindamycin phos-phate to that of the internal standard. Amount [ mg (potency)] of clindamycin (C18H33ClN2O5S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Clindamycin Phosphate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (3 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 10.54 g of potassium dihydrogen phosphate in 775 mL of water, adjust the pH to 2.5 with phosphoric acid, and add 225 mL of acetonitrile. Flow rate: Adjust so that the retention time of clindamy-cin phosphate is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, clindamycin phosphate and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of clindamycin phosphate to that of the in-ternal standard is not more than 2.5z.
Clinofibrate occurs as a white to yellowish white powder. It is odorless and has no taste. It is freely soluble in methanol, in ethanol (99.5), in ace-tone and in diethyl ether, and practically insoluble in water. A solution of Clinofibrate in methanol (1 in 20) shows no optical rotation. Melting point: about 1469C (with decomposition).
(1) Determine the absorption spectrum of a solution of Clinofibrate in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Clinofibrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Clinofibrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Clinofibrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Clinofibrate in 10 mL of acetone, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add acetone to make exactly 50 mL. Pipet 5 mL of this solution, add ace-tone to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, cyclohexane and acetic acid (100) (12:5:3) to a dis-tance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 30 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of hexane and 2-propanol (500:3). Flow rate: Adjust so that the retention time of the peak appearing first is about 35 minutes. Selection of column: Proceed with 5 mL of the sample so-lution under the above operating conditions. Use a column giving a complete separation of the three peaks.
Weigh accurately about 0.45 g of Clinofibrate, pre-viously dried, dissolve in 40 mL of ethanol (99.5), add 30 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydrox-ide VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 23.43 mg of C28H36O6
クロベタゾールプロピオン酸エステル
Clobetasol Propionate, when dried, contains not less than 97.
Clobetasol Propionate occurs as a white to pale yellow-white crystalline powder. It is soluble in methanol and in ethanol (99.5), and practi-cally insoluble in water. It gradually turns yellow by light. Melting point: about 1969C (with decomposition). ‌Identification Determine the infrared absorption spectra of Clobetasol Propionate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum or the spectrum of Clobetasol Propionate RS: both spectra exhibit similar in-tensities of absorbance at the same wave numbers. D Optical rotation <2.49> [a]20: ＋109 – ＋1159(after drying, 0.1 g, methanol, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Clobetasol Propionate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 10 mg of Clobetasol Propionate in 100 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 5 mL of the sample solu-tion, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area of these solutions by the automatic integration method: the area of the peak other than clobetasol propionate obtained from the sample solution is not larger than 2/5 times the peak area of clobetasol propionate from the standard solution. Furthermore, the total area of the peaks other than clobetasol propionate from the sample so-lution is not larger than the peak area of clobetasol propionate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of clobetasol propionate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of clobetasol propionate ob-tained with 10 mL of this solution is equivalent to 2.8 to 5.2z of that with 10 mL of the standard solution. System performance: Dissolve 20 mg of Clobetasol Propionate in 20 mL of methanol. To 5 mL of this solution add 10 mL of a solution of beclometasone dipropionate in methanol (1 in 1000), and then add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solu-tion under the above conditions, clobetasol propionate and beclometasone dipropionate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above condi-tions, the relative standard deviation of the peak area of clobetasol propionate is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 10 mg each of Clobetasol Propionate and Clobetasol Propionate RS, both previously dried, dissolve each in the mobile phase, add exactly 100 mL of the internal standard solution, add the mobile phase to make 250 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clobetasol propionate to that of the internal stand-ard. Amount (mg) of clobetasol propionate (C25H32ClFO5) ＝ MS × QT/QS MS: Amount (mg) of Clobetasol Propionate RS taken Internal standard solution—A solution of beclometasone dipropionate in the mobile phase (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.80 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust the pH to 2.5 with phosphoric acid, and then add water to make 1000 mL. To 425 mL of this solution add 475 mL of acetonitrile and 100 mL of methanol. Flow rate: Adjust so that the retention time of clobetasol propionate is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above conditions, clobetasol propionate and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of clobetasol propionate to that of the internal standard is not more than 1.0z.
Clocapramine Hydrochloride Hydrate, when dried, contains not less than 98.
Clocapramine Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in acetic acid (100), sparingly soluble in water and in methanol, slightly soluble in ethanol (95), in chloroform and in isopropylamine, and practically insoluble in acetic anhydride and in diethyl ether. It is gradually colored by light. Melting point: about 2609C (with decomposition, after drying).
(1) To 5 mL of a solution of Clocapramine Hydrochloride Hydrate (1 in 2500) add 1 mL of nitric acid: a blue color develops at first, and rapidly changes to deep blue, and then changes to green to yellow-green. Determine the absorption spectrum of a solution of Clocapramine Hydrochloride Hydrate in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Clocapramine Hydrochloride Hydrate as directed in the po-tassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.1 g of Clocapramine Hydrochloride Hy-drate in 10 mL of water by warming, and after cooling, add 2 mL of ammonia TS, and filter. Acidify the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride.
(1) Sulfate <1.14>—Dissolve 0.5 g of Clocapra-mine Hydrochloride Hydrate in 40 mL of water by warming, after cooling, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Clocapramine Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Clocapramine Hydrochloride Hydrate in 10 mL of a mixture of chloroform and isopropylamine (99:1), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add a mixture of chloroform and isopropyla-mine (99:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, ethyl acetate, methanol and ammonia solution (28) (100:70:40:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> 2.0 – 3.5z (0.5 g, in vacuum at a pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Clocapramine Hy-drochloride Hydrate, previously dried, dissolve in 70 mL of a mixture of acetic anhydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.70 mg of C28H37ClN4O.2HCl
Clofedanol Hydrochloride, when dried, contains not less than 98.
Clofedanol Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in methanol, in ethanol (95) and in acetic acid (100), sparingly soluble in water, and practically insoluble in diethyl ether. A solution of Clofedanol Hydrochloride in methanol (1 in 20) does not show optical rotation. Melting point: about 1909C (after drying, with decompo-sition).
(1) Determine the absorption spectrum of a solution of Clofedanol Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clofedanol Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Clofedanol Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Clofedanol Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.05 g of Clofedanol Hydrochloride in 25 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 3 mL ‌782 each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the total area of the peaks other than clofedanol obtained from the sample solution is not larger than the peak area of clofedanol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.34 g of potassium methanesul-fonate in diluted phosphoric acid (1 in 1000) to make 1000 mL, and to 650 mL of this solution add 350 mL of metha-nol. Flow rate: Adjust so that the retention time of clofedanol is about 9 minutes. Selection of column: Dissolve 0.01 g each of Clofedanol Hydrochloride and ethyl parahydroxybenzoate in methanol to make 100 mL. Proceed with 3 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of clofedanol and ethyl parahy-droxybenzoate in this order with the resolution of these peaks being not less than 4. Detection sensitivity: Adjust the detection sensitivity so that the peak height of clofedanol obtained from 3 mL of the standard solution composes between 20z and 50z of the full scale. Time span of measurement: About three times as long as the retention time of clofedanol, beginning after the solvent peak. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, silica gel, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Clofedanol Hydro-chloride, previously dried, dissolve in 15 mL of acetic acid (100), add 35 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS basis. Description Clofibrate occurs as a colorless or light yellow, clear, oily liquid. It has a characteristic odor and taste, which is bitter at first, and subsequently sweet. It is miscible with methanol, with ethanol (95), with ethanol (99.5), with diethyl ether and with hexane, and prac-tically insoluble in water. It is gradually decomposed by light. Identification (1) Determine the absorption spectrum of a solution of Clofibrate in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1 or the spectrum of a solution of Clofibrate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, determine the absorption spectrum of a solution of Clofibrate in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2 or the spectrum of a solution of Clofibrate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Clofibrate as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Clofibrate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.500 – 1.505 Specific gravity <2.56> d 20: 1.137 – 1.144 Purity (1) Acidity—Dissolve 2.0 g of Clofibrate in 100 mL of neutralized ethanol, and add 1 drop of phenolphthal-ein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: the solution is red in color. Heavy metals <1.07>—Proceed with 2.0 g of Clofibrate according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—To 5.0 g of Clofibrate add 20 mL of nitric acid and 5 mL of sulfuric acid, and heat until white fumes are evolved. After cooling, if necessary, add further 5 mL of nitric acid, heat until white fumes are evolved, and repeat this procedure until the solution is colorless to light yellow. After cooling, add 15 mL of saturated ammonium oxalate solution, and heat again until white fumes are ＝ 32.63 mg of C17H ClNO.HCl evolved. Cool, add water to make 25 mL, use 5 mL of this solution as the test solution, and perform the test.
Weigh accurately about 0.5 g of Clofibrate, add exactly 50 mL of 0.1 mol/L potassium hydroxide-ethanol VS, and heat in a water bath under a reflux condenser with a carbon dioxide absorbing tube (soda-lime) for 2 hours with frequent shaking. Cool, and titrate <2.50> immediately the excess potassium hydroxide with 0.1 mol/L hydrochloric acid VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 24.27 mg of C12H15ClO3
Clomifene Citrate, when dried, contains not less than 98.
Clomifene Citrate occurs as a white to pale yel-low-white powder. It is odorless. It is freely soluble in methanol and in acetic acid (100), sparingly soluble in ethanol (95), and practically insoluble in diethyl ether. It gradually changes in color by light. Melting point: about 1159C
(1) To 2 mL of a solution of Clomifene Citrate in methanol (1 in 200) add 2 mL of Reinecke salt TS: a light red precipitate is produced. Determine the absorption spectrum of a solution of Clomifene Citrate in 0.1 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Clomifene Citrate RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Clomifene Citrate in methanol (1 in 200) responds to Qualitative Tests <1.09> (1) and (2) for ci-trate salt.
(1) Clarity and color of solution—A solution of 1.0 g of Clomifene Citrate in 30 mL of methanol is clear and colorless. (2) Heavy metals <1.07>—Proceed with 2.0 g of Clomi- fene Citrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio To 10 mg of Clomifene Citrate add 10 mL of water and 1 mL of sodium hydroxide TS, and shake to uni-formly disperse. Add 10 mL of ethyl acetate, shake vigor-ously for 5 minutes, allow to stand for 5 minutes, and use the upper layer as the sample solution. Perform the test with 1 mL of the sample solution as directed under Gas Chroma-tography <2.02> according to the following conditions. De-termine the areas of two adjacent peaks, Aa and Ab, having the retention time of about 8 minutes, where Aa is the peak area of shorter retention time and Ab is the peak area of lon-ger retention time: Ab/(Aa ＋ Ab) is between 0.3 and 0.5. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.25 mm in inside diameter and 15 m in length, coated the inside surface with a layer about 0.1 mm thick of dimethylpolysiloxane for gas chroma-tography. Column temperature: A constant temperature of about 2309C. Injection port temperature: A constant temperature of about 2709C. Detector temperature: A constant temperature of about 3009C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of the first peak of clomifene citrate is about 7.5 minutes. Split ratio: 1:50. System suitability— System performance: When the procedure is run with 1 mL of the sample solution under the above operating conditions, the resolution between the two adjacent peaks having the retention time of about 8 minutes is not less than 5. System repeatability: When the test is repeated 6 times with 1 mL of the sample solution under the above operating conditions, the relative standard deviation of the result of Ab/(Aa ＋ Ab) is not more than 1.0z.
Weigh accurately about 1 g of Clomifene Citrate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 59.81 mg of C26H28ClNO.C6H8O7
Clomipramine Hydrochloride, when dried, contains not less than 98.
Clomipramine Hydrochloride occurs as a white to pale yellow, crystalline powder. It is odorless. It is very soluble in acetic acid (100), freely soluble in water, in methanol and in chloroform, soluble in ethanol (95), sparingly soluble in acetic anhydride, slightly soluble in acetone, and practically insoluble in ethyl acetate and in diethyl ether.
(1) Dissolve 3 mg of Clomipramine Hydro-chloride in 1 mL of nitric acid: a deep blue color develops. Determine the absorption spectrum of a solution of Clomipramine Hydrochloride in 0.1 mol/L hydrochloric acid TS (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Take 1 g of Clomipramine Hydrochloride in a sepa-rator, dissolve in 10 mL of water, add 5 mL of sodium hy-droxide TS, and extract with two 30-mL portions of diethyl ether [the water layer is used for Identification (4)]. Combine the diethyl ether extracts, add 20 mL of water, and shake. Take diethyl ether layer, dry with a small portion of anhy-drous sodium sulfate, and filter. Evaporate the combined ex-tracts by warming on a water bath, and proceed the test with the residue as directed under Flame Coloration Test <1.04> (2): a green color appears. The solution neutralized by adding dilute nitric acid to the water layer obtained in (3) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Clomipramine Hydrochloride in 10 mL of water: the pH of this solution is between 3.5 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Clomipramine Hydrochloride in 10 mL of water: the solu-tion is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 2.0 g of Clomipramine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Clomipramine Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Clomipra-mine Hydrochloride in 10 mL of methanol, and use this so- ‌786 lution as the sample solution. Separately, weigh 20 mg of Imipramine Hydrochloride, dissolve in methanol to make exactly 100 mL, and use this solution as the standard solu-tion (1). Then pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of the solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetone and ammonia solution (28) (15:5:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly po-tassium dichromate-sulfuric acid TS on the plate: the spot obtained from the sample solution, corresponding to that from the standard solution (1), is not more intense than the spot from the standard solution (1). Each of the spots other than the principal spot and the spot mentioned above from the sample solution is not more intense than the spot from the standard solution (2). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Clomipramine Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.13 mg of C19H23ClN2.HCl
Clonazepam, when dried, contains not less than 99.
Clonazepam occurs as white to light yellow, crystals or crystalline powder. It is sparingly soluble in acetic anhydride and in acetone, slightly soluble in methanol and in ethanol (95), very slightly soluble in diethyl ether, and practically insoluble in water. It is gradually colored by light. Melting point: about 2409C (with decomposition).
(1) Determine the absorption spectrum of a solution of Clonazepam in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clonazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Clonazepam as directed under Clonazepam Fine Granules
(1) Chloride <1.03>—To 1.0 g of Clonazepam add 50 mL of water, allow to stand for 1 hour with occasional shaking, and filter. Discard the first 20 mL portion of the fil-trate, take the subsequent 20 mL portion of the filtrate, and add 6 mL of dilute nitric acid and water to make 50 mL. Use this solution as the test solution, and perform the test. Pre-pare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.022z). Heavy metals <1.07>—Proceed with 1.0 g of Clonazepam according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.25 g of Clonazepam in 10 mL of acetone, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, then pipet 1 mL of this solution, add acetone to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of nitromethane and acetone (10:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Clonazepam, previ-ously dried, dissolve in 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.57 mg of C15H10ClN3O3
Clonidine Hydrochloride occurs as white, crys-tals or crystalline powder. It is freely soluble in methanol, soluble in water and in ethanol (95), slightly soluble in acetic acid (100), and practi- cally insoluble in acetic anhydride and in diethyl ether.
(1) To 5 mL of a solution of Clonidine Hy-drochloride (1 in 1000) add 6 drops of Dragendorff's TS: an orange precipitate is formed. Determine the absorption spectrum of a solution of Clonidine Hydrochloride in 0.01 mol/L hydrochloric acid TS (3 in 10,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clonidine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Clonidine Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Clonidine Hydrochloride in 20 mL of water: the pH of this solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Clonidine Hydrochloride in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Cloni- dine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.5 g of Clonidine Hydrochloride according to Method 3, and per-form the test (not more than 4 ppm). Related substances—Dissolve 0.20 g of Clonidine Hydrochloride in 2 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 1 mL and 2 mL of this solution, to each add methanol to make exactly 20 mL, and use these solutions as the standard solution (1) and the standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 2 mL each of the sample solution and stand-ard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of tolu-ene, 1,4-dioxane, ethanol (99.5) and ammonia solution (28) (10:8:2:1) to a distance of about 12 cm, air-dry the plate, and then dry at 1009C for 1 hour. Spray evenly sodium hypochlorite TS on the plate, air-dry the plate for 15 minutes, and then spray evenly potassium iodide starch TS on the plate: the spots other than the principal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solu-tion (2), and the numbers of spots other than the principal spot and the spot of the starting point, which are more in-tense than the spot from the standard solution (1), are not more than 3. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Clonidine Hydro-chloride, previously dried, and dissolve in 30 mL of acetic acid (100) by warming. After cooling, add 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. ‌790 Each mL of 0.1 mol/L perchloric acid VS ＝ 26.66 mg of C9H9Cl2N3.HCl
Cloperastine Fendizoate, when dried, contains not less than 99.
Cloperastine Fendizoate occurs as white, crys-tals or crystalline powder. It is freely soluble in isopropylamine, slightly soluble in methanol, in ethanol (99.5) and in acetic acid (100), and practically insoluble in water. A solution of Cloperastine Fendizoate in isopropylamine (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Cloperastine Fendizoate in methanol (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cloperastine Fendizoate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—To 2.0 g of Cloperastine Fen-dizoate add 50 mL of water, warm at 709C for 5 minutes, cool, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Cloper- astine Fendizoate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). 4-Chlorobenzophenone—Dissolve exactly 25 mg of Cloperastine Fendizoate in the mobile phase A to make ex-actly 50 mL, and use this solution as the sample solution. Separately, dissolve exactly 25 mg of 4-chlorobenzophenone in the mobile phase A to make exactly 200 mL. Pipet 1 mL of this solution, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas of 4-chlorobenzophenone by the automatic integration method: the peak area of 4-chlorobenzophenone obtained from the sample solution is not larger than that from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 226 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of 0.1 mol/L potassium dihy-drogen phosphate TS, acetonitrile for liquid chromatogra-phy and perchloric acid (400:320:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, 0.1 mol/L potassium dihydrogen phosphate TS and perchloric acid (1050:450:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 12 12 – 22 100 → 0 0 → 100 Flow rate: 1.2 mL per minute. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of 4-chlorobenzophenone obtained with 20 mL of this solution is equivalent to 14 to 26z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of 4-chlorobenzophenone are not less than 10,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 4-chlorobenzophenone is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of dried Cloperastine Fendizoate, add 100 mL of acetic acid (100), warm to dis-solve, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 64.82 mg of C20H24ClNO.C20H14O4
Cloperastine Hydrochloride, when dried, contains not less than 98.
Cloperastine Hydrochloride occurs as white, crystals or crystalline powder. It is very soluble in water, in methanol, in ethanol (95) and in acetic acid (100), and soluble in acetic anhydride. A solution of Cloperastine Hydrochloride (1 in 10) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Cloperastine Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Sepa-rately, determine the absorption spectrum of a solution of Cloperastine Hydrochloride in 0.1 mol/L hydrochloric acid TS (1 in 62,500) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cloperastine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Shake 10 mL of a solution of Cloperastine Hydro-chloride (1 in 100) with 2 mL of ammonia TS and 20 mL of diethyl ether, separate the water layer, wash the water layer with 20 mL of diethyl ether, and filter. Acidify the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Cloperastine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 40 mg of Cloperastine Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks, having the relative retention times of about 0.8 and about 3.0 to cloperastine obtained from the sample solu-tion are not larger than the peak area of cloperastine from the standard solution, and the area of the peak having the relative retention time about 2.0 from the sample solution is not larger than 5/3 times the peak area of cloperastine from the standard solution. The area of the peak other than cloperastine and the peaks mentioned above from the sample solution are not larger than 3/5 times the peak area of cloperastine from the standard solution, and the total area of these peaks is not larger than 2 times the peak area of cloperastine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, 0.1 mol/L potas-sium dihydrogen phosphate TS and perchloric acid (500:250:1). Flow rate: Adjust so that the retention time of cloper-astine is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of cloperastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, add the mobile phase to make exactly 20 mL. Con-ˆrm that the peak area of cloperastine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 30 mg of Cloperastine Hy-drochloride and 40 mg of benzophenone in 100 mL of the mobile phase. To 2 mL of this solution add the mobile phase to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, cloper-astine and benzophenone are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cloperastine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Cloperastine Hy-drochloride, previously dried, dissolve in 70 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.63 mg of C20H24ClNO.HCl ‌JP XVIII Official Monographs / Clopidogrel Sulfate 793
Clopidogrel Sulfate contains not less than 97.
Clopidogrel Sulfate occurs as a white to pale yellow-white, crystalline powder or powder. It is freely soluble in water and in methanol, and soluble in ethanol (99.5). It gradually develops a brown color on exposure to light. Melting point: about 1779C (with decomposition). 2.5 mL of this solution, add a mixture of acetonitrile for liquid chromatography and the mobile phase A (3:2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.5 and about 1.1 to clopidogrel, obtained from the sample solution is not larger than 2 times the peak area of clopidogrel from the standard solution, the area of the peak other than clopidogrel and the peaks men-tioned above from the sample solution is not larger than the peak area of clopidogrel from the standard solution, and the total area of the peaks other than clopidogrel from the sam-ple solution is not larger than 5 times the peak area of clopidogrel from the standard solution. Operating conditions— Detector, column and column temperature: Proceed as di-rected in the operating conditions in the Assay. Mobile phase A: Dissolve 0.87 g of sodium 1-pen-tanesufonate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 950 mL of this solution add 50 mL of methanol. Mobile phase B: A mixture of acetonitrile for liquid chro-matography and methanol (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) solution of Clopidogrel Sulfate in methanol (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Clopidogrel Sulfate RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Clopidogrel Sulfate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Clopidogrel Sulfate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Clopidogrel Sulfate, or each of Clopidogrel Sulfate and Clopidogrel Sulfate RS in ethanol (99.5), respectively. Then evaporate the ethanol to dryness, and repeat the test on the residues dried in vacuum. Perform the test with Clopidogrel Sulfate as directed under Flame Coloration Test <1.04> (2): a green color ap-pears. A solution of Clopidogrel Sulfate in a mixture of water and methanol (1:1) (1 in 100) responds to Qualitative Tests <1.09> (1) for sulfate.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Clopidogrel Sulfate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 65 mg of Clopidogrel Sulfate in 10 mL of a mixture of acetonitrile for liquid chro-matography and mobile phase A (3:2), and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add a mixture of acetonitrile for liquid chromatography and the mobile phase A (3:2) to make exactly 100 mL. Pipet 0 – 3 89.5 10.5 3 – 48 89.5 → 31.5 10.5 → 68.5 48 – 68 31.5 68.5 Flow rate: 1.0 mL per minute. Time span of measurement: For 68 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add a mixture of acetonitrile for liquid chromatography and the mobile phase A (3:2) to make ex-actly 20 mL. Confirm that the peak area of clopidogrel ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clopidogrel are not less than 60,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clopidogrel is not more than 2.0z. Enantiomer—Dissolve 0.10 g of Clopidogrel Sulfate in 25 mL of ethanol (99.5) for liquid chromatography, add heptane for liquid chromatography to make 50 mL, and use this solution as the sample solution. Pipet 2.5 mL of the sample solution, and add a mixture of ethanol (99.5) for liq-uid chromatography and heptane for liquid chromatography (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of ethanol (99.5) for liquid chromatography and heptane for liquid chromatography (1:1) to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution ‌794 and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the peak area of the enantiomer, having the relative reten-tion time of about 0.6 to clopidogrel, obtained from the sample solution is not larger than the peak area of clopidogrel from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with cellulose derivative-coated silica gel for liquid chromatography (10 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of heptane for liquid chromatog-raphy and ethanol (99.5) for liquid chromatography (17:3). Flow rate: Adjust so that the retention time of clopidogrel is about 18 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clopidogrel are not less than 3500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clopidogrel are not more than 2.0z. Water <2.48> Not more than 0.5z (1 g, coulometric titra-tion). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 45 mg each of Clopidogrel Sulfate and Clopidogrel Sulfate RS (separately, determine the water <2.48> in the same manner as Clopidogrel Sulfate), and dissolve them separately in the mobile phase to make ex-actly 50 mL. Take exactly 7 mL of each solution, add separately the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of clopidogrel in each solution. is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clopidogrel are not less than 4500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clopidogrel is not more than 1.0z.
Clotiazepam, when dried, contains not less than 98.
Clotiazepam occurs as white to light yellow-white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in chloroform, freely soluble in metha-nol, in ethanol (95), in acetone, in acetic acid (100) and in ethyl acetate, soluble in diethyl ether, and practically insolu-ble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. It is gradually colored by light. of Clotiazepam, according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Clotiazepam in 10 mL of acetone, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add acetone to make exactly 20 mL, pipet 2 mL of this solution, add ace-tone to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and acetone (5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 3 hours). Residue on ignition <2.44> not more than 0.1z (1 g).
(1) Dissolve 0.01 g of Clotiazepam in 3 mL of sulfuric acid: the solution shows a light yellow fluores-cence under ultraviolet light (main wavelength: 365 nm). Determine the absorption spectrum of a solution of Clotiazepam in 0.1 mol/L hydrochrolic acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Prepare the test solution with 0.01 g of Clotiazepam as directed under Oxygen Flask Combustion Method <1.06>, using 10 mL of diluted hydrogen peroxide (30) (1 in 5) as the absorbing liquid. Apply a small amount of water to the up-per part of the Apparatus A, pull out C carefully, wash C, B and the inner side of A with 15 mL of methanol, and use the obtained solution as the test solution. Add 0.5 mL of dilute nitric acid to 15 mL of the test solution: this solution re-sponds to Qualitative Tests <1.09> (2) for chloride. The remaining test solution responds to Qualitative Tests <1.09> for sulfate.
(1) Clarity and color of solution—Dissolve 1.0 g of Clotiazepam in 10 mL of ethanol (95): the solution is clear and is not more colored than the following control solution. Control solution: To 5 mL of Matching Fluid C add 0.01 mol/L hydrochloric acid TS to make 10 mL. Chloride <1.03>—To 1.0 g of Clotiazepam add 50 mL of water, shake for 30 minutes, and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.015z). Heavy metals <1.07>—Proceed with 2.0 g of Clotia- zepam according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g
Weigh accurately about 0.5 g of Clotiazepam, previ-ously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid (potentiometric titra- tion). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.88 mg of C H ClN OS 16 15 2 Identification (1) Dissolve 0.01 g of Clotiazepam in 3 mL of sulfuric acid: the solution shows a light yellow fluores-cence under ultraviolet light (main wavelength: 365 nm). Determine the absorption spectrum of a solution of Clotiazepam in 0.1 mol/L hydrochrolic acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Prepare the test solution with 0.01 g of Clotiazepam as directed under Oxygen Flask Combustion Method <1.06>, using 10 mL of diluted hydrogen peroxide (30) (1 in 5) as the absorbing liquid. Apply a small amount of water to the up-per part of the Apparatus A, pull out C carefully, wash C, B and the inner side of A with 15 mL of methanol, and use the obtained solution as the test solution. Add 0.5 mL of dilute nitric acid to 15 mL of the test solution: this solution re-sponds to Qualitative Tests <1.09> (2) for chloride. The remaining test solution responds to Qualitative Tests <1.09> for sulfate. Melting point <2.60> 106 – 1099C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Clotiazepam in 10 mL of ethanol (95): the solution is clear and is not more colored than the following control solution. Control solution: To 5 mL of Matching Fluid C add 0.01 mol/L hydrochloric acid TS to make 10 mL. Chloride <1.03>—To 1.0 g of Clotiazepam add 50 mL of water, shake for 30 minutes, and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.015z). Heavy metals <1.07>—Proceed with 2.0 g of Clotia- zepam according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g
Clotrimazole, when dried, contains not less than 98.
Clotrimazole occurs as a white, crystalline powder. It is odorless and tasteless. It is freely soluble in dichloromethane and in acetic acid (100), soluble in N,N-dimethylformamide, in methanol and in ethanol (95), slightly soluble in diethyl ether, and practi-cally insoluble in water.
(1) To 0.1 g of Clotrimazole add 10 mL of 5 mol/L hydrochloric acid TS, dissolve by heating, and cool. To this solution add 3 drops of Reinecke salt TS: a light red precipitate is produced. Determine the absorption spectrum of a solution of Clotrimazole in methanol (1 in 5000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clotrimazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Clotrimazole as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Clarity and color of solution—Dissolve 0.5 g of Clotrimazole in 10 mL of dichloromethane: the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Clotrimazole in 40 mL of N,N-dimethylformamide, add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.60 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of N,N-dimethylformamide, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate <1.14>—Dissolve 0.5 g of Clotrimazole in 10 mL of methanol, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.05 mL of 0.005 mol/L sulfuric acid VS, 10 mL of metha-nol, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Clotrimazole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Clotrimazole according to Method 3, and perform the test ‌800 (not more than 2 ppm). Imidazole—Dissolve 0.10 g of Clotrimazole in ex-actly 10 mL of dichloromethane, and use this solution as the sample solution. Separately, dissolve 25 mg of imidazole for thin-layer chromatography in dichloromethane to make exactly 50 mL. Pipet 5 mL of this solution, add dichloro-methane to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and chloroform (3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly sodium hypochlorite TS on the plate, and air-dry the plate for 15 minutes, then spray evenly potassium iodide-starch TS on the plate: the spot obtained from the sample so-lution, corresponding to that from the standard solution, is not more intense than that from the standard solution. (2-Chlorophenyl)-diphenylmethanol—Dissolve 0.20 g of Clotrimazole in exactly 10 mL of dichloromethane, and use this solution as the sample solution. Separately, dissolve 10 mg of (2-chlorophenyl)-diphenylmethanol for thin-layer chromatography in dichloromethane to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and ammonia solu-tion (28) (50:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution, cor-responding to that from the standard solution, is not more intense than that from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Clotrimazole, pre-viously dried, and dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.48 mg of C22H17ClN2
Cloxazolam, when dried, contains not less than 99.
Cloxazolam occurs as white, crystals or crys-talline powder. It is odorless and tasteless. It is freely soluble in acetic acid (100), sparingly soluble in dichloromethane, slightly soluble in ethanol (99.5) and in diethyl ether, very slightly soluble in ethanol (95), and practi-cally insoluble in water. It dissolves in dilute hydrochloric acid. It is gradually colored by light. Melting point: about 2009C (with decomposition).
(1) Dissolve 0.01 g of Cloxazolam in 10 mL of ethanol (99.5) by heating, and add 1 drop of hydrochloric acid: the solution shows a light yellow color and a yellow-green fluorescence under ultraviolet light (main wavelength: 365 nm). Add 1 mL of sodium hydroxide TS to this solution: the color and fluorescence of this solution disappear immedi-ately. Dissolve 0.01 g of Cloxazolam in 5 mL of dilute hy-drochloric acid by heating in a water bath for 10 minutes. After cooling, 1 mL of this solution responds to Qualitative Tests <1.09> for primary aromatic amines. Place 2 g of Cloxazolam in a 200-mL flask, add 50 mL of ethanol (95) and 25 mL of sodium hydroxide TS, and boil under a reflux condenser for 4 hours. After cooling, neutralize with dilute hydrochloric acid, and extract with 30 mL of dichloromethane. Dehydrate with 3 g of anhydrous sodium sulfate, filter, and evaporate the dichloromethane of the filtrate. Dissolve the residue in 5 mL of methanol by heating on a water bath, and cool immediately in an ice bath. Collect the crystals, and dry the crystals is vacuum at 609C for 1 hour: it melts <2.60> between 879C and 919C. Determine the absorption spectrum of a solution of Cloxazolam in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare ‌802 the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Proceed with Cloxazolam as directed under Flame Coloration Test <1.04> (2), and perform the test: a green color appears. Cocaine Hydrochloride
Cocaine Hydrochloride, when dried, contains not less than 98.
Cocaine Hydrochloride occurs as colorless crystals or a white crystalline powder. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), slightly soluble in acetic anhydride, and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Cocaine Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Sepa-rately, determine the absorption spectrum of a solution of Cocaine Hydrochloride in 0.01 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cocaine Hydrochloride, previously dried, as directed in the potassium bromide disk method under the Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Cocaine Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －70 – －739(after drying, 0.5 g, water, 20 mL, 100 mm).
(1) Chloride <1.03>—To 1.0 g of Cloxazolam add 50 mL of water, allow to stand for 1 hour with occasional shaking, and filter. To 25 mL of this filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Clox- azolam according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Place 1.0 g of Cloxazolam in a Kjel- dahl flask, add 5 mL of sulfuric acid and 5 mL of nitric acid, and heat gently. Repeat the addition of 2 to 3 mL of nitric acid at times, and continue heating until a colorless to light yellow solution is obtained. After cooling, add 15 mL of saturated ammonium oxalate solution, and heat the solution until dense white fumes are evolved, and evaporate to a volume of 2 to 3 mL. After cooling, dilute with water to 10 mL, and perform the test with this solution as the test solu-tion (not more than 2 ppm). Related substances—Dissolve 0.05 g of Cloxazolam in 10 mL of dichloromethane, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dichloromethane to make exactly 200 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Immediately after air-drying, develop the plate with a mixture of toluene and acetone (5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than that from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Cloxazolam, previ-ously dried, and dissolve in 50 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same menner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.92 mg of C17H14Cl2N2O2
Cod Liver Oil is the fatty oils obtained from fresh livers and pyloric appendages of Gadus macrocephalus Tilesius or Theragra chalcogramma Pallas (Gadidae). Cod Liver Oil contains not less than 2000 Vitamin A Units and not more than 5000 Vitamin A Units per g.
Cod Liver Oil is a yellow to orange oily liquid. It has a characteristic, slightly fishy odor and a mild taste. It is miscible with chloroform. It is slightly soluble in ethanol (95), and practically insolu-ble in water. It is decomposed by air or by light.
Dissolve 0.1 g of Cod Liver Oil in 10 mL of chloroform, and to 1 mL of this solution add 3 mL of an-timony (III) chloride TS: a blue color develops immediately, but the color fades rapidly. Specific gravity <1.13> d 20: 0.918 – 0.928 Acid value <1.13> Not more than 1.7. Saponification value <1.13> 180 – 192 Unsaponifiable matter <1.13> Not more than 3.0z. Iodine value <1.13> 130 – 170
Rancidity—No unpleasant odor of rancid oil is per-ceptible on warming Cod Liver Oil.
Proceed with about 0.5 g of Cod Liver Oil, accu-rately weighed, as directed in Method 2 under the Vitamin A Determination <2.55>, and perform the test.
Codeine Phosphate Hydrate occurs as white to yellowish white, crystals or crystalline powder. It is freely soluble in water and in acetic acid (100), slightly soluble in methanol and in ethanol (95), and practically in-soluble in diethyl ether. The pH of a solution of 1.0 g of Codeine Phosphate Hy-drate in 10 mL of water is between 3.0 and 5.0. It is affected by light.
(1) Determine the absorption spectrum of a solution of Codeine Phosphate Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Codeine Phosphate Hydrate, previously dried at 1059C for 4 hours, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Codeine Phosphate Hydrate (1 in 20) responds to Qualitative Tests <1.09> (1) for phosphate. D Optical rotation <2.49> [a]20: －98 – －1029(0.4 g calcu-lated on the anhydrous basis, water, 20 mL, 100 mm).
(1) Chloride <1.03>—Perform the test with 0.5 g of Codeine Phosphate Hydrate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.20 g of Codeine Phosphate Hydrate. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.240z). Related substances—Dissolve 0.20 g of Codeine Phosphate Hydrate in 10 mL of a mixture of 0.01 mol/L hydrochloric acid TS and ethanol (99.5) (4:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of 0.01 mol/L hydrochloric acid TS and ethanol (99.5) (4:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5), toluene, acetone and ammonia solution (28) (14:14:7:1) to a distance of about 10 cm, and air-dry the ‌804 plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 1.5 – 3.0z (0.5 g, volumetric titration, direct titration).
Dissolve about 0.5 g of Codeine Phosphate Hydrate, accurately weighed, in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to greenish blue (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.74 mg of C18H21NO3.H3PO4
Colchicine occurs as a yellowish white powder. It is very soluble in methanol, freely soluble in N,N-dimethylformamide, in ethanol (95) and in acetic anhydride, and sparingly soluble in water. It is colored by light.
(1) Determine the absorption spectrum of a solution of Colchicine in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) To 1 g of potassium bromide for infrared absorption spectrum add 0.5 mL of a solution of Colchicine in methanol (1 in 50), grind thoroughly, and dry in vacuum at 809C for 1 hour. Determine the infrared absorption spectrum of this powder as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －235 – －2509(0.1 g calcu-lated on the anhydrous basis and corrected by the amount of ethyl acetate, ethanol (95), 10 mL, 100 mm).
(1) Colchicine—Dissolve 0.10 g of Colchicine in 10 mL of water, and to 5 mL of this solution add 2 drops of iron (III) chloride TS: no definite green color develops. Chloroform and ethyl acetate—Weigh accurately about 0.6 g of Colchicine, dissolve in exactly 2 mL of the internal standard solution, add N,N-dimethylformamide to make 10 mL, and use this solution as the sample solution. Separately, weigh 0.30 g of chloroform using a 100-mL volu-metric flask containing about 20 mL of N,N-dimethylfor-mamide, and add N,N-dimethylformamide to make exactly 100 mL. Pipet 2 mL of this solution, add N,N-dimethylfor-mamide to make exactly 200 mL, and use this solution as the standard solution (1). Separately, weigh accurately about 1.8 g of ethyl acetate using a 100-mL volumetric flask con-taining about 20 mL of N,N-dimethylformamide, and add N,N-dimethylformamide to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal stand-ard solution and N,N-dimethylformamide to make 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 2 mL each of the sample solution and standard solutions (1) and (2) as directed under Gas Chro-matography <2.02> according to the following conditions: Column temperature: 609C for 7 minutes, then up to 1009C at a rate of 409C per minute if necessary, and hold at 1009C for 10 minutes. Injection port temperature: A constant temperature of about 1309C. Detector temperature: A constant temperature of about 2009C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of ethyl ace-tate is about 3 minutes. Split ratio: 1:20. System suitability— Test for required detectability: Pipet 2 mL of the standard solution (2), and add N,N-dimethylformamide to make exactly 25 mL. Pipet 1 mL of this solution, and add N,N-dimethylformamide to make exactly 50 mL. Confirm that the peak area of ethyl acetate obtained with 2 mL of this so-lution is equivalent to 0.11 to 0.21z of that with 2 mL of the standard solution (2). System performance: To 1 mL of chloroform add N,N-dimethylformamide to make 10 mL. To 1 mL of this solu-tion add 2 mL of ethyl acetate and N,N-dimethylformamide to make 100 mL. To 2 mL of this solution add 2 mL of the internal standard solution and N,N-dimethylformamide to make 10 mL. When the procedure is run with 2 mL of this so-lution under the above operating conditions, ethyl acetate, chloroform and the internal standard are eluted in this order with the resolution between the peaks of chloroform and the internal standard being not less than 2.0. System repeatability: When the test is repeated 3 times with 2 mL of the standard solution (2) under the above oper-ating conditions, the relative standard deviation of the ratio of the peak area of ethyl acetate to that of the internal stand-ard is not more than 3.0z. Related substances—Dissolve 60 mg of Colchicine in 100 mL of diluted methanol (1 in 2). To 1 mL of this solu-tion, add diluted methanol (1 in 2) to make 100 mL, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the total amount of the peaks other than colchicine by the area percentage method: not more than 5.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌808 Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 450 mL of 0.05 mol/L potassium dihy-drogen phosphate TS add methanol to make 1000 mL. Adjust the pH to 5.5 with diluted phosphoric acid (7 in 200). Flow rate: Adjust so that the retention time of colchicine is about 7 minutes. Time span of measurement: About 2 times as long as the retention time of colchicine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, and add diluted methanol (1 in 2) to make exactly 50 mL. Confirm that the peak area of colchicine obtained from 20 mL of this solution is equivalent to 1.4 to 2.6z of that obtained from 20 mL of the sample solution. System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of colchicine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of colchicine is not more than 2.0z. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, back titration).
Weigh accurately about 0.4 g of Colchicine, dissolve in 25 mL of acetic anhydride, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 19.97 mg of C22H25NO6
Colestimide is an anion exchange resin, composed of a copolymer of 2-methylimidazole and 1-chloro-2,3-epoxypropane. It contains not less than 18.
Colestimide occurs as a white to pale yellow-white powder. It is practically insoluble in water and in ethanol (99.5). It is hygroscopic.
Determine the infrared absorption spectrum of Colestimide, previously dried, as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Heavy metals <1.07>—Take 2.0 g of Coles-timide in a porcelain or platinum crucible, and carbonize by weakly heating. After cooling, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) and 5 mL of hydrogen peroxide (30), and ignite the ethanol. After cooling, add 1 mL of sulfuric acid, then, proceed ac-cording to Method 4, and perform the test. Prepare the con-trol solution as follows: To 10 mL of a solution of magne-sium nitrate hexahydrate in ethanol (95) (1 in 10) add 5 mL of hydrogen peroxide (30), and ignite the ethanol. After cooling, add 1 mL of sulfuric acid, then, proceed in the same manner as for the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 10 ppm). (2) Related substances—To exactly 0.50 g of Colestimide add exactly 20 mL of water, shake for 1 hour, centrifuge, and use the supernatant liquid as the sample solution. Deter-mine the absorbance of the sample solution at 210 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.50. Loss on drying <2.41> Not more than 10.0z (1 g, in vacu-um, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Degree of swelling Weigh accurately about 1 g of Coles-timide, put in a 25-mL glass stoppered measuring cylinder (about 11 mm in inside diameter), add 23 mL of water, shake for 2 minutes, and add water to make 25 mL. After standing for 2 hours, measure the volume of the resin layer, and determine the volume per g, calculated on the dried basis: the volume is 12 – 18 mL/g.
(1) Chlorine—Weigh accurately about 0.2 g of Colestimide, add 50 mL of water, and shake. Add 1 mL of nitric acid and 25 mg of potassium nitrate, shake, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titra- tion). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L silver nitrate VS ＝ 3.545 mg of Cl (2) Exchange capacity—Weigh accurately about 0.45 g of sodium cholate hydrate (separately determine the water), dissolve in water to make exactly 100 mL, and use this solu-tion as the sodium cholate standard stock solution. Sepa-rately, weigh accurately about 30 mg of Colestimide, add ex-actly 30 mL of the sodium cholate standard stock solution, shake for 1 hour, and centrifuge or filter through a mem-brane filter with a pore size not exceeding 0.8 mm. Pipet 5 mL of the supernatant liquid or the filtrate, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, pipet 5 mL of the sodium cholate standard stock solution, add exactly 5 mL of the in-ternal standard solution, and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of cholic acid to that of the internal standard. Exchanged amount (g) of cholic acid per g of Colestimide, calculated on the dried basis ＝ MS/MT × (QS － QT)/QS × 3/10 × 0.947 MS: Amount (mg) of sodium cholate hydrate taken, calcu- ‌JP XVIII Official Monographs / Colestimide Tablets 809‌ lated on the anhydrous basis MT: Amount (mg) of Colestimide taken, calculated on the dried basis Internal standard solution—A solution of butyl parahy-droxybenzoate in acetonitrile (1 in 80,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of cholic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cholic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cholic acid to that of the internal standard is not more than 1.0z.
Colistin Sulfate is the sulfate of a mixture of peptide substances having antibacterial activity produced by the growth of Bacillus polymyxa var. colistinus. It, when dried, contains not less than 16,000 units per mg.
Colistin Sulfate occurs as a white to light yel-low-white powder. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic.
(1) Dissolve 20 mg of Colistin Sulfate in 2 mL of water, add 0.5 mL of sodium hydroxide TS, then add 5 drops of copper (II) sulfate TS while shaking: a purple color develops. Dissolve 50 mg of Colistin Sulfate in 10 mL of diluted hydrochloric acid (1 in 2). Transfer 1 mL of this solu-tion in a tube for hydrolysis, seal, and heat at 1359C for 5 hours. After cooling, open the tube, and evaporate the con-tent to dryness until the odor of hydrochloric acid is no more perceptible. Dissolve the residue in 0.5 mL of water, and use this solution as the sample solution. Separately, dissolve 20 mg each of L-leucine, L-threonine, phenylalanine and L-ser-ine in 10 mL of water, and use these solutions as the stand-ard solution (1), (2), (3) and (4). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution (1), (2), (3) and (4) on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, acetic acid (100), water, pyridine and ethanol (99.5) (60:15:10:6:5) to a distance of about 10 cm, and dry the plate at 1059C for 10 minutes. Spray evenly ninhydrin TS on the plate, and heat at 1109C for 5 minutes: three principal spots are obtained from the sample solution, the Rf values of two spots of them are the same with those of the cor-responding spots obtained from the standard solution (1) and the standard solution (2), and the Rf value of the rest principal spot is about 0.1. No spot is observed at the posi-tion corresponding to the spots from the standard solution (3) and the standard solution (4). A solution of Colistin Sulfate (1 in 20) responds to Qualitative Tests <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: －63 – －739(1.25 g, after drying, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving Related substances—Dissolve 50 mg of Colistin Sul-fate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of pyridine, 1-butanol, water and acetic acid (100) (6:5:4:1) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly ninhydrin-butanol TS on the plate, and heat at 1009C for about 20 minutes: the spot other than the principal spot obtained from the sample solu-tion is not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 6.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
(1) Sulfuric acid—Weigh accurately about 0.25 g of previously dried Colistin Sulfate, dissolve in a suitable amount of water, adjust the pH to 11 with ammonia solution (28), and add water to make 100 mL. To this solution add exactly 10 mL of 0.1 mol/L barium chloride VS and 50 mL of ethanol (99.5), and titrate with <2.50> 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the blue-purple color of the solution disappears (indicator: 0.5 mg of phthalein purple): the amount of sulfuric acid (SO4) is 16.0 to 18.0z. Colistin A Sulfate C H N O .21/ H SO : 1414.66 Each mL of 0.1 mol/L barium chloride VS 53 100 16 13 2 2 4 ＝ 9.606 mg of SO4 Colistin B Sulfate C52H98N16O13.21/2H2SO4: 1400.63 [1264-72-8] Colistin Sulfate is the sulfate of a mixture of peptide substances having antibacterial activity produced by the growth of Bacillus polymyxa var. colistinus. It, when dried, contains not less than 16,000 units per mg. The potency of Colistin Sulfate is expressed as unit calculated from the amount of colistin A (C53H100N16O13: 1169.46). One unit of Colistin Sulfate is equivalent to 0.04 mg of colistin A (C53H100N16O13). Description Colistin Sulfate occurs as a white to light yel-low-white powder. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic. Identification (1) Dissolve 20 mg of Colistin Sulfate in 2 mL of water, add 0.5 mL of sodium hydroxide TS, then add 5 drops of copper (II) sulfate TS while shaking: a purple color develops. Dissolve 50 mg of Colistin Sulfate in 10 mL of diluted hydrochloric acid (1 in 2). Transfer 1 mL of this solu-tion in a tube for hydrolysis, seal, and heat at 1359C for 5 hours. After cooling, open the tube, and evaporate the con-tent to dryness until the odor of hydrochloric acid is no more perceptible. Dissolve the residue in 0.5 mL of water, and use this solution as the sample solution. Separately, dissolve 20 mg each of L-leucine, L-threonine, phenylalanine and L-ser-ine in 10 mL of water, and use these solutions as the stand-ard solution (1), (2), (3) and (4). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution (1), (2), (3) and (4) on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, acetic acid (100), water, pyridine and ethanol (99.5) (60:15:10:6:5) to a distance of about 10 cm, and dry the plate at 1059C for 10 minutes. Spray evenly ninhydrin TS on the plate, and heat at 1109C for 5 minutes: three principal spots are obtained from the sample solution, the Rf values of two spots of them are the same with those of the cor-responding spots obtained from the standard solution (1) and the standard solution (2), and the Rf value of the rest principal spot is about 0.1. No spot is observed at the posi-tion corresponding to the spots from the standard solution (3) and the standard solution (4). A solution of Colistin Sulfate (1 in 20) responds to Qualitative Tests <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: －63 – －739(1.25 g, after drying, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving Related substances—Dissolve 50 mg of Colistin Sul-fate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of pyridine, 1-butanol, water and acetic acid (100) (6:5:4:1) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly ninhydrin-butanol TS on the plate, and heat at 1009C for about 20 minutes: the spot other than the principal spot obtained from the sample solu-tion is not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 6.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Escherichia coli NIHJ Culture medium—Dissolve 10.0 g of peptone, 30.0 g of sodium chloride, 3.0 g of meat extract and 15.0 g of agar in 1000 mL of water, adjust the pH with sodium hydroxide TS so that the solution will be 6.5 to 6.6 after sterilization, and use as the agar media for seed layer and for base layer. Standard solutions—Weigh accurately an amount of Colistin Sulfate RS, previously dried, equivalent to about 1,000,000 units, dissolve in phosphate buffer solution (pH 6.0) to make exactly 10 mL, and use this solution as the standard stock solution. Keep the standard stock solution at not exceeding 109C, and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 10,000 units and 2500 units, and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Colistin Sulfate, previously dried, equivalent to about 1,000,000 units, and dissolve in phosphate buffer solution (pH 6.0) to make exactly 10 mL. Take exactly a suitable amount of this solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 10,000 units and 2500 units, and use these solutions as the high concen-tration sample solution and the low concentration sample so-lution, respectively.
Compound Iodine Glycerin contains not less than w/vz and not more than 1.
Compound Iodine Glycerin is a red-brown, viscous liquid. It has a characteristic odor. Specific gravity d 20: about 1.23
(1) The colored solution obtained in the Assay (1) acquires a red color. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 510 nm and 514 nm (iodine). The colored solution obtained in the Assay (2) ac-quires a red color. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 510 nm and 514 nm (potassium iodide). The colored solution obtained in the Assay (4) has a yellow color. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 401 nm and 405 nm (phenol). Take 1 mL of Compound Iodine Glycerin in a glass-stoppered test tube, add 10 mL of ethanol (95), and mix. Then add 2 mL of sodium hydroxide TS, add 1 mL of a so-lution of copper (II) chloride dihydrate in ethanol (95) (1 in 10), and shake: a blue color develops (glycerin).
(1) Iodine—Measure the specific gravity of Com-pound Iodine Glycerin according to Method 2 under Deter-mination of Specific gravity and density <2.56>. Weigh ex-actly about 7 mL of it, add ethanol (95) to make exactly 200 mL, and use this solution as the sample solution. On the other hand, weigh accurately about 80 mg of iodine for assay and about 0.17 g of potassium iodide for assay, previously dried at 1059C for 4 hours, dissolve in ethanol (95) to make exactly 200 mL, and use this solution as the standard solu- ‌1172 tion. Pipet 3 mL each of the sample solution and the stand-ard solution into 50-mL separators, to each add exactly 10 mL of a mixture of chloroform and hexane (2:1) and 15 mL of water successively, and shake immediately and vigor-ously. Separate the chloroform-hexane layers [use the water layers in (2)], and filter through a pledget of cotton. Deter-mine the absorbances of the filtrates, AT and AS, at 512 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a mixture of chloroform and hexane (2:1) as the blank. solution, and use so obtaind solution as the standard solu-tion. Pipet 3 mL each of the sample solution and standard solution, to each add 2 mL of dilute hydrochloric acid, and place in a water bath at 309C. Allow to stand for 10 minutes, and add exactly 2 mL of a solution of sodium nitrite (1 in 100), shake, and allow to stand at 309C for 60 minutes. Add dilute potassium hydroxide-ethanol TS to make exactly 25 mL, and determine the absorbances of these solutions, AT and AS, at 403 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using the solution prepared in the Amount (mg) of iodine (I) ＝ MS × AT/AS same manner with 3 mL of water instead of the sample solu- tion as the blank. MS: Amount (mg) of iodine for assay taken Potassium iodide—Separate the water layers of the sample solution and the standard solution obtained in (1), pipet 10 mL of each of the water layers, and to each add 1 mL of diluted dilute hydrochloric acid (1 in 2), 1 mL of so-dium nitrite TS and exactly 10 mL of a mixture of chlo-roform and hexane (2:1). Shake immediately and vigorously, separate the chloroform-hexane layers, and filter through a pledget of cotton. Determine the absorbances, AT and AS, of both solutions at 512 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a mixture of chloroform and hexane (2:1) as the blank. Amount (mg) of potassium iodide (KI) ＝ MS × AT/AS MS: Amount (mg) of potassium iodide for assay taken Total iodine—Measure the specific gravity of Com-pound Iodine Glycerin according to Method 2 under Deter-mination of Specific gravity and density <2.56>. Weigh ex-actly about 5 mL of it, and add water to make exactly 50 mL. Pipet 5 mL of this solution into a 50-mL flask, and add 0.5 g of zinc powder and 5 mL of acetic acid (100). Shake until the color of iodine disappears, and heat under a reflux condenser on a water bath for 30 minutes. Wash the con-denser with 10 mL of hot water, and filter through a glass filter (G3). Wash the flask with two 10-mL portions of warm water, and combine the filtrate and the washings. After cool-ing, add water to make exactly 50 mL, and use this solution as the sample solution. On the other hand, dissolve about 0.2 g of potassium iodide for assay, previously dried at 1059C for 4 hours and accurately weighed, in water to make exactly 50 mL. Pipet 5 mL of this solution, add 5 mL of ace-tic acid (100) and water to make exactly 50 mL, and use this solution as the standard solution. Pipet 4 mL each of the sample solution and standard solution into 30-mL separa-tors, and to each add 5 mL of water, 1 mL of diluted dilute hydrochloric acid (1 in 2), 1 mL of sodium nitrite TS and 10 mL of a mixture of chloroform and hexane (2:1). Shake well immediately, and proceed as directed in (2). Amount (mg) of total iodine (I) ＝ MS × AT/AS × 0.764 MS: Amount (mg) of potassium iodide for assay taken Phenol—Measure the specific gravity of Compound Iodine Glycerin according to Method 2 under Determination of Specific gravity and density <2.56>. Weigh exactly about 2 mL of it, add 3 mL of 0.1 mol/L sodium thiosulfate VS, and shake. Add 2 mL of dilute hydrochloric acid, and shake with two 10-mL portions of chloroform. Separate the chloroform layer, and shake with two 10-mL portions of 0.5 mol/L so-dium hydroxide TS. Separate the water layer, add water to make exactly 500 mL, and use this solution as the sample so-lution. Dissolve about 0.5 g of phenol for assay, accurately weighed, in ethanol (95) to make exactly 100 mL, pipet 2 mL of this solution, proceed in the same manner as the sample Amount (mg) of phenol (C6H6O) ＝ MS × AT/AS × 1/50 MS: Amount (mg) of phenol for assay taken
Compound Methyl Salicylate Spirit is a red-dish yellow liquid, having a characteristic odor and a burn-ing taste.
(1) Shake 1 mL of Compound Methyl Salicylate Spirit with 5 mL of dilute ethanol, and add 1 drop of iron (III) chloride TS: a purple color is produced (methyl salicylate). (2) Shake thoroughly 1 mL of Compound Methyl Salicy-late Spirit with 10 mL of chloroform, and use this solution as the sample solution. Dissolve 40 mg of methyl salicylate in 10 mL of chloroform, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on the plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of hexane and chlo-roform (4:1) to a distance of about 10 cm, air-dry the plate, and examine under ultraviolet light (main wavelength: 254 nm): the spots obtained from the sample solution and the standard solution show the same Rf value. Spray evenly iron (III) chloride TS upon the plate: the spot from the standard solution and the corresponding spot from the sample solu-tion reveal a purple color.
Compound Salicylic Acid Spirit contains not less than 1.
Compound Salicylic Acid Spirit is a clear, col-orless to light red liquid. Specific gravity d 20: about 0.88
(1) To 1 mL of Compound Salicylic Acid Spirit add hydrochloric acid-potassium chloride buffer solu-tion (pH 2.0) to make 200 mL, and to 5 mL of this solution add 5 mL of a solution of iron (III) nitrate enneahydrate (1 in 200): a red-purple color is produced (salicylic acid). To 1 mL of Compound Salicylic Acid Spirit add 20 mL of water and 5 mL of dilute hydrochloric acid, and ex-tract with 20 mL of diethyl ether. Wash the diethyl ether ex-tract with two 5-mL portions of sodium hydrogen carbonate TS, and extract with 10 mL of dilute sodium hydroxide TS. Shake 1 mL of the extract with 1 mL of sodium nitrite TS and 1 mL of dilute hydrochloric acid, allow to stand for 10 minutes, and add 3 mL of sodium hydroxide TS: a yellow color is produced (phenol). To 0.5 mL of Compound Salicylic Acid Spirit add 5 mL of dilute hydrochloric acid, extract with 5 mL of chlo-roform, and use the extract as the sample solution (1). To 2 mL of Compound Salicylic Acid Spirit add 5 mL of dilute hydrochloric acid, extract with 5 mL of chloroform, wash the extract with two 5-mL portions of sodium hydrogen car-bonate TS, and use the chloroform extract as the sample so- ＝ MS × AT/AS 7 6 3 lution (2). Separately, dissolve 0.01 g each of salicylic acid and phenol in 5 mL each of chloroform, and use both solu- MS: Amount (mg) of salicylic acid for assay taken
Concentrated Glycerin is a clear, colorless and viscous liquid. It has a sweet taste. It is miscible with water and with ethanol (99.5). It is hygroscopic.
Determine the infrared absorption spectrum of Concentrated Glycerin as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: Not less than 1.470. Specific gravity <2.56> d 20: Not less than 1.258.
(1) Color—Place 50 mL of Concentrated Glycerin in a Nessler tube, and observe downward: the solution has no more color than the following control solution. Control solution: Pipet 0.40 mL of Iron (III) Chloride CS into a Nessler tube, and add water to make 50 mL. Acidity or alkalinity—To 2 mL of Concentrated Glycerin add 8 mL of water and mix: the solution is neutral. Chloride <1.03>—Take 10.0 g of Concentrated Glyce- rin, and perform the test. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.001z). Sulfate <1.14>—Take 10.0 g of Concentrated Glyce- rin, and perform the test. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.002z). Ammonium—To 5 mL of Concentrated Glycerin add 5 mL of a solution of sodium hydroxide (1 in 10), and boil: the gas evolved does not change moistened red litmus paper to blue. Heavy metals <1.07>—Proceed with 5.0 g of Concen- trated Glycerin according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Calcium—To 5 mL of the solution obtained in (2) add 3 drops of ammonium oxalate TS: the solution remains unchanged. Arsenic <1.11>—Prepare the test solution with 1.0 g of Concentrated Glycerin according to Method 1, and per-form the test (not more than 2 ppm). Acrolein, glucose, or other reducing substances—To 1.0 g of Concentrated Glycerin add 1 mL of ammonia TS, mix, and warm in a water bath at 609C for 5 minutes: no yel-low color is produced. Take the solution out of the water bath, add 3 drops of silver nitrate TS immediately, and allow to stand in a dark place for 5 minutes: the color of the solu-tion does not change, and no turbidity is produced. Fatty acids and esters—Mix 50 g of Concentrated Glycerin with 50 mL of freshly boiled and cooled water, add 10 mL of 0.1 mol/L sodium hydroxide VS, accurately meas-ured, boil the mixture for 15 minutes, cool, and titrate <2.50> the excess sodium hydroxide with 0.1 mol/L hydrochloric acid VS: not more than 3.0 mL of 0.1 mol/L sodium hy-droxide VS is consumed (indicator: 3 drops of phenolphthal-ein TS). Perform a blank determination in the same manner. Ethylene glycol, diethylene glycol and related sub-stances—Weigh accurately about 5 g of Concentrated Glyce-rin, mix with methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g each of ethylene glycol and diethylene glycol, mix with methanol to make exactly 100 mL. Pipet 5 mL of this solution and transfer into a 100-mL volumetric flask. Separately, weigh 5.0 g of glycerin for gas chromatography, mix with a suitable amount of methanol and put in the volu-metric flask, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 1 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions, and determine the peak areas, AT1 and AS1, of ethylene glycol and, AT2 and AS2, of diethylene glycol by the automatic integration method. The amounts of ethylene glycol and diethylene glycol, calculated by the following equations, are not more than 0.1z, respec-tively. The amount of the peak other than glycerin, ethylene glycol and diethylene glycol obtained from the sample solu-tion, calculated by the area percentage method, is not more than 0.1z, and the total amount of the peaks other than glycerin is not more than 1.0z. ‌1082 Amount (z) of ethylene glycol ＝ MS1/MT × AT1/AS1 × 5 Amount (z) of diethylene glycol ＝ MS2/MT × AT2/AS2 × 5 ‌MS1: Amount (g) of ethylene glycol taken MS2: Amount (g) of diethylene glycol taken MT: Amount (g) of Concentrated Glycerin taken Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused-silica column 0.32 mm in inside diame-ter and 30 m in length, coated the inner surface with 14z cyanopropylphenyl-86z dimethyl silicone polymer for gas chromatography 1 mm in thickness. Column temperature: Inject at a constant temperature of about 1009C, raise the temperature at the rate of 7.59C per minute to 2209C, and maintain at a constant temperature of about 2209C. Injection port temperature: A constant temperature of about 2209C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: about 38 cm per second. Split ratio: 1:20. Time span of measurement: About 3 times as long as the retention time of glycerin, beginning after the solvent peak. System suitability— System performance: Mix 50 mg each of ethylene glycol, diethylene glycol and glycerin for gas chromatography with 100 mL of methanol. When the procedure is run with 1 mL of this solution under the above operating conditions, ethyl-ene glycol, diethylene glycol and glycerin are eluted in this order, and the resolution between the peaks of ethylene glycol and diethylene glycol is not less than 40, and between the peaks of diethylene glycol and glycerin is not less than 10. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviations of the peak area of ethylene glycol and diethylene glycol are not more than 10z, respectively. Readily carbonizable substances—To 5 mL of Con-centrated Glycerin add carefully 5 mL of sulfuric acid for readily carbonizable substances, mix gently at a temperature between 189C and 209C, and allow to stand for 1 hour be-tween 159C and 259C: the solution has no more color than Matching Fluid H. Water <2.48> Not more than 2.0z (6 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Weigh accurately about 10 g of Concentrated Glycerin in a tared crucible, heat to boiling, and fire to burn immediately. Cool, moisten the residue with 1 to 2 drops of sulfuric acid, and ignite cautiously to con-stant mass: the mass of the residue is not more than 0.01z.
Weigh accurately about 0.2 g of Concentrated Glycerin, transfer into a glass-stoppered flask, add 50 mL of water, mix, add exactly 50 mL of sodium periodate TS, shake, and allow to stand in a dark place at a room tempera-ture for about 30 minutes. Add 10 mL of a mixture of water and ethylene glycol (1:1), allow to stand for about 20 minutes, add 100 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 2 drops of phenol-phthalein TS). Perform a blank determination in the same manner, and make the necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 9.209 mg of C3H8O3
Condurango Fluidextract is a brown liquid. It has a characteristic odor and a bitter taste.
Mix 1 mL of Condurango Fluidextract with 5 mL of water, filter, if necessary, and heat the clear solu-tion: turbidity is produced. However, it becomes almost clear upon cooling.
Heavy metals <1.07>—Prepare the test solution with 1.0 g of Condurango Fluidextract as direct under the Fluidextracts (4), and perform the test (not more than 30 ppm).
Containers and storage Containers—Hermetic containers. Storage—Light-resistant.
To 0.5 g of 10z Ephedrine Hydrochloride Powder add 100 mL of water, shake for 20 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 249 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 265 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of 10z Ephedrine Hydrochloride Powder is not less than 85z. Start the test with about 0.25 g of 10z Ephedrine Hydro-chloride Powder, accurately weighed, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 28 mg of ephe-drine hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ephedrine in each solution. Dissolution rate (z) with respect to the labeled amount of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS/MT × AT/AS × 9/10 MS: Amount (mg) of ephedrine hydrochloride for assay taken MT: Amount (g) of 10z Ephedrine Hydrochloride Pow-der taken Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (4) under Ephedrin Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ephedrine are not less than 10,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌JP XVIII Official Monographs / Ephedrine Hydrochloride Tablets 937 ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 2.0z.
Weigh accurately about 0.4 g of 10z Ephedrine Hy-drochloride Powder, add 150 mL of water, and extract by sonicating for 10 minutes with occasional shaking. Shake more for 10 minutes, then add exactly 10 mL of the internal standard solution and water to make 200 mL, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, weigh accurately about 40 mg of ephedrine hydro-chloride for assay, previously dried at 1059C for 3 hours, add exactly 10 mL of the internal standard solution to dis-solve, add water to make 200 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ephedrine to that of the internal standard. Amount (mg) of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS × QT/QS MS: Amount (mg) of ephedrine hydrochloride for assay taken Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Perform as directed in the operating conditions in the Purity (4) under Ephedrine Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ephedrine are eluted in this order with the resolution between these peaks being not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ephedrine to that of the internal standard is not more than 1.0z.
Containers and storage Containers—Tight containers. Storage—Light-resistant.
Determine the absorption spectrum of a so-lution of 1z Dihydrocodeine Phosphate Powder (1 in 100) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 281 nm and 285 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of 1z Dihydrocodeine Phosphate Powder is not less than 85z. Start the test with about 1 g of 1z Dihydrocodeine Phos-phate Powder, accurately weighed, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 50 mg of dihydro-codeine phosphate for assay (separately determine the loss on drying <2.41> at 1059C for 4 hours), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of dihydrocodeine in each solution. ‌858 Dissolution rate (z) with respect to the labeled amount of dihydrocodeine phosphate (C18H23NO3.H3PO4) ＝ MS/MT × AT/AS × 9/5 MS: Amount (mg) of dihydrocodeine phosphate for assay taken, calculated on the dried basis MT: Amount (g) of 1z Dihydrocodeine Phosphate Pow-der taken Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry ing conditions, the relative standard deviation of the ratios of the peak area of dihydrocodeine to that of the internal standard is not more than 1.0z.
◆Containers and storage Containers—Tight containers.◆
10z Sodium Chloride Injection is a clear, col-orless liquid. It has a saline taste. It is neutral.
10z Sodium Chloride Injection responds to Qualitative Tests <1.09> for sodium salt and for chloride. Bacterial endotoxins <4.01> Less than 3.6 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet 10 mL of 10z Sodium Chloride Injection, and add water to make exactly 100 mL. Pipet 20 mL of this solu-tion, add 30 mL of water, and titrate <2.50>, with vigorous shaking, with 0.1 mol/L silver nitrate VS (indicator: 3 drops of fluorescein sodium TS). Each mL 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl
Containers—Hermetic containers. Plastic containers for aqueous injections may be used.
Containers and storage Containers—Well-closed contain-ers.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 238 nm and 242 nm. (2) To 6 g of 10z Phenobarbital Powder add 150 mL of ethanol, shake well, and filter. Condense the filtrate on a water bath to about 5 mL, add about 50 mL of water, filter to collect the formed crystals, and dry them at 1059C for 2 hours. Determine the infrared absorption spectrum of the crystals as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of 10z Phenobarbital Powder is not less than 80z. Start the test with an accurately weighted about 0.3 g of 10z Phenobarbital Powder, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add exactly 10 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) and use this solution as the sample solution. Separately, weigh accurately about 17 mg of phenobarbital for assay, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 10 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a mixture of boric acid-potassium chloride-sodium hy-droxide buffer solution (pH 9.6) and water (2:1) as the blank, and determine the absorbances, AT and AS, at 240 nm. Dissolution rate (z) with respect to the labeled amount of phenobarbital (C12H12N2O3) ＝ MS/MT × AT/AS × 1/C × 180 MS: Amount (mg) of phenobarbital for assay taken MT: Amount (g) of 10z Phenobarbital Powder taken C: Labeled amount (mg) of phenobarbital (C12H12N2O3) in 1 g
Weigh accurately about 0.2 g of 10z Phenobarbital ‌Powder, dissolve in a boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) to make exactly 100 mL. Pipet 5 mL of this solution, add a boric acid-potassium chlo-ride-sodium hydroxide buffer solution (pH 9.6) to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of phenobarbital for assay, previously dried at 1059C for 2 hours, and add a boric acid-potassium chloride-sodium hydroxide buffer solu-tion (pH 9.6) to make exactly 100 mL. Pipet 5 mL of this so-lution, add a boric acid-potassium chloride-sodium hydrox-ide buffer solution (pH 9.6) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) as the blank, and determine the absorbances, AT and AS, at 240 nm. minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 33 mg of phenobarbital (C12H12N2O3). Pipet 5 mL of this solution, add exactly 10 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6), and use this solution as the sample solution. Separately, weigh accurately about 17 mg of phenobarbital for assay, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 25 mL. Pipet 5 mL of this solu-tion, add exactly 10 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6), and use this so-lution as the standard solution. Determine the absorbances, AT and AS, at 240 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho- Amount (mg) of phenobarbital (C ＝ MS × AT/AS 12H12N2O3) tometry <2.24>, using a mixture of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) and water (2:1) as the blank. MS: Amount (mg) of phenobarbital for assay taken
Copovidone is a copolymer of 1-vinyl-2-pyrrolidone and vinyl acetate at the ratio by mass of 3:2. It contains not less than 35.
Copovidone occurs as a white to yellowish white powder. It is odorless or has a faint, characteristic odor. It is very soluble in methanol and in ethanol (95), and freely soluble in water. It is hygroscopic.◆
Determine the infrared absorption spectrum of Copovidone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp- mL, and use this solution as the standard solution. Measure exactly 0.5 mL each of the sample solution, standard solu-tion and water, transfer to separate 1-cm cells, add 2.5 mL of 0.05 mol/L pyrophosphate buffer solution (pH 9.0) and 0.2 mL of b-nicotinamide adenine dinucleotide TS to each of these cells, mix and stopper tightly. Allow to stand for 2 to 3 minutes at 22 ± 29C, and perform the test with these solu-tions as directed under Ultraviolet-visible Spectrophotome-try <2.24> using water as the control solution. Determine the absorbances, AT1, AS1 and AB1, of the subsequent solutions of the sample solution, the standard solution and water (blank) at 340 nm. Then, add 0.05 mL of aldehyde de-hydrogenase TS to each of the cells, stir, and stopper tightly. Allow to stand at 22 ± 29C for 5 minutes. Determine the ab-sorbances, AT2, AS2 and AB2, of these solutions in the same manner as above: the content of aldehyde is not more than 500 ppm. Content (ppm) of aldehydes [as acetaldehyde (CH3CHO)] ＝ C/M ×{(AT2 － AT1) － (AB2 － AB1)}/{(AS2 － AS1) － (AB2 － AB1)}× 100,000 M: Amount (g) of Copovidone taken, calculated on the dried basis C: Concentration (mg/mL) of acetaldehyde in the stand-ard solution, using 0.72 as conversion factor for acetal-dehyde ammonia trimer trihydrate to acetaldehyde 1-Vinyl-2-pyrrolidone and free vinyl acetate—Store the sample solution and the standard solution at 59C or below, and use within 8 hours. Weigh accurately about 0.25 g of Copovidone, dissolve in a mixture of water and aceto-nitrile (23:2) to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 50 mg each of 1-vinyl-2-pyrrolidone and vinyl acetate in methanol to make exactly 100 mL. Pipet 1 mL of this solution and add metha-nol to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of water and acetonitrile (23:2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, deter-mine the peak areas, ATa, ATb, ASa and ASb, of 1-vinyl-2-pyr-rolidone and free vinyl acetate in each solution, and calcu-late the content of 1-vinyl-2-pyrrolidone and vinyl acetate by the following equations: they are not more than 10 ppm. Content (ppm) of 1-vinyl-2-pyrrolidone tion at the same wave numbers. ＝ ATa/ASa × CSa/CT × 1000 pH <2.54> Dissolve 1.0 g of Copovidone in 10 mL of water: the pH of this solution is between 3.0 and 7.0. Content (ppm) of free vinyl acetate ＝ ATb/ASb × CSb/CT × 1000
(1) Clarity and color of solution—Dissolve 1.0 g of Copovidone in 10 mL of water: the solution is clear or slightly opalescent and colorless to pale yellow, or pale red. CSa : Concentration ( mg/mL) of 1-vinyl-2-pyrrolidone in the standard solution (2) Heavy metals <1.07>—Ignite 2.0 g of Copovidone as directed under Residue on Ignition Test <2.44>, add 2 mL of hydrochloric acid to the residue, then proceed according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◇ Aldehydes—Weigh accurately about 1 g of Copovi-done, dissolve in 0.05 mol/L pyrophosphate buffer solution (pH 9.0) to make exactly 100 mL. Stopper, heat at 609C for 60 minutes, allow to cool to room temperature, and use this solution as the sample solution. Separately, dissolve 0.140 g of acetaldehyde ammonia trimer trihydrate in water to make exactly 200 mL. Pipet 1 mL of this solution, add 0.05 mol/L pyrophosphate buffer solution (pH 9.0) to make exactly 100 CSb: Concentration (mg/mL) of vinyl acetate in the stand- ard solution CT: Concentration (mg/mL) of Copovidone in the sample solution, calculated on the dried basis Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 235 nm for 1-vinyl-2-pyrrolidone, 205 nm for vinyl acetate). Column: Two stainless steel columns, one is 4 mm in in-side diameter and 33 mm in length and the other is 4 mm in inside diameter and 250 mm in length, packed with oc-tadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter), and use them as the pre-column and the separation column, respectively. Column temperature: A constant temperature of about JP XVIII Official Monographs / Copovidone 813 409C. Mobile phase: A mixture of water and acetonitrile (23:2). Flow rate: 1.0 mL per minute (Retention times of 1-vinyl- pyrrolidone and vinyl acetate are about 17 and about 22 minutes, respectively). tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of 2-pyrrolidone in each solution. Calculate the content of 2-pyrrolidone by the following equation: not more than 0.5z. Time span of measurement: For 40 minutes. Washing of column: After each test with the sample solu- Content (z) of 2-pyrrolidone ＝ AT/AS × CS/CT × 100 tion, elute and wash away remaining sample by passing the mobile phase through the separation column or the pre-column backwards at the flow rate mentioned above for about 30 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions (wavelength: 205 nm), 1-vinyl-2-pyrrolidone and vinyl acetate are eluted in this order with the resolution be-tween these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak areas of 1-vinyl-2-pyrrolidone and vinyl acetate are not more than 2.0z, respectively. Peroxides—Weigh exactly an amount of Copovi-done, equivalent to 4.0 g calculated on the dried basis, dis-solve in water to make exactly 100 mL, and use this solution as the sample solution. To 25 mL of the sample solution add 2 mL of titanium (III) chloride-sulfuric acid TS, and mix. Allow to stand for 30 minutes, and perform the test with this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared by adding 2 mL of diluted sulfuric acid (13 in 100) to 25 mL of the sample solu-tion as a blank: the absorbance of the sample solution at 405 nm is not more than 0.35 (not more than 400 ppm, as hydro-gen peroxide). Hydrazine—Weigh exactly an amount of Copovi-done equivalent to 2.5 g calculated on the dried basis, trans-fer to a 50-mL centrifuge tube, add 25 mL of water, and stir to dissolve. Add 500 mL of a solution of salicylaldehyde in methanol (1 in 20), stir and warm at 609C for 15 minutes in a water bath. Allow to cool, add 2.0 mL of toluene, stopper tightly, shake vigorously for 2 minutes, centrifuge, and use the upper layer of the mixture as the sample solution. Sepa-rately, dissolve 90 mg of salicylaldazine in toluene to make exactly 100 mL. Pipet 1 mL of this solution, add toluene to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of dimethylsilanized silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol and water (2:1) to a distance of about three-fourths of the length of the plate, and air-dry the plate. Exa-mine under ultraviolet light (main wavelength: 365 nm): the fluorescence of the spot obtained from the sample solution corresponding to the spot having a Rf value of about 0.3 from the standard solution is not more intense than that of the spot from the standard solution (not more than 1 ppm). 2-Pyrrolidone—Weigh accurately about 1 g of Copovidone, add 5 mL of methanol for liquid chromatogra-phy, and sonicate to dissolve. Add water to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 0.150 g of 2-pyrrolidone in a mixture of water and methanol for liquid chromatography (19:1) to make exactly 100 mL. Pipet 3 mL of this solution, add a mixture of water and methanol for liquid chromatography (19:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu- CS: Concentration (mg/mL) of 2-pyrrolidone in the stand-ard solution CT: Concentration (mg/mL) of Copovidone in the sample solution, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: Two stainless steel columns, one is 4.0 mm in in-side diameter and 10 mm in length and the other is 4.6 mm in inside diameter and 150 mm in length, packed with oc-tadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter), and use them as the pre-column and the separation column, respectively. Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol for liquid chromatography (19:1). Flow rate: 0.8 mL per minute (retention time of 2-pyrroli-done is about 7 minutes). Time span of measurement: For 30 minutes. Washing of column: After each test with the sample solu-tion, elute and wash away remaining sample by passing the mobile phase through the separation column or the pre-column backwards at the flow rate mentioned above for about 30 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the symmetry factor of the peak of 2-pyrrolidone is not more than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 2-pyrrolidone is not more than 2.0z. Loss on drying <2.41> Not more than 5.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). K-value Weigh accurately an amount of Copovidone, equivalent to 1.00 g, calculated on the dried basis, dissolve in water to make exactly 100 mL, allow to stand for 60 minutes, and use this solution as the sample solution. Per-form the test with the sample solution and with water at 259C as directed in Method 1 under Viscosity Determination <2.53>, and calculate the K-value by the following formula: the K-value of Copovidone is not less than 90.0z and not more than 110.0z of the nominal K-value. K ＝ 1.5 lognrel. － 1 ＋ 300c lognrel. ＋ (c ＋ 1.5c lognrel.)2 0.15 ＋ 0.003c 0.15c ＋ 0.003c2 c: Mass (g) of Copovidone in 100 mL of the solution, cal-culated on the dried basis nrel.: Kinematic viscosity of the sample solution relative to that of water
(1) Vinyl acetate—Weigh accurately about 2 g of Copovidone, add exactly 25 mL of 0.5 mol/L potassium hy-droxide-ethanol VS and a few glass beads, heat under reflux for 30 min. Titrate immediately with 0.5 mol/L hydrochloric acid VS (indicator: 1 mL of phenolphthalein TS). Perform a blank determination in the same manner, and make any nec-essary correction. Amount (z) of vinyl acetate Description Cortisone Acetate occurs as white, crystals or crystalline powder. It is sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. Melting point: about 2409C (with decomposition). It shows crystal polymorphism. Identification (1) To 2 mg of Cortisone Acetate add 2 mL ‌＝ 0.1 × 86.09 × 56.11 28.05 (n2 – n1) M of sulfuric acid, and allow to stand for a while: a yellowish green color is produced, and it gradually changes to yellow-orange. Examine the solution under ultraviolet light: the so- M: Amount (g) of Copovidone taken, calculated on the dried basis n1: Volume (mL) of 0.5 mol/L hydrochloric acid VS con-sumed in the test n2: Volume (mL) of 0.5 mol/L hydrochloric acid VS con-sumed in the blank test (2) Nitrogen—Weigh accurately about 0.1 g of Copovi-done, and place in a Kjeldahl flask. Add 5 g of a decomposi-tion accelerator (a powdered mixture of 33 g of potassium sulfate, 1 g of copper (II) sulfate pentahydrate and 1 g of titanium (IV) oxide), and wash down any adhering sample from the neck of the flask with a small amount of water. Add 7 mL of sulfuric acid allowing to flow down the inside wall of the flask. Heat the flask gradually until the solution has a clear, yellow-green color, and the inside wall of the flask is free from a carbonized material, and then heat for further 45 minutes. After cooling, add cautiously 20 mL of water, and connect the flask to the distillation apparatus pre-viously washed by passing steam through it. To the absorp-tion flask add 30 mL of a solution of boric acid (1 in 25), 3 drops of bromocresol green-methyl red TS and sufficient water to immerse the lower end of the condenser tube. Add 30 mL of a solution of sodium hydroxide (2 in 5) through the funnel, rinse cautiously the funnel with 10 mL of water, im-mediately close the clamp attached to the rubber tube, then start the distillation with steam to obtain 80 to 100 mL of the distillate. Remove the absorption flask from the lower end of the condenser tube, rinsing the end part with a small quantity of water, and titrate <2.50> the distillate with 0.025 mol/L sulfuric acid VS until the color of the solution changes from green through pale grayish blue to pale grayish red-purple. Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.025 mol/L sulfuric acid VS ＝ 0.700 mg of N ◆Containers and storage Containers—Tight containers.◆ Cortisone Acetate
Corn Starch occurs as white to pale yellow-white, masses or powder. It is practically insoluble in water and in ethanol (99.5).◆
(1) Examined under a microscope <5.01>, using mixture of water and glycerin (1:1), Corn Starch ap-pears as either angular polyhedral granules of irregular sizes with diameters of 2 – 23 mm or as rounded or spheroidal granules of irregular sizes with diameters of 25 – 35 mm. The central hilum consists of a distinct cavity or two- to five-rayed cleft, and there are no concentric striations. Between orthogonally oriented polarizing plates or prisms, the starch granules show a distinct black cross intersecting at the hilum. To 1 g of Corn Starch add 50 mL of water, boil for 1 minute, and allow to cool: a thin, cloudy mucilage is formed. To 1 mL of the mucilage obtained in (2) add 0.05 mL of diluted iodine TS (1 in 10): an orange-red to deep blue color is formed and the color disappears by heating. pH <2.54> Put 5.0 g of Corn Starch in a non-metal vessel, add 25.0 mL of freshly boiled and cooled water, mix gently for 1 minute, and allow to stand for 15 minutes: the pH of the solution is between 4.0 and 7.0.
(1) Iron—To 1.5 g of Corn Starch add 15 mL of 2 mol/L hydrochloric acid TS, mix, filter, and use the filtrate as the test solution. To 2.0 mL of Standard Iron Solution add water to make 20 mL, and use this solution as the con-trol solution. Put 10 mL each of the test solution and the control solution in test tubes, add 2 mL of a solution of citric acid (1 in 5) and 0.1 mL of mercapto acetic acid, and mix. Alkalize with ammonia solution (28) to litmus paper, add water to make 20 mL, and mix. Transfer 10 mL each of these solutions into test tubes, allow to stand for 5 minutes, ‌and compare the color of these solutions against a white background: the color of the test solution is not more intense than that of the control solution (not more than 10 ppm). Oxidizing substances—To 4.0 g of Corn Starch add ppm. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 50.0 mL of water, shake for 5 minutes, and centrifuge. To 30.0 mL of the supernatant liquid add 1 mL of acetic acid (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow to stand for 25 to 30 minutes at a dark place. Add 1 mL of starch TS, and titrate <2.50> with 0.002 mol/L sodium thio-sulfate VS until the color of the solution disappears. Per-form a blank determination in the same manner, and make any necessary correction: the volume of 0.002 mol/L sodium thiosulfate VS consumed is not more than 1.4 mL (not more than 20 ppm, calculated as hydrogen peroxide). Sulfur dioxide— Apparatus Use as shown in the following figure. A: Three-necked round-bottom flask (500 mL) B: Cylindrical dropping funnel (100 mL) C: Condenser D: Test tube E: Tap Procedure Introduce 150 mL of water into the three-necked round-bottom flask, close the tap of the cylin-drical dropping funnel, and pass carbon dioxide through the whole system at a rate of 100 ± 5 mL per minute. Pass cool-ing water through the condenser, and place 10 mL of hydro-gen peroxide-sodium hydroxide TS in the test tube. After 15 minutes, remove the funnel without interrupting the stream of carbon dioxide, and introduce through the opening into the flask about 25 g of Corn Starch, accurately weighed, with the aid of 100 mL of water. Apply tap grease to the out-side of the connection part of the funnel, and load the fun-nel. Close the tap of the funnel, pour 80 mL of 2 mol/L hy-drochloric acid TS into the funnel, open the tap to introduce the hydrochloric acid into the flask, and close the tap while several mL of the hydrochloric acid remains, in order to avoid losing sulfur dioxide. Place the flask in a water bath, and heat the mixture for 1 hour. Transfer the contents of the test tube with the aid of a little water to a wide-necked coni-cal flask. Heat in a water bath for 15 minutes, and cool. Add 0.1 mL of bromophenol blue TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the amount of sulfur dioxide by applying the following formula: it is not more than 50 M: Amount (g) of Corn Starch taken V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed (4) Foreign matter—Under a microscope <5.01>, Corn Starch does not contain starch granules of any other origin. It may contain a minute quantity, if any, of fragments of the tissue of the original plant.◆ Loss on drying <2.41> Not more than 15.0z (1 g, 1309C, 90 minutes). Residue on ignition <2.44> Not more than 0.6z (1 g).
Cortisone Acetate, when dried, contains not less than 97.
Related substances—Dissolve 25 mg of Cortisone Acetate in 10 mL of a mixture of acetonitrile, water and ace-tic acid (100) (70:30:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution add the mixture of acetonitrile, water and acetic acid (100) (70:30:1) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: each peak area other than cortisone ace-tate obtained from the sample solution is not larger than 1/2 times the peak area of cortisone acetate from the standard solution, and the total area of the peaks other than cortisone acetate is not larger than 1.5 times the peak area of cortisone acetate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water and acetonitrile (7:3). Mobile phase B: A mixture of acetonitrile and water (7:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 17,21-Dihydroxypregn-4-ene-3,11,20-trione 21-acetate [50-04-4] Cortisone Acetate, when dried, contains not less than 97.0z and not more than 102.0z of cortisone acetate (C23H30O6). ‌JP XVIII Official Monographs / Absorptive Cream 815‌ Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 5 5 – 25 90 → 10 10 → 90 25 – 30 with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cortisone acetate to that of the internal standard is not more than 1.0z.
Dissolve about 10 mg each of Cortisone Acetate and Cortisone Acetate RS, previously dried and accurately weighed, in 50 mL of methanol, add exactly 5 mL each of the internal standard solution, then add methanol to make 100 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cortisone acetate to that of the internal standard. Amount (mg) of cortisone acetate (C23H30O6) ＝ MS × QT/QS MS: Amount (mg) of Cortisone Acetate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (13:7). Flow rate: Adjust so that the retention time of cortisone acetate is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cortisone acetate and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times Absorptive Cream
Wood Creosote is a colorless or pale yellow, clear liquid. It has a characteristic odor. It is slightly soluble in water. It is miscible with methanol and with ethanol (99.5). Its saturated solution is acidic. It is highly refractive. It gradually changes in color by light or by air.
Use the sample solution obtained in the Assay as the sample solution. Separately, dissolve 0.1 g of phenol, p-cresol, guaiacol, and 2-methoxy-4-methylphenol in methanol respectively, to make 100 mL. To 10 mL of each solution add methanol to make 50 mL, and use these solu-tions as standard solution (1), standard solution (2), stan-dard solution (3) and standard solution (4). Perform the test with 10 mL each of the sample solution, standard solution (1), (2), (3) and (4) as directed under Liquid Chromato-graphy <2.01> according to the following conditions: the main peaks obtained with the sample solution show the same retention times with those obtained with the standard solu-tions (1), (2), (3) and (4). Operating conditions— Proceed as directed in the operating conditions in the Assay. Wood Creosote / Crude Drugs and Related Drugs Specific gravity <2.56> d 20: not less than 1.076.
(1) Coal Creosote—Accurately measure 10 mL of Wood Creosote, add methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, to 1 mg each of benzo[a]pyrene, benz[a]anthracene and dibenz[a,h]anthracene add a small quantity of ethyl acetate, if necessary, and add methanol to make 100 mL. To 1 mL of this solution add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions: No peaks are detected with the sample solution for the retention times corresponding to benzo[a]pyrene, benz[a]anthracene and dibenz[a,h]anthra-cene of the standard solution. Change these conditions if any peak is detected for retention times that correspond to ben-zo[a]pyrene, benz[a]anthracene or dibenz[a,h]anthracene, to verify that such a peak does not belong to benzo[a]pyrene, benz[a]anthracene or dibenz[a,h]anthracene. Operating conditions— Detector: A mass spectrometer (EI). Monitored ions: Benz[a]anthracene: Molecular ion m/z 228, Fragment ion m/z 114 About 14 to 20 minutes Benzo[a]pyrene: Molecular ion m/z 252, Fragment ion m/z 125 About 20 to 25 minutes Dibenz[a,h]anthracene: Molecular ion m/z 278, Fragment ion m/z 139 About 25 to 30 minutes Column: A quartz tube 0.25 mm in inside diameter and 30 m in length, with internal coating 0.25 – 0.5 mm in thick-ness made of 5z diphenyl-95z dimethyl polysiloxane for gas chromatography. Column temperature: Inject sample at a constant tempera-ture in vicinity of 459C, then raise temperature to 2409C at the rate of 409C per minute, maintain the temperature at 2409C for 5 minutes, then raise temperature to 3009C at the rate of 49C per minute, then raise the temperature to 3209C at the rate of 109C per minute, then maintain temperature at 3209C for 3 minutes. Injection port temperature: A constant temperature in vicinity of 2509C. Interface temperature: A constant temperature in vicinity of 3009C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of benzo[a]py-rene is about 22 minutes. Splitless. System suitability— Test for required detectability: Accurately measure 1 mL of standard solution and add methanol to make exactly 10 mL, and use this solution as the solution for system suita-bility test. When the test is performed with conditions described above with 1 mL of the solution for system suita-bility test, the SN ratio of each substance is not less than 3. System performance: When the procedure is run with conditions described above with 1 mL of the solution for system suitability test, the elution takes place in order of benz[a]anthracene, benzo[a]pyrene and then dibenz[a,h]anthracene. System repeatability: When the test is repeated 6 times with 1 mL of the solution for system suitability test under the above conditions, the relative standard deviation of the peak area of benzo[a]pyrene, benz[a]anthracene and dibenz[a,h]anthracene is respectively not more than 10z. Acenaphthene—To 0.12 g of Wood Creosote add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of acenaphthene in methanol to make 50 mL. To 5 mL of this solution add methanol to make 20 mL. To 2 mL of this solution add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with exactly 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con-ditions: No peaks are detected with sample solution for the retention time corresponding to acenaphthene of the stand-ard solution. Change these conditions if any peak is detected for the retention time corresponding to acenaphthene, to verify that such a peak does not belong to athenaphthene. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica tube 0.25 mm inside diameter and 60 m in length, with internal coating 0.25 – 0.5 mm in thick-ness made of polymethylsiloxane for gas chromatography. Column temperature: Perform injection at a constant tem-perature in vicinity of 459C, then raise the temperature by 11.59C per minute until reaching 1609C, then raise the tem-perature by 49C per minute until reaching 1809C, then raise the temperature by 89C until reaching 2709C, then maintain temperature at 2709C for 3 minutes. Injection port temperature: 2509C. Detector temperature: 2509C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of acenaphthene is about 18 minutes. Splitless. System suitability— Test for required detectability: Accurately measure 1 mL of the standard solution, add methanol to make exactly 10 mL, and use this solution as the solution for system suita-bility test. When the procedure is run with conditions described above with 1 mL of solution for system suitability test, the SN ratio of acenaphthene is not less than 3. System repeatability: When the test is repeated 6 times with 1 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of acenaphthene is not more than 6.0z. Other impurities Add 2 mL of petroleum benzin to 1.0 mL of Wood Creo-sote, then add 2 mL of barium hydroxide test solution, agitate to mix and allow to stand. No blue or muddy brown color develops in the upper layer of the mixture. Further-more, no red color develops in the lower layer. Distilling range <2.57> 200 – 2209C, not less than 85 volz.
To about 0.1 g of Wood Creosote, accurately weighed, add methanol to make exactly 50 mL. Pipet 10 mL of this solution add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, add methanol to about 30 mg of accurately measured guaiacol for assay to make exactly 50 mL. Pipet 10 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of guaiacol in each solution. Amount (mg) of guaiacol ＝ MS × AT/AS JP XVIII Crude Drugs and Related Drugs / Yokukansan Extract 2173 ‌MS: Amount (mg) of guaiacol for assay taken‌ Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of guaiacol is about 9 minutes. System suitability— System performance: Dissolve 2 mg each of guaiacol and phenol in methanol to make 10 mL. The procedure is run with conditions described above with 10 mL of this solution, the elution takes place in order of phenol then guaiacol, with the degree in separation of not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of guaiacol is not more than 1.5z.
Cresol is a clear, colorless or yellow to yellow-brown liquid. It has a phenol-like odor. It is miscible with ethanol (95) and with diethyl ether. It is sparingly soluble in water. It dissolves in sodium hydroxide TS. A saturated solution of Cresol is neutral to bromocresol purple TS. It is a highly refractive liquid. It becomes dark brown by light or on aging.
To 5 mL of a saturated solution of Cresol add 1 to 2 drops of dilute iron (III) chloride TS: a blue-purple color develops. Specific gravity <2.56> d 20: 1.032 – 1.041
(1) Hydrocarbons—Dissolve 1.0 mL of Cresol in 60 mL of water: the solution shows no more turbidity than that produced in the following control solution. Control solution: To 54 mL of water add 6.0 mL of 0.005 mol/L sulfuric acid VS and 1.0 mL of barium chloride TS, and after thorough shaking, allow to stand for 5 minutes. (2) Sulfur compounds—Transfer 20 mL of Cresol in a 100-mL conical flask, place a piece of moistened lead (II) acetate paper on the mouth of the flask, and warm for 5 minutes on a water bath: the lead (II) acetate paper may de-velop a yellow color, but neither a brown nor a dark tint. Distilling range <2.57> 196 – 2069C, not less than 90 volz.
Croconazole Hydrochloride, when dried, contains not less than 98.
Croconazole Hydrochloride occurs as white to pale yellow-white, crystals or crystalline powder. It is very soluble in water, freely soluble in methanol, in ethanol (95) and in acetic acid (100), and practically insolu-ble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Croconazole Hydrochloride in methanol (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Croconazole Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.05 g of Croconazole Hydrochloride in 10 mL of water, add 2 mL of sodium hydroxide TS and 20 mL of diethyl ether, and shake. Wash the separated aqueous layer with two 10-mL portions of diethyl ether, and acidify the solution with 2 mL of dilute nitric acid: the solution re-sponds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Croconazole Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Croconazole Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, methanol and ammonia solution (28) (30:15:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Croconazole Hy-drochloride, previously dried, dissolve in 10 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS [indicator: 1 to 2 drops of a solution of malachite green oxalate in acetic acid (100) (1 in 100)] until the color of the solution changes from blue-green through green to yellow-green. Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.72 mg of C18H15ClN2O.HCl
Thin cord-like stigma, externally dark yellow-red to red-brown, 1.5 – 3.5 cm in length, tripartite or sepa-rate; the end of partite part widened and the other end nar-rowed gradually. Odor, strong and characteristic; taste, bitter; colors the saliva yellow on chewing. Under a microscope <5.01>, when softened by immersion in water, the upper end has numerous tubular protrusions about 150 mm in length, with a small number of pollen grains.
Use the sample solution obtained in the Purity (1) as the sample solution. Perform the test with this solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, water and acetic acid (100) (20:5:4:1) to a distance of about 10 cm, and air-dry the plate: three yellow spots appear at the Rf values of about 0.1, about 0.25 and about 0.4.
(1) Synthetic dye—To 0.10 g of pulverized Saffron add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol, water and acetic acid (100) (20:5:4:1) to a distance of about 10 cm, and air-dry the plate: any spot other than yellow spots does not appear upper in position than a yellow spot at an Rf value of about 0.4 and any yellow spot does not ap-pear between yellow spots at Rf values of about 0.25 and about 0.4. Spray evenly diluted sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: any clear yellow spot does not appear at an Rf value of less than 0.1, and any orange spot does not appear between blue-purple spots at Rf values of about 0.1 and about 0.25. Glycerol, sugar or honey—Saffron has no sweet taste. Press it between two pieces of paper: no spot is left on the paper. Yellow style—When perform the test of foreign matter <5.01>, the yellow style in Saffron does not exceed 10.0z. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 7.5z. Content of the active principle Crocin—Dry Saffron in a desiccator (silica gel) for 24 hours, and powder. To exactly 0.100 g of the powder add 150 mL of warm water, warm the mixture between 609C and 709C for 30 minutes with fre-quent shaking, cool, and filter. Pipet 1 mL of the filtrate, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve exactly 98 mg of carbazochrome sodium sulfonate trihydrate in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Determine the absorbances of the sample solu-tion and standard solution at 438 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>: the absorbance of the sample solution is larger than that of the standard so-lution.
Crospovidone is a cross-linked polymer of 1-vinyl-2-pyrrolidone. It contains not less than 11.
Crospovidone occurs as a white to pale yel-low-powder. It is practically insoluble in water, in methanol and in ethanol (99.5). It is hygroscopic.◆
(1) Suspend 1 g of Crospovidone in 10 mL of water, add 0.1 mL of iodine TS, shake for 30 seconds, then add 1 mL of starch TS, and shake: a blue color is not produced within 30 seconds. (2) When add 0.1 g of Crospovidone to 10 mL of water, shake to suspend, and allow the suspension to stand, a clear liquid is not produced within 15 minutes. Particle size Weigh accurately about 20 g of Crospovidone, place in a 1000-mL conical flask, add 500 mL of water, shake for 30 minutes, and pour onto an accurately tared No. 235 (63 mm) sieve, previously washed with hot water and dried at 1059C for a night, and wash the residue with water until the passing water is clear. Dry the residue together with the sieve in a drying machine at 1059C for 5 hours without air-circulation. After cooling down in a desiccator for 30 minutes, weigh the mass of the residue with sieve, and calcu-late the amount of the residue on the sieve by the following equation: Type A is more than 15z, and type B is not more than 15z. Amount (z) of the residue of Crospovidone on No. 235 (63 mm) sieve ＝ (M1 － M3)/M2 × 100 M1: The mass (g) of the residue with sieve after 5 hours drying M2: Amount (g) of Crospovidone taken, calculated on the dried basis M3: Mass (g) of the sieve
◆(1) Heavy metals <1.07>—Proceed with 2.0 g of Crospovidone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Water-soluble substances—Place 25.0 g of Crospovi-done in a 400-mL beaker, add 200 mL of water, and stir for 1 hour. Transfer the suspension to a 250-mL volumetric flask, rinsing with water, and dilute to volume with water. Allow the bulk of the solids to settle. Filter about 100 mL of the almost clear supernatant liquid through a 0.45 mm mem-brane filter, protected by superimposing a 3 mm membrane filter. Transfer exactly 50 mL of the clear filtrate to a tared 100-mL beaker, evaporate to dryness and dry at 105 – 1109C for 3 hours: the mass of the residue is not more than 75 mg. 1-Vinyl-2-pyrrolidone—To 1.250 g of Crospovidone add exactly 50 mL of methanol, and shake for 60 minutes. Leave bulk to settle, filter through a 0.2 mm membrane filter, and use the filtrate as the sample solution. Separately, dis-solve 50 mg of 1-vinyl-2-pyrrolidone in methanol to make exactly 100 mL. Pipet 1 mL of this solution, and add metha-nol to make exactly 100 mL. To exactly 5 mL of this solution add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: the peak area of 1-vinyl-2-pyrrolidone obtained from the sample solution is not larger than that from the standard solution (not more than 10 ppm). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: Two stainless steel columns, one is 4 mm in in-side diameter and 25 mm in length and the other is 4 mm in inside diameter and 250 mm in length, they are packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter), and used them as the pre-column and the separation column, respectively. Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (9:1). Flow rate: 1.0 mL per minute. Washing of pre-column: After each injection of the sam-ple solution, wash the pre-column by passing the mobile phase backwards, at the same flow rate as applied in the test, for 30 minutes. System suitability— System performance: Dissolve 10 mg of 1-vinyl-2-pyrroli-done and 0.50 g of vinyl acetate in methanol to make 100 mL. To 1 mL of this solution add the mobile phase to make 100 mL. When the procedure is run with 50 mL of this solu-tion under the above operating conditions, 1-vinyl-2-pyrroli-done and vinyl acetate are eluted in this order with the reso-lution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 1-vinyl-2-pyrrolidone is not more than 2.0z. Peroxides— Method 1: Apply to the sample labeled as type A. Suspend 4.0 g of Crospovidone in 100 mL of water, and use as the sample suspension. To 25 mL of the sample suspension add 2 mL of titanium (III) chloride-sulfuric acid TS, allow to stand for 30 minutes, and filter. Determine the absorbance of the filtrate at 405 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the control, prepared by filtrating the sample suspension and adding 2 mL of diluted sulfuric acid (13 in 100) to 25 mL of this filtrate: not more than 0.35 (not more than 400 ppm expressed as hydrogen peroxide). Method 2: Apply to the sample labeled as type B. Suspend 2.0 g of Crospovidone in 50 mL of water, and use as the sample suspension. To 10 mL of the sample suspension add water to make 25 mL, add 2 mL of titanium (III) chloride-sulfuric acid TS, allow to stand for 30 minutes, and filter. Determine the absorbance of the filtrate at 405 nm as di- JP XVIII Official Monographs / Cyanamide 819 rected under Ultraviolet-visible Spectrophotometry <2.24>, using the control, prepared by filtrating the sample suspen-sion, adding water to 10 mL of this filtrate to make 25 mL and 2 mL of diluted sulfuric acid (13 in 100): not more than 0.35 (not more than 1000 ppm expressed as hydrogen peroxide). Loss on drying <2.41> Not more than 5.0z (0.5 g, 1059C, constant mass). ‌Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Crospovidone, place in a Kjeldahl flask, add 5 g of a decomposition acceler-ator (a powdered mixture of 33 g of potassium sulfate, 1 g of copper (II) sulfate pentahydrate and 1 g of titanium (IV) oxide) and 3 glass beads. Wash any adhering particles from the neck into the flask with a small quantity of water. Add 7 mL of sulfuric acid, allowing it to run down the inside wall of the flask. Gradually heat the flask until the solution has a clear, yellowish-green color, and the inside wall of the flask is free from carbonized material, and then heat for a further 45 minutes. After cooling, cautiously add 20 mL of water, and connect the flask to the distillation apparatus previously washed by passing steam through it. To the absorption flask add 30 mL of a solution of boric acid (1 in 25), 3 drops of bromocresol green-methyl red TS and sufficient water to im-merse the lower end of the condenser tube. Add 30 mL of a solution of sodium hydroxide (21 in 50) through a funnel, cautiously rinse the funnel with 10 mL of water, immediately close the clamp attached to the rubber tube, then start the distillation with steam to obtain 80 – 100 mL of distillate. Remove the absorption flask from the lower end of the con-denser tube, rinsing the end part with a small quantity of water, and titrate <2.50> the distillate with 0.025 mol/L sul-furic acid VS until the color of the solution changes from green through pale grayish blue to pale grayish red-purple. Carry out a blank determination in the same manner, and make any necessary correction. Each mL of 0.025 mol/L sulfuric acid VS ＝ 0.7003 mg of N
Cyanamide occurs as white, crystals or crystal-line powder. It is very soluble in water, in methanol, in ethanol (99.5) and in acetone. The pH of a solution of 1.0 g of Cyanamide in 100 mL of water is between 5.0 and 6.5. It is hygroscopic. Melting point: about 469C
(1) To 1 mL of a solution of Cyanamide (1 in 100) add 1 mL of potassium 1,2-naphthoquinone-4- sulfonate TS and 0.2 mL of sodium hydroxide TS: a deep red color develops. (2) Drop one or two drops of a solution of Cyanamide in acetone (1 in 100) onto a potassium bromide disk prepared as directed in the potassium bromide disk method under In-frared Spectrophotometry <2.25>, and air-dry the disk. De-termine the infrared absorption spectrum of the disk as di-rected in the film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Cyanamide in 10 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 2.0 g of Cyanamide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Water <2.48> Not more than 1.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Cyanamide, and dis-solve in water to make exactly 250 mL. Pipet 15 mL of this solution, add 2 to 3 drops of dilute nitric acid, 10 mL of am-monia TS and exactly 50 mL of 0.1 mol/L silver nitrate VS, and allow to stand for 15 minutes with occasional shaking. Add water to make exactly 100 mL, filter, discard the first 20 mL of the filtrate, and pipet 50 mL of the subsequent filtrate. After neutralizing this solution with dilute nitric acid, add 3 mL of dilute nitric acid, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 2.102 mg of CH2N2
Cyclopentolate Hydrochloride occurs as a white crystalline powder. It is odorless, or has a characteris-tic odor. It is very soluble in water, freely soluble in ethanol (95), in acetic acid (100) and in chloroform, sparingly soluble in ace-tic anhydride, and practically insoluble in diethyl ether.
(1) To 1 mL of a solution of Cyclopento-late Hydrochloride (1 in 100) add 1 mL of Reinecke salt TS: a light red precipitate is formed. Dissolve 0.2 g of Cyclopentolate Hydrochloride in 2 mL of water, add 2 mL of sodium hydroxide TS, and boil for 1 minute. After cooling, add 2 drops of nitric acid: a phenylacetic acid-like odor is perceptible. Determine the infrared absorption spectrum of Cy-clopentolate Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Cyclopentolate Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 0.20 g of Cyclopentolate Hydrochlo-ride in 20 mL of water: the pH of this solution is between 4.5 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Cyclopentolate Hydrochloride in 10 mL of water: the so-lution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Cy- clopentolate Hydrochloride according to Method 1, and per- 63 88 14 14 form the test. Prepare the control solution with 2.0 mL of use this solution as the sample solution. Separately, weigh accurately about 20 mg of Cyanocobalamin RS (previously determine the loss on drying <2.41> under the same condi-tions as Cyanocobalamin), add water to make exactly 1000 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cyanocobalamin. Amount (mg) of cyanocobalamin (C63H88CoN14O14P) ＝ MS × AT/AS × 1/10 Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Cyclopento-late Hydrochloride in 10 mL of chloroform, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, and add chloroform to make exactly 20 mL. Pipet 1 mL of this solution, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro- ‌822‌ matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-propanol, n-butyl acetate, water and ammonia solution (28) (100:60:23:17) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of sulfuric acid in ethanol (99.5) (1 in 10) on the plate, and heat the plate at 1209C for 30 minutes. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.05z (1 g).
Weigh accurately about 0.5 g of Cyclopentolate Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (4:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-green to yel-low-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.79 mg of C17H25NO3.HCl
Cyclophosphamide Hydrate contains not less than 97.
Cyclophosphamide Hydrate occurs as white, crystals or crystalline powder. It is odorless. It is very soluble in acetic acid (100), freely soluble in ethanol (95), in acetic anhydride and in chloroform, and soluble in water and in diethyl ether. Melting point: 45 – 539C
(1) Dissolve 0.1 g of Cyclophosphamide Hydrate in 10 mL of water, and add 5 mL of silver nitrate TS: no precipitate is produced. Then boil this solution: a white precipitate is produced. Collect the precipitate, and add dilute nitric acid to a portion of this precipitate: it does not dissolve. Add excess ammonia TS to another portion of the precipitate: it dissolves. (2) Add 1 mL of diluted sulfuric acid (1 in 25) to 0.02 g of Cyclophosphamide Hydrate, and heat until white fumes are evolved. After cooling, add 5 mL of water, and shake. Neutralize with ammonia TS, then acidify with dilute nitric acid: this solution responds to Qualitative Tests <1.09> (2) for phosphate.
(1) Clarity and color of solution—Dissolve 0.20 g of Cyclophosphamide Hydrate in 10 mL of water: the solu-tion is clear and colorless. Chloride <1.03>—Perform the test with 0.40 g of Cy- clophosphamide Hydrate at a temperature not exceeding 209C. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 1.0 g of Cy- clophosphamide Hydrate according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Water <2.48> 5.5 – 7.0z (0.5 g, volumetric titration, direct titration).
Weigh accurately about 0.3 g of Cyclophosphamide Hydrate, add 15 mL of hydrogen chloride-ethanol TS, and heat in a water bath under a reflux condenser for 3.5 hours while protecting from moisture. Distil the ethanol under reduced pressure. Dissolve the residue in 40 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid-dioxane VS (indicator: 2 drops of crystal violet TS) until the color of the solution changes from blue through green to yellow. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid-dioxane VS ＝ 13.96 mg of C7H15Cl2N2O2P.H2O
Cyproheptadine Hydrochloride Hydrate, when dried, contains not less than 98.
Cyproheptadine Hydrochloride Hydrate oc-curs as a white to pale yellow crystalline powder. It is odor-less, and has a slightly bitter taste. It is freely soluble in methanol and in acetic acid (100), soluble in chloroform, sparingly soluble in ethanol (95), slightly soluble in water, and practically insoluble in diethyl ether.
(1) Dissolve 0.1 g of Cyproheptadine Hy-drochloride Hydrate in 10 mL of methanol, apply 1 drop of this solution on filter paper, air-dry, and examine under ul-traviolet light (main wavelength: 254 nm): the solution shows a pale blue fluorescence. Weigh 0.1 g of Cyproheptadine Hydrochloride Hy-drate, transfer to a separator, dissolve in 5 mL of chlo-roform, add 4 mL of water and 1 mL of sodium carbonate TS, and shake. Transfer the chloroform layer to another separator, and wash with 4 mL of water by shaking well. Filter the chloroform layer through absorbent cotton moistened previously with chloroform, and evaporate the filtrate to dryness. Dissolve the residue in 8 mL of dilute ethanol by warming at 659C. Rub the inner wall of the con-tainer with a glass rod while cooling until crystallization be-gins, and allow to stand for 30 minutes. Collect the crystals, and dry at 809C for 2 hours: the crystals melt <2.60> between 1119C and 1159C. Determine the absorption spectrum of a solution of Cyproheptadine Hydrochloride Hydrate in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. A saturated solution of Cyproheptadine Hydrochlo-ride Hydrate responds to Qualitative Tests <1.09> (2) for chloride.
(1) Acidity—Dissolve 2.0 g of Cyproheptadine Hydrochloride Hydrate in 25 mL of methanol, and add 1 drop of methyl red TS and 0.30 mL of 0.1 mol/L sodium hy-droxide VS: a yellow color develops. (2) Heavy metals <1.07>—Proceed with 1.0 g of Cypro- heptadine Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> 7.0 – 9.0z (1 g, in vacuum at a pres- ‌JP XVIII Official Monographs / L-Cysteine 825 sure not exceeding 0.67 kPa, 1009C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) Medium for test organism [5] under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 6.0 to 6.1 after sterilization. Standard solutions—Weigh accurately an amount of Cycloserine RS, previously dried at 609C for 3 hours under reduced pressure of not exceeding 0.67 kPa, equivalent to about 40 mg (potency), dissolve in water to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 24 hours. Take exactly a suitable amount of the standard stock solution before use, and add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 100 mg (potency) and 50 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Cycloserine equivalent to about 40 mg (potency), dissolve in water to make exactly 100 mL. Take exactly a suitable amount of this solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 100 mg (po-tency) and 50 mg (potency), and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
Cytarabine, when dried, contains not less than 98.
Cytarabine occurs as white, crystals or crystal-line powder. It is freely soluble in water, soluble in acetic acid (100), and very slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. Melting point: about 2149C (with decomposition).
(1) Determine the absorption spectrum of a solution of Cytarabine in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cytarabine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋154 – ＋1609(after drying, 0.1 g, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.20 g of Cytarabine in 20 mL of water: the pH of this solution is between 6.5 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Cytarabine in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 1.0 g of Cytarabine. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.009z). Heavy metals <1.07>—Proceed with 1.0 g of Cytara-bine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Cytarabine in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 200 mL, and use this solution as the standard solution (1). Pipet 10 mL of the standard solution (1), add water to make exactly 25 mL and use this solution as the standard so-lution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with 1-butanol saturated with water to a distance of about 12 cm, and air-dry the ‌828‌‌ plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and the number of them which are more intense than the spot from the standard solution (2) is not more than two. Spray evenly acidic potassium perman-ganate TS on the plate: any spot other than the principal spot does not appear. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
Weigh accurately about 0.2 g of Cytarabine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 12.16 mg of C9H13N3O5
d-Camphor occurs as colorless or white, trans-lucent crystals, crystalline powder or masses. It has a charac-teristic, agreeable odor, and a slightly bitter taste, followed by a pleasant, cooling sensation. It is freely soluble in ethanol (95), in diethyl ether and in carbon disulfide, and slightly soluble in water. It slowly volatilizes at room temperature.
Dissolve 0.1 g of d-Camphor in 2 mL of methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and heat for 5 minutes on a water bath: an orange-red precipitate is formed. D Optical rotation <2.49> [a]20: ＋41.0 – ＋43.09(5 g, ethanol System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of d-camphor to that of the internal standard is not more than 1.0z.
Weigh accurately about 0.1 g each of d-Camphor and d-Camphor RS, add exactly 5 mL each of the internal standard solution, dissolve in ethanol (99.5) to make 100 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of d-camphor to that of the internal standard. Amount (mg) of d-camphor (C10H16O) ＝ MS × QT/QS MS: Amount (mg) of d-Camphor RS taken Internal standard solution—A solution of methyl salicylate in ethanol (99.5) (1 in 25). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 3 m in length, which is packed with 10z of polyethylene glycol 20 M for gas chromatography supported on 180 to 250 mm mesh silanized siliceous earth for gas chromatography. Column temperature: A constant temperature of about 1609C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of d-camphor is about 6 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, d-camphor and the internal standard are eluted in this phor (C 10H16 O). order with the resolution between these peaks being not less than 7. Description d-Camphor occurs as colorless or white, trans-lucent crystals, crystalline powder or masses. It has a charac-teristic, agreeable odor, and a slightly bitter taste, followed by a pleasant, cooling sensation. It is freely soluble in ethanol (95), in diethyl ether and in carbon disulfide, and slightly soluble in water. It slowly volatilizes at room temperature. Identification Dissolve 0.1 g of d-Camphor in 2 mL of methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and heat for 5 minutes on a water bath: an orange-red precipitate is formed. D Optical rotation <2.49> [a]20: ＋41.0 – ＋43.09(5 g, ethanol System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of d-camphor to that of the internal standard is not more than 1.0z.
d-クロルフェニラミンマレイン酸塩
d-Chlorpheniramine Maleate, when dried, contains not less than 99.
d-Chlorpheniramine Maleate occurs as a white crystalline powder. It is very soluble in water, in methanol and in acetic acid (100), and freely soluble in N,N-dimethylformamide and in ethanol (99.5). It dissolves in dilute hydrochloric acid.
(1) Determine the absorption spectrum of a solution of d-Chlorpheniramine Maleate in 0.1 mol/L hy-drochloric acid TS (3 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the ‌744 spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of d-Chlorpheniramine Maleate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.10 g of d-Chlorpheniramine Maleate in 5 mL of methanol, and use this solution as the sample solu-tion. Separately, dissolve 56 mg of maleic acid in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, methanol, acetic acid (100) and water (70:20:7:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot among two of the spots obtained with the sample solution shows the same intense to the spot obtained with the standard solution, and its Rf value is about 0.4. D Optical rotation <2.49> [a]20: ＋39.5 – ＋43.09(after dry-ing, 0.5 g, N,N-dimethylformamide, 10 mL, 100 mm). pH <2.54> Dissolve 1.0 g of d-Chlorpheniramine Maleate in 100 mL of freshly boiled and cooled water: the pH of this so-lution is between 4.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of d-Chlorpheniramine Maleate in 50 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with l.0 g of d-Chlor- pheniramine Maleate according to Method 4 and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of d-Chlor-pheniramine Maleate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 3 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than maleic acid and d-chlorpheniramine obtained from the sample solution is not larger than 2/3 times the peak area of d-chlorpheniramine from the standard solution, and the total area of these peaks is not larger than the peak area of d-chlorpheniramine from the standard solution. Time span of measurement: About 4 times as long as the retention time of d-chlorpheniramine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2.5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of d-chlorpheniramine ob-tained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of d-chlorpheniramine are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of d-chlorpheniramine is not more than 4.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 659C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately the mass of not less than 20 Chlor-pheniramine Maleate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 4 mg of chlor-pheniramine maleate (C16H19ClN2.C4H4O4), add 70 mL of the internal standard solution, shake for 15 minutes, then add the internal standard solution to make exactly 100 mL, filter through a membrane filter with a pore size not exceed-ing 0.5 mm, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Chlorphenira-mine Maleate RS, previously dried at 1059C for 3 hours, and add the internal standard to make exactly 100 mL. Pipet 20 mL of this solution, add the internal standard solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of (3S )-3-(4-Chlorophenyl)-N,N-dimethyl-3-pyridin-2-ylpropylamine monomaleate [2438-32-6] d-Chlorpheniramine Maleate, when dried, contains not less than 99.0z and not more than 101.0z of d-chlorpheniramine maleate (C16H19ClN2.C4H4O4). Description d-Chlorpheniramine Maleate occurs as a white crystalline powder. It is very soluble in water, in methanol and in acetic acid (100), and freely soluble in N,N-dimethylformamide and in ethanol (99.5). It dissolves in dilute hydrochloric acid. Identification (1) Determine the absorption spectrum of a solution of d-Chlorpheniramine Maleate in 0.1 mol/L hy-drochloric acid TS (3 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the ‌744 spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of d-Chlorpheniramine Maleate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.10 g of d-Chlorpheniramine Maleate in 5 mL of methanol, and use this solution as the sample solu-tion. Separately, dissolve 56 mg of maleic acid in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, methanol, acetic acid (100) and water (70:20:7:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot among two of the spots obtained with the sample solution shows the same intense to the spot obtained with the standard solution, and its Rf value is about 0.4. D Optical rotation <2.49> [a]20: ＋39.5 – ＋43.09(after dry-ing, 0.5 g, N,N-dimethylformamide, 10 mL, 100 mm). pH <2.54> Dissolve 1.0 g of d-Chlorpheniramine Maleate in 100 mL of freshly boiled and cooled water: the pH of this so-lution is between 4.0 and 5.0. Melting point <2.60> 111 – 1159C Purity (1) Clarity and color of solution—Dissolve 1.0 g of d-Chlorpheniramine Maleate in 50 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with l.0 g of d-Chlor- pheniramine Maleate according to Method 4 and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of d-Chlor-pheniramine Maleate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 3 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than maleic acid and d-chlorpheniramine obtained from the sample solution is not larger than 2/3 times the peak area of d-chlorpheniramine from the standard solution, and the total area of these peaks is not larger than the peak area of d-chlorpheniramine from the standard solution. Time span of measurement: About 4 times as long as the retention time of d-chlorpheniramine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2.5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of d-chlorpheniramine ob-tained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of d-chlorpheniramine are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of d-chlorpheniramine is not more than 4.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 659C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.3 g of d-Chlorphenira-mine Maleate, previously dried, and dissolve in 20 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-green to green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.54 mg of C16H19ClN2.C4H4O4
D-Mannitol [69-65-8] MS: Amount (mg) of Manidipine Hydrochloride RS taken C: Labeled amount (mg) of manidipine hydrochloride (C35H38N4O6.2HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and a solution of potassium dihydrogen phosphate (681 in 100,000) (3:2). Flow rate: Adjust so that the retention time of manidipine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of manidipine are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times This monograph is harmonized with the European Phar-macopoeia and the U.S. Pharmacopeia. The parts of the text that are not harmonized are marked with symbols (◆ ◆). Information on the harmonization with the European Pharmacopoeia and the U.S. Pharmacopeia is available on the website of the Pharmaceuticals and Medical Devices Agency. D-Mannitol contains not less than 97.
D-Mannitol occurs as white, crystals, powder or grain. It has a sweet taste with a cold sensation. It is freely soluble in water, and practically insoluble in ethanol (99.5). It dissolves in sodium hydroxide TS. It shows crystal polymorphism.◆
Determine the infrared absorption spectrum of D-Mannitol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of D-Mannitol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, put 25 mg each of D-Mannitol and D-Mannitol RS in glass vessels, dissolve in 0.25 mL of water without heating, dry them in a 600 – 700 W microwave oven for 20 minutes or in a drying chamber at 1009C for 1 hour, then further dry by gradual reducing pres-sure, and perform the same test as above with so obtained non-sticky white to pale yellow powders: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 5.0 g of D-Mannitol in water to make 50 mL, and use this solution as the test solution. Perform the test with the test solution as directed under Turbidity Measurement <2.61>: the solution is clear. Perform the test with the test solution according to Method 2 under Methods for Color Matching <2.65>: the so-lution is colorless. (2) Heavy metals <1.07>—Proceed with 5.0 g of D-Man- nitol according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm)◆. Nickel—Shake 10.0 g of D-Mannitol with 30 mL of 2 mol/L acetic acid TS, and add water to make exactly 100 mL. Add 2.0 mL of a saturated solution of ammonium pyrrolidinedithiocarbamate (about 10 g/L) and 10.0 mL of water-saturated 4-methyl-2-pentanone, and shake for 30 sec-onds without exposure to light. Allow the layers to separate, and use the 4-methyl-2-pentanone layer as the sample solu-tion. Separately, put 10.0 g each of D-Mannitol in three ves-sels, add 30 mL of 2 mol/L acetic acid TS to them, shake, add a suitable amount of water and exactly 0.5 mL, 1.0 mL and 1.5 mL respectively of Standard Nickel Solution for Atomic Absorption Spectrophotometry, and add water to make them exactly 100 mL. Then, proceed in the same man-ner as the sample solution, and use so obtained three 4-methyl-2-pentanone layers as the standard solutions. Addi-tionally, prepare a 4-methyl-2-pentanone layer by proceeding in the same manner as the sample solution without using D-Mannitol, and use this layer as the blank solution. Perform the test with the sample solution and standard solutions as directed in the standard addition method under Atomic Ab-sorption Spectrophotometry <2.23> according to the follow-ing conditions. Set the zero of the instrument using the blank solution, and between each measurement, rinse with water and ascertain that the readings return to zero with the blank solution: amount of nickel is not more than 1 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Nickel hollow-cathode lamp. Wavelength: 232.0 nm. Related substances—Dissolve 0.50 g of D-Mannitol in water to make 10 mL, and use this solution as the sample so-lution. Pipet 2 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 0.5 mL of the standard solution (1), add water to make exactly 20 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> accord-ing to the following conditions. Determine each peak area by the automatic integration method: the peak area of D-sor-bitol, having the relative retention time of about 1.2 to D-mannitol, obtained from the sample solution is not larger than that of D-mannitol from the standard solution (1) (not more than 2.0z), the total peak area of maltitol, having the relative retention time of about 0.69, and isomalt, having the relative retention times of about 0.6 and about 0.73, is not larger than the peak area of D-mannitol from the standard solution (1) (not more than 2.0z), and the area of the peak other than D-mannitol and the peaks mentioned above is not larger than 2 times the peak area of D-mannitol from the standard solution (2) (not more than 0.1z). Furthermore, the total area of the peak other than D-mannitol from the sample solution is not larger than the peak area of D-man-nitol from the standard solution (1) (not more than 2.0z). For these calculations exclude the peak which area is not larger than the peak area of D-mannitol from the standard solution (2) (not more than 0.05z). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of D-mannitol. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Confirm that the peak area of D-mannitol obtained with 20 mL of the standard solu-tion (2) is equivalent to 1.75 to 3.25z of that with 20 mL of the standard solution (1). System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviation of the peak area of D-mannitol is not more than 1.0z.◆ Glucose—To 7.0 g of D-Mannitol add 13 mL of water and 40 mL of Fehling's TS, boil gently for 3 minutes, and allow to stand for 2 minutes to precipitate copper (I) oxide. Separate the supernatant liquid, filter through a sintered glass filter for cupric oxide filtration coated with siliceous earth or a sintered glass filter (G4). Wash the precipitates with 50 – 609C hot water until the washing no longer alkaline, and filter the washings through the filter de-scribed above. Discard all the filtrate at this step. Immedi-ately, dissolve the precipitate with 20 mL of iron (III) sulfate TS, filter through the filter described above in a clean flask, and wash the filter with 15 – 20 mL of water. Combine the filtrate and the washings, heat to 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS until the green color turns to light red and the color persists at least 10 sec-onds: not more than 3.2 mL is required to change the color of the solution (not more than 0.1z expressed as glucose). Conductivity <2.51> Dissolve 20.0 g of D-Mannitol in a fleshly boiled and cooled water prepared from distilled water by heating to 40 – 509C, add the same water to make 100 mL, and use this solution as the sample solution. After cool-ing, measure the conductivity of the sample solution at 25 ± 0.19C while gently stirring with a magnetic stirrer: not more than 20 mS･cm－1. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours).
Weigh accurately about 0.5 g each of D-Mannitol and D-Mannitol RS (separately determine the loss on drying <2.41> under the same conditions as D-Mannitol), dissolve separately in water to make exactly 10 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of D-mannitol in each solution. Amount (g) of D-mannitol (C6H14O6) ＝ MS × AT/AS MS: Amount (g) of D-Mannitol RS taken, calculated on the dried basis Operating conditions— ‌Detector: A differential refractometer maintained at a constant temperature (409C for example).‌ Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (calcium type) composed with a sulfonated polystyrene cross-linked with 8z of divinylbenzene (9 mm in particle diameter). Column temperature: 85 ± 29C. Mobile phase: water. Flow rate: 0.5 mL per minute (the retention time of D-mannitol is about 20 minutes). System suitability— System performance: Dissolve 0.25 g each of D-Mannitol and D-sorbitol in water to make 10 mL, and use this solution as the solution for system suitability test (1). Separately, dis-solve 0.5 g each of maltitol and isomalt in water to make 100 mL. To 2 mL of this solution add water to make 10 mL, and in 5), no precipitate is produced. pH <2.54> 4.5 – 7.0 Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Measure exactly a volume of D-Mannitol Injection, equivalent to about 5 g of D-mannitol (C6H14O6), and add water to make exactly 250 mL. To exactly 10 mL of this so-lution add water to make exactly 100 mL. Measure exactly 10 mL of this solution into an iodine flask, add exactly 50 mL of potassium periodate TS, and heat for 15 minutes in a water bath. After cooling, add 2.5 g of potassium iodide, stopper tightly, and shake well. Allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium thiosulfate VS use this solution as the solution for system suitability test (2). When proceed with 20 mL each of the solution for system ＝ 1.822 mg of C6H 14O6 suitability test (1) and the solution for system suitability test (2) as directed under the above operating conditions, isomalt (first peak), maltitol, isomalt (second peak), D-mannitol and D-sorbitol are eluted in this order, the relative retention time of isomalt (first peak), maltitol, isomalt (second peak) and D-sorbitol to D-mannitol is about 0.6, about 0.69, about 0.73 and about 1.2, respectively, and the resolution between the peaks of D-mannitol and D-sorbitol is not less than 2.0. Coe-lution of maltitol and the second peak of isomalt may be ob-served. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of D-mannitol is not more than 1.0z.◆
D-Sorbitol, when dried, contains not less than 97.
D-Sorbitol occurs as white, granules, powder, or crystalline masses. It is odorless, and has a sweet taste with a cold sensation. It is very soluble in water, sparingly soluble in ethanol (95), and practically insoluble in diethyl ether. It is hygroscopic.
(1) To 1 mL of a solution of D-Sorbitol (7 in 10) add 2 mL of iron (II) sulfate TS and 1 mL of a solu-tion of sodium hydroxide (1 in 5): a blue-green color devel-ops, but no turbidity is produced. Shake thoroughly 1 mL of a solution of D-Sorbitol (1 in 20) with 1 mL of a freshly prepared solution of catechol (1 in 10), add rapidly 2 mL of sulfuric acid, and shake: a reddish purple to red-purple color immediately develops. Boil 0.5 g of D-Sorbitol with 10 mL of acetic anhy-dride and 1 mL of pyridine under a reflux condenser for 10 minutes, cool, shake with 25 mL of water, and allow to stand in a cold place. Transfer the solution to a separator, extract with 30 mL of chloroform, and evaporate the extract on a water bath. Add 80 mL of water to the oily residue, heat for 10 minutes on a water bath, then filter the hot mix-ture. After cooling, collect the produced precipitate through a glass filter (G3), wash with water, recrystallize once from ethanol (95), and dry in a desiccator (in vacuum, silica gel) for 4 hours: the precipitate melts <2.60> between 979C and 1019C.
(1) Clarity and color of solution, and acidity or alkalinity—Dissolve 5 g of D-Sorbitol in 20 mL of water by warming with shaking: the solution is clear, colorless, and neutral. Chloride <1.03>—Perform the test with 2.0 g of D- Sorbitol. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.005z). Sulfate <1.14>—Perform the test with 4.0 g of D- Sorbitol. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.006z). Heavy metals <1.07>—Proceed with 5.0 g of D-Sorbitol according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Nickel—Dissolve 0.5 g of D-Sorbitol in 5 mL of water, add 3 drops of dimethylglyoxime TS and 3 drops of ammonia TS, and allow to stand for 5 minutes: no red color develops. Arsenic <1.11>—Prepare the test solution with 1.5 g of D-Sorbitol according to Method 1, and perform the test (not more than 1.3 ppm). Glucose—Dissolve 20.0 g of D-Sorbitol in 25 mL of water, and boil gently with 40 mL of Fehling's TS for 3 minutes. After cooling, filter the supernatant liquid cau-tiously through a glass filter (G4), leaving the precipitate in the flask as much as possible, wash the precipitate with hot water until the last washings no longer show an alkali reac-tion, and filter the washings through the glass filter. Dissolve the precipitate in the flask in 20 mL of iron (III) sulfate TS, filter through the glass filter, and wash with water. Combine the filtrate and the washings, heat at 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS: not more than 6.3 mL of volume for titration consumed or consump-tion is required. Sugars—Dissolve 20.0 g of D-Sorbitol in 25 mL of water, and heat with 8 mL of dilute hydrochloric acid under a reflux condenser in a water bath for 3 hours. After cool-ing, add 2 drops of methyl orange TS, followed by sodium hydroxide TS until an orange color develops, and add water to make 100 mL. Boil gently 10 mL of this solution with 10 mL of water and 40 mL of Fehling's TS for 3 minutes and proceed as directed in (7). Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.02z (5 g).
Weigh accurately about 0.2 g of D-Sorbitol, previ-ously dried, dissolve in water and add water to make exactly 100 mL. Pipet 10 mL of the solution into an iodine flask, add exactly 50 mL of potassium periodate TS, and heat for 15 minutes in a water bath. Cool, add 2.5 g of potassium iodide, immediately stopper tightly, and shake well. Allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 1.822 mg of C6H14O6
Actinomycin D occurs as an orange-red to red crystalline powder. It is freely soluble in acetone, sparingly soluble in aceto-nitrile and in methanol, slightly soluble in ethanol (99.5), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Actinomycin D in methanol (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Actinomycin D RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve 0.1 g each of Actinomycin D and Actinomy-cin D RS in 10 mL of acetone, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and methanol (4:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the principal spot from the sample solution is the same as that from the standard solution. D Optical rotation <2.49> [a]20: －293 – －3299(after drying, 10 mg, methanol, 10 mL, 100 mm). Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, 609C, 3 hours).
Weigh accurately an amount of Actinomycin D and Actinomycin D RS, previously dried, equivalent to about 60 mg (potency), dissolve each in the mobile phase to make ex-actly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak area of actino- mycin D, AT and AS, in each solution. Amount [ mg (potency)] of actinomycin D (C62H86N12O16) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Actinomycin D RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L acetic acid-sodi-um acetate TS and acetonitrile (25:23). Flow rate: Adjust so that the retention time of actinomy-cin D is about 23 minutes. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of actinomycin D are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of actinomycin D is not more than 2.0z.
Daiokanzoto Extract contains not less than 3.
Daiokanzoto Extract occurs as a brown pow-der. It has a characteristic odor and an astringent first then slightly sweet taste.
(1) To 1.0 g of Daiokanzoto Extract add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solu-tion. Separately, dissolve 1 mg of rhein for thin-layer chro-matography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange fluo-rescent spot from the standard solution (Rhubarb). (2) To 0.5 g of Daiokanzoto Extract add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a acetate, shake for 10 minutes, centrifuge, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of meth-anol, shake for 30 minutes, centrifuge, and take the superna-tant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the superna-tant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 5 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of sennoside A in each solution. Amount (mg) of sennoside A (C42H38O20) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (2460:540:1). Flow rate: 1.0 mL per minute (the retention time of senno-side A is about 14 minutes.) System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sennoside A are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z. (2) Glycyrrhizic acid—Weigh accurately about 0.2 g of ‌1996 Daisaikoto Extract / Crude Drugs and Related Drugs Daiokanzoto Extract, add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re- Method of preparation 1) 2) 3) 4) 5) move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove Bupleurum Root 6 g 6 g 6 g 6 g 6 g the ethyl acetate layer. To the aqueous layer add 10 mL of Pinellia Tuber 4 g 4 g 4 g 3 g 4 g methanol, shake for 30 minutes, centrifuge, and take the Scutellaria Root 3 g 3 g 3 g 3 g 3 g supernatant liquid. To the residue add 20 mL of diluted Peony Root 3 g 3 g 3 g 3 g 3 g methanol (1 in 2), shake for 5 minutes, centrifuge, and take Jujube 3 g 3 g 3 g 3 g 3 g the supernatant liquid. Combine these supernatant liquids, Immature Orange 2 g 2 g 2 g 2 g 2 g add diluted methanol (1 in 2) to make exactly 50 mL, and use Ginger 1 g 1 g 2 g 1 g 1.5 g this solution as the sample solution. Separately, weigh accu- Rhubarb 1 g 2 g 1 g 1 g 2 g rately about 10 mg of Glycyrrhizic Acid RS (separately de- termine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Daiokanzoto Extract as directed under Extract (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of Daiokanzoto Extract according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z.
(1) Sennoside A—Weigh accurately about 0.2 g of Daiokanzoto Extract, add 20 mL of ethyl acetate and 10 mL of sennoside A (C42 H38 O20 : 862.74), and not less than 7 of water, shake for 10 minutes, centrifuge, and remove the ethyl acetate layer. To the aqueous layer add 20 mL of ethyl mg and not more than 21 mg (for preparation prescribed 1 g of Glycyrrhiza) or not less than 14 mg and not more than 42 mg (for preparation prescribed 2 g of Glycyrrhiza) of glycyrrhizic acid (C42H62O16: 822.93), per extract prepared with the amount speci-fied in the Method of preparation. Method of preparation 1) 2) Rhubarb 4 g 4 g Glycyrrhiza 1 g 2 g Prepare a dry extract as directed under Extracts, according to the prescription 1) or 2), using the crude drugs shown above. Description Daiokanzoto Extract occurs as a brown pow-der. It has a characteristic odor and an astringent first then slightly sweet taste. Identification (1) To 1.0 g of Daiokanzoto Extract add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solu-tion. Separately, dissolve 1 mg of rhein for thin-layer chro-matography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange fluo-rescent spot from the standard solution (Rhubarb). (2) To 0.5 g of Daiokanzoto Extract add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a acetate, shake for 10 minutes, centrifuge, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of meth-anol, shake for 30 minutes, centrifuge, and take the superna-tant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the superna-tant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 5 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of sennoside A in each solution. Amount (mg) of sennoside A (C42H38O20) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (2460:540:1). Flow rate: 1.0 mL per minute (the retention time of senno-side A is about 14 minutes.) System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sennoside A are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z. (2) Glycyrrhizic acid—Weigh accurately about 0.2 g of ‌1996 Daisaikoto Extract / Crude Drugs and Related Drugs Daiokanzoto Extract, add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re- Method of preparation 1) 2) 3) 4) 5) move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove Bupleurum Root 6 g 6 g 6 g 6 g 6 g the ethyl acetate layer. To the aqueous layer add 10 mL of Pinellia Tuber 4 g 4 g 4 g 3 g 4 g methanol, shake for 30 minutes, centrifuge, and take the Scutellaria Root 3 g 3 g 3 g 3 g 3 g supernatant liquid. To the residue add 20 mL of diluted Peony Root 3 g 3 g 3 g 3 g 3 g methanol (1 in 2), shake for 5 minutes, centrifuge, and take Jujube 3 g 3 g 3 g 3 g 3 g the supernatant liquid. Combine these supernatant liquids, Immature Orange 2 g 2 g 2 g 2 g 2 g add diluted methanol (1 in 2) to make exactly 50 mL, and use Ginger 1 g 1 g 2 g 1 g 1.5 g this solution as the sample solution. Separately, weigh accu- Rhubarb 1 g 2 g 1 g 1 g 2 g rately about 10 mg of Glycyrrhizic Acid RS (separately de- termine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Daisaikoto Extract contains not less than 1.
Daisaikoto Extract occurs as light yellow-brown to brown powder or black-brown viscous extract, having a slightly order, and a hot first, then a bitter taste.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes. Examine under ul-traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown to grayish brown spot from the standard solution (Scutellaria Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) JP XVIII Crude Drugs and Related Drugs / Daisaikoto Extract 1997 to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzoaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 2 minutes: one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the red-purple to purple spot from the standard solution (Peony Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 1.0 g of pulverized immature orange add 10 mL of methanol, shake, centrifuge, and use the su-pernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and allow to stand in an ammonia gas: two con-secutive spots at Rf values of about 0.7 obtained from the sample solution have respectively the same color tone and Rf value with the blue-green spot and blue spot underneath from the standard solution (Immature Orange). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of rhein for thin-layer chromatography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the orange fluorescent spot from the standard solution (Rhubarb).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 11.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same man-ner as described above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, separate the supernatant liquid, com-bine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of saikosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 CS: Concentration (mg/mL) of saikosaponin b2 in sai-kosaponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of saikosaponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly ‌1998 Digenea / Crude Drugs and Related Drugs 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, flow through in a column packed with 2 g of polyamide for column chromatography, elute with 20 mL of water, add 1 mL of acetic acid (100), to the effluent, then add water to make exactly 25 mL, and use this as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z.
Danazol, when dried, contains not less than 98.
Danazol occurs as a white to pale yellow crys-talline powder. It is soluble in acetone, sparingly soluble in ethanol (99.5), and practically insoluble in water. Melting point: about 2259C (with decomposition).
(1) Determine the absorption spectrum of a solution of Danazol in ethanol (95) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Danazol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Danazol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Danazol RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋8 – ＋119 (after drying, 0.25 g, ethanol (99.5), 50 mL, 100 mm).
(1) Chloride <1.03>—To 2.0 g of Danazol add 80 mL of water, shake well, and boil for 5 minutes. After cool-ing, add water to make 100 mL, and filter through a glass filter (G4). Discard the first 30 mL of the filtrate, take 40 mL of the subsequent filtrate, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu- Amount (mg) of danazol (C22H27NO2) ＝ MS × AT/AS MS: Amount (mg) of Danazol RS taken
Weigh accurately about 25 mg each of Danazol and Danazol RS, previously dried, dissolve separately in ethanol (95) to make exactly 50 mL. Pipet 2 mL each of these solu-tions, add ethanol (95) to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and determine the absorbances, AT and AS, at 285 nm. Danazol, when dried, contains not less than 98.5z and not more than 101.0z of danazol (C22H27NO2). Description Danazol occurs as a white to pale yellow crys-talline powder. It is soluble in acetone, sparingly soluble in ethanol (99.5), and practically insoluble in water. Melting point: about 2259C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Danazol in ethanol (95) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Danazol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Danazol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Danazol RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋8 – ＋119 (after drying, 0.25 g, ethanol (99.5), 50 mL, 100 mm). Purity (1) Chloride <1.03>—To 2.0 g of Danazol add 80 mL of water, shake well, and boil for 5 minutes. After cool-ing, add water to make 100 mL, and filter through a glass filter (G4). Discard the first 30 mL of the filtrate, take 40 mL of the subsequent filtrate, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu- Amount (mg) of danazol (C22H27NO2) ＝ MS × AT/AS MS: Amount (mg) of Danazol RS taken
Daunorubicin Hydrochloride is the hydrochloride of an anthracycline substance having antitumor activity produced by the growth of Streptomyces peucetius or Streptomyces coeruleorubidus. It contains not less than 940 mg (potency) and not more than 1050 mg (potency) per mg, calculated on the dried basis.
Daunorubicin Hydrochloride occurs as a red powder. It is soluble in water and in methanol, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Daunorubicin Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Daunorubi-cin Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Daunorubicin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Daunorubicin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Daunorubicin Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: ＋250 – ＋2759(15 mg calcu-lated on the dried basis, methanol, 10 mL, 100 mm). pH <2.54> Dissolve 0.15 g of Daunorubicin Hydrochloride in 30 mL of water: the pH of the solution is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve 20 mg of Daunorubicin Hydrochloride in 10 mL of water: the solu-tion is clear and red. Heavy metals <1.07>—Proceed with 1.0 g of Daunorubicin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Weigh accurately about 50 mg of Daunorubicin Hydrochloride, dissolve in diluted aceto-nitrile (43 in 100) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg of Daunorubicin Hydrochloride RS, and dis-solve in diluted acetonitrile (43 in 100) to make exactly 50 mL. Pipet 1 mL of this solution, add diluted acetonitrile (43 in 100) to make exactly 200 mL, and use this solution as the standard solution (1). Separately, weigh accurately about 5 mg of Doxorubicin Hydrochloride RS, and dissolve in diluted acetonitrile (43 in 100) to make exactly 100 mL. Pipet 1 mL of this solution, add diluted acetonitrile (43 in 100) to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 5 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of related substances by the following equations: each amount of each peak, having a relative retention time of about 0.3, about 0.6, about 0.7, about 0.8, about 1.7 and about 2.0 to daunorubicin, is not more than 1.3z, not more than 1.0z, not more than 0.3z, not more than 0.5z, not more than 0.4z and not more than 0.5z, respectively, and the amount of doxorubicin is not more than 0.4z. Furthermore, the total amount of the peaks, other than daunorubicin and the peaks mentioned above, is not more than 0.4z. For the area of the peak, having a relative retention time of about 0.3 to daunorubicin, multiply the correction factor, 0.7. Each amount (z) of related substances other than doxorubicin ＝ MS1/MT × AT/AS1 × 1/2 MS1: Amount (mg) of Daunorubicin Hydrochloride RS taken MT: Amount (mg) of Daunorubicin Hydrochloride taken AS1: Peak area of daunorubicin obtained from the stand-ard solution (1) AT: Peak area of each related substance obtained from the Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.88 g of sodium lauryl sulfate and 2.25 g of phosphoric acid in water to make 1000 mL. To 570 mL of this solution add 430 mL of acetonitrile. Flow rate: Adjust so that the retention time of daunorubi-cin is about 26 minutes. Time span of measurement: About 2 times as long as the retention time of daunorubicin. System suitability— Test for required detectability: To exactly 1 mL of the standard solution (1) add diluted acetonitrile (43 in 100) to make exactly 10 mL. Confirm that the peak area of daunorubicin obtained with 5 mL of this solution is equiva-lent to 7 to 13z of that with 5 mL of the standard solution (1). System performance: Dissolve 5 mg each of Daunorubicin Hydrochloride and doxorubicin hydrochloride in 25 mL of diluted acetonitrile (43 in 100). To 1 mL of this solution add diluted acetonitrile (43 in 100) to make 10 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, doxorubicin and daunorubicin are eluted in this order with the resolution between these peaks being not less than 13. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution (1) under the above oper-ating conditions, the relative standard deviation of the peak area of daunorubicin is not more than 3.0z. Loss on drying <2.41> Not more than 7.5z (0.1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours).
Weigh accurately an amount of Daunorubicin Hy-drochloride and Daunorubicin Hydrochloride RS, equiva-lent to about 20 mg (potency), dissolve each in a suitable amount of the mobile phase, add exactly 4 mL of the in-ternal standard solution and the mobile phase to make 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of daunorubicin to that of the internal standard. Amount [ mg (potency)] of daunorubicin hydrochloride (C27H29NO10.HCl) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Daunorubicin Hydrochlo-ride RS taken sample solution Amount (z) of doxorubicin ＝ M S2/MT × AT/AS2 × 5 Internal standard solution—A solution of 2-naphthalenesul-fonic acid in the mobile phase (1 in 100). Operating conditions— MS2: Amount (mg) of Doxorubicin Hydrochloride RS taken MT: Amount (mg) of Daunorubicin Hydrochloride taken AS2: Peak area of doxorubicin obtained from the standard solution (2) AT: Peak area of doxorubicin obtained from the sample solution Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust the pH of a mixture of water and ‌JP XVIII Official Monographs / Deferoxamine Mesilate 831 acetonitrile (31:19) to 2.2 with phosphoric acid. Flow rate: Adjust so that the retention time of daunorubi-cin is about 9 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and daunorubicin are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of daunorubicin to that of the internal stand-ard is not more than 2.0z.
Deferoxamine Mesilate contains not less than 98.
Deferoxamine Mesilate occurs as a white to pale yellow-white crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (99.5), in 2-propanol and in diethyl ether. Melting point: about 1479C (with decomposition).
(1) To 5 mL of a solution of Deferoxamine Mesilate (1 in 500) add 1 drop of iron (III) chloride TS: a deep red color develops. A 50 mg portion of Deferoxamine Mesilate responds to Qualitative Tests <1.09> (1) for mesilate. Determine the infrared absorption spectrum of Deferoxamine Mesilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Deferoxamine Mesilate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Deferoxamine Mesilate in 10 mL of water: the pH of this solution is between 3.5 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Deferoxamine Mesilate in 10 mL of water: the solution is clear and colorless to pale yellow. Chloride <1.03>—Perform the test with 1.0 g of Deferoxamine Mesilate. Prepare the control solution with 0.90 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.032z). Sulfate <1.14>—Perform the test with 0.6 g of Deferoxamine Mesilate. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.040z). Heavy metals <1.07>—Proceed with 2.0 g of Deferox- amine Mesilate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Deferoxamine Mesilate according to Method 3, and per-form the test. Use a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 10) (not more than 2 ppm). Related substances—Dissolve 50 mg of Deferoxa-mine Mesilata in 50 mL of the mobile phase, and use this so-lution as the sample solution. Pipet 3 mL of the sample solu-tion, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of peaks other than deferoxamine obtained from the sample solution is not larger than the peak area of deferoxamine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4 mm in inside diameter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.32 g of diammonium hydrogen phosphate, 0.37 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate and 1.08 g of sodium 1-heptanesul-fonate in 950 mL of water, and adjust the pH of this solu-tion to 2.8 with phosphoric acid. To 800 mL of this solution add 100 mL of 2-propanol. Flow rate: Adjust so that the retention time of deferoxa-mine is about 15 minutes. Time span of measurement: About two times as long as the retention time of deferoxamine, beginning after the sol-vent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of deferoxamine obtained with 20 mL of this solution is equivalent to 1.5 to 2.5z of that with 20 mL of the standard solution. System performance: Dissolve 16 mg of Deferoxamine Mesilate and 4 mg of methyl parahydroxybenzoate in 50 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, deferoxamine and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of deferoxamine is not more than 3.0z. Water <2.48> Not more than 2.0z (0.2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 60 mg of Deferoxamine Mesilate and Deferoxamine Mesilate RS (previously deter-mine the water <2.48> in the same manner as Deferoxamine ‌832 ‌Mesilate), dissolve each in 20 mL of water, add exactly 10 mL of 0.05 mol/L sulfuric acid TS, and add water to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of 0.05 mol/L sulfuric acid TS and exactly 0.2 mL of iron (III) chloride TS, then add water to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared by adding 0.05 mol/L sulfuric acid TS to 0.2 mL of iron (III) chloride TS to make exactly 50 mL as the blank, and determine the absor-bances, AT and AS, of each solution from the sample solu-tion and the standard solution at 430 nm. Amount (mg) of deferoxamine mesilate (C25H48N6O8.CH4O3S) ＝ MS × AT/AS MS: Amount (mg) of Deferoxamine Mesilate RS taken, calculated on the anhydrous basis
Anhydrous Ethanol is a clear, colorless liquid. It is miscible with water. It is flammable and burns with a light blue flame on igni-tion. It is volatile. Boiling point: 78 – 799C◆
Determine the infrared absorption spectrum of Anhydrous Ethanol as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 15: 0.79422 – 0.79679
(1) Clarity and color of solution—Anhydrouse Ethanol is clear and colorless. To 1.0 mL of Anhydrous Ethanol add water to make 20 mL, and allow to stand for 5 minutes: the resulting liquid is clear. Control solution: water ‌966 Acidity or alkalinity—To 20 mL of Anhydrous Ethanol add 20 mL of freshly boiled and cooled water and 0.1 mL of a solution obtained by addition of 7.0 mL of ethanol (95) and 2.0 mL of water to 1.0 mL of phenolphthal-ein TS: no color develops. Add 1.0 mL of 0.01 mol/L so-dium hydroxide VS to this solution: pink color develops. Volatile impurities—Pipet 500 mL of Anhydrous Ethanol, add 150 mL of 4-methylpentan-2-ol, and use this so-lution as the sample solution. Separately, to 100 mL of anhy-drous methanol add Anhydrous Ethanol to make exactly 50 mL. Pipet 5 mL of this solution, add Anhydrous Ethanol to make exactly 50 mL, and use this solution as the standard solution (1). Separately, to 50 mL each of anhydrous metha-nol and acetaldehyde add Anhydrous Ethanol to make ex-actly 50 mL. To 100 mL of this solution add Anhydrous Ethanol to make exactly 10 mL, and use this solution as the standard solution (2). Separately, to 150 mL of acetal add Anhydrous Ethanol to make exactly 50 mL. To 100 mL of this solution add Anhydrous Ethanol to make exactly 10 mL, and use this solution as the standard solution (3). Sepa-rately, to 100 mL of benzene add Anhydrous Ethanol to make exactly 100 mL. To 100 mL of this solution add Anhy-drous Ethanol to make exactly 50 mL, and use this solution as the standard solution (4). Perform the test with exactly 1 mL each of Anhydrous Ethanol, the sample solution and standard solutions (1), (2), (3) and (4) as directed under Gas Chromatography <2.02> according to the following condi-tions, and determine the peak areas of acetaldehyde, AE, benzene, BE and acetal, CE obtained with Anhydrous Ethanol, and the peak area of methanol with the standard solution (1), the peak area of acetaldehyde, AT with the standard solution (2), the peak area of acetal, CT with the standard solution (3) and the peak area of benzene, BT with the standard solution (4) by the automatic integration method: the peak area of methanol obtained with Anhy-drous Ethanol is not larger than 1/2 times the peak area of Flow rate: 35 cm per second. Split ratio: 1:20. System suitability— System performance: When the procedure is run with 1 mL of the standard solution (2) under the above operating condi-tions, acetaldehyde and methanol are eluted in this order with the resolution between these peaks being not less than 1.5. Other impurities (absorbance)—Determine the ab-sorption spectrum of Anhydrous Ethanol between 235 nm and 340 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, in a 5-cm cell using water as a blank: the absorbances at 240 nm, between 250 nm and 260 nm and between 270 nm and 340 nm are not more than 0.40, 0.30, and 0.10, respectively, and the spectrum shows a steadily descending curve with no observable peaks or shoulders. Residue on evaporation—Evaporate 100 mL of An-hydrous Ethanol, exactly measured, in a tared dish on a water bath, and dry at 1059C for 1 hour: the mass of the residue does not exceed 2.5 mg.
Dehydrocholic Acid, when dried, contains not less lution is clear and colorless.
Dehydrocholic Acid occurs as a white crystal-line powder. It is odorless, and has a bitter taste. It is sparingly soluble in acetone, slightly soluble in ethanol (95), and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Dissolve 5 mg of Dehydrocholic Acid in 1 mL of sulfuric acid and 1 drop of formaldehyde solution, and allow to stand for 5 minutes. Add 5 mL of water to the solution: the solution shows a yellow color and a blue-green fluorescence. (2) To 0.02 g of Dehydrocholic Acid add 1 mL of ethanol (95), shake, add 5 drops of 1,3-dinitrobenzene TS and 0.5 mL of a solution of sodium hydroxide (1 in 8), and allow to stand: a purple to red-purple color develops, and gradually changes to brown. D Optical rotation <2.49> [a]25: ＋20 – ＋269(after drying, 0.2 g, acetone, 10 mL, 100 mm).
(1) Odor—To 2.0 g of Dehydrocholic Acid add 100 mL of water, and boil for 2 minutes: the solution is Each mL of 0.1 mol/L sodium hydroxide VS ＝ 40.25 mg of C24H34O5
Weigh accurately about 0.5 g of Dehydrocholic Acid, previously dried, add 40 mL of neutralized ethanol and 20 mL of water, and dissolve by warming. Add 2 drops of phenolphthalein TS, titrate <2.50> with 0.1 mol/L sodium hydroxide VS, adding 100 mL of freshly boiled and cooled water as the end point is approached, and continue the titra- than 98.5z of dehydrocholic acid (C24 H34 O5). tion. Description Dehydrocholic Acid occurs as a white crystal-line powder. It is odorless, and has a bitter taste. It is sparingly soluble in acetone, slightly soluble in ethanol (95), and practically insoluble in water. It dissolves in sodium hydroxide TS. Identification (1) Dissolve 5 mg of Dehydrocholic Acid in 1 mL of sulfuric acid and 1 drop of formaldehyde solution, and allow to stand for 5 minutes. Add 5 mL of water to the solution: the solution shows a yellow color and a blue-green fluorescence. (2) To 0.02 g of Dehydrocholic Acid add 1 mL of ethanol (95), shake, add 5 drops of 1,3-dinitrobenzene TS and 0.5 mL of a solution of sodium hydroxide (1 in 8), and allow to stand: a purple to red-purple color develops, and gradually changes to brown. D Optical rotation <2.49> [a]25: ＋20 – ＋269(after drying, 0.2 g, acetone, 10 mL, 100 mm). Melting point <2.60> 233 – 2429C Purity (1) Odor—To 2.0 g of Dehydrocholic Acid add 100 mL of water, and boil for 2 minutes: the solution is Each mL of 0.1 mol/L sodium hydroxide VS ＝ 40.25 mg of C24H34O5
デメチルクロルテトラサイクリン塩酸塩
Demethylchlortetracycline Hydrochloride is the hy-drochloride of a tetracycline substance having antibac-terial activity produced by the growth of the mutant of Streptomyces aureofaciens. It contains not less than 900 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the dried basis.
Demethylchlortetracycline Hydrochloride oc-curs as a yellow crystalline powder. It is soluble in water, and slightly soluble in ethanol (99.5).
(1) Dissolve 40 mg of Demethylchlortetra-cycline Hydrochloride in 250 mL of water. To 10 mL of this solution add 85 mL of water and 5 mL of a solution of so-dium hydroxide (1 in 5). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of a solution of Demethyl-chlortetracycline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Demethylchlortetracycline Hydrochloride as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Demethylchlortetracycline Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Demethylchlortetracycline Hydrochlo-ride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －248 – －2639(0.25 g calcu-lated on the dried basis, 0.1 mol/L hydrochloric acid TS, 25 mL, 100 mm). (2) Related substances—Dissolve 25 mg of Demethyl-chlortetracycline Hydrochloride in 50 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Pipet 5 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than demethylchlortetracycline obtained from the sample solution is not larger than 1.2 times the peak area of demethylchlortetracycline from the standard solution, and the total area of the peaks other than demethylchlortetra-cycline from the sample solution is not larger than 2 times the peak area of demethylchlortetracycline from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of demethylchlortetracycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 10 mL of the standard solution add 0.01 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the solution for system suitabil-ity test. Pipet 5 mL of the solution for system suitability test, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of demethylchlortetra-cycline obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitabil-ity test. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of demethylchlortetracycline is not more than 1.0z. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Demethylchlortetracycline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Demethylchlortetracycline Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene ‌JP XVIII Official Monographs / Deslanoside 835 copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase: Dissolve 3.5 g of dipotassium hydrogen phosphate, 1.5 g of tetrabutylammonium hydrogensulfate and 0.4 g of disodium dihydrogen ethylenediamine tetra-acetate dihydrate in 300 mL of water, and adjust the pH to 8.5 with sodium hydroxide TS. To this solution add 75.0 g of t-butanol and water to make 1000 mL. Flow rate: Adjust so that the retention time of demethyl-chlortetracycline is about 8 minutes. System suitability— System performance: Heat 10 mL of the standard solution on a water bath for 60 minutes. When the procedure is run with 20 mL of this solution so obtained under the above operating conditions, 4-epidemethylchlortetracycline and demethylchlortetracycline are eluted in this order with the resolution between these peaks being not less than 3. The relative retention time of 4-epidemethylchlortetracycline to demethylchlortetracycline is about 0.7. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of demethylchlortetracycline is not more than 1.0z.
Weigh accurately an amount of Demethylchlortetra-cycline Hydrochloride and Demethylchlortetracycline Hy-drochloride RS, equivalent to about 25 mg (potency), dis-solve each in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of demethylchlortetracycline in each solution. Amount [ mg (potency)] of demethylchlortetracycline hydrochloride (C H ClN O .HCl) 21 21 2 8 pH <2.54> Dissolve 1.0 g of Demethylchlortetracycline Hy- drochloride in 100 mL of water: the pH of the solution is be-tween 2.0 and 3.0. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Demethylchlortetracycline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Demethylchlortetracycline Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene ‌JP XVIII Official Monographs / Deslanoside 835 copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase: Dissolve 3.5 g of dipotassium hydrogen phosphate, 1.5 g of tetrabutylammonium hydrogensulfate and 0.4 g of disodium dihydrogen ethylenediamine tetra-acetate dihydrate in 300 mL of water, and adjust the pH to 8.5 with sodium hydroxide TS. To this solution add 75.0 g of t-butanol and water to make 1000 mL. Flow rate: Adjust so that the retention time of demethyl-chlortetracycline is about 8 minutes. System suitability— System performance: Heat 10 mL of the standard solution on a water bath for 60 minutes. When the procedure is run with 20 mL of this solution so obtained under the above operating conditions, 4-epidemethylchlortetracycline and demethylchlortetracycline are eluted in this order with the resolution between these peaks being not less than 3. The relative retention time of 4-epidemethylchlortetracycline to demethylchlortetracycline is about 0.7. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of demethylchlortetracycline is not more than 1.0z.
Dental Iodine Glycerin contains not less than 9.
Dental Iodine Glycerin is a dark red-brown liq-uid, having the odor of iodine.
(1) The colored solution obtained in the Assay (1) acquires a red color. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 510 nm and 514 nm (iodine). The colored solution obtained in the Assay (2) acquires a red color. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 510 nm and 514 nm (potassium iodide). Put 1 mL of Dental Iodine Glycerin in a glass-stoppered, test tube, add 10 mL of ethanol (95), and mix. Then add 2 mL of sodium hydroxide TS, add 1 mL of a so-lution of copper (II) chloride dihydrate in ethanol (95) (1 in 10), and shake: a blue color develops (glycerin). The colored solution obtained in the Assay (3) acquires a red-purple to purple color. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 618 nm and 622 nm (zinc sulfate hydrate).
(1) Iodine—Pipet 5 mL of Dental Iodine Glycerin, ‌JP XVIII Official Monographs / Iodine, Salicylic Acid and Phenol Spirit 1173 and add diluted ethanol (3 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL, and use this solution as the sample solution. On the other hand, weigh accurately about 0.5 g of iodine for assay and about 0.4 g of potassium iodide for assay, previously dried at 1059C for 4 hours, and dissolve in diluted ethanol (3 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample solu-tion and standard solution, to each add exactly 20 mL of a mixture of chloroform and hexane (2:1), shake immediately, and separate the chloroform-hexane layer [use the water layer in (2)]. Filter through a pledget of cotton. Determine the absorbances, AT and AS, of the filtrates obtained from the sample solution and standard solution, respectively, at 512 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a mixture of chloroform and hexane (2:1) as the blank. Amount (mg) of iodine (I) ＝ MS × AT/AS MS: Amount (mg) of iodine for assay taken Potassium iodide—Separate the water layers of the sample solution and standard solution obtained in (1), pipet 7 mL each of the water layers, and to each add exactly 1 mL of diluted hydrochloric acid (1 in 2), 1 mL of sodium nitrite TS and 10 mL of a mixture of chloroform and hexane (2:1), and shake immediately. Separate the chloroform-hexane layer, and filter through a pledget of cotton. Determine the absorbances, AT and AS, of the filtrates obtained from the sample solution and standard solution, respectively, at 512 nm as directed under Ultraviolet-visible Spectrophotometry, using a mixture of chloroform and hexane (2:1) as the blank. Amount (mg) of potassium iodide (KI) ＝ MS × AT/AS MS: Amount (mg) of potassium iodide for assay taken Zinc sulfate hydrate—Pipet 5 mL of Dental Iodine Glycerin, and add diluted ethanol (3 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the sample solution. On the other hand, pipet 10 mL of Standard Zinc Stock So-lution, add diluted ethanol (3 in 200) to make exactly 1000 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample solution and standard solution, to each add 10 mL of a mixture of chloroform and hexane (2:1), shake, and allow to stand. Pipet 3 mL each of the water layers, and to each add 2 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 10.0), 2 mL of zincon TS and water to make exactly 25 mL. Determine the absorbances, AT and AS, obtained from the sample solu-tion and standard solution, respectively, at 620 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using the solution prepared in the same manner with 3 mL of water as the blank. Amount (mg) of zinc sulfate hydrate (ZnSO4.7H2O) ＝ MS × AT/AS × 4.398 MS: Amount (mg) of zinc in 10 mL of Standard Zinc Stock Solution
Dental Paraformaldehyde Paste is yellowish white in color. It has a characteristic odor.
(1) To 0.15 g of Dental Paraformaldehyde Paste add 20 mL of diethyl ether and 20 mL of 0.5 mol/L sodium hydroxide TS, shake well, separate the water layer, and dilute with water to make 100 mL. To 1 mL of this solu-tion add 10 mL of acetylacetone TS, and heat on a water bath for 10 minutes: a yellow color is produced (paraformal-dehyde). To the diethyl ether layer obtained in (1) add 5 mL of dilute hydrochloric acid and 20 mL of water, shake well, and separate the water layer: the solution responds to Qualitative Tests <1.09> for primary aromatic amines (procaine hydro-chloride). To 0.15 g of Dental Paraformaldehyde Paste add 25 mL of diethyl ether and 25 mL of water, shake, separate the water layer, filter, and use the filtrate as the sample solution. Separately, dissolve 0.01 g of procaine hydrochloride in 5 mL of water, and use this solution as standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia solution (28) (50:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): spots obtained from the sample solution and standard solution show the same Rf value.
Dental Phenol with Camphor is a colorless or light red liquid. It has a characteristic odor.
Deslanoside, when dried, contains not less than 90.
Deslanoside occurs as colorless or white crys-tals or a white, crystalline powder. It is odorless. It is freely soluble in dehydrated pyridine, sparingly solu-ble in methanol, slightly soluble in ethanol (95), and practi- MS: Amount (mg) of Deslanoside RS taken
Transfer 1 mg of Deslanoside to a small test tube about 10 mm in inside diameter, dissolve in 1 mL of a solution of iron (III) chloride hexahydrate in acetic acid (100) (1 in 1000), and underlay gently with 1 mL of sulfuric acid: at the zone of contact of two liquids a brown ring is produced, and the color of the upper layer near to the con-tact zone changes gradually to blue through purple, and the entire acetic acid layer shows a blue-green color through a deep blue color.
(1) Clarity and color of solution—Dissolve 20 mg of Deslanoside in 10 mL of ethanol (95) and 3 mL of water by warming, cool, and dilute to 100 mL with water: the solu-tion is clear and colorless. (2) Related substances—Dissolve 10 mg of Deslanoside in exactly 5 mL of methanol, and use this solution as the sample solution. Dissolve 1.0 mg of Deslanoside RS in exactly 5 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, methanol and water (84:15:1) to a distance of about 13 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1109C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not larger and not more intense than the spot from the standard solution. D Optical rotation <2.49> [a]20: ＋6.5 – ＋8.59(after drying, 0.5 g, dehydrated pyridine, 25 mL, 100 mm). Loss on drying <2.41> Not more than 8.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.5z (0.1 g).
Dissolve about 12 mg each of Deslanoside and Des-lanoside RS, previously dried and accurately weighed, in 20 mL each of methanol, add water to make exactly 100 mL, and use these solutions as the sample solution and the stand-ard solution, respectively. Pipet 5 mL each of these solu-tions, transfer to light-resistant, 25-mL volumetric flasks, shake well with 5 mL each of 2,4,6-trinitrophenol TS and 0.5 mL each of a solution of sodium hydroxide (1 in 10), add diluted methanol (1 in 4) to make 25 mL, and allow to stand at a temperature between 189C and 229C for 25 minutes. De-termine the absorbances, AT and AS, of the subsequent solu-tions of the sample solution and the standard solution, re-spectively, at 485 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of diluted methanol (1 in 5) in the same manner as the blank. hexopyranosyl-(1→4)-2,6-dideoxy-b-D-ribo-hexopyranosyloxy]-12b,14-dihydroxy-5b,14b-card-20(22)- Amount (mg) of deslanoside (C 47H 74O 19) ＝ MS × AT/AS enolide [17598-65-1] Deslanoside, when dried, contains not less than 90.0z and not more than 102.0z of deslanoside (C47H74O19). Description Deslanoside occurs as colorless or white crys-tals or a white, crystalline powder. It is odorless. It is freely soluble in dehydrated pyridine, sparingly solu-ble in methanol, slightly soluble in ethanol (95), and practi- MS: Amount (mg) of Deslanoside RS taken
Dexamethasone, when dried, contains not less than 97.
Dexamethasone occurs as white to pale yellow, crystals or crystalline powder. It is sparingly soluble in methanol, in ethanol (95) and in acetone, slightly soluble in acetonitrile, and practically in-soluble in water. Melting point: about 2459C (with decomposition). It shows crystal polymorphism.
(1) Place a volume of Deslanoside Injec-tion, equivalent to 2 mg of Deslanoside, in a separator, add sodium chloride in the ratio of 0.2 g to each mL of this solu-tion, and extract with three 10-mL portions of chloroform. Combine the chloroform extracts, mix uniformly, pipet 15 mL of this solution, and evaporate the chloroform under reduced pressure. Proceed with the residue as directed in the Identification under Deslanoside. (2) Evaporate the remaining chloroform extract obtained in (1) under reduced pressure, dissolve the residue in 5 mL of methanol, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of Deslanoside RS in 5 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of these solutions on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of dichloromethane, methanol and water (84:15:1) to a distance of about 13 cm, and air-dry the plate. Spray evenly dilute sulfuric acid upon the plate, and heat the plate at 1109C for 10 minutes: the spots from the sample solution and standard solution show a black color and have the same Rf value. Bacterial endotoxins <4.01> Less than 500 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Dexamethasone according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard MS: Amount (mg) of Deslanoside RS taken
Measure exactly a volume of Deslanoside Injection, equivalent to about 3 mg of deslanoside (C47H74O19). Add 5 mL of methanol and water to make exactly 25 mL. Use this solution as the sample solution, and proceed as directed in the Assay under Deslanoside. Description Dexamethasone occurs as white to pale yellow, crystals or crystalline powder. It is sparingly soluble in methanol, in ethanol (95) and in acetone, slightly soluble in acetonitrile, and practically in-soluble in water. Melting point: about 2459C (with decomposition). It shows crystal polymorphism. Identification (1) Proceed with 10 mg of Dexamethasone as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution obtained responds to Qualitative Tests <1.09> for fluoride. Dissolve 1 mg of Dexamethasone in 10 mL of ethanol (95). Mix 2 mL of the solution with 10 mL of phenyl-hydrazinium chloride TS, heat in a water bath at 609C for 20 minutes, and cool the solution. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using as the blank the solution prepared with 2 mL of ethanol (95) in the same manner as the former solution, and compare the spectrum with the Ref-erence Spectrum or the spectrum of a solution of Dexa-methasone RS prepared in the same manner as the former solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Dexa-methasone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Dexamethasone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Dexamethasone and Dexamethasone RS in acetone, respectively, then evaporate the acetone to dry-ness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋86 – ＋949(after drying, 0.1 g, methanol, 10 mL, 100 mm). Amount (mg) of deslanoside (C ＝ MS × AT/AS × 1/4 47H 74O 19) Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Dexamethasone according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard MS: Amount (mg) of Deslanoside RS taken
Dextrin occurs as a white or light yellow, amorphous powder or granules. It has a slight, characteristic odor and a sweet taste. It does not irritate the tongue. Dextrin is freely soluble in boiling water, soluble in water, and practically insoluble in ethanol (95) and in diethyl ether.
To 0.1 g of Dextrin add 100 mL of water, shake, and filter if necessary. To 5 mL of the filtrate add 1 drop of iodine TS: a light red-brown or light red-purple color develops.
(1) Clarity and color of solution—Take 2.0 g of Dextrin in a Nessler tube, add 40 mL of water, dissolve by heating, cool, and add water to make 50 mL: the solution is colorless or light yellow. It is clear, and even if turbid, the turbidity is not more than that of the following control solu-tion. Control solution: To 1.0 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid, 46 mL of water and 2 mL of barium chloride TS, allow to stand for 10 minutes, and shake before use. Acidity—To 1.0 g of Dextrin add 5 mL of water, dis-solve by heating, cool, and add 1 drop of phenolphthalein TS and 0.50 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Chloride <1.03>—To 2.0 g of Dextrin add 80 mL of water, dissolve by heating, cool, add water to make 100 mL, and filter. Take 40 mL of the filtrate, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.013z). Sulfate <1.14>—To 45 mL of the filtrate obtained in (3) add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Oxalate—To 1.0 g of Dextrin add 20 mL of water, dissolve by heating, cool, add 1 mL of acetic acid (31), and filter. To 5 mL of the filtrate add 5 drops of calcium chloride TS: no turbidity is produced immediately. Calcium—To a 5-mL portion of the filtrate obtained in (5) add 5 drops of ammonium oxalate TS: no turbidity is immediately produced. Heavy metals <1.07>—Proceed with 0.5 g of Dextrin according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 50 ppm). Loss on drying <2.41> Not more than 10z (0.5 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.5z (0.5 g).
デキストロメトルファン臭化水素酸塩水和物
Dextromethorphan Hydrobromide Hydrate occurs as white, crystals or crystalline powder. It is very soluble in methanol, freely soluble in ethanol (95) and in acetic acid (100), and sparingly soluble in water. Melting point: about 1269C (Insert the capillary tube into the bath preheated to 1169C, and continue the heating so that the temperature rises at a rate of about 39C per minute.)
(1) Determine the absorption spectrum of a solution of Dextromethorphan Hydrobromide Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Dex-tromethorphan Hydrobromide Hydrate as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. To 50 mL of a solution of Dextromethorphan Hydrobromide Hydrate (1 in 100) add 2 drops of phenol-phalein TS and sodium hydroxide TS until a red color de-velops. Add 50 mL of chloroform, shake, and add 5 mL of dilute nitric acid to 40 mL of the water layer. This solution responds to Qualitative Tests <1.09> for bromide. D Optical rotation <2.49> [a]20: ＋26 – ＋309(0.34 g calcu-lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Dextromethorphan Hydro-bromide Hydrate in 100 mL of water: the pH of this solution is between 5.2 and 6.5.
(1) Clarity and color of solution—Dissolve 0.20 g of Dextromethorphan Hydrobromide Hydrate in 20 mL of water: the solution is clear and colorless. N,N-dimethylaniline—To 0.50 g of Dextromethor-phan Hydrobromide Hydrate add 20 mL of water, and dis-solve by heating on a water bath. After cooling, add 2 mL of dilute acetic acid, 1 mL of sodium nitrite TS and water to make 25 mL: the solution has no more color than the follow-ing control solution. Control solution: Dissolve 0.10 g of N,N-dimethylaniline in 400 mL of water by warming on a water bath, cool, and add water to make 500 mL. Pipet 5 mL of this solution, and add water to make 200 mL. To 1.0 mL of this solution add 2 mL of dilute acetic acid, 1 mL of sodium nitrite TS and water to make 25 mL. Heavy metals <1.07>—Proceed with 1.0 g of Dex- tromethorphan Hydrobromide Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Phenolic compounds—Dissolve 5 mg of Dextro-methorphan Hydrobromide Hydrate in 1 drop of dilute hy-drochloric acid and 1 mL of water, add 2 drops of iron (III) chloride TS and 2 drops of potassium hexacyanoferrate (III) TS, shake, and allow to stand for 15 minutes: no blue-green color develops. Related substances—Dissolve 0.25 g of Dextro-methorphan Hydrobromide Hydrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of tolu- ene, ethyl acetate, methanol, dichloromethane and 13.5 mol/L ammonia TS (55:20:13:10:2) to a distance of about 15 cm, and air-dry the plate. Spray evenly bismuth potassium iodide TS on the plate, and then spray evenly hydrogen peroxide TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 4.0 – 5.5z (0.2 g, volumetric titration, back titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Dextromethorphan Hydrobromide Hydrate, dissolve in 10 mL of acetic acid (100) and add 40 mL of acetic anhydride. Titrate <2.50> with mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.23 mg of C18H25NO.HBr
Diastase is an enzyme drug mainly prepared from malt. It has amylolytic activity. It contains not less than 440 starch saccharifying activity units per g.
Diastase occurs as a light yellow to light brown powder. It is hygroscopic.
Rancidity—Diastase has no unpleasant or rancid odor, and has no unpleasant or rancid taste. Loss on drying <2.41> Not more than 4.0z (1 g, 1059C, 5 hours).
(i) Substrate solution—Use potato starch TS for amylolytic activity test. Sample solution—Weigh accurately about 0.1 g of Diastase, and dissolve in water to make exactly 100 mL. Procedure—Proceed as directed in 1.1. Measure-ment of starch saccharifying activity of 1. Assay for starch digestive activity under Digestion Test <4.03>.
Diazepam, when dried, contains not less than 98.
Diazepam occurs as a white to light yellow crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in acetone, soluble in acetic anhydride and in ethanol (95), sparingly soluble in diethyl ether, slightly soluble in ethanol (99.5), and practically insoluble in water.
(1) Dissolve 10 mg of Diazepam in 3 mL of sulfuric acid, and observe under ultraviolet light (main wave-length: 365 nm): the solution shows a yellow-green fluores-cence. Dissolve 2 mg of Diazepam in 200 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the absorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dia-zepam, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Diazepam as directed under Flame Coloration Test <1.04> (2): a blue to blue-green color appears.
(1) Clarity of solution—Dissolve 0.10 g of Diaze-pam in 20 mL of ethanol (95): the solution is clear. Chloride <1.03>—To 1.0 g of Diazepam add 50 mL of water, allow to stand for 1 hour, with occasional shaking, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Diaze- pam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 1.0 g of Diazepam in 10 mL of acetone, and use this solution as the sample solu-tion. Pipept 1 mL of the sample solution, and add acetone to make exactly 100 mL. Pipet 1 mL of this solution, add ace-tone to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Diazepam, previ-ously dried, dissolve in 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. ‌844 Each mL of 0.1 mol/L perchloric acid VS ＝ 28.47 mg of C16H13ClN2O
Dibekacin Sulfate is the sulfate of a derivative of bekanamycin. It contains not less than 640 mg (potency) and not more than 740 mg (potency) per mg, calculated on the dried basis.
Dibekacin Sulfate occurs as a white to yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 20 mg each of Dibekacin Sul-fate and Dibekacin Sulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia solution (28) and methanol (1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS, and heat at 1009C for 10 minutes: the principal spots obtained from the sample solution and the spot from the standard solution show a purple-brown color and the same Rf value. (2) To 5 mL of a solution of Dibekacin Sulfate (1 in 50) add 1 drop of barium chloride TS: a white precipitate is pro-duced. D Optical rotation <2.49> [a]20: ＋96 – ＋1069(0.25 g calcu-lated on the dried basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Dibekacin Sulfate in 20 mL of water is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 3.0 g of Dibekacin Sulfate in 10 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.15. Heavy metals <1.07>—Proceed with 1.0 g of Dibeka- cin Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer having pH 6.5 to 6.6 after sterilization. Standard solutions—Weigh accurately an amount of Dibekacin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solu-tion (pH 6.0) (1 in 2) to make exactly 50 mL, and use this so-lution as the standard stock solution. Keep the standard stock solution at 5 to 159C and use within 30 days. Take exactly a suitable amount of the standard stock solution be-fore use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Dibekacin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these pressed as mass (potency) of dibekacin (C 451.52). 18H37N5O8: solutions as the high concentration sample solution and the low concentration sample solution, respectively. Description Dibekacin Sulfate occurs as a white to yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5). Identification (1) Dissolve 20 mg each of Dibekacin Sul-fate and Dibekacin Sulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia solution (28) and methanol (1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS, and heat at 1009C for 10 minutes: the principal spots obtained from the sample solution and the spot from the standard solution show a purple-brown color and the same Rf value. (2) To 5 mL of a solution of Dibekacin Sulfate (1 in 50) add 1 drop of barium chloride TS: a white precipitate is pro-duced. D Optical rotation <2.49> [a]20: ＋96 – ＋1069(0.25 g calcu-lated on the dried basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Dibekacin Sulfate in 20 mL of water is between 6.0 and 8.0. Purity (1) Clarity and color of solution—Dissolve 3.0 g of Dibekacin Sulfate in 10 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.15. Heavy metals <1.07>—Proceed with 1.0 g of Dibeka- cin Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead
ジエチルカルバマジンクエン酸塩
Diethylcarbamazine Citrate, when dried, contains not less than 98.
Difenidol Hydrochloride, when dried, contains not less than 98.
Difenidol Hydrochloride occurs as white, crys-tals or crystalline powder. It is odorless. It is freely soluble in methanol, soluble in ethanol (95), sparingly soluble in water and in acetic acid (100), and prac-tically insoluble in diethyl ether. Melting point: about 2179C (with decomposition).
(1) Dissolve 0.01 g of Difenidol Hydro-chloride in 1 mL of sulfuric acid: an orange-red color de-velops. To this solution add carefully 3 drops of water: the solution becomes yellowish brown, and colorless on the ad-dition of 10 mL of water. To 5 mL of a solution of Difenidol Hydrochloride (1 in 100) add 2 mL of Reinecke salt TS: a light red precipitate is formed. To 10 mL of a solution of Difenidol Hydrochloride (1 in 100) add 2 mL of sodium hydroxide TS, and extract with two 15-mL portions of chloroform. Combine the ex-tracts, wash with three 10-mL portions of water, evaporate the chloroform on a water bath, and dry the residue in a desiccator (in vacuum, silica gel, 559C) for 5 hours: the residue melts <2.60> between 1039C and 1069C. A solution of Difenidol Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Difenidol Hydrochloride in 100 mL of freshly boiled and cooled water: the pH of this solu-tion is between 4.7 and 6.5. JP XVIII Official Monographs / Diflorasone Diacetate 851 ‌Purity (1) Clarity and color of solution—Dissolve 1.0 g of Difenidol Hydrochloride in 10 mL of methanol: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Difenidol Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Difenidol Hydrochloride according to Method 3, and per-form the test (not more than 1 ppm). Related substances—Dissolve 0.10 g of Difenidol Hydrochloride in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of 1,1-diphenyl-4-piperidino-1-butene hydrochloride for thin-layer chromatography in methanol to make exactly 20 mL, pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of toluene, methanol and acetic acid (100) (10:2:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Difenidol Hydro-chloride, previously dried, dissolve in 30 mL of acetic acid (100) by warming if necessary, cool, add 30 mL of acetic an-hydride, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 17.30 mg of C21H27NO.HCl
Diflorasone Diacetate, when dried, contains not less than 97.
Diflorasone Diacetate occurs as a white to pale yellow, crystals or crystalline powder. It is soluble in acetonitrile, slightly soluble in ethanol (99.5), and practically insoluble in water. Melting point: about 2229C (with decomposition).
(1) Determine the infrared absorption spec-trum of Diflorasone Diacetate, previously dried, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Diflorasone Diacetate RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers. (2) Prepare the test solution with 10 mg of Diflorasone Diacetate as directed under Oxygen Flask Combustion Method <1.06>, using 20 mL of diluted 0.01 mol/L sodium hydroxide VS (1 in 40) as the absorbing liquid: the test solu-tion responds to Qualitative Tests <1.09> for fluoride. D Optical rotation <2.49> [a]20: ＋ 88 – ＋ 939(after drying, 0.1 g, acetonitrile, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Diflorasone Diacetate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Diflorasone Diacetate in 20 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the areas of the peaks, having a relative retention time of about 0.5, about 0.7, about 0.9 and about 1.1 to diflorasone diacetate, obtained from the sample solution are respectively not larger than 1/4 times, 1/4 times, 1/2 times and 3/4 times the peak area of diflorasone dia-cetate from the standard solution, and the total area of the peaks other than diflorasone diacetate and the peaks men-tioned above from the sample solutions is not larger than 1/5 times the peak area of diflorasone diacetate from the stand-ard solution. Furthermore, the total area of the peaks other than diflorasone diacetate from the sample solution is not larger than 1.5 times the peak area of diflorasone diacetate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.4 times as long as the retention time of diflorasone diacetate, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, and add acetonitrile to make exactly 20 mL. Con-firm that the peak area of diflorasone diacetate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution, the relative standard de-viation of the peak area of diflorasone diacetate is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (0.2 g, in vacu-um, phosphorus (V) oxide, 609C, 4 hours). ‌852 Residue on ignition <2.44> Not more than 0.2z (0.5 g, platinum crucible).
Weigh accurately about 20 mg each of Diflorasone Diacetate and Diflorasone Diacetate RS, both previously dried, dissolve in exactly 4 mL each of the internal standard solution, add acetonitrile to make them 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of diflorasone diacetate to that of the internal standard. Amount (mg) of diflorasone diacetate (C26H23F2O7) ＝ MS × QT/QS MS: Amount (mg) of Diflorasone Diacetate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 4.0 with diluted phosphoric acid (1 in 200). To 550 mL of this solu-tion add 400 mL of acetonitrile and 100 mL of tetrahydrofu-ran. Flow rate: Adjust so that the retention time of diflorasone diacetate is about 15 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and diflorasone diacetate are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of diflorasone diacetate to that of the internal standard is not more than 1.0z.
ジフルコルトロン吉草酸エステル
Diflucortolone Valerate contains not less than 98.
Diflucortolone Valerate occurs as white, crys-tals or crystalline powder. It is sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Prepare the test solution by proceeding with 10 mg of Diflucortolone Valerate according to Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution responds to Qualita-tive Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Diflucortolone Valerate in methanol (3 in 200,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Diflucortolone Valerate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Diflucortolone Valerate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Diflucortolone Valerate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋110 – ＋1159(0.1 g calcu-lated on the dried basis, ethanol (99.5), 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Diflucortolone Valerate in a platinum crucible according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Carbonize and incinerate as directed under Residue on Ignition <2.44>. (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of sample solution by the automatic integration method, and calculate the amounts of these peaks by the area percentage method: the amount of each peak of flucortolone valerate, 12a diflucortolone valer-ate and D4 diflucortolone valerate, having the relative reten-tion times of about 0.97, 1.03 and 1.05 to diflucortolone valerate, respectively, is not more than 0.6z, respectively; the amount of the peak of clocortolone valerate, having the relative retention time of about 1.09, is not more than 0.3z; and the amount of each peak other than those mentioned above is not more than 0.1z. Furthermore, the total amount of the peaks other than diflucortolone valerate is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.4 times as long as the retention time of diflucortolone valerate, beginning after the solvent peak. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 0.1 mL of the sample ‌JP XVIII Official Monographs / Digoxin 853 solution, add a mixture of water and acetonitrile (1:1) to make 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suita-bility test, and add a mixture of water and acetonitrile (1:1) to make exactly 20 mL. Confirm that the peak area of diflucortolone valerate obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solu-tion for system suitability test. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 5 mg each of Diflucortolone Valerate and Diflucortolone Valerate RS (separately, deter-mine the loss on drying <2.41> under the same conditions as Diflucortolone Valerate), dissolve each in a mixture of water and acetonitrile (1:1) to make exactly 10 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the Digoxin
Rounded, string-like algae, 2 – 3 mm in diame-ter; externally, dark red-purple to dark grayish red or grayish brown; a few branched rods irregularly forked, covered with short hairy twigs; calcified weeds and other small algae often attached. Odor, seaweed-like; taste, disagreeable and slightly salty.
To 2 g of pulverized Digenea add 10 mL of dilute ethanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, dissolve 5 mg of kainic acid in 10 mL of dilute ethanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl formate, water and formic acid (5:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the stand-ard solution.
Foreign matter <5.01>—The amount of other algae in Digenea does not exceed 20.0z. Loss on drying <5.01> Not more than 22.0z. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 5/8 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos- Acid-insoluble ash <5.01> Not more than 8.0z.
Digoxin, when dried, contains not less than 96.
Digoxin occurs as colorless or white crystals or a white crystalline powder. It is freely soluble in pyridine, slightly soluble in ethanol (95), very slightly soluble in acetic acid (100), and practically insoluble in water.
(1) Transfer 1 mg of Digoxin to a small test tube about 10 mm in inside diameter, dissolve in 1 mL of a solution of iron (III) chloride hexahydrate in acetic acid (100) (1 in 10,000), and underlay gently with 1 mL of sulfuric acid: at the zone of contact of the two liquids a brown ring free from a reddish color is produced, and the color of the upper layer near the contact zone changes to green through purple. Finally the entire acetic acid layer shows a green color through a deep blue color. (2) Determine the infrared absorption spectrum of Digoxin, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋10.0 – ＋13.09(after dry-ing, 0.20 g, dehydratead pyridine, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.10 g of Digoxin in 15 mL of diluted ethanol (95) (4 in 5) by warm-ing at 709C: the solution is clear and colorless. (2) Related substances—Dissolve 25.0 mg of Digoxin in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, add 10 mL of water and dilute ethanol to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve exactly 5.0 mg of Gitoxin for Purity RS, previously dried under reduced pressure at 1059C for 1 hour, in a mixture of acetonitrile and water (7:3) to make exactly 200 mL. Pipet 2 mL of this solution, add dilute ethanol to make exactly 50 mL, and use this solution as the standard solution. Perform ‌the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of gitoxin: AT is not larger than AS, and the total area of the peaks other than digoxin and gitoxin from the sample solution, obtained by the area per-centage method, is not more than 3z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of digoxin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 25 mg of Digoxin in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make 100 mL. To 10 mL of this solution add 10 mL of water and dilute ethanol to make 50 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the so-lution for system suitability test, and add dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, and add dilute ethanol to make exactly 100 mL. Confirm that the peak area of digoxin obtained with 10 mL of this solution is equivalent to 0.07 to 0.13z of that with 10 mL of the solu-tion for system suitability test. System performance: Dissolve 25 mg of Digoxin in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make 100 mL. To 10 mL of this solution add 5 mL of a solution of propyl parahydroxybenzoate in ethanol (95) (1 in 4000), 10 mL of water and dilute ethanol to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, digoxin and propyl parahydroxyben-zoate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of digoxin is not more than 2.5z. Loss on drying <2.41> Not more than 1.0z (0.5 g, in vacu-um, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.5z (0.1 g).
Weigh accurately about 25 mg each of Digoxin and Digoxin RS, previously dried, dissolve in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make exactly 100 mL. Pipet 10 mL of these solutions, add exactly 5 mL of the internal standard solution, 10 mL of water and dilute ethanol to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of digoxin to that of the internal stand-ard. 309C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of digoxin is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, digoxin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of digoxin to that of the internal standard is not more than 1.0z.
Dihydrocodeine Phosphate contains not less than 98.
Dihydrocodeine Phosphate occurs as a white to yellowish white crystalline powder. It is freely soluble in water and in acetic acid (100), slightly soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Dihydrocodeine Phos-phate in 10 mL of water is between 3.0 and 5.0. It is affected by light.
(1) Determine the absorption spectrum of a solution of Dihydrocodeine Phosphate (1 in 10,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared spectrum of Dihydrocodeine Phosphate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Dihydrocodeine Phosphate (1 in 20) responds to Qualitative Tests <1.09> (1) for phosphate.
(1) Chloride <1.03>—Perform the test with 0.5 g of Dihydrocodeine Phosphate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.20 g of Dihy- drocodeine Phosphate. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.240z). Related substances—Dissolve 0.20 g of Dihydro-codeine Phosphate in 10 mL of diluted ethanol (1 in 2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ethanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-chromatography. Develop the plate with a mixture of ethanol (99.5), toluene, acetone and ammonia solution (28) (14:14:7:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 4 hours).
Weigh accurately about 0.5 g of Dihydrocodeine Phosphate, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to greenish blue (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.94 mg of C18H23NO3.H3PO4
ジヒドロエルゴタミンメシル酸塩
Dihydroergotamine Mesilate contains not less than 97.
Dihydroergotamine Mesilate occurs as a white to yellowish white or grayish white to reddish white powder. It is freely soluble in acetic acid (100), sparingly soluble in methanol and in chloroform, slightly soluble in water and in ethanol (95), and practically insoluble in acetic anhydride and in diethyl ether. It is gradually colored by light. Melting point: about 2149C (with decomposition).
(1) Dissolve 1 mg of Dihydroergotamine Mesilate in 5 mL of a solution of L-tartaric acid (1 in 100). To 1 mL of this solution add 2 mL of 4-dimethylaminoben-zaldehyde-ferric chloride TS, and shake: a blue color de-velops. To 0.1 g of Dihydroergotamine Mesilate add 0.4 g of sodium hydroxide, stir well, and incinerate by gradual igni-tion. After cooling, add 10 mL of water to the residue, heat to boiling, cool, and filter. To the filtrate add 0.5 mL of hy-drochloric acid: the solution responds to Qualitative Tests <1.09> for sulfate. Separately, to 0.1 g of Dihydroergotamine Mesilate add 5 mL of dilute hydrochloric acid, shake for 5 minutes, filter, and to the filtrate add 1 mL of barium chlo-ride TS: the solution is clear. Determine the absorption spectrum of a solution of Dihydroergotamine Mesilate in methanol (1 in 20,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Di-hydroergotamine Mesilate, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －16.7 – －22.79[0.5 g, calcu-lated on the dried basis, a mixture of ethanol (99.5), chlo-roform and ammonia solution (28) (10:10:1), 20 mL, 100 mm]. pH <2.54> Dissolve 0.05 g of Dihydroergotamine Mesilate in 50 mL of water: the pH of this solution is between 4.4 and 5.4.
(1) Clarity and color of solution—Dissolve 0.10 g of Dihydroergotamine Mesilate in 0.1 mL of a solution of ‌860 methanesulfonic acid (7 in 100) and 50 mL of water: the solution is clear, and has no more color than the following control solutions [1] or [2]. Control solution [1]: Pipet 0.6 mL of Iron (III) Chloride CS and 0.15 mL of Cobalt (II) Chloride CS, mix, and add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. Control solution [2]: Pipet 0.6 mL of Iron (III) Chloride CS, 0.25 mL of Cobalt (II) Chloride CS and 0.1 mL of Cop-per (II) Sulfate CS, mix, and add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Dihydroergotamine Mesilate in 5 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (9:1) to make exactly 200 mL, and use this solution as the standard solution (1). Pipet 10 mL of the standard solution (1), add a mixture of chloroform and methanol (9:1) to make exactly 25 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, ethyl acetate, methanol and ammonia solution (28) (50:50:6:1) to a distance of about 15 cm, and dry the plate with cold wind within 1 minute. Develop the plate again immediately with a freshly prepared mixture of dichloromethane, ethyl acetate, methanol and ammonia so-lution (28) (50:50:6:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and dry the plate with warm wind: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and the spots, which are more in-tense than the spot from the standard solution (2), are not more than two. Loss on drying <2.41> Not more than 4.0z (0.5 g, in vacu-um at a pressure not exceeding 0.67 kPa, 1009C, 6 hours).
Weigh accurately about 0.2 g of Dihydroergotamine Mesilate, dissolve in 170 mL of a mixture of acetic anhydride and acetic acid (100) (10:1), and titrate <2.50> with 0.02 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L perchloric acid VS ＝ 13.60 mg of C33H37N5O5.CH4O3S
Dilazep Hydrochloride Hydrate occurs as a white crystalline powder. It is odorless. It is freely soluble in acetic acid (100) and in chloroform, soluble in water, slightly soluble in ethanol (95) and in acetic anhydride, and practically insoluble in diethyl ether. Melting point: 200 – 2049C Immerse the sample in a bath of 1109C, and raise the temperature at the rate of about 39C per minute from 1409C to 1509C, about 109C per minute from 1609C to 1959C and about 19C per minute from 1959C. trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Dilazep Hydrochloride Hy-drate in 100 mL of water: the pH of this solution is between 3.0 and 4.0.
(1) To 1 mL of a solution of Dilazep Hy-drochloride Hydrate (1 in 100) add 0.1 mL of a solution of hydroxylammonium chloride (1 in 10) and 0.1 mL of 8 mol/L potassium hydroxide TS, and warm in a water bath of 709C for 10 minutes. After cooling, add 0.5 mL of dilute hydrochloric acid and 0.1 mL of iron (III) chloride TS: a purple color develops.
(1) Clarity and color of solution—Dissolve 1.0 g of Dilazep Hydrochloride Hydrate in 20 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.5 g of Dilazep Hydrochloride Hydrate. Prepare the control solu-tion with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Dilazep Hydrochloride Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dilazep Hydrochloride Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.40 g of Dilazep Hy-drochloride Hydrate in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chroma-tography. Develop the plate with a mixture of methanol, ethyl acetate, dichloromethane and hydrochloric acid (500:200:100:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> 2.0 – 3.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Dilazep Hydrochlo-ride Hydrate, dissolve in 40 mL of a mixture of acetic anhy-dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 33.88 mg of C H N O .2HCl 31 44 2 10 Identification (1) To 1 mL of a solution of Dilazep Hy-drochloride Hydrate (1 in 100) add 0.1 mL of a solution of hydroxylammonium chloride (1 in 10) and 0.1 mL of 8 mol/L potassium hydroxide TS, and warm in a water bath of 709C for 10 minutes. After cooling, add 0.5 mL of dilute hydrochloric acid and 0.1 mL of iron (III) chloride TS: a purple color develops.
Diltiazem Hydrochloride occurs as white, crys-tals or crystalline powder. It is odorless. It is very soluble in formic acid, freely soluble in water, in methanol and in chloroform, sparingly soluble in aceto-nitrile, slightly soluble in acetic anhydride and in ethanol (99.5), and practically insoluble in diethyl ether.
(1) Dissolve 0.05 g of Diltiazem Hydro-chloride in 1 mL of 1 mol/L hydrochloric acid TS, add 2 mL of ammonium thiocyanate-cobalt (II) nitrate TS and 5 mL of chloroform, shake well, and allow to stand: a blue color de-velops in the chloroform layer. Proceed as directed under Oxygen Flask Combustion Method <1.06> with 0.03 g of Diltiazem Hydrochloride, using 20 mL of water as the absorbing liquid, and prepare the test solution: the test solution responds to Qualitative Tests <1.09> (1) for sulfate. Dissolve 0.01 g of Diltiazem Hydrchloride in 0.01 mol/L hydrochloric acid TS to make 100 mL. To 2 mL of the solution add 0.01 mol/L hydrochloric acid TS to make 20 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dil-tiazem Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>: it exhibits absorption at the wave numbers of about 1741 cm－1, 1678 cm－1, 1252 cm－1 and 1025 cm－1. A solution of Diltiazem Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: ＋115 – ＋1209(after drying, 0.20 g, water, 20 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Diltiazem Hydrochloride in 20 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 1.0 g of Diltia-zem Hydrochloride. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Diltia- zem Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Place 1.0 g of Diltiazem Hydrochlo- ride in a decomposition flask, add 5 mL of nitric acid and 2 mL of sulfuric acid, put a small funnel on the neck of the flask, and heat cautiously until white fumes are evolved. After cooling, add 2 mL of nitric acid, heat, and repeat this procedure twice, add several 2-mL portions of hydrogen peroxide (30), and heat until the solution becomes colorless to pale yellow. After cooling, add 2 mL of saturated solution of ammonium oxalate monohydrate, and heat again until white fumes are evolved. After cooling, add water to make 5 mL, use this solution as the test solution, and perform the test: the test solution has no more color than the following control solution (not more than 2 ppm). Control solution: Proceed in the same manner as the test solution without Diltiazem Hydrochloride, add 2.0 mL of Standard Arsenic Solution and water to make 5 mL, and proceed in the same manner as the test solution. Related substances—Dissolve 50 mg of Diltiazem Hy-drochloride in 50 mL of diluted ethanol (4 in 5), and use this solution as the sample solution. Measure exactly 1 mL of the sample solution, add diluted ethanol (4 in 5) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area of both solutions by automatic integra-tion method: the total area of peaks other than the peak of diltiazem obtained from the sample solution is not larger than 3/5 times the peak area of diltiazem from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 8 g of sodium acetate trihydrate and 1.5 g of d-camphorsulfonic acid in 500 mL of water, and filter using a membrane filter (0.4 mm in pore size). Add 250 mL each of acetonitrile and methanol to the filtrate. Flow rate: Adjust so that the retention time of diltiazem is about 9 minutes. Time span of measurement: About twice as long as the retention time of diltiazem, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add diluted ethanol (4 in 5) to make exactly 10 mL. Confirm that the peak area of diltiazem ob-tained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: Dissolve 0.03 g of Diltiazem Hydrochloride, 0.02 g of d-3-hydroxy-cis-2,3-dihydro-5-[2-(dimethylamino)ethyl]-2-(4-methoxyphenyl)-1,5-benzothia-zepin-4-(5H )-one hydrochloride (hereinafter referred to as de-acetyl substance) and 0.02 g of phenylbenzoate in 160 mL of ethanol (99.5), and add water to make 200 mL. When the ‌864 procedure is run with 20 mL of this solution under the above operating conditions, de-acetyl substance, diltiazem and phenyl benzoate are eluted in this order with the resolutions between the peaks of de-acetyl substance and diltiazem and between the peaks of diltiazem and phenyl benzoate being not less than 2.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diltiazem is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Diltiazem Hydro-chloride, previously dried, dissolve in 2.0 mL of formic acid, add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 45.10 mg of C22H26N2O4S.HCl
Dilute Hydrochloric Acid contains not less than 9.
Dilute Hydrochloric Acid is a colorless liquid. It is odorless and has a strong acid taste. Specific gravity d 20: about 1.05.
A solution of Dilute Hydrochloric Acid (1 in 30) changes blue litmus paper to red and responds to Quali-tative Tests <1.09> for chloride.
(1) Sulfate—To 3.0 mL of Dilute Hydrochloric Acid add 5 mL of water and 5 drops of barium chloride TS, and allow to stand for 1 hour: no turbidity is produced. Sulfite—To 3.0 mL of Dilute Hydrochloric Acid add 5 mL of water and 1 drop of iodine TS: the color of iodine TS does not disappear. Bromide or iodide—Place 10 mL of Dilute Hydro-chloric Acid in a glass-stoppered test tube, add 1 mL of chlo- ‌1112‌ roform and 1 drop of 0.002 mol/L potassium permanganate VS, and shake well: the chloroform layer remains colorless. Bromine or chlorine—Place 10 mL of Dilute Hydro-chloric Acid in a glass-stoppered test tube, add 5 drops of potassium iodide TS and 1 mL of chloroform, and shake for 1 minute: the chloroform layer remains free from a purple color. Heavy metals <1.07>—Evaporate 9.5 mL of Dilute Hydrochloric Acid on a water bath to dryness, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution as follows: to 3.0 mL of Standard Lead Solu-tion add 2 mL of dilute acetic acid and water to make 50 mL (not more than 3 ppm). Arsenic <1.11>—Prepare the test solution with 4.0 mL of Dilute Hydrochloric Acid according to Method 1, and perform the test (not more than 0.5 ppm). Mercury—Dilute 80 mL of Dilute Hydrochloric Acid with water to make exactly 100 mL, and use this solution as the sample solution. Perform the test with the sample solu-tion as directed under Atomic Absorption Spectrophotome-try <2.23> (cold vapor type). Place the sample solution in a sample bottle of the atomic absorption spectrophotometer, add 10 mL of tin (II) chloride-sulfuric acid TS, connect the bottle immediately to the spectrophotometer, circulate air, and determine the absorbance AT of the sample solution after the recorder reading has risen rapidly and become con-stant at a wavelength of 253.7 nm. On the other hand, to 8 mL of Standard Mercury Solution add water to make exactly 100 mL, and determine the absorbance AS of the solution obtained by the same procedure as used for the sample solu-tion: AT is smaller than AS (not more than 0.01 ppm). Residue on ignition <2.44> Pipet 10 mL of Dilute Hydro-chloric Acid, add 2 drops of sulfuric acid, evaporate to dry-ness, and ignite: the mass of the residue is not more than 1.0 mg.
Measure exactly 10 mL of Dilute Hydrochloric Acid, and dilute with 20 mL of water. Titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 to 3 drops of methyl red TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 36.46 mg of HCl
Dilute Iodine Tincture contains not less than 2.
Dilute Iodine Tincture is a dark red-brown liq-uid, and has a characteristic odor. Specific gravity d 20: about 0.93
(1) To a mixture of 1 mL of starch TS and 9 mL of water add 1 drop of Dilute Iodine Tincture: a dark blue-purple color develops. (2) Evaporate 3 mL of Diluted Iodine Tincture to dry-ness on a water bath, and heat gently over a free flame: a white residue is formed which responds to the Qualitative Tests <1.09> for potassium salt and iodide. Alcohol number <1.01> Not less than 6.7 (Method 2). Per-form the pretreatment (ii) in the Method 1.
(1) Iodine—Pipet exactly 10 mL of Dilute Iodine Tincture, add 0.5 g of potassium iodide, 20 mL of water and 1 mL of dilute hydrochloric acid, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 2 mL of starch TS). Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 12.69 mg of I (2) Potassium iodide—Pipet 10 mL of Dilute Iodine Tincture into an iodine flask, add 20 mL of water, 50 mL of hydrochloric acid and 5 mL of chloroform. Cool to room temperature, and titrate <2.50> with 0.05 mol/L potassium iodate VS until the red-purple color in the chloroform layer disappears while agitating vigorously and continuously. After the chloroform layer has been decolorized, allow the mixture to stand for 5 minutes. If the color reappears, the mixture should be titrated <2.50> further with 0.05 mol/L potassium iodate VS. Calculate the amount (mg) of potas-sium iodide from the volume (a mL) of 0.05 mol/L potas-sium iodate VS consumed as above and the volume ( b mL) of 0.1 mol/L sodium thiosulfate VS consumed in the titra-tion under Assay (1). Amount (mg) of potassium iodide (KI) ＝ 16.60 ×{a － (b/2)}
Dimemorfan Phosphate, when dried, contains not less than 98.
Dimemorfan Phosphate occurs as white to pale yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in water and in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 2659C (with decomposition).
(1) Determine the absorption spectrum of a solution of Dimemorfan Phosphate (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dimemorfan Phosphate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibits similar intensities of absorption at the same wave numbers. To 2 mL of a solution of Dimemorfan Phosphate (1 in 100) add 2 to 3 drops of silver nitrate TS: a yellow precipi-tate is formed, and it dissolves on the addition of dilute nitric acid. D Optical rotation <2.49> [a]20: ＋25 – ＋279(after drying, 1 g, methanol, 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Dimemorfan Phosphate in 100 mL of water: the pH of this solution is between 4.0 and 5.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Dimemorfan Phosphate according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dimemorfan Phosphate according to Method 3, and per-form the test. Use 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) (not more than 2 ppm). Related substances—Dissolve 0.10 g of Dimemorfan Phosphate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, chloroform and am-monia solution (28) (150:150:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly the plate with Dragen-dorff's TS for spraying: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately about 0.6 g of Dimemorfan Phos-phate, previously dried, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.34 mg of C18H25N.H3PO4
Dimenhydrinate, when dried, contains not less than 53.
Dimenhydrinate occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is very soluble in chloroform, freely soluble in ethanol (95), and slightly soluble in water and in diethyl ether.
(1) Dissolve 0.5 g of Dimenhydrinate in 30 mL of dilute ethanol, add 30 mL of water, and use this solu-tion as the sample solution. Transfer 30 mL of the sample solution to a separator, and add 2 mL of ammonia solution (28). Extract with two 10-mL portions of diethyl ether, com-bine the diethyl ether extracts, wash the combined extracts with 5 mL of water, and then extract the combined extracts with 15 mL of diluted hydrochloric acid (1 in 100). With this acid extract perform the following tests. To 5 mL of this acid extract add 5 drops of Reinecke salt TS: a light red precipitate is produced. To 10 mL of this acid extract add 10 mL of 2,4,6-trinitrophenol TS dropwise, and allow to stand for 30 minutes. Collect the precipitate by filtrating, recrystallize from dilute ethanol, and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1289C and 1339C. To 30 mL of the sample solution obtained in (1) add 2 mL of dilute sulfuric acid, and cool for 30 minutes. Scratch the inside wall of the container frequently to facilitate crystallization. Filter, and wash the white crystals with a small amount of ice-cooled water. Dry the crystals for 1 hour at 1059C: the crystals melt <2.60> between 3009C and 3059C with decomposition. To 0.01 g of the crystals obtained in (2) add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, and evaporate on a water bath to dryness: the residue shows a yellow-red color. When the dish containing the residue is held over a vessel containing 2 to 3 drops of ammonia TS, the color changes to red-purple, which is discharged on the addition of 2 to 3 drops of sodium hydroxide TS. Mix well 0.05 g of the crystals obtained in (2) with 0.5 g of sodium peroxide in a nickel crucible, and heat until the mass melts. Cool, dissolve the melted mass in 20 mL of water, and acidify with dilute nitric acid: the solution re-sponds to Qualitative Tests <1.09> for chloride.
(1) Chloride <1.03>—Transfer 50 mL of the fil-trate obtained in the Assay (2) to a Nessler tube, add 1 mL of nitric acid, and allow to stand for 5 minutes: the turbidity of the solution is not greater than that of the following control ether extracts, and wash the combined diethyl ether extracts with three 50-mL portions of water. To the diethyl ether ex-tracts add exactly 25 mL of 0.05 mol/L sulfuric acid VS, and add 25 mL of water. Shake thoroughly, and evaporate the diethyl ether gently. Cool, and titrate the excess sulfuric acid with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of methyl red TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sulfuric acid VS ＝ 25.54 mg C17H21NO (2) 8-Chlorotheophylline—Weigh accurately about 0.8 g of Dimenhydrinate, previously dried, transfer to a 200-mL volumetric flask, add 50 mL of water, 3 mL of ammonia TS and 6 mL of a solution of ammonium nitrate (1 in 10), and heat on a water bath for 5 minutes. Add exactly 25 mL of 0.1 mol/L silver nitrate VS, heat on a water bath for 15 minutes with occasional shaking, cool, and add water to make exactly 200 mL. Allow to stand overnight to settle the precipitate, and filter through a dry filter paper, discarding the first 20 mL of the filtrate. Measure exactly 100 mL of the subsequent filtrate, acidify with nitric acid, add 3 mL of nitric acid, and titrate the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L silver nitrate VS ＝ 21.46 mg of C7H7ClN4O2
(1) Diphenhydramine—Weigh accurately about 0.5 g of Dimenhydrinate, previously dried, transfer to a 250-mL separator, and add 50 mL of water, 3 mL of ammo- mine (C 17H21 NO: 255.36), and not less than 44.0z nia TS and 10 g of sodium chloride. Extract with six 15-mL portions of diethyl ether with shaking, combine the diethyl and not more than 47.0z of 8-chlorotheophylline (C7H7ClN4O2: 214.61). Description Dimenhydrinate occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is very soluble in chloroform, freely soluble in ethanol (95), and slightly soluble in water and in diethyl ether. Identification (1) Dissolve 0.5 g of Dimenhydrinate in 30 mL of dilute ethanol, add 30 mL of water, and use this solu-tion as the sample solution. Transfer 30 mL of the sample solution to a separator, and add 2 mL of ammonia solution (28). Extract with two 10-mL portions of diethyl ether, com-bine the diethyl ether extracts, wash the combined extracts with 5 mL of water, and then extract the combined extracts with 15 mL of diluted hydrochloric acid (1 in 100). With this acid extract perform the following tests. To 5 mL of this acid extract add 5 drops of Reinecke salt TS: a light red precipitate is produced. To 10 mL of this acid extract add 10 mL of 2,4,6-trinitrophenol TS dropwise, and allow to stand for 30 minutes. Collect the precipitate by filtrating, recrystallize from dilute ethanol, and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1289C and 1339C. To 30 mL of the sample solution obtained in (1) add 2 mL of dilute sulfuric acid, and cool for 30 minutes. Scratch the inside wall of the container frequently to facilitate crystallization. Filter, and wash the white crystals with a small amount of ice-cooled water. Dry the crystals for 1 hour at 1059C: the crystals melt <2.60> between 3009C and 3059C with decomposition. To 0.01 g of the crystals obtained in (2) add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, and evaporate on a water bath to dryness: the residue shows a yellow-red color. When the dish containing the residue is held over a vessel containing 2 to 3 drops of ammonia TS, the color changes to red-purple, which is discharged on the addition of 2 to 3 drops of sodium hydroxide TS. Mix well 0.05 g of the crystals obtained in (2) with 0.5 g of sodium peroxide in a nickel crucible, and heat until the mass melts. Cool, dissolve the melted mass in 20 mL of water, and acidify with dilute nitric acid: the solution re-sponds to Qualitative Tests <1.09> for chloride. Melting point <2.60> 102 – 1079C Purity (1) Chloride <1.03>—Transfer 50 mL of the fil-trate obtained in the Assay (2) to a Nessler tube, add 1 mL of nitric acid, and allow to stand for 5 minutes: the turbidity of the solution is not greater than that of the following control ether extracts, and wash the combined diethyl ether extracts with three 50-mL portions of water. To the diethyl ether ex-tracts add exactly 25 mL of 0.05 mol/L sulfuric acid VS, and add 25 mL of water. Shake thoroughly, and evaporate the diethyl ether gently. Cool, and titrate the excess sulfuric acid with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of methyl red TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sulfuric acid VS ＝ 25.54 mg C17H21NO (2) 8-Chlorotheophylline—Weigh accurately about 0.8 g of Dimenhydrinate, previously dried, transfer to a 200-mL volumetric flask, add 50 mL of water, 3 mL of ammonia TS and 6 mL of a solution of ammonium nitrate (1 in 10), and heat on a water bath for 5 minutes. Add exactly 25 mL of 0.1 mol/L silver nitrate VS, heat on a water bath for 15 minutes with occasional shaking, cool, and add water to make exactly 200 mL. Allow to stand overnight to settle the precipitate, and filter through a dry filter paper, discarding the first 20 mL of the filtrate. Measure exactly 100 mL of the subsequent filtrate, acidify with nitric acid, add 3 mL of nitric acid, and titrate the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L silver nitrate VS ＝ 21.46 mg of C7H7ClN4O2
Dimercaprol is a colorless or pale yellow liq-uid. It has a mercaptan-like, disagreeable odor. It is miscible with methanol and with ethanol (99.5) It is soluble in peanut oil, and sparingly soluble in water. It shows no optical rotation.
(1) Add 1 drop of Dimercaprol to a mix-ture of 1 drop of a solution of cobalt (II) chloride hexahy-drate (1 in 200) and 5 mL of water: a yellow-brown color de-velops. (2) Determine the infrared absorption spectrum of Dimercaprol as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.570 – 1.575 Specific gravity <2.56> d 20: 1.238 – 1.248
(1) Clarity and color of solution—Dissolve 1.0 mL of Dimercaprol in 20 mL of peanut oil: the solution is clear and colorless to pale yellow. Bromide—To 2.0 g of Dimercaprol add 25 mL of dilute potassium hydroxide-ethanol TS, and heat in a water bath under a reflux condenser for 2 hours. Evaporate the ethanol in a current of warm air, add 20 mL of water, and cool. Add a mixture of 10 mL of hydrogen peroxide (30) and 17 21 7 7 4 2 40 mL of water, boil gently under a reflux condenser for 10 ethanol (95), and heat with swirling on a water bath until the solution just boils. Continue to heat for 30 seconds, and filter through a glass filter (G4). Wash the filter with warm ethanol (95), transfer the filtrate and washings to a flask, and evaporate the ethanol on a water bath to make 5 mL. Add 50 mL of water, 3 mL of ammonia TS and 6 mL of a solution of ammonium nitrate (1 in 10), heat the mixture on a water bath for 5 minutes, add exactly 25 mL of 0.1 mol/L silver nitrate VS, and heat on a water bath for 15 minutes with occasional shaking. Transfer the mixture to a 200-mL volumetric flask, using water to rinse the flask, cool, add water to make exactly 200 mL, and proceed as directed in the Assay (2) under Dimenhydrinate. Each mL of 0.1 mol/L silver nitrate VS ＝ 47.00 mg of C17H21NO.C7H7ClN4O2
Weigh accurately about 0.15 g of Dimercaprol into a glass-stoppered flask, dissolve in 10 mL of methanol, and titrate <2.50> immediately with 0.05 mol/L iodine VS until a pale yellow color is produced. Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.05 mol/L iodine VS ＝ 6.212 mg of C3H8OS2
Dimorpholamine, when dried, contains not less than 98.
Dimorpholamine is a white to light yellow, crystalline powder, masses or syrupy liquid. It is very soluble in ethanol (99.5) and in acetic anhydride, and soluble in water. The pH of a solution prepared by dissolving 1.0 g of Dimorpholamine in 10 mL of water is between 6.0 and 7.0. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Dimorpholamine (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. (2) Determine the infrared absorption spectrum of Dimorpholamine, previously dried, as directed in the potas-sium bromide disk method under the Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Dimorpholamine in 50 mL of water: the solution is clear and colorless to pale yellow. Chloride <1.03>—To 20 mL of the solution obtained in (1) add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.036z). Sulfate <1.14>—To 10 mL of the solution obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.096z). Heavy metals <1.07>—Proceed with 2.0 g of Dimor- pholamine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of Dimorphola-mine in 10 mL of ethanol (99.5), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5) and water (4:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 8 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Dimorpholamine, previously dried, dissolve in 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.85 mg of C20H38N4O4
Dinoprost occurs as white, waxy masses or powder, or a clear, colorless to light yellow and viscous liquid. It is odorless. It is very soluble in N,N-dimethylformamide, freely solu-ble in methanol, in ethanol (99.5) and in diethyl ether, and very slightly soluble in water.
(1) To 5 mg of Dinoprost add 2 mL of sul-furic acid, and dissolve by shaking for 5 minutes: a dark red color develops. To this solution add 30 mL of sulfuric acid: an orange color develops with a green fluorescence. Dissolve 1 mg of Dinoprost in 50 mL of diluted sulfu-ric acid (7 in 10), and warm in a water bath warmed at 509C for 40 minutes. After cooling, determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Amount (mg) of dimorpholamine (C ＝ MS × QT/QS × 1/5 20H38N4O4) Warm Dinoprost at 409C to effect a liquid, and de-termine the infrared absorption spectrum of the liquid as directed in the liquid film method under Infrared Spectro- MS: Amount (mg) of dimorpholamine for assay taken Internal standard solution—A solution of butyl parahy-droxybenzoate in acetonitrile (1 in 25,000). Operating conditions— photometry <2.25>, and compare the spectrum with the Ref- erence Spectrum: both spectra exhibits similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋24 – ＋319(0.2 g, ethanol Detector: An ultraviolet absorption photometer (wave- (99.5), 10 mL, 100 mm). ‌870
(1) Clarity and color of solution—Dissolve 0.20 g of Dinoprost in 5 mL of ethanol (99.5): the solution is clear and colorless to pale yellow. (2) Related substances—Dissolve 10 mg of Dinoprost in 2 mL of methanol, add water to make 10 mL, and use this solution as the sample solution. Pipet 3 mL of the sample so-lution, add diluted methanol (1 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the total area of the peaks other than dinoprost ob-tained from the sample solution is not larger than the peak area of dinoprost from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column about 5 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogenphosphate TS and acetonitrile (5:2). Flow rate: Adjust so that the retention time of dinoprost is about 20 minutes. Selection of column: Dissolve 0.01 g each of isopropyl parahydroxybenzoate and propyl parahydroxybenzoate in 2 mL of methanol, and add water to make 10 mL. To 1 mL of this solution add diluted methanol (1 in 5) to make 30 mL, proceed with 10 mL of this solution under the above operat-ing conditions, and calculate the resolution. Use a column giving elution of isopropyl parahydroxybenzoate and propyl parahydroxybenzoate in this order with the resolution be-tween these peaks being not less than 2.5. Detection sensitivity: Adjust the detection sensitivity so that the peak height of dinoprost from the standard solution composes 5z to 15z of the full scale. Time span of measurement: About 1.5 times as long as the retention time of dinoprost, beginning after the solvent peak. Water <2.48> Not more than 0.5z (0.3 g, volumetric titra-tion, direct titration).
Weigh accurately about 50 mg of Dinoprost, dis-solve in 30 mL of N,N-dimethylformamide, and titrate <2.50> with 0.02 mol/L tetramethylammonium hydroxide VS under a stream of nitrogen (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L tetramethylammonium hydroxide VS Diphenhydramine
Diphenhydramine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a bitter taste, followed by a sensation of numbness on the ton-gue. It is very soluble in methanol and in acetic acid (100), freely soluble in water and in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in diethyl ether. It is gradually affected by light.
(1) Determine the absorption spectrum of a solution of Diphenhydramine Hydrochloride in methanol (1 in 2000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Dipenhydramine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Diphenhydramine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Diphenhydramine Hydrochlo-ride in 10 mL of water: the pH of this solution is between 4.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Diphenhydramine Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Diphen- hydramine Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Diphenhydra-mine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard
Weigh accurately about 0.4 g of Diphenhydramine Hydrochloride, previously dried, dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3). Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS [147-24-0] ＝ 29.18 mg of C 17H 21NO.HCl Diphenhydramine Hydrochloride, when dried, contains not less than 98.0z of diphenhydramine hydrochloride (C17H21NO.HCl). Description Diphenhydramine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a bitter taste, followed by a sensation of numbness on the ton-gue. It is very soluble in methanol and in acetic acid (100), freely soluble in water and in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in diethyl ether. It is gradually affected by light. Identification (1) Determine the absorption spectrum of a solution of Diphenhydramine Hydrochloride in methanol (1 in 2000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Dipenhydramine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Diphenhydramine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Diphenhydramine Hydrochlo-ride in 10 mL of water: the pH of this solution is between 4.0 and 5.0. Melting point <2.60> 166 – 1709C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Diphenhydramine Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Diphen- hydramine Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Diphenhydra-mine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard
Diphenhydramine Tannate occurs as a grayish white to light brown powder. It is odorless or has a slight, characteristic odor. It is tasteless. It is slightly soluble in ethanol (95), and practically insolu-ble in water and in diethyl ether.
(1) To 1 g of Diphenhydramine Tannate add 15 mL of water and 0.3 mL of dilute hydrochloric acid, shake thoroughly for 1 minute, filter, and use this filtrate as the sample solution. Transfer 10 mL of the sample solution to a separator, extract with two 20-mL portions of chlo-roform, combine the chloroform extracts, and evaporate on a water bath to dryness. To 5 mL of a solution of the residue (1 in 100) add 5 drops of Reinecke salt TS: a light red pre-cipitate is produced. To 10 mL of a solution of the residue obtained in (1) (1 in 100) add 10 mL of 2,4,6-trinitrophenol TS dropwise, and allow to stand for 30 minutes. Collect the precipitate by filtration, recrystallize from dilute ethanol, and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1289C and 1339C. To 1 mL of the sample solution obtained in (1) add 1 drop of iron (III) chloride TS: a dark blue-purple color de-velops.
Heavy metals <1.07>—Proceed with 1.0 g of Diphenhydramine Tannate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 5 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
Transfer about 1.7 g of Diphenhydramine Tannate, accurately weighed, to a separator, dissolve in 20 mL of water and 3.0 mL of dilute hydrochloric acid with thorough shaking, add 20 mL of a solution of sodium hydroxide (1 in 10) and exactly 25 mL of isooctane, shake vigorously for 5 minutes, dissolve 2 g of sodium chloride with shaking, and allow to stand. To 20 mL of the isooctane layer add exactly 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 25.54 mg of C17H21NO
Diphenhydramine contains not less than 96.
Diphenhydramine is a clear, light yellow to yellow liquid. It has a characteristic odor, and has a burning taste at first, followed by a slight sensation of numbness on the tongue. It is miscible with acetic anhydride, with acetic acid (100), with ethanol (95) and with diethyl ether. D It is very slightly soluble in water. It is gradually affected by light. Refractive index n20: about 1.55 Boiling point: about 1629C (in vacuum, 0.67 kPa).
(1) To 50 mg of Diphenhydramine add 2 mL of sulfuric acid: an orange-red precipitate is produced immediately, and its color changes to red-brown on stand-ing. Add carefully 2 mL of water to this solution: the inten-sity of the color changes, but the color tone does not change. (2) Dissolve 0.1 g of Diphenhydramine in 10 mL of dilute ethanol, add an excess of a saturated solution of 2,4,6-trinitrophenol in dilute ethanol with stirring, and cool in ice. Collect the produced crystals, recrystallize from dilute ethanol, and dry at 1059C for 30 minutes: the crystals melt <2.60> between 1289C and 1339C. Specific gravity <2.56> d 20: 1.013 – 1.020
(1) b-Dimethylaminoethanol—Dissolve 1.0 g of Diphenhydramine in 20 mL of diethyl ether, and extract with two 10-mL portions of water with thorough shaking. Com-bine the water extracts, and add 2 drops of phenolphthalein TS and 1.0 mL of 0.05 mol/L sulfuric acid VS: no red color develops. Benzohydrol—Transfer 1.0 g of Diphenhydramine to a separator, dissolve in 20 mL of diethyl ether, and extract with two 25-mL portions of diluted hydrochloric acid (1 in 15) with thorough shaking. Separate the diethyl ether layer, evaporate slowly on a water bath, and dry in a desiccator (in vacuum, silica gel) for 2 hours: the mass of the residue is not more than 20 mg. ＝ 7.090 mg of C 20H 34O5 Heavy metals <1.07>—Proceed with 1.0 g of Diphen- hydramine according to Method 2, and perform the test.
Weigh accurately about 0.5 g of Diphenhydramine, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. ‌JP XVIII Official Monographs / Diphenhydramine and Bromovalerylurea Powder 871‌ Each mL of 0.1 mol/L perchloric acid VS ＝ 25.54 mg of C17H21NO
Containers—Tight containers. Storage—Light-resistant, and in a place not exceeding
Diphtheria Toxoid is a clear, colorless to light yellow-brown liquid. Adsorbed Diphtheria Toxoid for Adult Use
Dipyridamole, when dried, contains not less than 98.
Dipyridamole occurs as yellow, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in chloroform, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water and in diethyl ether.
(1) Dissolve 5 mg of Dipyridamole in 2 mL of sulfuric acid, add 2 drops of nitric acid, and shake: a deep purple color develops. Determine the absorption spectrum of a solution of Dipyridamole in a mixture of methanol and hydrochloric acid (99:1) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dipyridamole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Dipyridamole in 10 mL of chloroform: the solution is clear, and shows a yellow color. Heavy metals <1.07>—Proceed with 2.0 g of Dipyridamole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dipyridamole according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Dipyridamole in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than dipyridamole obtained from the sample solution is not larger than the peak area of dipyridamole from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.2 g of potassium dihydrogen phosphate in 200 mL of water, and add 800 mL of metha-nol. Flow rate: Adjust so that the retention time of dipyrida-mole is about 4 minutes. Time span of measurement: About 5 times as long as the retention time of dipyridamole. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of dipyridamole obtained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: Dissolve 7 mg of Dipyridamole and 3 mg of terphenyl in 50 mL of methanol. When the proce-dure is run with 20 mL of this solution under the above oper-ating conditions, dipyridamole and terphenyl are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of dipyridamole is not more than 1.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Dipyridamole, pre-viously dried, dissolve in 70 mL of methanol, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 50.46 mg of C24H40N8O4
Disopyramide occurs as white, crystals or crys-talline powder. It is very soluble in methanol and in ethanol (95), freely soluble in acetic anhydride, in acetic acid (100) and in diethyl ether, and slightly soluble in water.
(1) To 1 mL of a solution of Disopyramide in ethanol (95) (1 in 20) add 10 mL of 2,4,6-trinitrophenol TS, and warm: a yellow precipitate is formed. Filter this pre-cipitate, wash with water, and dry at 1059C for 1 hour: the residue melts <2.60> between 1729C and 1769C. Determine the absorption spectrum of a solution of Disopyramide in 0.05 mol/L sulfuric acid-methanol TS (1 in 25,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab- pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, 809C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.25 g of Disopyramide, dis-solve in 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.97 mg of C21H29N3O
Distigmine Bromide contains not less than 98.
Distigmine Bromide occurs as a white crystal-line powder. It is very soluble in water, freely soluble in methanol, in ethanol (95) and in acetic acid (100), and slightly soluble in acetic anhydride. The pH of a solution of Distigmine Bromide (1 in 100) is between 5.0 and 5.5. It is slightly hygroscopic. It is gradually colored by light. Melting point: about 1509C (with decomposition).
(1) Determine the absorption spectrum of a solution of Distigmine Bromide (1 in 25,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dis-tigmine Bromide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Distigmine Bromide (1 in 10) add 2 mL of dilute nitric acid: the solution responds to Qualitative Tests <1.09> (1) for bromide.
(1) Heavy metals <1.07>—Dissolve 1.0 g of Dis-opyramide in 10 mL of ethanol (95), and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 10 mL of ethanol (95), 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Disopyramide according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.40 g of Disopyramide in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 400 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 1-butanol, water and ammonia solution (28) (45:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi- Description Distigmine Bromide occurs as a white crystal-line powder. It is very soluble in water, freely soluble in methanol, in ethanol (95) and in acetic acid (100), and slightly soluble in acetic anhydride. The pH of a solution of Distigmine Bromide (1 in 100) is between 5.0 and 5.5. It is slightly hygroscopic. It is gradually colored by light. Melting point: about 1509C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Distigmine Bromide (1 in 25,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dis-tigmine Bromide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Distigmine Bromide (1 in 10) add 2 mL of dilute nitric acid: the solution responds to Qualitative Tests <1.09> (1) for bromide. Purity (1) Clarity and color of solution—Dissolve 0.25 g of Distigmine Bromide in 5 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.40 g of Dis- ‌876 tigmine Bromide. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Distig- TS to make exactly V? mL so that each mL contains about 30 mg of distigmine bromide (C22H32Br2N4O4), and use this solu-tion as the sample solution. Proceed as directed in the Assay. mine Bromide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Amount (mg) of distigmine bromide (C 22H32Br2N4O4) Solution (not more than 10 ppm). Related substances—Dissolve 40 mg of Distigmine Bromide in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of cellulose with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water, ethanol (99.5) and acetic acid (100) (8:3:2:1) to a distance of about 13 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Water <2.48> Not more than 1.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Distigmine Bromide, dissolve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (8:1), and titrate <2.50> with 0.1 mol/L ＝ MS × (AT2 － AT1)/(AS2 － AS1) × V?/V × 1/20 MS: Amount (mg) of distigmine bromide for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 500 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Distigmine Bromide Tablets is not less than 80z. Start the test with 1 tablet of Distigmine Bromide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 10 mg of distigmine bromide (C22H32Br2N4O 4), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of distigmine bromide for assay (separately determine the water <2.48> in the same manner as Distigmine Bromide), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 500 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT1 and AS1, at 270 nm, and AT2 and AS2, at 350 nm. Dissolution rate (z) with respect to the labeled amount perchloric acid VS (potentiometric titration with platinum electrode). Perform a blank determination in the same man- of distigmine bromide (C22H32 Br2N4O4) ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 10 MS: Amount (mg) of distigmine bromide for assay taken, ＝ 28.82 mg of C22H32 Br2N4O4 calculated on the anhydrous basis C: Labeled amount (mg) of distigmine bromide
Disulfiram occurs as white to yellowish white, crystals or crystalline powder. It is freely soluble in acetone and in toluene, sparingly soluble in methanol and in ethanol (95), and practically in-soluble in water.
(1) Determine the absorption spectrum of a solution of Disulfiram in ethanol (95) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Disul-firam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Disulfiram according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Disulfiram according to Method 4, and perform the test (not more than 2 ppm). Diethyldithiocarbamic acid—Dissolve 0.10 g of Dis-ulfiram in 10 mL of toluene, and shake with 10 mL of diluted sodium carbonate TS (1 in 20). Discard the toluene layer, wash the water layer with 10 mL of toluene, shake with 5 drops of a solution of cupric sulfate (1 in 250) and 2 mL of toluene, and allow to stand: no light yellow color de-velops in the toluene layer. Related substances—Dissolve 50 mg of Disulfiram in 40 mL of methanol, add water to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and the stand-ard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the total area of the peaks other than disulfiram ob-tained from the sample solution is not larger than the peak area of disulfiram from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 5 mm in inside di-ameter and about 15 cm in length, packed with octadecyl- silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (7:3). Flow rate: Adjust so that the retention time of disulfiram is about 8 minutes. Selection of column: Dissolve 50 mg of Disulfiram and 50 mg of benzophenone in 40 mL of methanol, and add water to make 50 mL. To 1 mL of this solution add the mobile phase to make 200 mL. Proceed with 10 mL of this solution under the above operating conditions, and calculate the reso-lution. Use a column giving elution of benzophenone and disulfiram in this order with the resolution between these peaks being not less than 4. Detection sensitivity: Adjust so that the peak height of dis-ulfiram obtained from 10 mL of the standard solution is 15 – 30 mm. Time span of measurement: About 3.5 times of the reten-tion time of disulfiram. Loss on drying <2.41> Not more than 0.20z (2 g, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (2 g).
Weigh accurately about 0.2 g of Disulfiram, previ-ously dried, in an iodine bottle, dissolve in 20 mL of ace-tone, add 1.5 mL of water and 1.0 g of potassium iodide, and dissolve by shaking thoroughly. To this solution add 3.0 mL of hydrochloric acid, stopper the bottle tightly, shake, and allow to stand in a dark place for 3 minutes. Add 70 mL of water, and titrate <2.50> with 0.1 mol/L sodium thiosul-fate VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 14.83 mg of C10H20N2S4
dl-Camphor occurs as colorless or white, translucent crystals, crystalline powder or masses. It has a characteristic, agreeable odor, and has a slightly bitter taste followed by a pleasant, cooling sensation. It is freely soluble in ethanol (95), in diethyl ether and in carbon disulfide, and slightly soluble in water. It slowly volatilizes at room temperature.
Dissolve 0.1 g of dl-Camphor in 2 mL of methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and heat for 5 minutes on a water bath: an orange-red precipitate is formed. D Optical rotation <2.49> [a]20: －1.5 – ＋1.59(5 g, ethanol System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of dl-camphor to that of the internal standard is not more than 1.0z.
(1) Water—Shake 1.0 g of d-Camphor with 10 mL of carbon disulfide: the solution is clear. (2) Chlorinated compounds—Mix 0.20 g of finely pow- C10 H16O: 152.23 dered d-Camphor with 0.4 g of sodium peroxide in a dried porcelain crucible. Heat the crucible gently by the open flame until the incineration is complete. Dissolve the residue in 20 mL of warm water, acidify with 12 mL of dilute nitric acid, and filter the solution into a Nessler tube. Wash the (1RS,4RS )-1,7,7-Trimethylbicyclo[2.2.1]heptan-2-one [76-22-2] dl-Camphor contains not less than 96.0z of dl-cam-phor (C10H16O). ‌612 Description dl-Camphor occurs as colorless or white, translucent crystals, crystalline powder or masses. It has a characteristic, agreeable odor, and has a slightly bitter taste followed by a pleasant, cooling sensation. It is freely soluble in ethanol (95), in diethyl ether and in carbon disulfide, and slightly soluble in water. It slowly volatilizes at room temperature. Identification Dissolve 0.1 g of dl-Camphor in 2 mL of methanol, add 1 mL of 2,4-dinitrophenylhydradine TS, and heat for 5 minutes on a water bath: an orange-red precipitate is formed. D Optical rotation <2.49> [a]20: －1.5 – ＋1.59(5 g, ethanol System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of dl-camphor to that of the internal standard is not more than 1.0z.
dl-Menthol occurs as colorless crystals. It has a characteristic and refreshing odor and a burning taste, fol-lowed by a cool taste. It is very soluble in ethanol (95) and in diethyl ether, and very slightly soluble in water. It sublimes gradually at room temperature.
(1) Triturate dl-Menthol with an equal amount of camphor, chloral hydrate or thymol: the mixture liquefies. (2) Shake 1 g of dl-Menthol with 20 mL of sulfuric acid: the mixture becomes turbid with a yellow-red color. Allow to stand for 3 hours: a clear, oily layer possesses no aroma of menthol is separated. Congealing point <2.42> 27 – 289C D Optical rotation <2.49> [a]20: －2.0 – ＋2.09(2.5 g, ethanol (95), 25 mL, 100 mm).
(1) Non-volatile residue—Volatilize 2.0 g of dl-Menthol on a water bath, and dry the residue at 1059C for 2 hours: the residue weighs not more than 1.0 mg. Thymol—Add 0.20 g of dl-Menthol to a cold mixture of 2 mL of acetic acid (100), 6 drops of sulfuric acid and 2 drops of nitric acid: no green to blue-green color immedi-ately develops. Nitromethane or nitroethane—To 0.5 g of dl-Men-thol placed in a flask add 2 mL of a solution of sodium hy-droxide (1 in 2) and 1 mL of hydrogen peroxide (30), connect a reflux condenser to the flask, and boil the mixture gently for 10 minutes. After cooling, add water to make exactly 20 mL, and filter. Take 1 mL of the filtrate in a Nessler tube, add water to make 10 mL, neutralize with dilute hydrochlo-ric acid, then add 1 mL of dilute hydrochloric acid, and cool. To the mixture add 1 mL of a solution of sulfanilic acid (1 in 100), allow to stand for 2 minutes, and then add 1 mL of a solution of N,N-diethyl-N?-1-naphthylethylenedia-mine oxalate (1 in 1000) and water to make 25 mL: no red-purple color immediately develops.
Weigh accurately about 2 g of dl-Menthol, add ex-actly 20 mL of a mixture of dehydrated pyridine and acetic anhydride (8:1), connect a reflux condenser, and heat on a water bath for 2 hours. Wash down the condenser with 20 mL of water, and titrate <2.50> with 1 mol/L sodium hy-droxide VS (indicator: 5 drops of phenolphthalein TS). Per-form a blank determination in the same manner. Each mL of 1 mol/L sodium hydroxide VS ＝ 156.3 mg of C10H20O
dl-Methylephedrine Hydrochloride occurs as colorless crystals or a white crystalline powder. It is freely soluble in water, sparingly soluble in ethanol (99.5), slightly soluble in acetic acid (100), and practically insoluble in acetic anhydride. A solution of dl-Methylephedrine Hydrochloride (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of dl-Methylephedrine Hydrochloride (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of dl-Methylephedrine Hydrochloride, previously dried, as di-rected in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. A solution of dl-Methylephedrine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution prepared by dissolving 1.0 g of dl-Methylephedrine Hydrochloride in 20 mL of water is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of dl-Methylephedrine Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of dl- Methylephedrine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of dl-Methyl-ephedrine Hydrochloride in 20 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than the peak of methylephedrine ob-tained from the sample solution is not larger than the peak area of methylephedrine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 257 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 13.6 g of potassium dihydrogen phosphate and 3 g of sodium 1-heptane sulfonate in 1000 mL of water, and adjust the pH to 2.5 with phosphoric acid. To 900 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of methylephedrine is about 10 minutes. Time span of measurement: About 2 times as long as the retention time of methylephedrine, beginning after the sol-vent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add water to make exactly 20 mL. Con-firm that the peak area of methylephedrine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 50 mg of dl-Methylephe-drine Hydrochloride and 0.4 mg of methyl parahydroxyben-zoate in 50 mL of water. When the procedure is run with 20 mL of this solution under the above operating conditions, methylephedrine and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methylephedrine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of dl-Methylephedrine Hydrochloride, previously dried, dissolve in 80 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.57 mg of C11H17NO.HCl
Dobutamine Hydrochloride, when dried, contains not less than 98.
Dobutamine Hydrochloride occurs as white to very pale orange, crystalline powder or grains. It is freely soluble in methanol, sparingly soluble in water and in ethanol (95), and practically insoluble in diethyl ether. A solution of Dobutamine Hydrochloride (1 in 100) shows no optical rotation.
(1) Determine the infrared absorption spec- ‌878 tra of Dobutamine Hydrochloride, previously dried, as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Dobuta-mine Hydrochloride RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. (2) A solution of Dobutamine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Dobutamine Hydrochloride in 100 mL of water: the pH of this solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Dobutamine Hydrochloride in 30 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Dissolve 1.0 g of Dobutamine Hydrochloride in 40 mL of water by warming, cool, and add 2 mL of dilute acetic acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add water to make 50 mL (not more than 20 ppm). Related substances—Dissolve 0.10 g of Dobutamine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and formic acid (78:22:5) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand for 5 minutes in iodine vapor: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g each of Dobutamine Hydrochloride and Dobutamine Hydrochloride RS, each previously dried, dissolve separately in exactly 10 mL of the internal standard solution, add diluted methanol (1 in 2) to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of dobutamine to that of the internal standard, respectively. Amount (mg) of dobutamine hydrochloride (C18H23NO3.HCl) ＝ MS × QT/QS MS: Amount (mg) of Dobutamine Hydrochloride RS taken Internal standard solution—A solution of salicylamide in diluted methanol (1 in 2) (1 in 125). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of tartrate buffer solution (pH 3.0) and methanol (7:3). Flow rate: Adjust so that the retention time of dobuta-mine is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, dobutamine and internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of dobutamine to that of the internal standard is not more than 1.0z.
Docetaxel Hydrate contains not less than 97.
Docetaxel Hydrate occurs as a white crystalline powder. It is freely soluble in N,N-dimethylformamide and in ethanol (99.5), soluble in methanol and in dichloromethane, and practically insoluble in water. It decomposes on exposure to light.
(1) Determine the absorption spectrum of a solution of Docetaxel Hydrate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Docetaxel Hydrate RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. (2) Dissolve 60 mg of Docetaxel Hydrate in 1 mL of dichloromethane. Perform the test with this solution as di-rected in the solution method under Infrared Spectropho-tometry <2.25> using a fixed cell composed of potassium bromide optical plates with the cell length of 0.1 mm, and compare the spectrum with the Reference Spectrum or the spectrum of Docetaxel Hydrate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. ‌JP XVIII Official Monographs / Docetaxel Injection 879 Optical rotation <2.49> [a]20: －39 – －419(0.2 g calculated Amount (mg) of docetaxel (C H NO ) ＝ M × A /A D 43 53 14 S T S on the anhydrous and residual solvent-free basis, methanol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Docetaxel Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Perform the test with 10 mL of the sample solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of them by the area percentage method: the amount of each peak, having the relative retention time of about 0.97, about 1.08, and about 1.13 to docetaxel, is not more than 0.50z, not more than 0.30z, and not more than 0.30z, respectively, the amount of each peak other than docetaxel and the peaks MS: Amount (mg) of Docetaxel RS taken, calculated on the anhydrous and residual solvent-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase A: Water. Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. mentioned above is not more than 0.10z, and the total amount of the peaks other than docetaxel is not more than Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 1.0z. For the area of the peak, having the relative retention time of about 0.97 to docetaxel, multiply the correction factor 1.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 39 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water, acetonitrile for liquid chroma-tography and acetic acid (100) (1000:1000:1) to make 100 mL. To 1 mL of this solution add a mixture of water, aceto-nitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make 10 mL, and use this solution as the so-lution for system suitability test. Pipet 5 mL of the solution for system suitability test, add a mixture of water, aceto-nitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 10 mL. Confirm that the peak area of docetaxel obtained with 10 mL of this solution is equivalent to 35 to 65z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of docetaxel is not more than 2.0z. Water <2.48> 5.0 – 7.0z (50 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Docetaxel Hydrate and Docetaxel RS (separately determine the water <2.48> and the residual solvent in the same manner as Docetaxel Hydrate), dissolve them separately in 2.5 mL of ethanol (99.5), add a mixture of water, acetonitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of docetaxel in each solution. 0 – 9 72 28 9 – 39 72 → 28 28 → 72 Flow rate: 1.2 mL per minute. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of docetaxel is not more than 1.0z.
Domperidone, when dried, contains not less than 99.
Domperidone occurs as a white to pale yellow, crystalline powder or powder. It is freely soluble in acetic acid (100), slightly soluble in methanol and in ethanol (99.5), very slightly soluble in 2-propanol, and practically insoluble in water. Melting point: about 2439C (with decomposition).
(1) Determine the absorption spectrum of a solution of Domperidone in a mixture of 2-propanol and 0.1 mol/L hydrochloric acid TS (9:1) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Domperidone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave ‌882 numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Domperidone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Domperidone in 100 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area of each solution by the automatic integration method: the area of each peak other than domperidone obtained from the sample solution is not larger than 1/2 times the peak area of domperidone from the standard solution. Furthermore, the total area of the peaks other than domperidone is not larger than the peak area of domperidone from the standard solution. ers. Storage—Light-resistant. Donepezil Hydrochloride
Donepezil Hydrochloride contains not less than 98.
Donepezil Hydrochloride occurs as a white crystalline powder. It is soluble in water, and slightly soluble in ethanol (99.5). A solution of Donepezil Hydrochloride (1 in 100) shows no optical rotation. Donepezil Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Donepezil Hydrochloride (1 in 50,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Donepezil Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Donepezil Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Donepezil Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the RS according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals. A solution of Donepezil Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—To 1.0 g of Donepezil Hydrochloride in a porcelain or platinum crucible add 5 mL of sulfuric acid, incinerate by heating gradually, then incin-erate by ignition between 500 and 6009C. If a carbonized residue still retains, moisten the residue with a little amount of sulfuric acid, and incinerate again by ignition between 500 and 6009C. After cooling, dissolve the residue with 3 mL of hydrochloric acid, then evaporate to dryness on a water bath or hot plate, and dissolve the residue with 10 mL of water by warming. Then, proceed as directed in Method 4, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Donepezil Hydrochloride in 25 mL of the mobile phase. To 10 mL of this solution add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sam-ple solution, and add the mobile phase to make exactly 100 ‌JP XVIII Official Monographs / Donepezil Hydrochloride Fine Granules 883 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak other than donepezil obtained from the sample solution is not larger than the peak area of donepezil from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of donepezil, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of donepezil is not more than 2.0z. Water <2.48> Not more than 0.2z (0.2 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Domperidone, pre-viously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 42.59 mg of C22H24ClN5O2
Dopamine Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water and in formic acid, and spar-ingly soluble in ethanol (95). Melting point: about 2489C (with decomposition).
(1) Determine the absorption spectrum of a solution of Dopamine Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 25,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dopamine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Dopamine Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> (1) for chloride. pH <2.54> Dissolve 1.0 g of Dopamine Hydrochloride in 50 mL of water: the pH of this solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Dopamine Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.8 g of Dopa- mine Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.021z). Heavy metals <1.07>—Proceed with 1.0 g of Dopa- mine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dopamine Hydrochloride according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.1 g of Dopamine Hydrochloride in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 250 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, water and acetic acid (100) (16:8:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solu-tion of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 909C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Dopamine Hydro-chloride, previously dried, dissolve in 5 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat on a water bath for 15 minutes. After cooling, add 50 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.96 mg of C8H11NO2.HCl
Doripenem Hydrate contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Doripenem Hydrate occurs as a white to pale yellow-brown-white crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, and practically insoluble in ethanol (99.5). It is gradually colored to pale yellow-brown-white by light. Identiˆcation (1) Determine the absorption spectrum of a Doripenem Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Doripenem RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋33 – ＋389(0.25 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.3 g of Doripenem Hydrate in 30 mL of water: the pH of the solution is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve 0.2 g of Doripenem Hydrate in 20 mL of water, and perform the test with this solution as directed under Turbidity Meas-urement <2.61>: the solution is clear. Perform the test with this solution according to Method 2 under Methods for Color Matching <2.65>: the solution is not more colored than Matching Fluid Y4. Heavy metals <1.07>—Moisten 1.0 g of Doripenem Hydrate with sulfuric acid, cover loosely, and heat gently to carbonize. Then proceed according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances (i)—Dissolve 20 mg of Doripen-em Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak areas of related sub-stance A, having the relative retention time of about 2.2 to doripenem, related substance B, having the relative retention time of about 2.5, and related substance C, having the rela-tive retention time of about 3.2, obtained from the sample solution, are not larger than 1/10 times the peak area of doripenem from the standard solution, and the area of the peak other than doripenem, the peaks mentioned above and the peak having the relative retention time of about 2.1, from the sample solution, is not larger than 1/20 times the peak area of doripenem from the standard solution. Further-more, the total area of the peaks other than doripenem and the peak having the relative retention time of about 2.1 from the sample solution is not larger than 1/2 times the peak area of doripenem from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.6 – 5.7 with a solution prepared by dissolving 2.61 g of dipotas-sium hydrogen phosphate in water to make 1000 mL. To 970 mL of this solution add 30 mL of acetonitrile for liquid chromatography. Mobile phase B: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.6 – 5.7 with a solution prepared by dissolving 2.61 g of dipotas-sium hydrogen phosphate in water to make 1000 mL. To 700 mL of this solution add 300 mL of acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 15 – 45 100 → 50 0 → 50 45 – 50 50 → 0 50 → 100 50 – 55 Flow rate: 1.0 mL per minute. Time span of measurement: For 55 minutes after injec-tion, beginning after the peak having the relative retention time of about 0.2 to doripenem. System suitability— Test for required detectability: Pipet 1.5 mL of the stand-ard solution, and add water to make exactly 50 mL. Conˆrm that the peak area of doripenem obtained with 20 mL of this solution is equivalent to 2.1 to 3.9z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of doripenem are not less than 5000 and not more than 1.3, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 0.95z. Dissolve 20 mg of Doripenem Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the peak area of related substance D, having the relative retention time of about 0.5 to doripenem, obtained from the sample solution is not larger than 2/5 times the peak area of doripenem from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecyl-strong anion exchange-silanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 9 mL of phosphoric acid add 200 mL of water, add 20 mL of triethylamine, and add water to make 2000 mL. Adjust to pH 5.7 – 5.9 with phosphoric acid. To 950 mL of this solution add 50 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of doripenem is about 10 minutes. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add water to make exactly 20 mL. Conˆrm that the peak area of doripenem obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: To 1 mL of the sample solution add 1 mL of 0.1 mol/L hydrochloric acid TS, allow to stand at 25 ± 59C for 15 minutes, and add water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, related substance D and doripenem are eluted in this order with the resolution be-tween these peaks being not less than 5. The number of theo-retical plates and the symmetry factor of the peak of related substance D are not less than 300 and 0.7 to 1.3, respec-tively, and those of the peak of doripenem are not less than 5000 and 0.7 to 1.3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 2.0z. Dissolve 20 mg of Doripenem Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the areas of the peaks, having the relative retention time of about 1.8, about 2.2 and about 2.3 to doripenem, obtained from the sample solution are not larger than 1/20, 7/100 and 1/20 times the peak area of doripenem from the standard solu-tion, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 310 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: To 11 mL of perchloric acid add water to make 500 mL. To 100 mL of this solution add water to make 1000 mL. To 600 mL of this solution add 100 mL of water, and adjust to pH 1.9 – 2.0 with a solution prepared by add-ing water to 2.81 g of sodium perchlorate monohydrate to make 1000 mL. To 900 mL of this solution add 100 mL of acetonitrile. Mobile phase B: To 11 mL of perchloric acid add water to make 500 mL. To 100 mL of this solution add water to make 1000 mL. To 600 mL of this solution add 100 mL of water, and adjust to pH 1.9 – 2.0 with a solution prepared by add-ing water to 2.81 g of sodium perchlorate monohydrate to make 1000 mL. To 300 mL of this solution add 200 mL of acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 25 25 – 55 100 → 0 0 → 100 55 – 60 Flow rate: 0.8 mL per minute. JP XVIII Official Monographs / Doripenem for Injection 889 System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, and add water to make exactly 50 mL. Conˆrm that the peak area of doripenem obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of doripenem are not less than 15,000 and not more than 1.3, respectively. ‌System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 0.95z. Water <2.48> 4.0 – 5.0z (0.3 g, volumetric titration, back titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately amounts of Doripenem Hydrate and Doripenem RS (separately determine the water <2.48> in the same manner as Doripenem Hydrate), equivalent to about 25 mg (potency), dissolve each in water to make ex-actly 200 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of doripenem in each solution. Amount [mg (potency)] of doripenem (C15H24N4O6S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Doripenem RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 300 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust the pH of 90 mL of 0.02 mol/L potassium dihydrogen phosphate TS to pH 5.6 – 5.7 with a solution prepared by dissolving 3.48 g of dipotassium hydro-gen phosphate in water to make 1000 mL. To 100 mL of this solution add water to make exactly 1000 mL. To 970 mL of this solution add 30 mL of acetonitrile. Flow rate: Adjust so that the retention time of doripenem is about 15 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of doripenem are not less than 5000 and not more than 1.3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 1.0z.
Containers—Tight containers. Storage—At a temperature between 29C and 89C.
Dorzolamide Hydrochloride occurs as a white crystalline powder. It is soluble in water, sparingly soluble in methanol, and very slightly soluble in ethanol (99.5). It dissolves in diluted ammonia solution (28) (13 in 400). Time span of measurement: About 3 times as long as the retention time of dorzolamide, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the sample solution, and add a mixture of water and methanol (4:1) to make exactly 100 mL. Pipet 1 mL of this solution, add a mixture of water and methanol (4:1) to make exactly 20 mL, and use this solution as the solution for system suitability test. Confirm that the peak area of dorzolamide obtained with 10 mL of the solution for system suitability test is equivalent to 0.07 to 0.13z of that with 10 mL of the sample solution. System performance: To 1 mL of the sample solution add 2 mL of a mixture of water and methanol (4:1). When the procedure is run with 10 mL of this solution under the above 404.7 Optical rotation [a]25 : －16.0 – －17.59(0.25 g calcu- operating conditions, the number of theoretical plates and lated on the anhydrous basis, water, 25 mL, 100 mm). Dorzolamide Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Dorzolamide Hydrochloride in a solution of hy-drochloric acid in methanol (9 in 1000) (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Dorzolamide Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dor-zolamide Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Dorzolamide Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. A solution of Dorzolamide Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Dorzolamide Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 30 mg of Dorzolamide Hydrochloride in 50 mL of a mixture of water and methanol (4:1), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate their amounts by the area percentage method: the amount of the peaks other than dor-zolamide is not more than 0.1z. Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase A: Adjust to pH 4.5 of a mixture of water and acetic acid (100) (1000:1) with triethylamine. the symmetry factor of the peak of dorzolamide are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of dorzolamide is not more than 7z. Enantiomer—Dissolve 20 mg of Dorzolamide Hydro-chloride in 4 mL of diluted ammonia solution (28) (13 in 400), and extract this solution with two 4-mL portions of ethyl acetate. Combine the extracts, and evaporate the ethyl acetate at 509C under a current of nitrogen. Dissolve the residue in 3 mL of acetonitrile, add 3 drops of (S)-1-phenylethyl isocyanate, and allow to stand at 509C for 10 minutes. Evaporate at 509C under a current of nitrogen, dis-solve the residue in 10 mL of a mixture of tert-butylmethyl ether, acetic acid (100) and acetonitrile (873:100:27), and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas of dorzolamide, A2, and that of the enantiomer, having the relative retention time of about 1.5 to dorzolamide, A1, by the automatic integration method: the result of A1/(A1 ＋ A2) is not more than 0.005. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To a mixture of 30 mL of acetonitrile and 3 mL of water add tert-butylmethyl ether to make 1000 mL. To 650 mL of this solution add 350 mL of heptane. Flow rate: Adjust so that the retention time of dorzola-mide is about 8 minutes. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add a mixture of tert-butylmethyl ether, acetic acid ‌892 (100) and acetonitrile (873:100:27) to make exactly 200 mL, and use this solution as the solution for system suitability test. Confirm that the peak area of dorzolamide obtained with 5 mL of the solution for system suitability test is equiva-lent to 0.4 to 0.6z of that with 5 mL of the sample solution. System performance: When the procedure is run with 5 mL of the sample solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of dorzolamide are not less than 4000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of dorzolamide is not more than 7z. Water <2.48> Not more than 0.5z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Dorzolamide Hydrochloride and Dorzolamide Hydrochloride RS (separately, determine the water <2.48> in the same manner as Dorzolamide Hydrochloride), dissolve in a mixture of water and methanol (4:1) to make exactly 100 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of dorzolamide in each solution. Amount (mg) of dorzolamide hydrochloride (C10H16N2O4S3.HCl) ＝ MS × AT/AS MS: Amount (mg) of Dorzolamide Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8.3 cm in length, packed with octylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust to pH 4.5 of a mixture of water and acetic acid (100) (1000:1) with triethylamine. Flow rate: Adjust so that the retention time of dorzola-mide is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of dorzolamide are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of dorzolamide is not more than 1.0z.
Doxapram Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is freely soluble in methanol and in acetic acid (100), sparingly soluble in water, in ethanol (95) and in acetic anhy-dride, and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Doxapram Hydrochloride Hydrate (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dox-apram Hydrochloride Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Doxapram Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Doxapram Hydrochloride Hy-drate in 50 mL of water: the pH of this solution is between 3.5 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Doxapram Hydrochloride Hydrate in 50 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 1.0 g of Dox- apram Hydrochloride Hydrate. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Dox- apram Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Doxapram Hydrochloride Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.5 g of Doxapram Hy-drochloride Hydrate in 10 mL of methanol, and use this so-lution as the sample solution. Pipet 3 mL of the sample solu-tion, and add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test ‌896 with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 6 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of chlo-roform, formic acid, ethyl formate and methanol (8:3:3:2) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the spots other than the prin-cipal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 3.5 – 4.5z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.8 g of Doxapram Hydro-chloride Hydrate, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.50 mg of C24H30N2O2.HCl
Doxazosin Mesilate, when dried, contains not less than 98.
Doxazosin Mesilate occurs as a white to yel-lowish white crystalline powder. It is freely soluble in dimethylsulfoxide, slightly soluble in water and in methanol, and very slightly soluble in ethanol (99.5). A solution of Doxazosin Mesilate in dimethylsulfoxide solution (1 in 20) shows no optical rotation. Melting point: about 2729C (with decomposition).
(1) Determine the absorption spectrum of a solution of Doxazosin Mesilate in 0.01 mol/L hydrochloric acid-methanol TS (1 in 200,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Doxazosin Mesilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Dox-azosin Mesilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec- trum of Doxazosin Mesilate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. 30 mg of Doxazosin Mesilate responds to Qualitative Tests <1.09> (2) for mesilate.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Doxazosin Mesilate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Doxazosin Mesilate in 5 mL of a mixture of methanol and acetic acid (100) (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of metha-nol and acetic acid (100) (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of methanol and acetic acid (100) (1:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with an upper layer of a mixture, prepared by adding 1 volume of water and 1 volume of acetic acid (100) to 2 volumes of 4-methyl-2-pen-tanon and shaking, to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot at the Rf value about 0.15 obtained from the sample solution is not more intense than the spot from the standard solution, and no spots other than the principal spot and other than the spots mentioned above appear from the sample solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 25 mg each of Doxazosin Mesilate and Doxazosin Mesilate RS, previously dried, dis-solve separately in methanol to make exactly 50 mL. Pipet 3 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of doxazosin in each solution. Amount (mg) of doxazosin mesilate (C23H25N5O5.CH4O3S) ＝ MS × AT/AS MS: Amount (mg) of Doxazosin Mesilate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 246 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0), methanol and acetonitrile (12:8:3). Flow rate: Adjust so that the retention time of doxazosin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of doxazosin are not less than 2000 and ‌JP XVIII Official Monographs / Doxazosin Mesilate Tablets 897 not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doxazosin is not more than 1.0z.
Doxifluridine, when dried, contains not less than 98.
Doxifluridine occurs as a white crystalline powder. It is freely soluble in N,N-dimethylformamide, soluble in water and in methanol, and slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS and in 0.01 mol/L sodium hydroxide TS. Melting point: about 1919C (with decomposition).
(1) Determine the absorption spectrum of a solution of Doxifluridine in 0.1 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Doxifluridine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Optical rotation <2.49> [a]20 : ＋160 – ＋1749(after drying, 0.1 g, water, 10 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Doxifluridine in 10 mL of water is between 4.2 and 5.2.
(1) Fluoride—Dissolve 0.10 g of Doxifluridine in 10.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20). Transfer 5.0 mL of this solution into a 20-mL volumet-ric flask, add 5 mL of a mixture of acetone and lanthanum-alizarin complexone TS (2:1) and water to make 20 mL, allow to stand for 1 hour, and use this solution as the sample solution. Separately, put 1.0 mL of Standard Fluorine Solu-tion in a 20-mL volumetric flask, add 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20) and 5 mL of the mix-ture of acetone and alizarin complexone TS (2:1), then proceed in the same manner as for preparation of the sample solution, and use the solution so obtained as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 620 nm as directed under Ultraviolet-visible Spectrophotometory <2.24>, using a solution obtained in the same way with 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20) as a blank: AT is not larger than AS. Chloride <1.03>—Perform the test with 0.30 g of Doxifluridine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.035z). Heavy metals <1.07>—Proceed with 1.0 g of Doxi-fluridine according to Method 1, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Related substances—Dissolve 20 mg of Doxifluridine in 2 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 25 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetic acid (100) and water (17:2:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the number of the spot other than the principal spot obtained from the sample solu-tion is not more than three, and they are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.25 g of Doxifluridine, pre-viously dried, dissolve in 50 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hy-droxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 24.62 mg of C9H11FN2O5
Doxorubicin Hydrochloride is the hydrochloride of a derivative of daunorubicin. It contains not less than 980 mg (potency) and not more than 1080 mg (potency) per mg, calculated on the anhydrous basis.
Doxorubicin Hydrochloride occurs as a red-orange crystalline powder. It is sparingly soluble in water, slightly soluble in metha-nol, very slightly soluble in ethanol (99.5), and practically in-soluble in acetonitrile.
(1) Determine the absorption spectrum of a solution of Doxorubicin Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Doxorubicin Hydrochloride RS prepared in the same manner as the sam-ple solution: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Doxorubicin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry ‌900 <2.25>, and compare the spectrum with the Reference Spec-trum or the spectrum of Doxorubicin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. to the following conditions, and calculate the ratios, QT and QS, of the peak area of doxorubicin to that of the internal standard. Amount [mg (potency)] of doxorubicin hydrochloride A solution of Doxorubicin Hydrochloride (1 in 200) responds to Qualitative Tests <1.09> (1) for chloride. (C27H 29NO11 .HCl) D Optical rotation <2.49> [a]20: ＋240 – ＋2909(20 mg calcu-lated on the anhydrous basis, methanol, 20 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 50 mg of Doxorubicin Hydrochloride in 10 mL of water is be-tween 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 50 mg of Doxorubicin Hydrochloride in 10 mL of water: the solu-tion is clear and red. (2) Related substances—Dissolve 25 mg of Doxorubicin Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than doxorubicin obtained from the sample solution is not larger than 1/4 times the peak area of doxoru-bicin from the standard solution, and the total area of the peaks other than doxorubicin from the sample solution is not larger than the peak area of doxorubicin from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of doxorubicin. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of doxorubicin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: Dissolve 5 mg of Doxorubicin Hy-drochloride in 20 mL of water, add 1.5 mL of phosphoric acid, and allow to stand at room temperature for 30 minutes. Adjust the pH of this solution to 2.5 with 2 mol/L sodium hydroxide TS. When the procedure is run with 20 mL of this solution under the above operating conditions, doxoru-bicinone, having the relative retention time of about 0.6 to ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Doxorubicin Hydrochlo-ride RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3 g of sodium lauryl sulfate in 1000 mL of diluted phosphoric acid (7 in 5000), and add 1000 mL of acetonitrile. Flow rate: Adjust so that the retention time of doxoru-bicin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, doxorubicin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5, and the symmetry factor of the peak of doxoru-bicin is 0.8 to 1.2. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of doxorubicin to that of the internal standard is not more than 1.0z.
Doxycycline Hydrochloride Hydrate is the hydro-chloride of a derivative of oxytetracycline. It contains not less than 880 mg (potency) and not more than 943 mg (potency) per mg, calculated on the anhydrous and residual ethanol-free basis.
Doxycycline Hydrochloride Hydrate occurs as yellow to dark yellow, crystals or crystalline powder. It is freely soluble in water and in methanol, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Doxycycline Hydrochloride Hydrate in 0.01 mol/L hydrochloric acid-methanol TS (1 in 74,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Doxycycline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption Spectrum of Dox-ycycline Hydrochloride Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Doxycycline Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. 1 cm Dissolve 10 mg of Doxycycline Hydrochloride Hy-drate in 10 mL of water, and add silver nitrate TS: a white turbidity is produced. System performance: When the procedure is run with 10 Absorbance <2.24> E 1z (349 nm): 285 – 315 (10 mg, 0.01 mL of the standard solution under the above operating con-ditions, doxorubicin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5, and the symmetry factor of the peak of doxoru- mol/L hydrochloric acid-methanol TS, 500 mL). D Optical rotation <2.49> [a]20: －105 – －1209(0.25 g calcu-lated on the anhydrous and residual ethanol-free basis, 0.01 mol/L hydrochloric acid-methanol TS, 25 mL, 100 mm). Determine within 5 minutes after the sample solution is pre-pared.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Doxycycline Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 5.0 mL of Standard Lead Solution (not more than 50 ppm). (2) Related substance—Dissolve 20 mg of Doxycycline Hydrochloride Hydrate in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the sample solution. Separately, dissolve 20 mg of 6-epidoxycycline hy-drochloride in 0.01 mol/L hydrochloric acid TS to make ex-actly 25 mL, and use this solution as the 6-epidoxycycline hy-drochloride stock solution. Separately, dissolve 20 mg of metacycline hydrochloride in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the metacycline hydrochloride stock solution. Pipet 2 mL each of the 6-epidoxycycline hydrochloride stock solution and the metacycline hydrochloride stock solution, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak areas of metacycline and 6-epidoxycycline obtained from the sample solution are not larger than the peak areas of them from the standard solution, respectively, and the areas of the two peaks, appeared between the solvent peak and metacy- 2 mL of the metacycline hydrochloride stock solution add 0.01 mol/L hydrochloric acid TS to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, metacycline, 6-epidoxycycline and doxycycline are eluted in this order with the resolutions be-tween the peaks, metacycline and 6-epidoxycycline, and 6-epidoxycycline and doxycycline, being not less than 1.3 and not less than 2.0, respectively, and the symmetry factor of the peak of doxycycline is not more than 1.3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak areas of metacycline and 6-epidoxycycline are not more than 3.0z and not more than 2.0z, respectively. Ethanol Weigh accurately about 0.1 g of Doxycycline Hy-drochloride Hydrate, dissolve in the internal standard solu-tion to make exactly 10 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 0.4 g of ethanol (99.5), and add the internal standard solution to make exactly 100 mL. Pipet 1 mL of this solution, add the internal standard solution to make exactly 10 mL, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethanol to that of the internal standard: the amount of ethanol is not less than 4.3z and not more than 6.0z. cline and behind of doxycycline, from the sample solution are not larger than 1/4 times the peak area of 6-epidoxycy- Amount (z) of ethanol ＝ MS/MT × QT/QS cline from the standard solution, and the total area of the peaks other than doxycycline from the sample solution is not larger than 1.5 times the peak area of 6-epidoxycycline from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (8 mm in particle di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase: To 125 mL of 0.2 mol/L potassium dihy-drogen phosphate TS and 117 mL of 0.2 mol/L sodium hy-droxide TS add water to make 500 mL. To 400 mL of this solution add 50 mL of a solution of tetrabutylammonium hydrogensulfate (1 in 100), 10 mL of a solution of disodium dihydrogen ethylenediamine tetraacetate dihydrate (1 in 25), 60 g of t-butyl alcohol and 200 mL of water, adjust to pH 8.0 with 2 mol/L sodium hydroxide TS, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of doxycycline is about 19 minutes. Time span of measurement: About 2.4 times as long as the retention time of doxycycline, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Confirm that the peak areas of 6-epidoxycy-cline and metacycline obtained with 20 mL of this solution are equivalent to 3.5 to 6.5z of them with 20 mL of the standard solution, respectively. System performance: To 8 mL of the sample solution, 3 mL of the 6-epidoxycycline hydrochloride stock solution and MS: Amount (mg) of ethanol (99.5) taken MT: Amount (mg) of Doxycycline Hydrochloride Hydrate taken Internal standard solution—A solution of 1-propanol (1 in 2000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3.2 mm in inside diameter and 1.5 m in length, packed with porous ethylvinylbenzene-divinylbenzene copolymer for gas chromatography (0.0075 mm in average pore size, 500 – 600 m2/g in specific surface area) (150 – 180 mm in particle diameter). Column temperature: A constant temperature of about 1359C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethanol is about 5 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, ethanol and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 5 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of ethanol to that of the internal standard is not more than 2.0z. Water <2.48> Not less than 1.4z and not more than 2.8z (0.6 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately an amount of Doxycycline Hydro-chloride Hydrate and Doxycycline Hydrochloride RS, equivalent to about 50 mg (potency), dissolve each in water ‌JP XVIII Official Monographs / Doxycycline Hydrochloride Tablets 903 to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of doxycycline in each solution. ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having a relative retention time of about 0.6 to doxycycline, obtained from the sample solution is not larger than 1.5 times the peak area of doxycycline from the standard solution. Operating conditions— Amount [ mg (potency)] of doxycycline (C ＝ MS × AT/AS × 1000 22H24N2O8) Proceed as directed in the operating conditions in the Assay. System suitability— MS: Amount [mg (potency)] of Doxycycline Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 7.0 g of sodium dihydrogen phos-phate dihydrate in 450 mL of water, add 553 mL of a mix-ture of methanol and N,N-dimethyl-n-octylamine (550:3), and adjust the pH to 8.0 with a solution of sodium hydrox-ide (43 in 200). Flow rate: Adjust so that the retention time of doxycycline is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the theoretical plates and the symmetry factor of the peak of doxycycline are not less than 1000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doxycycline is not more than 1.0z.
Dried Yeast is dried and powdered cells of yeast be-longing to Saccharomyces. It contains not less than 400 mg of protein and not less than 100 mg of thiamine compounds [as thiamine chloride hydrochloride (C12H17ClN4OS.
Dried Yeast occurs as a light yellow-white to brown powder. It has a characteristic odor and taste.
Dried Yeast, when examined under a micro-scope <5.01>, shows isolated cells, spheroidal or oval in shape, and 6 to 12 mm in length.
(1) Rancidity—Dried Yeast is free from any un-pleasant or rancid odor or taste. (2) Starch—Add iodine TS to Dried Yeast, and examine microscopically <5.01>: no or only a few granules are tinted black-purple. Loss on drying <2.41> Not more than 8.0z (1 g, 1009C, 8 hours). Total ash <5.01> Not more than 9.0z (1 g).
(1) Protein—Weigh accurately about 50 mg of Dried Yeast and perform the test as directed under Nitrogen Determination <1.08>. Amount (mg) of protein in 1 g of Dried Yeast ＝ N × 6.25 × 1/M N: Amount (mg) of nitrogen (N) M: Amount (g) of Dried Yeast taken (2) Thiamine—Weigh accurately about 1 g of Dried Yeast, add 1 mL of dilute hydrochloric acid and 80 mL of water, and heat in a water bath at 80 to 859C for 30 minutes with occasional shaking. After cooling, add water to make exactly 100 mL, and centrifuge for 10 minutes. Pipet 4 mL of the supernatant liquid, add exactly 5 mL of acetic acid-sodium acetate TS and exactly 1 mL of enzyme TS, and allow to stand at 45 to 509C for 3 hours. Place exactly 2 mL of this solution onto a chromatographic column prepared by pouring 2.5 mL of a weakly acidic CM-bridged cellulose cation exchanger (H type) (40 to 110 mm in particle diameter) into a chromatographic tube about 1 cm in inside diameter and about 17 cm in length, and elute at the flow rate of about 0.5 mL per minute. Wash the upper part of the column with a small amount of water, and wash the column with two 10-mL portions of water at the flow rate of about 1 mL per minute. Elute the column with two 2.5-mL portions of diluted phosphoric acid (1 in 50) at the flow rate of about 0.5 mL per minute, and combine the eluate. To the eluate add exactly 1 mL of the internal standard solution and 0.01 g of sodium 1-octanesulfonate, and after dissolving, use this solution as the sample solution. Separately, weigh accurately about 15 mg of Thiamine Chloride Hydrochloride RS (previ-ously determine the water <2.48> in the same manner as Thiamine Chloride Hydrochloride), dissolve in 0.001 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 1 mL of this solution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add exactly 1 mL of the in-ternal standard solution and 3 mL of the mobile phase, and use this solution as the standard solution. Perform the test with 200 mL each of the sample solution and standard solu- ‌tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of thiamine to that of the inter-nal standard.‌ Amount ( mg) of thiamine in 1 g of Dried Yeast ＝ MS/MT × QT/QS × 12.5 MS: Amount (mg) of Thiamine Chloride Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (g) of the Dried Yeast taken Internal standard solution—Dissolve 0.01 g of phenacetin in acetonitrile to make 100 mL, and to 1 mL of this solution add diluted acetonitrile (1 in 5) to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside diameter and 15 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.7 g of potassium dihydrogen-phosphate in 1000 mL of water, and adjust the pH to 3.5 with diluted phosphoric acid (1 in 10). Dissolve 1.6 g of sodi-um 1-octanesulfonate in 800 mL of this solution, and add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of thiamine is about 8 minutes. Selection of column: Proceed with 200 mL of the standard solution under the above operating conditions, and calculate the resolution. Use a column giving elution of thiamine and the internal standard in this order with the resolution be-tween these peaks being not less than 8.
Droperidol, when dried, contains not less than 98.
Droperidol occurs as a white to light yellow powder. It is freely soluble in acetic acid (100), soluble in dichloro-methane, slightly soluble in ethanol (99.5), and practically insoluble in water. MS: Amount [mg (potency)] of Doxycycline Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 7.0 g of sodium dihydrogen phos-phate dihydrate in 450 mL of water. Add to this solution 553 mL of a mixture of methanol and N,N-dimethyl-n-octyla-mine (550:3), and adjust to pH 8.0 with sodium hydroxide solution (43 in 200). Flow rate: Adjust so that the retention time of doxycycline is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of doxycycline are not less than 2200 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doxycycline is not more than 1.0z.
(1) Put 30 mg of Droperidol in a brown volumetric flask, and dissolve in 10 mL of 0.1 mol/L hydro-chloric acid TS and ethanol (95) to make 100 mL. Transfer 5 mL of the solution to a brown volumetric flask, and add 10 mL of 0.1 mol/L hydrochloric acid TS and ethanol (95) to make 100 mL. Determine the absorption spectrum of the so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Droperidol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Droperidol in acetone, evaporate the acetone, dry the residue in a desiccator (in vacuum, silica gel, 709C) for 4 hours, and perform the test with the residue.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Droperidol in a platinum crucible according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 50 mg of Droperidol in 5 mL of dichloromethane, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dichloromethane to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, chloroform, methanol and acetic acid-sodium acetate buffer solution (pH 4.7) (54:23:18:5) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 ‌JP XVIII Official Monographs / Droxidopa 905 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 3.0z (0.5 g, in vacu-um, silica gel, 709C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g, plati-num crucible).
Weigh accurately about 0.5 g of Droperidol, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.94 mg of C22H22FN3O2
Droxidopa, when dried, contains not less than 99.
Droxidopa occurs as white to light brown, crystals or crystalline powder. It is slightly soluble in water and practically insoluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS.
(1) Determine the absorption spectrum of a solution of Droxidopa in 0.1 mol/L hydrochloric acid TS (1 in 25,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Drox-idopa as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －38 – －439(after drying, 0.1 g, 0.1 mol/L hydrochloric acid TS, 20 mL, 100 mm).
(1) Chloride <1.03>—Dissolve 0.40 g of Droxidopa in 6 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 2.0 g of Droxido- pa according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Droxidopa according to Method 3, and perform the test (not more than 2 ppm). Related substances—To 0.10 g of Droxidopa add 50 mL of 0.1 mol/L hydrochloric acid TS, dissolve by shaking while cooling in an ice bath, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard so-lution. Perform the test with exactly 10 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of both solutions by the au-tomatic integration method: the area of each peak other than droxidopa obtained from the sample solution is not larger than the peak area of droxidopa from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.0 g of sodium 1-heptanesul-fonate and 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 2.0 with phosphoric acid. To 930 mL of this solution add 70 mL of acetonitrile. Flow rate: Adjust so that the retention time of droxidopa is about 5 minutes. Time span of measurement: About 12 times as long as the retention time of droxidopa, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of droxidopa are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of droxidopa is not more than 2.0z. Loss on drying <2.41> Not more than 0.1z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Droxidopa, previ-ously dried, dissolve in exactly 20 mL of 0.1 mol/L perchlo-ric acid VS, add 50 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.32 mg of C9H11NO5
Dydrogesterone, when dried, contains not less than 98.
Dydrogesterone occurs as white to light yel-low-white, crystals or crystalline powder. It is odorless. It is freely soluble in chloroform, soluble in acetonitrile, sparingly soluble in methanol and in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water.
(1) To 5 mg of Dydrogesterone add 5 mL of 4-methoxybenzaldehyde-acetic acid TS and 2 to 3 drops of sulfuric acid, and heat in a water bath for 2 minutes: an orange-red color develops. Determine the absorption spectrum of a solution of Dydrogesterone in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Dydrogesterone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: －470 – －5009(after drying, 0.1 g, chloroform, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Dydrogesterone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 10 mg of Dydroges-terone in 200 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of these solutions by the automatic integration method: the total area of peaks other than dydrogesterone obtained from the sample solution is not larger than the peak area of dydrogesterone from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (3 mm in parti- ‌cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, ethanol (95) and aceto-nitrile (53:26:21). mL of the filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 5 mg of dydrogesterone (C21H28O2), and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. Flow rate: Adjust so that the retention time of dydroges-terone is about 12 minutes. Selection of column: Dissolve 1 mg each of Dydroges- Amount (mg) of dydrogesterone (C21 ＝ MS × AT/AS × V?/V × 1/20 H28O2) terone and progesterone in 20 mL of the mobile phase. Pro-ceed with 10 mL of the solution under the above operating conditions, and calculate the resolution. Use a column giving elution of dydrogesterone and progesterone in this order with the resolution between these peaks being not less than 8. Wavelength is 265 nm. Detection sensitivity: Adjust the detection sensitivity so that the peak height of dydrogesterone obtained with 10 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About twice as long as the retention time of dydrogesterone, beginning after the solvent peak. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg of Dydrogesterone, previously dried, and dissolve in methanol to make exactly 100 mL. Pipet 1 mL of this solution, and add methanol to make exactly 100 mL. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 286 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Amount (mg) of dydrogesterone (C21H28O2) ＝ A/845 × 100,000
Ebastine, when dried, contains not less than 99.
Ebastine occurs as white, crystals or crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in ethanol (95), and practically insoluble in water. It gradually becomes yellowish white on exposure to light.
(1) Dissolve 20 mg of Ebastine in 5 mL of ethanol (95), add 2 mL of 1,3-dinitrobenzene TS and 2 mL of sodium hydroxide TS, and allow to stand: the color of the solution is purple to red-purple, which gradually changes to blown. Determine the absorption spectrum of a solution of Ebastine in methanol (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ebastine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ebastine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). A platinum crucible may be used. (2) Related substances—Dissolve 0.10 g of Ebastine in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 2 mL of this so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust to pH 3.0 with diluted phosphoric acid (1 in 5), and add water to make 1000 mL. To 375 mL of this solution add 625 mL of acetonitrile for liquid chromatography, and dissolve 0.72 g of sodium lauryl sulfate in this solution. Flow rate: Adjust so that the retention time of ebastine is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of ebastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of ebastine obtained with 10 mL of this solution is equivalent to 35 to 65z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ebastine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ebastine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Ebastine, previ-ously dried, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 46.97 mg of C32H39NO2
Ecothiopate Iodide contains not less than 95.
Ecothiopate Iodide occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Dissolve 0.1 g of Ecothiopate Iodide in 2 mL of water, and add 1 mL of nitric acid: a brown precipi- tate is formed. To 1 drop of the turbid solution containing this precipitate add 1 mL of hexane, and shake: a light red color develops in the hexane layer. Heat the suspension of the precipitate obtained in (1) until it becomes colorless, cool, add 10 mL of water, and use this solution as the sample solution. Two mL of the sample solution responds to Qualitative Tests <1.09> (2) for phos-phate. Two mL of the sample solution obtained in (2) re-sponds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 0.1 g of Ecothiopate Iodide in 40 mL of water: the pH of this solution is between 3.0 and 5.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Ecothiopate Iodide in 5 mL of water: the solution is clear and colorless. Heavy metals <1.07>—To 1.0 g of Ecothiopate Iodide in a Kjeldahl flask add 5 mL of nitric acid and 2 mL of sul-furic acid, put a small funnel on the mouth of the flask, and heat carefully until white fumes are evolved. After cooling, add 2 mL of nitric acid, and heat. Repeat this procedure twice, add several 2-mL portions of hydrogen peroxide (30), and heat until the solution becomes colorless, and white fumes are evolved. After cooling, transfer the solution together with a small quantity of water to a Nessler tube, and add water to make about 20 mL. Adjust the solution with ammonia solution (28) and ammonia TS to a pH be-tween 3.0 and 3.5, add water to make 50 mL, and use this solution as the test solution. Prepare the control solution as follows: proceed in the same manner as the preparation of the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). Related substances—Dissolve 0.20 g of Ecothiopate Iodide in 10 mL of methanol, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 509C, 3 hours).
Weigh accurately about 0.125 g of Ecothiopate Iodide, and dissolve in water to make exactly 100 mL. Pipet 10 mL of of this solution, add 30 mL of water, then add exactly 10 mL of phosphate buffer solution (pH 12), stopper the container, and allow to stand at 25 ± 39C for 20 minutes. To this solution add quickly 2 mL of acetic acid (100), and titrate <2.50> with 0.002 mol/L iodine VS (poten-tiometric titration). Perform the test in the same manner without phosphate buffer solution (pH 12), and make any necessary correction. Each mL of 0.002 mol/L iodine VS ＝ 1.533 mg of C9H23INO3PS
Containers—Tight containers. Storage—Light-resistant, and not exceeding 09C.
Edaravone, when dried, contains not less than 99.
Edaravone occurs as white to pale yellow-white, crystals or crystalline powder. It is freely soluble in ethanol (99.5) and in acetic acid (100), and slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Edaravone (1 in 200,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Edaravone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> The pH of a solution obtained by dissolving 20 mg of Edaravone in 20 mL of water is between 4.0 and 5.5.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Edaravone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Edaravone in 25 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than edaravone obtained from the sample solu-tion is not larger than the peak area of edaravone from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, methanol and acetic acid (100) (100:100:1). Flow rate: Adjust so that the retention time of edaravone is about 4 minutes. Time span of measurement: About 7 times as long as the retention time of edaravone, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of edaravone are not less than 1500 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of edaravone is not more than 2.0z. Loss on drying <2.41> Not more than 0.1z (1 g, in vacu-um, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Edaravone, previ-ously dried, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 17.42 mg of C10H10N2O
Edrophonium Chloride occurs as white, crys-tals or crystalline powder. It is odorless. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), and practically insoluble in acetic anhydride and in diethyl ether. It is hygroscopic. It is gradually colored by light.
(1) To 5 mL of a solution of Edrophonium Chloride (1 in 100) add 1 drop of iron (III) chloride TS: a light red-purple color develops. Determine the absorption spectrum of a solution of Edrophonium Chloride in 0.1 mol/L hydrochloric acid TS (1 in 20,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of a solution of Edropho-nium Chloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Edrophonium Chloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride. ‌918 pH <2.54> Dissolve 1.0 g of Edrophonium Chloride in 10 mL of water: the pH of this solution is between 3.5 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Edrophonium Chloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Edrophonium Chloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Edrophonium Chloride according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Edrophonium Chloride in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add ethanol (95) to make exactly 100 mL. Pipet 3 mL of this solution, add ethanol (95) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with Edrophonium Chloride. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 272 nm and 276 nm. pH <2.54> 6.5 – 8.0 Bacterial endotoxins <4.01> Less than 15 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Conduct this procedure without exposure to light, using light-resistant vessels. Measure exactly a volume of Edrophonium Chloride Injection, equivalent to about 50 mg these solutions as directed under Thin-layer Chromatogra- phy <2.03>. Spot 10 mL each of the sample solution and of edrophonium chloride (C 10H 16ClNO), place in a chro- standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of methanol, chloroform and ammonia solu-tion (28) (16:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, in vacu-um, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Edrophonium Chloride, previously dried, and dissolve in 100 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3). Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS matographic column prepared by pouring 10 mL of weakly basic DEAE-bridged dextran anion exchanger (Cl type) (50 to 150 mm in particle diameter) into a chromatographic tube about 2 cm in inside diameter and about 10 cm in length, add 25 mL of water, and elute at the flow rate of 1 to 2 mL per minute. Wash the column with two 25-mL portions of water at the flow rate of 1 to 2 mL per minute. Combine the wash-ings with above effluent solutions, and add water to make exactly 100 mL. Measure exactly 10 mL of this solution, and add 10 mL of phosphate buffer solution (pH 8.0) and 5 g of sodium chloride. Wash this solution with four 20-mL por-tions of a mixture of diethyl ether and hexane (1:1), collect the water layer, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Edrophonium Chloride RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 3 hours, and dissolve in water to make exactly 100 mL. Measure exactly 10 mL of this solu-tion, and prepare the standard solution in the same manner as the sample solution. Determine the absorbances, AT and ＝ 20.17 mg of C10H ClNO AS, of the sample solution and standard solution at 273 nm as directed under Ultraviolet-visible Spectrophotometry
Elcatonin contains not less than 5000 Elcatonin Units and not more than 7000 Elcatonin Units per mg of peptide, calculated on the anhydrous and residual acetic acid-free basis.
Elcatonin is a white powder. It is very soluble in water, freely soluble in ethanol (95), and practically insoluble in acetonitrile. Chemical reaction vessel temperature: A constant temper-ature of about 1309C. Color developing time: About 1 minute. Mobile phase: Buffer solutions A, B, C and D, with so-dium ion concentrations of 0.10 mol/L, 0.135 mol/L, 1.26 mol/L and 0.20 mol/L, respectively. The ion concentration of the mobile phase is changed stepwise from 0.10 mol/L to 1.26 mol/L by using these buffer solutions. Components of buffer solutions Buffer solution: A B C D Citric acid monohydrate 8.85 g 7.72 g 6.10 g — Trisodium citrate dihydrate 3.87 g 10.05 g 26.67 g — Sodium hydroxide — — 2.50 g 8.00 g Sodium chloride 3.54 g 1.87 g 54.35 g — Ethanol (95) 60.0 mL — — 60.0 mL Thiodiglycol 5.0 mL 5.0 mL — — It is hygroscopic. The pH of a solution of Elcatonin (1 in 500) is between 4.5 Purified water a sufficient amount a sufficient amount a sufficient amount a sufficient amount and 7.0.
Dissolve 5 mg of Elcatonin in 5 mL of water. Determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Constituent amino acids Put about 1 mg of Elcatonin in a test tube for hydrolysis, add phenol-hydrochloric acid TS to dissolve, replace the air inside with Nitrogen, seal the tube under reduced pressure, and heat at 110 ± 29C for 24 hours. After cooling, open the tube, evaporate the hydrolyzate to dryness under reduced pressure, dissolve the residue in about 1 mL of 0.02 mol/L hydrochloric acid TS, and use this solu-tion as the sample solution. Separately, weigh exactly 1.33 mg of L-aspartic acid, 1.19 mg of L-threonine, 1.05 mg of L-serine, 1.47 mg of L-glutamic acid, 1.15 mg of L-proline, 0.75 mg of glycine, 0.89 mg of L-alanine, 1.17 mg of L-va-line, 1.89 mg of L-2-aminosuberic acid, 1.31 mg of L-leucine, 1.81 mg of L-tyrosine, 1.83 mg of L-lysine hydrochloride, 2.10 mg of L-histidine hydrochloride monohydrate and 2.11 mg of L-arginine hydrochloride, dissolve them in 0.02 mol/L hydrochloric acid TS to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: 14 peaks of amino acids appear on the chromatogram obtained from the sample solution, and their respective molar ratios against alanine are 1.7 – 2.2 for aspartic acid, 3.5 – 4.2 for threonine, 2.4 – 3.0 for serine, 2.7 – 3.2 for glutamic acid, 1.7 – 2.2 for proline, 2.7 – 3.2 for glycine, 1.6 – 2.2 for valine, 0.8 – 1.2 for 2-aminosuberic acid, 4.5– 5.2 for leucine, 0.7– 1.2 for tyrosine, 1.7– 2.2 for lysine, 0.8 – 1.2 for histidine and 0.7 – 1.2 for arginine. Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 nm and 570 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with a sulfonated styrene-divinylbenzene copolymer (3 mm in particle diameter). Column temperature: Varied between 509C and 659C. Total amount 1000 mL 1000 mL 1000 mL 1000 mL Reaction reagent: Mix 407 g of lithium acetate dihydrate, 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for about 20 minutes while passing Nitrogen, and use this solution as so-lution A. Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for about 20 minutes while passing Nitrogen, and use this solution as solution B. Mix solution A and solution B before use. Flow rate of mobile phase: Adjust so that the retention time of arginine is about 75 minutes. Flow rate of reaction reagent: About 0.2 mL per minute. Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions. Use a column from which aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, 2-aminosuberic acid, leu-cine, tyrosine, lysine, histidine and arginine are eluted in this order, with complete separation of each peak.
(1) Acetic acid—Weigh accurately 3 – 6 mg of Elcatonin quickly under conditions of 25 ± 29C and 50 ± 5z relative humidity, add exactly 1 mL of the internal standard solution to dissolve it, and use this solution as the sample solution. Separately, weigh accurately about 0.5 g of acetic acid (100), and add the internal standard solution to make exactly 100 mL. Pipet 5 mL of this solution, add the internal standard solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of acetic acid to that of the internal standard: the amount of acetic acid is not more than 7.0z. Amount (z) of acetic acid (CH3COOH) ＝ MST/MSA × QT/QS × 50 MST: Amount (g) of acetic acid (100) taken MSA: Amount (mg) of Elcatonin taken Internal standard solution—A solution of citric acid mono-hydrate (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 13.2 g of diammonium hydrogen phosphate in 900 mL of water, add phosphoric acid to adjust the pH to 2.5, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of acetic acid is about 4 minutes. Selection of column: Proceed with 20 mL of the standard solution under the above operating conditions. Use a column from which acetic acid and citric acid are eluted in this order with the resolution between their peaks being not less than 2.0. Related substances—Dissolve 1.0 mg of Elcatonin in 1 mL of a mixture of trifluoroacetic acid TS and acetonitrile (2:1), and use this solution as the sample solution. Take exactly 0.3 mL of the sample solution, add a mixture of trifluoroacetic acid TS and acetonitrile (2:1) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: each peak area other than elcatonin obtained from the sam-ple solution is not larger than 1/3 times the peak area of elcatonin from the standard solution, and the total of the peak areas other than elcatonin is not larger than the peak area of elcatonin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of trifluoroacetic acid TS and acetonitrile (change the ratio linearly from 85:15 to 55:45 in 30 minutes). Flow rate: Adjust so that the retention time of elcatonin is about 25 minutes. Selection of column: Dissolve 2 mg of Elcatonin in 200 mL of trypsin TS for test of elcatonin, warm at 379C for 1 hour, then add 1 drop of acetic acid (100), and heat at 959C for 1 minute. To 10 mL of this solution add 50 mL of the sample solution, and mix. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column such that the resolution between the peak of elcatonin and the peak which appears immediately before the peak of elcatonin is not less than 2.0, and the retention time of elcatonin is about 25 minutes. Detection sensitivity: Adjust the detection sensitivity so that the peak height of elcatonin from 10 mL of the standard solution is between 50 mm and 200 mm. Time span of measurement: Continue measurement until the regularly changing base-line of the chromatogram disap-pears, beginning after the solvent peak. Water <2.48> Weigh accurately 1 – 3 mg of Elcatonin quickly under conditions of 25 ± 29C and 50 ± 5z relative humidity, and perform the test as directed in Coulometric titration: not more than 8.0z. Nitrogen content Weigh accurately 0.015 – 0.02 g of Elcatonin quickly under conditions of 25 ± 29C and 50 ± 5z relative humidity, and perform the test as directed under Nitrogen Determination <1.08>: it contains not less than 16.1z and not more than 18.7z of nitrogen (N: 14.01) in the peptide, calculated on the anhydrous and residual acetic acid-free basis.
(i) Animals: Select healthy male Sprague-Dawley rats each weighing between 90 g and 110 g. Keep the rats for not less than 3 days before use, providing an appropriate uniform diet and water. Diluent for elcatonin: Dissolve 2.72 g of sodium ace-tate trihydrate in water to make 200 mL, add 0.2 g of bovine serum albumin, and adjust the pH to 6.0 with acetic acid (100). Prepare before use. Standard solution: Dissolve Elcatonin RS in the dil-uent for elcatonin to make two standard solutions, one to contain exactly 0.075 Unit in each mL which is designated as the high-dose standard solution, SH, and the other to contain exactly 0.0375 Unit in each mL which is designated as the low-dose standard solution, SL. Sample solution: Weigh accurately 0.5 – 2.0 mg of Elcatonin quickly under conditions of 25 ± 29C and 50 ± 5z relative humidity, and dissolve in the diluent for elcatonin to make two sample solutions, the high-dose sam-ple solution, TH, which contains the Units per mL equivalent to SH and the low-dose sample solution, TL, which contains the Units per mL equivalent to SL. Deproteinizing solution for elcatonin: Dissolve 160 g of trichloroacetic acid and 30.6 g of strontium chloride in water to make 3600 mL. Procedure: Divide the animals into 4 equal groups of not less than 10 animals each. Withhold all food, but not water, for 18 to 24 hours before the injections, and withhold water during the assay until the final blood sample is taken. Handle the animals with care in order to avoid undue excite-ment. Inject exactly 0.2 mL each of the standard solutions and the sample solutions into the tail vein of each animal as indi-cated in the following design: First group SH Third group TH Second group SL Fourth group TL At 1 hour after the injection, take a sufficient blood sam-ple to perform the test from the carotid artery and vein of each animal under ether anesthesia, centrifuge the blood samples to separate serum, and determine the serum calcium according to the following (vii). Serum calcium determination: Take exactly 0.3 mL of the serum, add the deproteinizing solution for elcatonin to make exactly 3 mL, mix well, centrifuge, and use the super-natant liquid as the sample solution for calcium determina-tion. Separately, pipet 1 mL of Standard Calcium Solution for Atomic Absorption Spectrophotometry, and add a solu-tion of sodium chloride (17 in 2000) to make exactly 10 mL. Pipet 5 mL of this solution, add the deproteinizing solution for elcatonin to make exactly 50 mL, and use this solution as the standard solution for calcium determination. Determine the absorbances, AT and AS, of the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions. Determine the absorbance, A0, of a solution obtained in the same manner used for preparation of the standard so-lution, but with 1 mL of water instead of the standard solu-tion. ‌JP XVIII Official Monographs / Emedastine Fumarate 921 Amount (mg) of calcium (Ca) in 100 mL of the serum ＝ 0.01 × (AT － A0)/(AS － A0) × 10 × 100 Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Calcium hollow-cathode lamp. Wavelength: 422.7 nm. Calculation: Amounts of calcium in 100 mL of the serum obtained with SH, SL, TH and TL in (vii) are symbo-lized as y1, y2, y3 and y4, respectively. Sum up individual y1, y2, y3 and y4 to obtain Y1, Y2, Y3 and Y4, respectively. Units per mg of peptide, calculated on the anhydrous and residual acetic acid-free basis ＝ antilog M × units per mL of SH × b/a M ＝ 0.3010 × Ya/Yb Ya ＝ －Y1 － Y2 ＋ Y3 ＋ Y4 Yb ＝ Y1 － Y2 ＋ Y3 － Y4 a: Amount (mg) of Elcatonin taken × [100 － {water content (z) ＋ acetic acid content (z)}/100] b: Total volume (mL) of the high-dose sample solution prepared by dissolving Elcatonin with diluent for elcatonin F? computed by the following equation should be smaller than F shown in the table against n with which s2 is calcu-lated. Calculate L (P ＝ 0.95) by use of the following equa-tion: L should be not more than 0.20. If F? exceeds F, or if L exceeds 0.20, repeat the test, increasing the number of animals or arranging the assay conditions so that F? is not more than F and L is not more than 0.20. Emedastine Fumarate
Emedastine Fumarate, when dried, contains not less than 98.
Emedastine Fumarate occurs as a white to pale yellow crystalline powder. It is freely soluble in water, soluble in methanol, sparingly soluble in ethanol (99.5), and slightly soluble in acetic acid (100). It shows crystal polymorphism.
(1) Dissolve 10 mg of Emedastine Fuma-rate in 10 mL of water. To 2 mL of this solution add 1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit F? ＝ (－ Y1 ＋ Y2 ＋ Y3 － Y4)2/4 fs2 similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of f: Number of the animals of each group s2 ＝ {Sy2 － (Y/f )}/n Sy2: The sum of squares of y1, y2, y3 and y4 in each group Y ＝ Y12 ＋ Y22 ＋ Y32 ＋ Y42 n ＝ 4 ( f － 1) (C － 1)(CM2 ＋ 0.09062) L ＝ 2 C ＝ Yb2/(Yb2 － 4 fs2t2) t2: Value shown in the following table against n used to calculate s 2 n t2 ＝ F n t2 ＝ F n t2 ＝ F Emedastine Fumarate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Dissolve 30 mg of Emedastine Fumarate in 5 mL of methanol, and use this solution as the sample solution. Sepa-rately, dissolve 10 mg of fumaric acid for thin-layer chroma-tography in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of isopropyl ether, formic acid and water (90:7:3) to a distance 161.45 4.667 4.242 of about 10 cm, and air-dry the plate. Examine under ultra- 18.51 4.600 4.225 violet light (main wavelength: 254 nm): the spots other than 10.129 4.543 4.210 the spot on the starting point obtained from the sample solu- 7.709 4.494 4.196 tion and the spot from the standard solution show the same 6.608 4.451 4.183 Rf value. 6 5.987 18 4.414 30 4.171 7 5.591 19 4.381 40 4.085 8 5.318 20 4.351 60 4.001 9 5.117 21 4.325 120 3.920 10 4.965 22 4.301 ∞ 3.841 11 4.844 23 4.279 12 4.747 24 4.260
(1) Heavy metals <1.07>—Proceed with 2.0 g of Emedastine Fumarate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 10 mg of Emedastine Fumarate in 10 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu- ‌tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the area of the peak other than emedastine and fumaric acid obtained from the sample solution is not larger than the peak area of emedastine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.9 g of sodium dihydrogen phos-phate dihydrate and 2.5 g of sodium lauryl sulfate in 1000 mL of water, and adjust to pH 2.4 with phosphoric acid. To 550 mL of this solution add 450 mL of acetonitrile. Flow rate: Adjust so that the retention time of emedastine is about 18 minutes. Time span of measurement: About 2 times as long as the retention time of emedastine, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of emedastine are not less than 10,000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of emedastine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Emedastine Fumarate, previously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.73 mg of C17H26N4O.2C4H4O4
Containers—Tight containers. Storage—Not exceeding 89C.
Emorfazone, when dried, contains not less than 98.
Emorfazone occurs as colorless crystals or a white to light yellow crystalline powder. It is very soluble in ethanol (99.5), and freely soluble in water and in acetic anhydride. It dissolves in 1 mol/L hydrochloric acid TS. It gradually turns yellow and decomposes on exposure to light.
(1) Dissolve 20 mg of Emorfazone in 2 mL of 1 mol/L hydrochloric acid TS, and add 5 drops of Reinecke's TS: light red floating matters are formed. Determine the absorption spectrum of a solution of Emorfazone (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Emorfazone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Perform the test with 1.0 g of Emorfazone. Prepare the control solution with 0.50 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Emorfa-zone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).
Weigh accurately about 0.2 g of Emorfazone, previ-ously dried, dissolve in 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 23.93 mg of C H N O 11 17 3 3 Purity (1) Chloride <1.03>—Perform the test with 1.0 g of Emorfazone. Prepare the control solution with 0.50 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Emorfa-zone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).
Enalapril Maleate, when dried, contains not less than 98.
Enalapril Maleate occurs as white, crystals or crystalline powder. It is freely soluble in methanol, sparingly soluble in water and in ethanol (99.5), and slightly soluble in acetonitrile. Melting point: about 1459C (with decomposition).
(1) Determine the infrared absorption spec-tra of Enalapril Maleate as directed in the potassium bro-mide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Enalapril Maleate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) To 20 mg of Enalapril Maleate add 5 mL of 1 mol/L hydrochloric acid TS, shake, add 5 mL of diethyl ether, and shake for 5 minutes. Take 3 mL of the upper layer, distil off the diethyl ether on a water bath, add 5 mL of water to the residue with shaking, and add 1 drop of potassium perman-ganate TS: the red color of the test solution immediately dis-appears. D Optical rotation <2.49> [a]20: －41.0 – －43.59(after dry-ing, 0.25 g, methanol, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Enalapril Maleate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Enalapril Maleate in 100 mL of a mixture of sodium dihydrogen phos-phate TS (pH 2.5) and acetonitrile (19:1), and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mixture of sodium dihydrogen phosphate TS (pH 2.5) and acetonitrile (19:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> acccording to the following conditions, and determine each peak area of these solutions by the automatic integration method: the area of the peak other than maleic acid and enalapril obtained from the sample solution is not larger than the peak area of enalapril from the standard solution. Furthermore, the total area of the peaks other than maleic acid and enalapril from the sample solution is not larger than 2 times the peak area of enalapril from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phases, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of enalapril, beginning after the peak of maleic acid. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of sodium dihydrogen phos-phate TS (pH 2.5) and acetonitrile (19:1) to make exactly 10 mL. Confirm that the peak area of enalapril obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of enalapril are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
To 10 tablets of Emorfazone Tablets add 200 mL of methanol, shake well to disintegrate, add methanol to make acid monomaleate [76095-16-4] Enalapril Maleate, when dried, contains not less than 98.0z and not more than 102.0z of enalapril maleate (C20H28N2O5.C4H4O4). JP XVIII Official Monographs / Enalapril Maleate 925 Description Enalapril Maleate occurs as white, crystals or crystalline powder. It is freely soluble in methanol, sparingly soluble in water and in ethanol (99.5), and slightly soluble in acetonitrile. Melting point: about 1459C (with decomposition). Identification (1) Determine the infrared absorption spec-tra of Enalapril Maleate as directed in the potassium bro-mide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Enalapril Maleate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) To 20 mg of Enalapril Maleate add 5 mL of 1 mol/L hydrochloric acid TS, shake, add 5 mL of diethyl ether, and shake for 5 minutes. Take 3 mL of the upper layer, distil off the diethyl ether on a water bath, add 5 mL of water to the residue with shaking, and add 1 drop of potassium perman-ganate TS: the red color of the test solution immediately dis-appears. D Optical rotation <2.49> [a]20: －41.0 – －43.59(after dry-ing, 0.25 g, methanol, 25 mL, 100 mm). Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Enalapril Maleate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Enalapril Maleate in 100 mL of a mixture of sodium dihydrogen phos-phate TS (pH 2.5) and acetonitrile (19:1), and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mixture of sodium dihydrogen phosphate TS (pH 2.5) and acetonitrile (19:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> acccording to the following conditions, and determine each peak area of these solutions by the automatic integration method: the area of the peak other than maleic acid and enalapril obtained from the sample solution is not larger than the peak area of enalapril from the standard solution. Furthermore, the total area of the peaks other than maleic acid and enalapril from the sample solution is not larger than 2 times the peak area of enalapril from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phases, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of enalapril, beginning after the peak of maleic acid. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of sodium dihydrogen phos-phate TS (pH 2.5) and acetonitrile (19:1) to make exactly 10 mL. Confirm that the peak area of enalapril obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of enalapril are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 30 mg each of Enalapril Maleate and Enalapril Maleate RS, both previously dried, and dissolve in a mixture of sodium dihydrogen phosphate TS (pH 2.5) and acetonitrile (19:1) to make exactly 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of enalapril in each solution. Amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4) ＝ MS × AT/AS MS: Amount (mg) of Enalapril Maleate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 15 cm in length, packed with porous styrene-divinyl-benzene copolymer for liquid chromatography (5 mm in par-ticle diameter). Column temperature: A constant temperature of about 709C. Mobile phase A: Dissolve 3.1 g of sodium dihydrogen phosphate dihydrate in 900 mL of water, adjust the pH to 6.8 with a solution of sodium hydroxide (1 in 4), and add water to make 1000 mL. To 950 mL of this solution, add 50 mL of acetonitrile for liquid chromatography. Mobile phase B: Dissolve 3.1 g of sodium dihydrogen phosphate dihydrate in 900 mL of water, adjust the pH to 6.8 with a solution of sodium hydroxide (1 in 4), and add water to make 1000 mL. To 340 mL of this solution, add 660 mL of acetonitrile for liquid chromatography. Flowing of mobile phase: Control the concentration gradient by changing the ratio of the mobile phases A and B as follows. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 20 95 → 40 5 → 60 20 – 25 Flow rate: 1.4 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of enalapril are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 1.0z.
Enflurane is a clear, colorless liquid. It is slightly soluble in water. It is miscible with ethanol (95) and with diethyl ether. It is a volatile, and not an inflammable. It shows no optical rotation. Boiling point: 54 – 579C
(1) Take 50 mL of Enflurane, and prepare the test solution as directed to the Oxygen Flask Combustion Method <1.06> using 40 mL of water as the absorbing liquid. The test solution responds to Qualitative Tests <1.09> for chloride and fluoride. (2) Determine the infrared absorption spectrum of En-flurane as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.302 – 1.304 Specific gravity <2.56> d 20: 1.520 – 1.540
(1) Acidity or alkalinity—To 60 mL of Enflurane add 60 mL of freshly boiled and cooled water, shake for 3 minutes, separate the water later, and use the layer as the sample solution. To 20 mL of the sample solution add one drop of bromocresol purple TS and 0.10 mL of 0.01 mol/L sodium hydroxide VS: the color of the solution is purple. To 20 mL of the sample solution add one drop of bromocresol purple TS and 0.06 mL of 0.01 mol/L hydrochloric acid VS: the color of the solution is yellow. Chloride <1.03>—To 20 g of Enflurane add 20 mL of water, shake well, and separate the water layer. Take 10 mL of the water layer add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.001z). Related substances—Proceed the test with 5 mL of Enflurane as directed under Gas chromatography <2.02> ac-cording to the following conditions. Determine each peak area other than the peak of air which appears soon after in-jection of the sample by the automatic integration method, and calculate the amount of each peak by the area percen-tage method: the amount of the substances other than enflu-rane is not more than 0.10z. Operating conditions— Detector: A thermal conductivity detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with siliceous earth for gas chromatography, 180 to 250 mm in particle diameter, coated with diethylene glycol succinate ester for gas chromatography in the ratio of 20z. Column temperature: A constant temperature of about 809C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of enflurane is about 3 minutes. Time span of measurement: About 3 times as long as the retention time of enflurane. System suitability— Test for required detectability: To exactly 1 mL of enflu-rane add 2-propanol to make exactly 100 mL. To exactly 2 mL of this solution add 2-propanol to make exactly 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 2-propanol to make exactly 10 mL. Confirm that the peak area of enflurane obtained with 5 mL of this solu-tion is equivalent to 7 to 13z of that with 5 mL of the solu-tion for system suitability test. System performance: Mix 5 mL of Enflurane and 5 mL of 2-propanol. When the procedure is run with 5 mL of this mixture under the above operating conditions, enflurane and 2-propanol are eluted in this order with the resolution be-tween these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of enflurane is not more than 2.0z. ‌928 ‌Nonvolatile residue—Evaporate exactly 65 mL of En-flurane on a water bath to dryness, and dry the residue at 1059C for 1 hour: the residue is not more than 1.0 mg. Water <2.48> Not more than 0.10z (10 g, volumetric titra-tion, direct titration).
Enoxacin Hydrate, when dried, contains not less than 98.
Enoxacin Hydrate occurs as white to pale yel-low-brown, crystals or crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in methanol, very slightly soluble in chloroform, and practi-cally insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in dilute sodium hydroxide TS. It is gradually colored by light.
(1) Place 0.02 g of Enoxacin Hydrate and 0.05 g of sodium in a test tube, and heat gradually to ignition with precaution. After cooling, add 0.5 mL of methanol and then 5 mL of water, and heat to boiling. To this solution add 2 mL of dilute acetic acid, and filter: the filtrate responds to Qualitative Tests <1.09> (2) for fluoride. Dissolve 0.05 g of Enoxacin Hydrate in dilute sodium hydroxide TS to make 100 mL. To 1 mL of the solution add water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Enox-acin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com- Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Enoxacin Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Enoxacin Hy-drate in 25 mL of a mixture of chloroform and methanol (7:3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (7:3) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> 7.0 – 9.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.3 g of Enoxacin Hydrate, previously dried, dissolve in 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.03 mg of C15H17FN4O3
Entacapone occurs as a yellow to greenish yel-low crystalline powder. It is sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It shows crystal polymorphism.
(1) Dissolve 35 mg of Entacapone in 200 mL of methanol. To 7 mL of this solution add 0.1 mol/L hy-drochloric acid TS to make 100 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared by adding 0.1 mol/L hydrochloric acid TS to 7 mL of meth-anol to make 100 mL as the blank, and compare the spec- JP XVIII Official Monographs / Entacapone 929 trum with the Reference Spectrum or the spectrum of a solu-tion of Entacapone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of En-tacapone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Entacapone RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—Dissolve 1.0 g of Enoxacin Hy-drate in 50 mL of dilute sodium hydroxide TS, shake with 10 mL of dilute hydrochloric acid, and centrifuge. Filter the su-pernatant liquid, and to 30 mL of the filtrate add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.50 mL of 0.005 mol/L sulfuric acid VS add 25 mL of dilute sodium hydroxide TS, 5 mL of dilute hydrochloric acid TS and water to make 50 mL (not more than 0.048z). (2) Heavy metals <1.07>—Proceed with 1.0 g of Enoxa- cin Hydrate according to Method 2, and perform the test. Description Entacapone occurs as a yellow to greenish yel-low crystalline powder. It is sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It shows crystal polymorphism. Identification (1) Dissolve 35 mg of Entacapone in 200 mL of methanol. To 7 mL of this solution add 0.1 mol/L hy-drochloric acid TS to make 100 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared by adding 0.1 mol/L hydrochloric acid TS to 7 mL of meth-anol to make 100 mL as the blank, and compare the spec- JP XVIII Official Monographs / Entacapone 929 trum with the Reference Spectrum or the spectrum of a solu-tion of Entacapone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of En-tacapone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Entacapone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Purity (1) Heavy metals—Dissolve 1.0 g of Entacapone in 20 mL of a mixture of methanol and N,N-dimethylfor-mamide (3:1), and use this solution as the sample solution. Separately, weigh exactly 0.400 g of lead (II) nitrate, dissolve in water to make exactly 250 mL. Before use, dilute this so-lution with water to make exactly 10 times the initial volume, then dilute this solution with water to make exactly 10 times the initial volume. Pipet 1 mL of this solution, add a mixture of methanol and N,N-dimethylformamide (3:1) to make ex-actly 20 mL, and use this solution as the standard solution. To each of the sample solution and standard solution add 2 mL of acetate buffer solution (pH 3.5), mix, add 1.2 mL of thioacetamide TS, and mix immediately. Allow them to stand for 2 minutes, filter separately all the amount of each solution through a membrane filter with a pore size of 0.45 mm, wash the membrane filters with not less than 20 mL of methanol, and compare the colors on the membrane filters: the color obtained from the sample solution is not darker than that obtained from the standard solution (not more than 10 ppm). peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, add a mixture of methanol and tetrahydrofuran (7:3) to make exactly 10 mL. Confirm that the peak area of entacapone obtained with 10 mL of this solution is equivalent to 35 to 65z of that obtained with 10 mL of the standard so-lution. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of entacapone is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Entacapone and Entacapone RS (separately determine the loss on drying <2.41> under the same conditions as Entacapone), dissolve each in a mixture of methanol and tetrahydrofuran (7:3) to make exactly 50 mL. Pipet 5 mL each of these solutions, add a mixture of methanol and tetrahydrofuran (7:3) to make ex-actly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of entacapone in each solution. Halide—Being specified separately when the drug is granted approval based on the Law. Related substances—Dissolve 50 mg of Entacapone Amount (mg) of entacapone (C ＝ MS × AT/AS 14H15N3O5) in 50 mL of a mixture of methanol and tetrahydrofuran (7:3), and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add a mixture of methanol and tetrahydrofuran (7:3) to make exactly 50 mL. Pipet 5 mL of this solution, and add a mixture of methanol and tetrahydrofuran (7:3) to make exactly 50 mL. Pipet 1 mL of this solution, add a mixture of methanol and tetrahydrofu-ran (7:3) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak area of the related sub-stance A, having the relative retention time of about 0.8 to entacapone, obtained from the sample solution is not larger than 1.5 times the peak area of entacapone from the stand-ard solution, the area of the peak other than entacapone and the peak mentioned above from the sample solution is not larger than the peak area of entacapone from the standard solution, and the total area of the peaks other than entaca-pone and the related substance A, having the relative reten-tion time of about 0.8 to entacapone, from the sample solu-tion is not larger than 2 times the peak area of entacapone from the standard solution. For the areas of the peaks of related substances B and C, having the relative retention times of about 0.6 and about 1.4 to entacapone, multiply their correction factors, 1.7 and 2.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of entacapone, beginning after the solvent MS: Amount (mg) of Entacapone RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 300 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.34 g of sodium dihydrogen phos-phate dihydrate in water to make 1000 mL, and adjust to pH 2.1 with phosphoric acid. To 540 mL of this solution add 440 mL of methanol and 20 mL of tetrahydrofuran. Flow rate: 1 mL per minute. System suitability— System performance: Dissolve 5 mg of Entacapone Relat-ed Substance A for System Suitability RS in a mixture of methanol and tetrahydrofuran (7:3) to make 25 mL. To 1 mL of this solution add a mixture of methanol and tetra-hydrofuran (7:3) to make 20 mL, and use this solution as the solution for system suitability test. Separately, to 5 mL of the standard solution add a mixture of methanol and tetra-hydrofuran (7:3) to make 50 mL. To 1 mL of this solution and 1 mL of the solution for system suitability test add a mixture of methanol and tetrahydrofuran (7:3) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the related substance A and entacapone are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌930 area of entacapone is not more than 1.0z.
Enviomycin Sulfate is the sulfate of a mixture of peptide substances having antibacterial activity pro-duced by the growth of Streptomyces griseoverticilla-tus var. tuberacticus. It contains not less than 770 mg (potency) and not more than 920 mg (potency) per mg, calculated on the dried basis.
Enviomycin Sulfate occurs as a white powder. It is very soluble in water, and practically insoluble in ethanol (99.5).
(1) To 5 mL of a solution of Enviomycin Sulfate (1 in 200) add 1.5 mL of sodium hydroxide TS, and add 1 drop of a mixture of 0.01 mol/L citric acid TS and copper (II) sulfate TS (97:3) : a blue-purple color develops. Determine the absorption spectrum of a solution of Enviomycin Sulfate (1 in 100,000) as directed under Ultravi-olet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. To 2 mL of a solution of Enviomycin Sulfate (1 in 20) add 1 drop of barium chloride TS: a white precipitate is produced. D Optical rotation <2.49> [a]20: －16 – －229(0.5 g calculated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 2.0 g of Enviomycin Sulfate in 20 mL of water is between 5.5 and 7.5. Content ratio of the active principle Dissolve 0.1 g of En-viomycin Sulfate in water to make 100 mL, and use this solu-tion as the sample solution. Perform the test with 3 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT1 and AT2, of tuberactinomycin N and tuberactinomycin O, having the relative retention time, 1.4 ± 0.4, to tuberactinomycin N, by the automatic integra-tion method: AT2/(AT1 ＋ AT2) is between 0.090 and 0.150. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of ammonium acetate TS, 1,4-dioxane, tetrahydrofuran, water and ammonia solution (28) (100:75:50:23:2). Flow rate: Adjust so that the retention time of tuberac-tinomycin N is about 9 minutes. System suitability— System performance: When the procedure is run with 3 mL of the sample solution under the above operating conditions, tuberactinomycin N and tuberactinomycin O are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 3 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of tuberactinomycin N is not more than 2.0z.
(1) Clarity and color of solution—Dissolve 1.0 g of Enviomycin Sulfate in 10 mL of water: the solution is clear and colorless. ‌Heavy metals <1.07>—Proceed with 2.0 g of Envio-mycin Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Enviomycin Sulfate according to Method 1, and perform the test (not more than 1 ppm). Loss on drying <2.41> Not more than 4.0z (0.2 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillius subtilis ATCC 6633 Culture medium—Use the medium i in 1) Medium for test organism [5] under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Enviomycin Sulfate RS, equivalent to about 20 mg (po-tency), dissolve in water to make exactly 20 mL, and use this solution as the standard stock solution. Keep the standard stock solution at a temperature not exceeding 59C and use within 10 days. Take exactly a suitable amount of the stand-ard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 400 mg (potency) and 100 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Enviomycin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 20 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 400 mg (potency) and 100 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Epalrestat, when dried, contains not less than 98.
Epalrestat occurs as yellow to orange, crystals or crystalline powder. It is soluble in N,N-dimethylformamide, slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water. It gradually fades the color and decomposes on exposure to light. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Epalrestat in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Epalrestat RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Epal-restat as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Epalrestat RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. If any difference ap-pears between the spectra, proceed as follows, using a light-resistant vessel. To 0.1 g of Epalrestat add 40 mL of methanol, dissolve the sample by warming in a water bath, and filter while hot, and cool in ice. Collect the crystals formed, dry, and perform the test.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Epalrestat according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve about 20 mg of Epalrestat in 8 mL of N,N-dimethylformamide, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add N,N-dimethylformamide to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 3 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than epalrestat obtained from the sample solution is not larger than 1/5 times the peak area of epalrestat from the standard solution, and the total area of the peaks other than epalrestat from the sample solution is not larger than the peak area of epalrestat from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of epalrestat, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add N,N-dimethylformamide to make exactly 10 mL. Confirm that the peak area of epalrestat obtained with 3 mL of this solution is equivalent to 7 to 13z of that with 3 mL of the standard solution. System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of epalrestat are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of epalrestat is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, in vacu-um, silica gel, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 20 mg each of Epalrestat and Epal- ‌JP XVIII Official Monographs / Epalrestat Tablets 933 restat RS, both previously dried, and separately dissolve in 8 mL of N,N-dimethylformamide, and add exactly 2 mL of the internal standard solution. To 2 mL each of these solu-tions add N,N-dimethylformamide to make 20 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of epalrestat to that of the internal standard. Conduct this procedure using light-resistant vessels. To 1 tablet of Epalrestat Tablets add exactly 30 mL of N,N-dimethylformamide, shake thoroughly to completely disinte-grate the tablet, and centrifuge. Pipet 1 mL of the superna-tant liquid, and add N,N-dimethylformamide to make exactly 100 mL. Pipet V mL of this solution, add exactly V? mL of N,N-dimethylformamide so that each mL contains about 4.2 mg of epalrestat (C15H13NO3S2), and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg of Epalrestat RS, previously dried in vacuum at Amount (mg) of epalrestat (C15H13NO3S2) ＝ M S × QT/QS 609C for 3 hours with silica gel as a desiccant, and dissolve in exactly 30 mL of N,N-dimethylformamide. Pipet 1 mL of MS: Amount (mg) of Epalrestat RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in N,N-dimethylformamide (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 0.05 mol/L potassium dihydrogen phos-phate TS add 0.05 mol/L disodium hydrogen phosphate TS so that the pH of this mixture is 6.5. To 2 volumes of this mixture add 1 volume of acetonitrile. Flow rate: Adjust so that the retention time of epalrestat is about 12 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, epalrestat and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of epalrestat to that of the internal standard is not more than 1.0z.
Eperisone Hydrochloride contains not less than 98.
Eperisone Hydrochloride occurs as a white crystalline powder. It is freely soluble in water, in methanol and in acetic acid (100), and soluble in ethanol (99.5). Melting point: about 1679C (with decomposition). A solution of Eperisone Hydrochloride in methanol (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Eperisone Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Eperi-sone Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Eperisone Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Eperisone Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Piperidine hydrochloride—Dissolve 1.0 g of Eperi-sone Hydrochloride in 20 mL of water, add 2.0 mL of diluted hydrochloric acid (1 in 2), 2.0 mL of a solution of copper (II) sulfate pentahydrate (1 in 20) and 1.5 mL of am-monia solution (28), and use this solution as the sample solu-tion. Separately, to 2.0 mL of a solution of piperidine hydro-chloride (1 in 1000) add 18 mL of water, 2.0 mL of diluted hydrochloric acid (1 in 2), 2.0 mL of a solution of copper (II) sulfate pentahydrate (1 in 20) and 1.5 mL of ammonia solu-tion (28), and use this solution as the standard solution. To retention time of eperisone. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of eperisone obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of eperisone are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of eperisone is not more than 3.0z. Water <2.48> Not more than 0.20z (0.1 g, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.6 g of Eperisone Hydro-chloride, dissolve in 20 mL of acetic acid (100), add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.59 mg of C17H25NO.HCl
Ephedrine Hydrochloride, when dried, contains not less than 99.
Ephedrine Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water, soluble in ethanol (95), slightly soluble in acetic acid (100), and practically insoluble in acetonitrile and in acetic anhydride.
(1) Determine the absorption spectrum of a solution of Ephedrine Hydrochloride (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ephe-drine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Ephedrine Hydrochloride (1 in 15) responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: －33.0 – －36.09(after dry-ing, 1 g, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Ephedrine Hydrochloride in 20 mL of water: the pH of this solution is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Ephedrine Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate—Dissolve 0.05 g of Ephedrine Hydrochloride in 40 mL of water, add 1 mL of dilute hydrochloric acid and 1 mL of barium chloride TS, and allow to stand for 10 minutes: no turbidity is produced. Heavy metals <1.07>—Proceed with 1.0 g of Ephe- drine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.05 g of Ephedrine Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas of each peak by the automatic integration method: the total area of the peaks other than ephedrine obtained from the sample solution is not larger than the peak area of ephe-drine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of a solution of sodium lauryl sulfate (1 in 128), acetonitrile and phosphoric acid (640:360:1). Flow rate: Adjust so that the retention time of ephedrine is about 14 minutes. Time span of measurement: About 3 times as long as the retention time of ephedrine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of ephedrine obtained with 10 mL of this solution is equivalent to 4 to 6z of that with 10 mL of the standard solution. System performance: Dissolve 1 mg of ephedrine hydro-chloride for assay and 4 mg of atropine sulfate hydrate in 100 mL of diluted methanol (1 in 2). When the procedure is run with 10 mL of this solution under the above operating conditions, ephedrine and atropine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of ephedrine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Ephedrine Hydro-chloride, previously dried, and dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3) by warm-ing. Cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.17 mg of C10H15NO.HCl
Adrenaline occurs as a white to grayish white crystalline powder. It is freely soluble in formic acid and in acetic acid (100), very slightly soluble in water, and practically insoluble in methanol and in ethanol (99.5). It dissolves in dilute hydrochloric acid. It gradually changes to brown in color by air and by light.
(1) Determine the absorption spectrum of a solution of Adrenaline in 0.01 mol/L hydrochloric acid TS (1 in 25,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Adrenaline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com- ‌pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.‌ D Optical rotation <2.49> [a]20: －50.0 – －53.59(after dry-ing, 1 g, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.10 g of Adrenaline in 10 mL of dilute hydrochloric acid: the solu-tion is clear, and is not more colored than Matching Fluid A. Heavy metals <1.07>—Proceed with 1.0 g of Adrena- line according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Adrenalone—Dissolve 50 mg of Adrenaline in 0.05 mol/L hydrochloric acid TS to make exactly 25 mL, and de-termine the absorbance of this solution at 310 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.2. Noradrenaline—Dissolve 0.20 g of Adrenaline in 1 mL of formic acid, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dis-solve 8.0 mg of Noradrenaline Bitartrate RS in methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of 1-butanol, water and formic acid (7:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Folin's TS on the plate: the spot obtained from the sample solution, corresponding to the spot from the standard solution, is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, silica gel, 18 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Adrenaline, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.32 mg of C9H13NO3
Containers—Tight containers. Storage—Light-resistant, and under Nitrogen atmosphere.
Epirizole, when dried, contains not less than 99.
Epirizole occurs as white, crystals or crystal-line powder. It is odorless, and has a bitter taste. It is very soluble in methanol and in acetic acid (100), freely soluble in ethanol (95), and sparingly soluble in water and in diethyl ether. It dissolves in dilute hydrochloric acid and in sulfuric acid. The pH of a solution of 1.0 g of Epirizole in 100 mL of water is between 6.0 and 7.0.
(1) To 0.1 g of Epirizole add 0.1 g of vanil-lin, 5 mL of water and 2 mL of sulfuric acid, and mix with shaking for a while: a yellow precipitate is formed. Dissolve 0.1 g of Epirizole in 10 mL of water, and add 10 mL of 2,4,6-trinitrophenol TS: a yellow precipitate is produced. Collect the precipitate by filtration, wash with 50 mL of water, and dry at 1059C for 1 hour: it melts <2.60> between 1639C and 1699C. Determine the absorption spectrum of a solution of Epirizole in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 0.20 g of Epirizole in 20 mL of water: the solution is clear and col-orless. Chloride <1.03>—Add 0.5 g of Epirizole to a ground mixture of 0.7 g of potassium nitrate and 1.2 g of anhydrous sodium carbonate, mix well, transfer little by little to a plati-num crucible, previously heated, and heat until the reaction is completed. After cooling, add 15 mL of dilute sulfuric acid and 5 mL of water to the residue, boil for 5 minutes, filter, wash the insoluble matter with 10 mL of water, and to the combined filtrate and washings add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test with this so-lution as the test solution. Prepare the control solution as follows: proceed with the same quantities of the same rea-gents as directed for the preparation of the test solution, and add 0.25 mL of 0.01 mol/L hydrochloric acid VS and water to make 50 mL (not more than 0.018z). Heavy metals <1.07>—Proceed with 2.0 g of Epirizole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Epirizole according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 1.0 g of Epirizole in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of isopropyl ether, ethanol (95) and water (23:10:2) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Place this plate in a chamber filled with iodine vapor: the spots other than the principal spot from the sample solu-tion are not more intense than the spot from the standard so-lution. Readily carbonizable substances <1.15>—Perform the test with 0.10 g of Epirizole: the solution has no more color than Matching Fluid A. Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 4 hours). ‌JP XVIII Official Monographs / Epirubicin Hydrochloride 939 Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Epirizole, previ-ously dried, dissolve in 40 mL of acetic acid (100) and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS) until the color of the solution changes from purple through blue-green to green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 23.43 mg of C11H14N4O2
Epirubicin Hydrochloride is the hydrochloride of a derivative of daunorubicin. It contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous and residual solvent-free basis.
Epirubicin Hydrochloride occurs as a pale yel-lowish red to brownish red powder. It is soluble in water and in methanol, slightly soluble in ethanol (95), and practically insoluble in acetonitrile. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Epirubicin Hydrochloride in methanol (3 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Epirubicin Hydrochloride and Epirubicin Hydrochloride RS as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare these spec-tra: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋310 – ＋3409(10 mg calcu-lated on the anhydrous and residual solvent-free basis, meth-anol, 20 mL, 100 mm). pH <2.54> Dissolve 10 mg of Epirubicin Hydrochloride in 2 mL of water: the pH of the solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 50 mg of Epirubicin Hydrochloride in 5 mL of water: the solution is clear and dark red. Heavy metals <1.07>—Proceed with 1.0 g of Epirubi- cin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Perform the test with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic in-tegration method, and calculate the total amount of the peaks other than epirubicin and 2-naphthalenesulfonic acid by the area percentage method: not more than 5.0z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of epirubicin, beginning after the solvent peak. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of epirubicin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. Water <2.48> Not more than 8.0z (0.1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.5z (0.1 g).
Weigh accurately an amount of Epirubicin Hydro-chloride and Epirubicin Hydrochloride RS, equivalent to about 50 mg (potency), dissolve each in the internal standard solution to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of epirubicin to that of the internal standard. Amount [ mg (potency)] of epirubicin hydrochloride (C27H29NO11.HCl) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Epirubicin Hydrochloride RS taken Internal standard solution—A solution of sodium 2-naphthalene sulfonate in a mixture of water, acetonitrile, methanol and phosphoric acid (540:290:170:1) (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with trimethylsilanized silica gel for liquid chromatography (6 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 2 g of sodium lauryl sulfate in a ‌940 mixture of water, acetonitrile, methanol and phosphoric acid (540:290:170:1) to make 1000 mL. Flow rate: Adjust so that the retention time of epirubicin is about 9.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and epirubicin are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of epirubicin to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage—At a temperature between 09C and 59C.
Eplerenone occurs as a white crystalline pow-der. It is freely soluble in acetonitrile, sparingly soluble in methanol, and very slightly soluble in water and in ethanol (99.5). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Eplerenone in methanol (1 in 77,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Eplerenone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Eplerenone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Eplerenone RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －14.0 – －16.09(0.25 g cal-culated on the dried basis, acetonitrile, 25 mL, 100 mm).
(1) Heavy metals—Take 1.0 g of Eplerenone in a crucible, wet the sample with a suitable amount of sulfuric acid, cover loosely, and heat gently to carbonize. After cool-ing, add 2 mL of nitric acid, and 5 drops of sulfuric acid to the carbonized residue, and heat gently until white fumes are no longer evolved. Then, incinerate by ignition at 500 – 6009C. After cooling, add 4 mL of 6 mol/L hydrochloric acid TS, cover the crucible, warm on a water bath for 15 minutes, then remove the cover from the crucible, and slowly evaporate to dryness on a water bath. Wet the residue with 1 drop of hydrochloric acid, add 10 mL of hot water, and warm for 2 minutes. After cooling, add ammonia TS until the solution shows alkalinity to litmus paper, add 15 mL of water, and adjust to pH 3.0 – 4.0 with dilute acetic acid. Filter, if necessary, wash the crucible and filter paper with 10 mL of water, put the filtrate and the washings in a Nessler tube, add water to make 40 mL, and use this solution as the sample solution. Separately, take 2.0 mL of Standard Lead Solution in a Nessler tube, and add water to make 25 mL. Adjust to pH 3.0 – 4.0 of this solution with dilute acetic acid or ammonia TS, add water to make 40 mL, and use this solution as the control solution. To the sample solution and the control solution add 2 mL of acetate buffer solution (pH 3.5) and 1.2 mL of thioacetamide-alkaline glycerin TS, then add water to make 50 mL, allow them to stand for 2 minutes, and observe vertically against a white background: the solution obtained from the sample solution is not more colored than that obtained from the control solution (not more than 20 ppm). (2) Related substances—Dissolve 25 mg of Eplerenone in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having a relative retention time of about 0.58, about 0.85, about 0.90, about 1.2 and about 1.6 to eplerenone, obtained from the sample solution is respectively not larger than 1/5, 3/10, 3/10, 3/10 and 3/10 times the peak area of eplerenone from the standard solution, and the area of the peak other than eplerenone and the peak mentioned above from the sample solution is not larger than 7/50 times the peak area of eplere-none from the standard solution. Furthermore, the total area of the peaks other than eplerenone from the sample solution is not larger than 1.2 times the peak area of eplerenone from the standard solution. For the area of the peak, having the relative retention time of about 0.85 to eplerenon, multiply the correction factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of eplerenone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add the mobile phase to make exactly 10 mL. Con-firm that the peak area of eplerenone obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of eplerenone are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌JP XVIII Official Monographs / Eplerenone Tablets 941 area of eplerenone is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 25 mg each of Eplerenone and Eplerenone RS (separately determine the loss on drying <2.41> under the same conditions as Eplerenone), separately dissolve in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of eplerenone in each solution. Amount (mg) of eplerenone (C24H30O6) ＝ MS × AT/AS MS: Amount (mg) of Eplerenone RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.4 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.0 with phosphoric acid. To 580 mL of this solution add 360 mL of acetonitrile for liquid chromatography and 60 mL of metha-nol. Flow rate: Adjust so that the retention time of eplerenone is about 12 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of eplerenone are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of eplerenone is not more than 1.0z.
エポエチン アルファ（遺伝子組換え）
Epoetin Alfa (Genetical Recombination) is an aque-ous solution in which a desired product is a recom-binant human erythropoietin produced in Chinese hamster ovary cells. It is a glycoprotein (molecular mass: ca. 37,000 to 42,000) consisting of 165 amino acid residues. It contains not less than 1.
Epoetin Alfa (Genetical Recombination) oc-curs as a clear and colorless liquid.
(1) Dilute a suitable volume of Epoetin Alfa (Genetical Recombination) and Epoetin Alfa RS with water. To 3 volume of these solutions add 1 volume each of buffer solution for epoetin alfa sample, heat at 1009C for 5 minutes, and use these solutions as the sample solution and the standard solution, respectively. Transfer a volume of the sample solution and the standard solution, equivalent to 0.7 mg of protein, into each sample well of the polyacrylamide gel for epoetin alfa, and start the SDS-polyacrylamide gel electrophoresis using a vertical discontinuous buffer solution system. After the electrophoresis, immerse the gel, a poly-vinylidene fluoride membrane and a filter paper in the blot-ting TS. Set them on a semi-dry blotting apparatus, and transcribe for about 1 hour with a constant electric current of 0.7 – 0.9 mA/cm2 depending on the dimension of the filter paper. Then, immerse the polyvinylidene fluoride membrane in the blocking TS for epoetin alfa for more than 1 hour while shaking, remove the blocking TS for epoetin alfa and add the primary antibody TS, then shake for a night or allow to stand at 49C for 3 nights. Remove the primary antibody TS, wash the membrane with phosphate-buffered sodium chloride TS, add the secondary antibody TS, and shake for more than 1 hour. Remove the secondary antibody TS, wash the membrane with phosphate-buffered sodium chloride TS, add the avidin-biotin TS, and shake for more than 1 hour. Remove the avidin-biotin TS, wash the mem- JP XVIII Official Monographs / Epoetin Alfa (Genetical Recombination) 943 brane with phosphate-buffered sodium chloride TS, and add the substrate TS for epoetin alfa for developing the color im-age: the main stained bands obtained from the sample solu-tion appear as similar migrating image as those from the standard solution. (2) Evaporate to dryness under reduced pressure a minic acid, AT1 and AT2, obtained from the sample solution, and the peak areas of those, AS1 and AS2, from the standard solution. Calculate the content of sialic acid in Epoetin Alfa (Genetical Recombination) by the following equation: be-tween 10 mol/mol and 12 mol/mol. Content (mol/mol) of sialic acid volume of Epoetin Alfa (Genetical Recombination) and Epoetin Alfa RS, equivalent to about 35 mg of protein, and ＝ (A T1/AS1 × 10 ＋ A T2/AS2 × 1/5)/a dissolve these residues in 100 mL of 0.1 mol/L tris buffer so- lution (pH 7.3). To these solutions add 5 mL of trypsin TS for epoetin alfa, warm at 379C for 6 hours, then cool in ice, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with 45 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and compare the chromatograms ob-tained from these solutions: both chromatograms show the similar peaks at the corresponding retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase A: A mixture of water and trifluoroacetic acid (5000:3). Mobile phase B: A mixture of acetonitrile, water and trifluoroacetic acid (4000:1000:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. a: Number (nmol) of moles of Epoetin Alfa (Genetical Recombination) where, molar concentration (mmol/L) of Epoetin Alfa (Genetical Recombination) is calculated by the following equation, using the absorbance A at 280 nm obtained in the Assay (1). Molar concentration (mmol/L) of Epoetin Alfa (Genetical Recombination) ＝ A × 103/22,430 22,430: Molar absorbance coefficient e Operating conditions— Detector: A fluorophotometer (excitation wavelength: 373 nm, fluorescence wavelength: 448 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile and methanol (84:9:7). Mobile phase B: A mixture of water and methanol (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) 0 – 5 98 2 5 – 95 98 → 35 2 → 65 Flow rate: 0.75 mL per minute. System suitability— System performance: When the procedure is run with 45 mL of the standard solution under the above conditions, the chromatogram shows the similar pattern with the chromato-gram of Epoetin Alfa RS obtained in the Peptide mapping. Sialic acid content To an exact volume of Epoetin Alfa (Genetical Recombination), equivalent to about 1 nmol of protein, add water to make exactly 45 mL. Add exactly 5 mL of sodium hydroxide TS, allow to stand in ice water for 90 minutes, and add exactly 5 mL of dilute acetic acid. Add exactly 45 mL of water and exactly 100 mL of a mixture of water and acetic acid (100) (27:8), and warm at 809C for 210 minutes. After cooling, add exactly 200 mL of the fluores-cence TS, and warm at 609C for 2 hours avoiding exposure to light. After cooling, add exactly 200 mL of sodium hy-droxide TS, and use this solution as the sample solution. Separately, just before starting the test, to exactly 250 mL of 0.4 mmol/L N-acetylneuraminic acid TS add exactly 20 mL of 0.1 mmol/L N-glycolylneuraminic acid TS and exactly 180 mL of water. Proceed with exactly 45 mL of this solution in the same manner as for the sample solution, and use the solution so obtained as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine the peak areas of N-acetylneuraminic acid and N-glycolylneura- 0 – 20 100 0 20 – 20.1 100 → 0 0 → 100 20.1 – 27 Flow rate: 0.6 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, N-glycolylneuraminic acid and N-acetylneuraminic acid are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of N-glycolylneuraminic acid and N-acetylneuraminic acid are not more than 2.0z, respectively. Oligosaccharide profile Being specified separately when the drug is granted approval based on the Law. Molecular mass Use the sample solution obtained in the Identification (1) as the sample solution. Separately, to 20 mL of molecular mass standard stock solution add 6.7 mL of the buffer solution for epoetin alfa sample, heat at 1009C for 5 minutes, and use this solution as the molecular mass standard solution. Transfer a volume of the sample solution, equivalent to 3.5 mg of protein and the total volume of the molecular mass standard solution into each sample well of the vertical discontinuous buffer solution system SDS-polyacrylamide gel, composed with resolving and stacking gels, and perform the electrophoresis. After the electropho-resis, immerse the gel in a solution of Coomassie brilliant blue R-250, containing 1.25 g in a mixture of 450 mL of methanol, 100 mL of acetic acid (100) and sufficient amount of water making up to 1000 mL. Determine the relative mobilities of the stained bands of egg albumin (molecular mass: about 45,000), carbonic anhydrase (molecular mass: about 31,000), soybean trypsin inhibitor (molecular mass: about 21,500) and lysozyme (molecular mass: 14,400), and prepare a calibration curve by linear regression against the logarithm of the molecular masses. Determine the relative mobility of the center of the main band obtained from the sample solution, and calculate the molecular mass of Epoe-tin Alfa (Genetical Recombination) from the calibration curve: it is between 37,000 and 42,000. pH <2.54> 5.7 – 6.7
(1) Multimers—Perform the test with a volume of Epoetin Alfa (Genetical Recombination), equivalent to 50 mg of protein, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the total amount of the peaks other than epoetin alfa is not more than 2z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with hydrophilic silica gel for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 91 mg of disodium hydrogen phos-phate dodecahydrate, 0.27 g of sodium dihydrogen phos-phate dihydrate and 8.77 g of sodium chloride in water to make 1000 mL. Flow rate: Adjust so that the retention time of epoetin alfa is about 16 minutes. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column until the elution of epoetin alfa is finished. System suitability— Test for required detectability: To 1 volume of Epoetin Alfa (Genetical Recombination) add 49 volumes of the mo-bile phase, and use this solution as the solution for system suitability test. Confirm that the peak area of epoetin alfa obtained with a volume, equivalent to 1 mg of protein, of the solution for system suitability test is equivalent to 1.5 to 2.5z of that with the same volume of Epoetin Alfa (Geneti-cal Recombination). System performance: Dissolve 40 mg of bovine serum al-bumin for gel filtration molecular mass marker and 20 mg of chymotrypsinogen for gel filtration molecular mass marker in 100 mL of the mobile phase. When the procedure is run with 50 mL of this solution under the above operating condi-tions, bovine serum albumin and chymotrypsinogen are eluted in this order with the resolution between these peaks being not less than 4. drug is granted approval based on the Law.
(1) Protein content—Take a suitable amount of Epoetin Alfa (Genetical Recombination), dilute with phos-phate buffer solution for epoetin alfa, if necessary, so that each mL contains 0.5 – 0.8 mg protein and use the solution as the sample solution. Determine the absorbance, A, at 280 nm of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the phosphate buffer solution for epoetin alfa as the blank. Amount (mg) of protein in 1 mL of Epoetin Alfa (Genetical Recombination) ＝ A × d × 0.909 d: Dilution factor for the sample solution 1 cm 0.909: Reciprocal number of absorption coefficient (E 0.1z) of epoetin alfa protein Specific activity Animals: Select healthy 6 to 8 weeks female mice (B6D2F1, etc.). Keep the mice for not less than a week be-fore use, providing an appropriate uniform diet and water. Standard solutions: To Epoetin Alfa RS add the bo-vine serum albumin-saline solution so that each mL contains exactly 10 – 40 units, and designate this solution as the high-dose standard solution, SH. Dilute SH exactly 4 times with the bovine serum albumin-saline solution, and designate this solution as the low-dose standard solution, SL. Sample solutions: To Epoetin Alfa (Genetical Recombination) add the bovine serum albumin-saline solu-tion to make two sample solutions, the high-dose sample so-lution, TH, which contains the Units per mL equivalent to SH and the low-dose sample solution, TL, which contains the Units per mL equivalent to SL. Procedure: Divide the animals into 4 equal groups of not less than 5 animals each. On the 1st, 2nd and 3rd days, inject exactly 0.2 mL each of the standard solutions and the sample solutions into each animal subcutaneously as indi-cated in the following design: First group SH Third group TH Second group SL Fourth group TL On the 4th day, take a sufficient blood sample to perform the test from each animal. To 10 mL of the dilution fluid for particle counter add exactly 20 mL of the blood sample, mix, add 100 mL of the appropriate hemolysis agent, stir for 5 minutes, and determine the count of particles derived from hemolytic-resistant erythroid cells. Calculation: Logarithmic converted counts of the fine particles obtained with SH, SL, TH and TL in (iv) are symbolized as y1, y2, y3 and y4, respectively. Sum up individ-ual y1, y2, y3 and y4 to obtain Y1, Y2, Y3 and Y4, respectively. Specific activity (unit/mg protein) of Epoetin Alfa (Geneti-cal Recombination) ＝ activity (unit/mL) of Epoetin Alfa (Genetical Recombination)/C Activity (unit/mL) of Epoetin Alfa (Genetical Recombina-tion) ＝ antilog M × unit in 1 mL of SH × d M ＝ log 4 × Y /Y System repeatability: When the test is repeated 6 times a b with a volume of Epoetin Alfa (Genetical Recombination), equivalent to 50 mg of protein, under the above operating conditions, the relative standard deviation of the area of the principal peak of epoetin alfa is not more than 2.0z. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the Ya ＝ －Y1 － Y2 ＋ Y3 ＋ Y4 Yb ＝ Y1 － Y2 ＋ Y3 － Y4 d: Dilution factor for TH C: Concentration (mg/mL) of protein obtained in Assay (1) F? computed by the following equation should be smaller ‌JP XVIII Official Monographs / Epoetin Beta (Genetical Recombination) 945 than F shown in the table against n with which s2 is calcu-lated. Calculate L (p ＝ 0.95) by use of the following equa-tion: L should be not more than 0.3. If F? exceeds F, or if L exceeds 0.3, repeat the test, arranging the assay conditions. F? ＝ (Y1 － Y2 － Y3 ＋ Y4)2/4fs2 f: Number of animals per group, which should be the same for each group and not less than 5. s2 ＝ (Sy 2 － Y/f )/n Sy 2: The sum of the squares of each y1, y2, y3 and y4. Y ＝ Y12 ＋ Y22 ＋ Y32 ＋ Y42 n ＝ 4 ( f － 1) (C － 1){CM 2 ＋ (log 4)2} L ＝ 2 C ＝ Yb2/(Yb2 － 4fs2t 2) F (＝ t 2) values against n n t2 ＝ F n t 2 ＝ F n t 2 ＝ F 161.45 4.667 4.242 18.51 4.600 4.225 10.129 4.543 4.210 7.709 4.494 4.196 6.608 4.451 4.183 5.987 4.414 4.171 5.591 4.381 4.085 5.318 4.351 4.001 5.117 4.325 3.920 4.965 4.301 / 3.841 4.844 4.279 4.747 4.260
Containers—Tight containers. Storage—Not exceeding －709C.
Epoetin Beta (Genetical Recombination) is an aque-ous solution in which a desired product is a recom-binant human erythropoietin produced in Chinese hamster ovary cells. It is a glycoprotein (molecular mass: ca. 30,000) consisting of 165 amino acid residues. It contains not less than 0.
Epoetin Beta (Genetical Recombination) oc-curs as a clear and colorless liquid.
(1) Use Epoetin Beta (Genetical Recombi-nation) and Epoetin Beta RS as the sample solution and the standard solution, respectively. When perform a capillary electrophoresis with the sample solution and standard solu-tion according to the following conditions, the mobility of each peak obtained from both solutions is the same and their migrating images are similar each other. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 200 nm). Column: A silica capillary tube 50 mm in inside diameter and about 50 cm in length, chemically coated inner surface with amino groups (about 40 cm in effective length). Electrolyte solution: Dissolve 32.8 g of sodium dihydrogen phosphate dihydrate in water to make 1000 mL, and adjust to pH 4.5 with a solution, prepared by dissolving 75.2 g of disodium hydrogen phosphate dodecahydrate in water to make 1000 mL. To 19 volumes of this solution add 1 volume of ethanol (99.5). Running temperature: A constant temperature of about 209C. Running conditions: Migration current (a constant current of about 45 mA), migration time (30 minutes). Injection of sample and standard solutions: 5 seconds (pressurization: 0.5 psi). Time span of measurement: From 10 minutes to 30 minutes after injection (excluding the peak of solvent ori- gin). System suitability— System performance: When the procedure is run with the standard solution under the above operating conditions, more than 4 major peaks of epoetin beta are detected, and the resolution between the first and second eluted major peaks is not less than 0.8. System repeatability: When the test is repeated 3 times with the standard solution under the above operating condi-tions, the relative standard deviation of the migration time of the first eluted major peak is not more than 2.0z. (2) Desalt a volume each of Epoetin Beta (Genetical Recombination) and Epoetin Beta RS, equivalent to 600 mg of protein, by a suitable method, and term them as the desalted sample and the desalted reference standard, respec-tively. Dissolve the desalted sample and the desalted refer-ence standard in 600 mL each of a solution, prepared by dis-solving 2.3 g of N-ethylmorpholine in 100 mL of water and adjusting to pH 8.0 with acetic acid (100), and use these so-lutions as the desalted sample solution and the desalted ref-erence standard solution, respectively. To 500 mL each of the desalted sample solution and the desalted reference standard solution add 3.3 mL of triethylamine for epoetin beta and 1.5 mL of 2-mercaptoethanol for epoetin beta, and react at 379C for 1 hour. After cooling, add 5.5 mL of 4-vinylpyridine to them, and react at 259C for 1 hour. To these solutions add 50 mL of diluted trifluoroacetic acid for epoetin beta (1 in 10) to stop the reaction, remove the reagents by a suitable method, and use the substances so obtained as the pyridylethylated sample and the pyridylethylated reference substance, respectively. Dissolve the pyridylethylated sample and the pyridylethylated reference substance separately in 500 mL of sodium hydrogen carbonate solution (21 in 2500). To 400 mL each of these solutions add 16 mL of a solution of lysyl endopeptidase in sodium hydrogen carbonate solution (21 in 2500) (1 in 50,000), and react at 379C for 24 hours. While this reaction, additional two 16-mL portions of a solu-tion of lysyl endopeptidase in sodium hydrogen carbonate (21 in 2500) (1 in 50,000) are added at 4 hours and 20 hours after starting the reaction. Then, stop the reaction by adding 100 mL of diluted trifluoroacetic acid for epoetin beta (1 in 10), and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and compare the chromatograms ob-tained from these solutions: both chromatograms show the similar peaks at the corresponding retention times. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water and trifluoroacetic acid for epoetin beta (1000:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, water, and trifluoroacetic acid for epoetin beta (900:100:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 10 – 30 90 → 80 10 → 20 30 – 50 50 – 130 80 → 40 20 → 60 130 – 140 40 → 10 60 → 90 140 – 150 Flow rate: Adjust so that the retention time of the first peak, which appears after the solvent peak, is about 17 minutes. System suitability— System performance: When the procedure is run with the standard solution under the above operating conditions, 9 major peptide peaks are appeared after the solvent peak, and the resolution between the peaks eluted at the fifth and the sixth is not less than 3. Sialic acid content To exactly 100 mL of Epoetin Beta (Genetical Recombination) add 1 mL of resorcinol-copper (II) sulfate TS, and heat on a water bath for 30 minutes. After ice-cooling, add 2 mL of a mixture of n-butyl acetate and 1-butanol (4:1), shake vigorously, and use the upper layer as the sample solution. Separately, dissolve N-acetyl-neuraminic acid in water to make three solutions, containing 0.1 mg, 0.2 mg and 0.3 mg of N-acetylneuraminic acid in each mL, and use these solutions as the standard stock solu-tion (1), the standard stock solution (2) and the standard stock solution (3), respectively. Pipet 100 mL each of these standard stock solutions, add 1 mL of resorcinol-copper (II) sulfate TS to them, then proceed in the same way as for the sample solution, and use these solutions so obtained as the standard solution (1), the standard solution (2) and the standard solution (3), respectively. Determine the absor-bances of the sample solution and the standard solutions (1), (2) and (3) at 625 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Calculate the amount of sialic acid (mg/mL) in the sample solution, by using the calibra-tion curve obtained from the standard solutions, and calcu-late the amount of sialic acid in Epoetin Beta (Genetical Recombination) by the following equation: between 10 mol/mol and 13 mol/mol. Amount of sialic acid (mol/mol of epoetin beta protein) ＝ A/C × 18,236/309.27 A: Amount (mg/mL) of sialic acid in the sample solution C: Amount (mg/mL) of protein in Epoetin Beta (Geneti-cal Recombination) 18,236: Molecular mass of protein moiety of epoetin beta 309.27: Molecular mass of N-acetylneuraminic acid Oligosaccharide profile Being specified separately when the drug is granted approval based on the Law. pH <2.54> 7.0 – 8.0
(1) Related substances—Perform the test with 20 mL of Epoetin Beta (Genetical Recombination) as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amount of these peaks other than the solvent peak by the area percentage method: the total area of the peaks other than epoetin beta is not more than 1.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave- JP XVIII Official Monographs / Epoetin Beta (Genetical Recombination) 947 length: 214 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with porous silica gel for liq-uid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.6 g of sodium dihydrogen phos-phate dihydrate and 16.1 g of sodium sulfate decahydrate in water to make 1000 mL, and adjust to pH 6.8 with a solu-tion, prepared by dissolving 16.1 g of sodium sulfate decahy-drate in 0.01 mol/L sodium hydroxide TS to make 1000 mL. Flow rate: Adjust so that the retention time of epoetin beta is about 18 minutes. Time span of measurement: About 2 times as long as the retention time of epoetin beta. System suitability— Test for required detectability: When the procedure is run with 20 mL of diluted Epoetin Beta RS with water containing 0.05 volz polysorbate 20 for epoetin beta (1 in 1000) under the above conditions, the peak of epoetin beta is detectable. System performance: When the procedure is run with Epoetin Beta RS under the above conditions, the number of theoretical plates of the peak of epoetin beta is not less than 600. System repeatability: When the test is repeated 6 times with 20 mL of Epoetin Beta RS under the above operating conditions, the relative standard deviation of the peak area of epoetin beta is not more than 1.0z. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law.
(1) Protein content—Use Epoetin Beta (Genetical Recombination) and Epoetin Beta RS as the sample solution and the standard solution, respectively. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the total area, AT and AS, of the main peak and the sub-peak of epoetin beta in each solution. Amount (mg) of protein in 1 mL of Epoetin Beta (Genetical Recombination) ＝ CS × AT/AS CS: Protein concentration (mg/mL) of Epoetin Beta RS Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with butylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile for liq-uid chromatography and trifluoroacetic acid for epoetin beta (400:100:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, water and trifluoroacetic acid for epoetin beta (400:100:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 18 65 → 50 35 → 50 18 – 33 50 → 0 50 → 100 33 – 43 Flow rate: Adjust so that the retention time of the main peak of epoetin beta is about 22 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the main peak and the sub-peak of epoetin beta are eluted in this order, and the number of theoretical plates of the main peak is not less than 600. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the total area of the main peak and the sub-peak of epoetin beta is not more than 4.0z. (2) Specific activity—To Epoetin Beta (Genetical Recombination) add 0.1 w/vz bovine serum albumin-sodium chloride-phosphate buffer solution to make three so-lutions so that each mL contains epoetin beta equivalent to 5, 10 and 20 units (estimate), and use these solutions as the sample solutions (1), (2) and (3), respectively. Separately, to Epoetin Beta RS add 0.1 w/vz bovine serum albumin-sodium chloride-phosphate buffer solution to make three so-lutions so that each mL contains epoetin beta equivalent to 5, 10 and 20 units, and use these solutions as the standard solutions (1), (2) and (3), respectively. Divide ICR strain mice into 6 equal groups of not less than 5 mice. Inject ex-actly 0.2 mL each of the sample solutions and the standard solutions to ICR strain mice of each group subcutaneously on the 1st, 2nd and 3rd days. On the 4th day, collect the blood from the mice, put 20 mL each of the collected blood in 9.94 mL of blood dilution liquid, mix, and use these mix-tures as the dilute blood solution. To each of the dilute blood solution add 100 mL of a hemolytic agent, mix gently to hemolyze, and count the particles of hemolytic agent-resistant red cell by using a particle counter. Determine the potency ratio ( Pr) of the sample solution to the standard solution, and calculate the unit per mg protein of Epoetin Beta (Genetical Recombination) by the following equation. Pr ＝ 10M M ＝ 4/3 × i × Ta/Tb i ＝ log 2 Ta ＝ －S1 － S2 － S3 ＋ U1 ＋ U2 ＋ U3 Tb ＝ －S1 ＋ S3 － U1 ＋ U3 U1: Sum of the responses obtained from the sample solu-tion (1) U2: Sum of the responses obtained from the sample solu-tion (2) U3: Sum of the responses obtained from the sample solu-tion (3) S1: Sum of the responses obtained from the standard solu-tion (1) S2: Sum of the responses obtained from the standard solu-tion (2) S3: Sum of the responses obtained from the standard solu-tion (3) ‌948 Specific activity (unit/mg of protein) of Epoetin Beta (Genetical Recombination) ＝ S × Pr × DT/DS/C S: Potency (unit/mL) of Epoetin Beta RS DT: Dilution factor for the sample solution (3) DS: Dilution factor for the standard solution (3) C: Protein amount (mg/mL) of Epoetin Beta (Genetical Recombination)
Freeze-dried Mamushi Antivenom, Equine, becomes a colorless or light yellow-brown, clear liquid, or a slightly white-turbid liquid on addition of solvent. Manidipine Hydrochloride
Ergometrine Maleate, when dried, contains not Loss on drying <2.
Ergometrine Maleate occurs as a white to pale yellow crystalline powder. It is odorless. It is sparingly soluble in water, slightly soluble in metha-nol and in ethanol (95), and practically insoluble in diethyl ether. It gradually changes to yellow in color on exposure to light. Melting point: about 1859C (with decomposition).
(1) Prepare a solution of Ergometrine Maleate (1 in 50): the solution shows a blue fluorescence. Dissolve 1 mg of Ergometrine Maleate in 5 mL of water. To 1 mL of this solution add 2 mL of 4-dimethyl-aminobenzaldehyde-ferric chloride TS, shake, and allow to stand for 5 to 10 minutes: a deep blue color develops. To 5 mL of a solution of Ergometrine Maleate (1 in 500) add 1 drop of potassium permanganate TS: the red color of the solution disappears immediately. MS: Amount (mg) of Ergometrine Maleate RS taken
Weigh accurately about 10 mg each of Ergometrine Maleate and Ergometrine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, dissolve in water to make exactly 250 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Pipet 2 mL of each solution into a separate brown glass-stoppered tube. To each tube add 4 mL of 4-dimethylaminobenzaldehyde-iron (III) chloride TS, exactly measured, while cooling in an ice bath, then warm at 459C for 10 minutes. Allow to stand at room temperature for 20 minutes, and perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution, prepared with 2 mL of water in the same manner, as the blank. Determine the ab-sorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 550 nm, respec-tively. less than 98.0z of ergometrine maleate (C19H23N3O2. Amount (mg) of ergometrine maleate (C H N O .C H O ) C4H4O4). ＝ MS × AT/AS 19 23 3 2 4 4 4 Description Ergometrine Maleate occurs as a white to pale yellow crystalline powder. It is odorless. It is sparingly soluble in water, slightly soluble in metha-nol and in ethanol (95), and practically insoluble in diethyl ether. It gradually changes to yellow in color on exposure to light. Melting point: about 1859C (with decomposition). Identification (1) Prepare a solution of Ergometrine Maleate (1 in 50): the solution shows a blue fluorescence. Dissolve 1 mg of Ergometrine Maleate in 5 mL of water. To 1 mL of this solution add 2 mL of 4-dimethyl-aminobenzaldehyde-ferric chloride TS, shake, and allow to stand for 5 to 10 minutes: a deep blue color develops. To 5 mL of a solution of Ergometrine Maleate (1 in 500) add 1 drop of potassium permanganate TS: the red color of the solution disappears immediately. MS: Amount (mg) of Ergometrine Maleate RS taken
Ergotamine Tartrate contains not less than 98.
Ergotamine Tartrate occurs as colorless crys-tals, or a white to pale yellow-white or grayish white crystal-line powder. It is slightly soluble in water and in ethanol (95). Melting point: about 1809C (with decomposition).
(1) Dissolve 1 mg of Ergotamine Tartrate in 10 mL of a mixture of acetic acid (100) and ethyl acetate (1:1). To 0.5 mL of this solution add slowly 0.5 mL of sulfu-ric acid, with shaking in cold water, and allow to stand: a ‌JP XVIII Official Monographs / Eribulin Mesilate 951 purple color develops. To this solution add 0.1 mL of diluted iron (III) chloride TS (1 in 12): the color of the solution changes to blue to blue-purple. (2) Dissolve 1 mg of Ergotamine Tartrate in 5 mL of a solution of L-tartaric acid (1 in 100). To 1 mL of this solu-tion add 2 mL of 4-dimethylaminobenzaldehyde-iron (III) chloride TS, and shake: a blue color develops. D Optical rotation <2.49> Ergotamine base [a]20: －155 – －1659. Dissolve 0.35 g of Ergotamine Tartrate in 25 mL of a solution of L-tartaric acid (1 in 100), add 0.5 g of sodium hydrogen carbonate, shake gently and sufficiently, and ex-tract with four 10-mL portions of ethanol-free chloroform. Filter the extracts successively through a small filter paper, moistened with ethanol-free chloroform, into a 50-mL volu-metric flask. Allow the flask to stand in a water bath at 209C for 10 minutes, and determine the optical rotation in a 100-mm cell. Separately, pipet 25 mL of this solution, evaporate to dryness under reduced pressure at a tempera-ture not higher than 459C, dissolve the residue in 25 mL of acetic acid (100), and titrate <2.50> with 0.05 mol/L perchlo-ric acid VS (indicator: 1 drop of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Calculate the specific rotation of the ergotamine base from the consumed volume of 0.05 mol/L perchloric acid VS and the optical rotation. Each mL of 0.05 mol/L perchloric acid VS ＝ 29.08 mg of C33H35N5O5
Related substances—Conduct this procedure with-out exposure to light, using light-resistant vessels. To 40 mg of Ergotamine Tartrate add 10 mL of a solution of L-tartaric acid in diluted methanol (1 in 2) (1 in 1000), dissolve with thorough shaking, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add a solution of L-tartaric acid in diluted methanol (1 in 2) (1 in 1000) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and methanol (9:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminoben-zaldehyde TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.1 g, in vacu-um, 609C, 4 hours).
Weigh accurately about 0.2 g of Ergotamine Tar-trate, dissolve in 15 mL of a mixture of acetic acid (100) and acetic anhydride (50:3), and titrate <2.50> with 0.05 mol/L perchloric acid VS (indicator: 1 drop of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 32.84 mg of (C33H35N5O5)2.C4H6O6
Eribulin Mesilate contains not less than 95.
Eribulin Mesilate occurs as a white powder. It is freely soluble in water, in methanol, in ethanol (99.5) and in dimethylsulfoxide. It is hygroscopic.
(1) Determine the 1H spectrum of a solu-tion of Eribulin Mesilate in deuterated methanol for nuclear magnetic resonance spectroscopy (1 in 200), as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using light hydrogen contaminated in deuterated methanol for nuclear magnetic resonance spectroscopy as an internal reference compound and the chemical shift of methyl group of deuterated methanol as d 3.3 ppm: it exhibits a doublet signal A at around d 1.1 ppm, a multiplet signal B at around d 2.7 ppm and a singlet signal C at around d 2.7 ppm, a singlet signal D at around d 3.4 ppm, a doublet signal E at around d 3.7 ppm, a doublet signal F at around d 4.5 ppm, a triplet signal G at around d 4.6 ppm, and a triplet signal H at around d 4.7 ppm. The ratio of integrated intensity of these signals, A:B ＋ C:D:E:F:G:H, is about 3:5:3:1:1:1:1. (When measured with a nuclear magnetic resonance spectrometer having 1H resonance frequency of not less than or equal to 400 MHz.) (2) Perform the test with 15 mL each of the sample solu-tion and standard solution obtained in the Assay (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions: the retention times of the peaks of methanesulfonic acid in the chromatograms obtained from the sample solution and standard solution are the same. Operating conditions— Proceed as directed in the operating conditions in the Assay (2). System suitability— Proceed as directed in the system suitability in the Assay (2). Optical rotation <2.49> [a]20 : －160 – －2109(50 mg calcu-lated on the anhydrous and solvent-free basis, dimethylsul-foxide, 10 mL, 100 mm).
(1) Heavy metals—Being specified separately when the drug is granted approval based on the Law. Related substances—Weigh accurately about 20 mg of Eribulin Mesilate, dissolve in the dissolving solution to make exactly 5 mL, and use this solution as the sample solu-tion. Separately, weigh accurately about 20 mg of Eribulin Mesilate RS (separately determine the water <2.48> in the same manner as Eribulin Mesilate), dissolve in the dissolving solution to make exactly 5 mL. Pipet 1 mL of this solution, add the dissolving solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area, AT, of the related substances in the sample solution and the peak area, AS, of eribulin in the standard solution by the automatic integration method, and calculate the amounts of the related substances by the following formula: the amounts of related substance A having the relative reten-tion time of about 0.29 to eribulin, related substance B hav-ing the relative retention time of about 0.87, related sub-stance C having the relative retention time of about 1.07, related substance D having the relative retention time of about 1.29, related substance E having the relative retention time of about 1.37, and related substance F having the rela-tive retention time of about 1.67, are not more than 0.15z, 0.22z, 0.68z, 0.50z, 0.15z, and 0.19z, respectively, and other related substances are not more than 0.10z. Further-more, the total amount of these related substances is not more than 3.0z. Amount (z) of related substance ＝ MS/MT × AT/AS MS: Amount (mg) of Eribulin Mesilate RS taken, calcu- lated on the anhydrous basis MT: Amount (mg) of Eribulin Mesilate taken, calculated on the anhydrous basis Dissolving solution—A mixture of water, acetonitrile for liq-uid chromatography and phosphoric acid (6500:3500:7) adjusted to pH 6.9 – 7.1 with diluted ammonium water (28) (1 in 5) or 1 mol/L hydrochloric acid TS. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (1). Time span of measurement: For 85 minutes after injec-tion, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). Test for required detectability: Pipet 1 mL of the standard solution, and add the dissolving solution to make exactly 20 mL. Confirm that the peak area of eribulin obtained with 5 mL of this solution is equivalent to 3.5 to 6.5z of that with 5 mL of the standard solution. System repeatability: When the test is repeated 5 times with 5 mL of the solution for system suitability test obtained in the Assay (1) under the above operating conditions, the relative standard deviation of the peak area of eribulin is not more than 1.0z. Residual solvent—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 3.0z (Weigh accurately 15-25 mg of Eribulin Mesilate, dissolve in 2.5 mL of methanol for water determination, and perform the test with exactly 1 mL of this solution; coulometric titration).
(1) Eribulin Mesilate—Weigh accurately about 20 mg each of Eribulin Mesilate and Eribulin Mesilate RS (separately determine the water <2.48> in the same manner as Eribulin Mesilate), dissolve each in the dissolving solution to make exactly 5 mL, and use these solutions as the sample so-lution and the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, JP XVIII Official Monographs / Eribulin Mesilate 953 AT and AS, of eribulin in each solution. Amount (mg) of eribulin mesilate (C H NO .CH O S) eribulin are not less than 13,500 and not more than 1.5, re-spectively. ＝ MS × AT/AS 40 59 11 4 3 System repeatability: When the test is repeated 5 times with 5 mL of the solution for system suitability test under the MS: Amount (mg) of Eribulin Mesilate RS taken, calcu-lated on the anhydrous basis Dissolving solution—A mixture of water, acetonitrile for liq-uid chromatography and phosphoric acid (6500:3500:7) adjusted to pH 6.9 – 7.1 with diluted ammonia water (28) (1 in 5) or 1 mol/L hydrochloric acid TS. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 200 nm). Column: A stainless steel column 3 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 7.0 g of ammonium trifluoromethanesulfonate in 760 mL of water, add 3.0 mL of a solution of tetrabutylammonium dihydrogen phosphate (17 in 50) and 240 mL of acetonitrile for liquid chromatogra-phy, and adjust to pH 6.9 – 7.1 with diluted ammonium water (28) (1 in 5) or 1 mol/L hydrochloric acid TS. Mobile phase B: Dissolve 7.0 g of ammonium trifluoromethanesulfonate in 300 mL of water, add 3.0 mL of a solution of tetrabutylammonium dihydrogen phosphate (17 in 50), 700 mL of acetonitrile for liquid chromatography and 20 mL of 2-propanol for liquid chromatography, and adjust to pH 6.9 – 7.1 with diluted ammonium water (28) (1 in 5) or 1 mol/L hydrochloric acid TS. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. above operating conditions, the relative standard deviation of the peak area of eribulin is not more than 1.0z. (2) Methanesulfonic acid—Weigh accurately about 50 mg of Eribulin Mesilate, dissolve in a mixture of the mobile phase and acetonitrile for liquid chromatography (13:7) to make exactly 10 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of methanesulfonic acid, dissolve in a mixture of the mobile phase and acetonitrile for liquid chromatography (13:7) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 15 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of methanesulfon-ic acid in each solution. Calculate the content of methanesul-fonic acid by the following formula. Content (z) of methanesulfonic acid (CH4O3S) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of methanesulfonic acid taken Operating conditions— Detector: An electric conductivity detector. Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with aminopropylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.8 g of sodium dihydrogen phos-phate monohydrate in 950 mL of water, add 11 mL of phos- 0 – 55 Flow rate: Adjust so that the retention time of 55 – 75 100 → 0 0 → 100 methanesulfonic acid is about 6.5 minutes. 75 – 85 System suitability— 85 – 86 0 → 100 100 → 0 System performance: When the procedure is run with 15 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) phoric acid, adjust to pH 4.2 – 4.3 with phosphoric acid if necessary, and add 50 mL of acetonitrile for liquid chroma-tography. 86 – 105 100 0 Flow rate: Control the flow rate as directed in the follow-ing table. mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of methanesulfonic acid are not less than 12,000 and not more than 0.7 – 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat- Time after injection of sample (min) Flow rate (mL/minute) ing conditions, the relative standard deviation of the peak area of methanesulfonic acid is not more than 3.0z. 0 – 55 0.50 55 – 75 0.50 → 0.63 75 – 105 0.63 System suitability— System performance: Dissolve 2 mg of Eribulin Mesilate Related Substance C for System Suitability RS in the dissolv-ing solution to make exactly 50 mL. To 0.2 mL of this solu-tion add 4 mg of Eribulin Mesilate RS, dissolve in the dis-solving solution to make exactly 1 mL, and use this solution as the solution for system suitability test. When the proce-dure is run with 5 mL of the solution for system suitability test under the above operating conditions, eribulin and the related substance C are eluted in this order with the resolu-tion between these peaks being not less than 1.5. The number of theoretical plates and the symmetry factor of the peak of
エリスロマイシンエチルコハク酸エステル
Erythromycin Ethylsuccinate is a derivative of erythromycin. It contains not less than 780 mg (potency) and not more than 900 mg (potency) per mg, calculated on the anhydrous basis.
Erythromycin Ethylsuccinate occurs as a white powder. It is freely soluble in methanol and in acetone, soluble in ethanol (95), and practically insoluble in water.
(1) Dissolve 3 mg of Erythromycin Ethyl-succinate in 2 mL of acetone, and add 2 mL of hydrochloric acid: an orange color develops and is immediately changed to red to deep purple. Determine the infrared absorption spectrum of Erythromycin Ethylsuccinate, previously dried in a desicca-tor (reduced pressure, silica gel) for 24 hours, as directed in the potassium bromide disk method under Infrared Spectro- ‌JP XVIII Official Monographs / Erythromycin Lactobionate 957 photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Culture medium—Use the medium i in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Erythromycin RS, equivalent to about 50 mg (potency), dis-solve in 50 mL of methanol, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 59C or below and use within 7 days. Take exactly a suitable amount of the standard stock solu-tion before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solution and the low concentra-tion standard solution, respectively. Sample solutions—Weigh accurately an amount of Erythromycin Ethylsuccinate, equivalent to about 50 mg (potency), dissolve in 50 mL of methanol, and add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL. Take exactly a suitable amount of the solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solu-tions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
エリスロマイシンラクトビオン酸塩
Erythromycin Lactobionate is the lactobionate of erythromycin. It contains not less than 590 mg (potency) and not more than 700 mg (potency) per mg, calculated on the anhydrous basis.
Erythromycin Lactobionate occurs as a white powder. It is freely soluble in water, in methanol and in ethanol (99.5), and very slightly soluble in acetone.
(1) To 3 mg of Erythromycin Lactobionate add 2 mL of acetone, and add 2 mL of hydrochloric acid: an orange color is produced, and it changes immediately to red to deep purple. Transfer about 0.3 g of Erythromycin Lactobionate to a separator, add 15 mL of ammonia TS and 15 mL of chloroform, shake, and take the separated aqueous layer. Wash the aqueous layer with three 15-mL portions of chlo-roform, and evaporate the aqueous liquid on a water bath to dryness. Dissolve the residue in 10 mL of a mixture of meth-anol and water (3:2), and use this solution as the sample so-lution. Separately, dissolve 0.10 g of lactobionic acid in 10 mL of a mixture of methanol and water (3:2), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromato-graphy. Develop the plate with the upper layer obtained from a mixture of water, 1-butanol and acetic acid (100) (3:3:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute sulfuric acid, and heat at 1059C for 20 minutes: the principal spot obtained from the sample solu-tion shows a deep brown and the Rf value which are the same as those of the principal spot from the standard solu-tion. pH <2.54> The pH of a solution obtained by dissolving 0.5 g of Erythromycin Lactobionate in 10 mL of water is be-tween 5.0 and 7.5. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Culture medium—Use the medium i in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Erythromycin RS, equivalent to about 50 mg (potency), dissolve in 50 mL of methanol, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at not exceeding 59C and use within 7 C H NO .C H O : 1092.22 days. Take exactly a suitable amount of the standard stock 37 67 13 12 22 12 solution before use, add 0.1 mol/L phosphate buffer solu- (2R,3S,4S,5R,6R,8R,10R,11R,12S,13R)-5-(3,4,6- Trideoxy-3-dimethylamino-b-D-xylo-hexopyranosyloxy)-3-(2,6-dideoxy-3-C-methyl-3-O-methyl-a-L-ribo-hexopyranosyloxy)-6,11,12-trihydroxy-2,4,6,8,10,12-hexamethyl-9-oxopentadecan-13-olide mono(4-O-b- D-galactopyranosyl-D-gluconate) [3847-29-8] Erythromycin Lactobionate is the lactobionate of erythromycin. It contains not less than 590 mg (potency) and not more than 700 mg (potency) per mg, calculated on the anhydrous basis. The potency of Erythromycin Lac- tion (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Erythromycin Lactobionate, equivalent to about 50 mg (po-tency), dissolve in 50 mL of methanol, and add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample ‌958‌ solution and the low concentration sample solution, respec-tively.
エリスロマイシンステアリン酸塩
Erythromycin Stearate is the stearate of erythromy-cin. It contains not less than 600 mg (potency) and not more than 720 mg (potency) per mg, calculated on the anhydrous basis.
Erythromycin is a macrolide substance having anti-bacterial activity produced by the growth of Saccharo-polyspora erythraea. It contains not less than 930 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Erythromycin occurs as a white to light yellow-white powder. It is freely soluble in methanol and in ethanol (95), and very slightly soluble in water.
(1) Determine the infrared absorption spec-trum of Erythromycin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Erythromycin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Dissolve 10 mg each of Erythromycin and Erythro-mycin RS in 1 mL of methanol, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of methanol and ammonia solution (28) (50:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-methoxybenzalde-hyde-sulfuric acid TS on the plate, and heat at 1009C for 15 minutes: the principal spot from the sample solution and the spot from the standard solution are dark purple in color, and their Rf values are the same. D Optical rotation <2.49> [a]20: －71 – －789(1 g calculated on the anhydrous basis, ethanol (95), 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Erythromycin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 40 mg of Erythromycin in 2 mL of methanol, add a mixture of phosphate buffer so-lution (pH 7.0) and methanol (15:1) to make exactly 10 mL, and use this solution as the sample solution. Separately, dis-solve 16 mg of Erythromycin RS in 2 mL of methanol, add a mixture of phosphate buffer solution (pH 7.0) and methanol (15:1) to make exactly 10 mL, and use this solution as the standard stock solution. Dissolve 5 mg each of erythromycin B and erythromycin C in 2 mL of methanol, add exactly 2 mL of the standard stock solution, add a mixture of phos-phate buffer solution (pH 7.0) and methanol (15:1) to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak areas of erythromycin B and erythromycin C obtained from the sample solution are not larger than those of erythromycin B and erythromycin C from the stand-ard solution, respectively, and the area of the peaks other than erythromycin, erythromycin B and erythromycin C from the sample solution is not larger than the peak area of erythromycin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (8 mm in particle di-ameter). Column temperature: A constant temperature of about 709C. Mobile phase: Dissolve 3.5 g of dipotassium hydrogen phosphate in water to make 100 mL, and adjust the pH to 9.0 with diluted phosphoric acid (1 in 10). To 50 mL of this solution add 190 mL of t-butyl alcohol, 30 mL of acetonitrile and water to make 1000 mL. Flow rate: Adjust so that the retention time of erythromy-cin is about 20 minutes. Time span of measurement: About 4 times as long as the retention time of erythromycin, beginning after the solvent peak. System suitability— System performance: Dissolve 2 mg of N-demethylery-thromycin in 10 mL of the standard solution. When the procedure is run with 100 mL of this solution under the above operating conditions, N-demethylerythromycin, erythromycin C, erythromycin and erythromycin B are eluted in this order, with the resolution between the peaks of N-demethylerythromycin and erythromycin C being not less than 0.8 and with the resolution between the peaks of N-demethylerythromycin and erythromycin being not less than 5.5. System repeatability: When the test is repeated 3 times with 100 mL of the standard solution under the above operat- ‌956‌ ing conditions, the relative standard deviation of the peak area of erythromycin is not more than 3.0z. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Culture medium—Use the medium i in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Erythromycin RS, equivalent to about 25 mg (potency), dissolve in 25 mL of methanol, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 59C or below, and use within 7 days. Take exactly a suitable amount of the standard stock solu-tion before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solution and the low concentra-tion standard solution, respectively. Sample solutions—Weigh accurately an amount of Erythromycin, equivalent to about 25 mg (potency), dissolve in 25 mL of methanol, and add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Estazolam, when dried, contains not less than 98.
Erythromycin Stearate occurs as a white pow-der. It is freely soluble in ethanol (95) and in acetone, soluble in methanol, and practically insoluble in water.
(1) Dissolve 3 mg of Erythromycin Stearate in 2 mL of acetone, and add 2 mL of hydrochloric acid: an orange color develops and is immediately changed to red to deep purple. Determine the infrared absorption spectrum of Erythromycin Stearate, previously dried in a desiccator (reduced pressure, silica gel) for 24 hours, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration).
(1) Clarity and color of solution—Dissolve 0.10 g of Estazolam in 10 mL of ethanol (95): the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Estazolam in 10 mL of ethanol (95) by heating, add 40 mL of water, cool ‌JP XVIII Official Monographs / Estradiol Benzoate 959 with shaking in ice water, allow to stand to attain ordinary temperature, and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS and 6 mL of ethanol (95) (not more than 0.015z). Heavy metals <1.07>—Proceed with 1.0 g of Estazol- am according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Estazolam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Estazolam in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of hexane, chlo-roform and methanol (5:3:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the principal spot from the standard solu-tion. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (2 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Culture medium—Use the medium i in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Estazolam, when dried, contains not less than 98.5z of estazolam (C16H11ClN4). Description Estazolam occurs as white to pale yellow-white, crystals or crystalline powder. It is odorless, and has a bitter taste. It is soluble in methanol and in acetic anhydride, sparingly soluble in ethanol (95), and practically insoluble in water and in diethyl ether. Identification (1) Dissolve 0.01 g of Estazolam in 3 mL of sulfuric acid: the solution shows a yellow-green fluorescence under ultraviolet light (main wavelength: 365 nm). Determine the absorption spectrum of a solution of Estazolam in 1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Perform the test with Estazolam as directed under Flame Coloration Test <1.04> (2): a green color appears. Melting point <2.60> 229 – 2339C Purity (1) Clarity and color of solution—Dissolve 0.10 g of Estazolam in 10 mL of ethanol (95): the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Estazolam in 10 mL of ethanol (95) by heating, add 40 mL of water, cool ‌JP XVIII Official Monographs / Estradiol Benzoate 959 with shaking in ice water, allow to stand to attain ordinary temperature, and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS and 6 mL of ethanol (95) (not more than 0.015z). Heavy metals <1.07>—Proceed with 1.0 g of Estazol- am according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Estazolam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Estazolam in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of hexane, chlo-roform and methanol (5:3:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the principal spot from the standard solu-tion. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (2 g). Assay Weigh accurately about 0.25 g of Estazolam, previ-ously dried, dissolve in 100 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration), until the solution changes to the second equivalence point. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 14.74 mg of C16H11ClN4
エストラジオール安息香酸エステル
Estradiol Benzoate, when dried, contains not less than 97.
Estradiol Benzoate occurs as a white crystal-line powder. It is odorless. It is sparingly soluble in acetone, slightly soluble in metha-nol, in ethanol (95) and in diethyl ether, and practically in- soluble in water.
(1) To 2 mg of Estradiol Benzoate add 2 mL of sulfuric acid: a yellowish green color with a blue fluorescence is produced, and the color of the solution changes to light orange on the careful addition of 2 mL of water. (2) Determine the infrared absorption spectrum of Estra-diol Benzoate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of dried Estradiol Benzoate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋54 – ＋589(after drying, 0.1 g, acetone, 10 mL, 100 mm).
(1) 3,17a-Estradiol—Dissolve 5.0 mg each of Estradiol Benzoate and Estradiol Benzoate RS in acetone to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Place exactly 2 mL each of the sample solution and standard solu-tion in separate glass-stoppered test tube, add boiling stones, evaporate the acetone by heating in a water bath, and dry the residue in a desiccator (in vacuum, phosphorus (V) oxide) for 1 hour. Add 1.0 mL of dilute iron-phenol TS to each test tube. Stopper the test tubes loosely, heat for 30 seconds in a water bath, shake in a water bath for several seconds, and heat for 2 minutes. Cool the solutions in ice for 2 minutes, add 4.0 mL of diluted sulfuric acid (7 in 20), and mix well: the solution obtained from the sample solution has no more color than that from the standard solution. (2) Related substances—Dissolve 40 mg of Estradiol Benzoate in 2 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.2z (0.1 g).
Weigh accurately about 10 mg each of Estradiol Benzoate and Estradiol Benzoate RS, previously dried, and dissolve each in methanol to make exactly 20 mL. Pipet 5 mL each of these solutions, add 5 mL of the internal stand-ard solution, then add methanol to make 20 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 5 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of estradiol benzoate to that of the internal standard. Amount (mg) of estradiol benzoate (C25H28O3) ＝ MS × QT/QS MS: Amount (mg) of Estradiol Benzoate RS taken ‌960‌ Internal standard solution—A solution of progesterone in methanol (13 in 80,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile and water (7:3). Flow rate: Adjust so that the retention time of estradiol benzoate is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL Assay Measure exactly a volume of well-mixed Estradiol Benzoate Injection (Aqueous Suspension), equivalent to about 2 mg of estradiol benzoate (C25H28O3), dissolve the crystals with an appropriate quantity of methanol, and add methanol to make exactly 20 mL. Pipet 10 mL of this solu-tion, add exactly 10 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Estradiol Benzoate RS, previously dried in desiccator (reduced pressure, phosphorus (V) oxide) for 4 hours, and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and methanol to make 100 mL, and use this solu-tion as the standard solution. Proceed with these solutions as directed in the Assay under Estradiol Benzoate. of the standard solution under the above operating condi-tions, the internal standard and estradiol benzoate are eluted in this order with the resolution between these peaks being Amount (mg) of estradiol benzoate (C ＝ MS × QT/QS × 1/5 25H 28O3) not less than 9. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of estradiol benzoate to that of the internal standard is not more than 1.0z.
Estriol, when dried, contains not less than 97.
Estriol occurs as a white crystalline powder. It is odorless. It is sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water.
(1) Dissolve 0.01 g of Estriol in 100 mL of ethanol (95) by warming, and use this solution as the sample solution. Evaporate 1 mL of the sample solution on a water bath to dryness, add 5 mL of a solution of sodium p-phenol-sulfonate in diluted phosphoric acid (1 in 50), heat at 1509C for 10 minutes, and cool: a red-purple color develops. Determine the absorption spectrum of the sample solution obtained in (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Estriol RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Estriol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Estriol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋55 – ＋659(after drying, 40 ‌JP XVIII Official Monographs / Estriol Tablets 961 mg, ethanol (99.5), 10 mL, 100 mm).
Weigh accurately about 25 mg each of Estriol and Estriol RS, previously dried, and dissolve each in methanol to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 5 mL of the internal standard solution, add methanol to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of estriol to that of the internal standard. Estriol Injection (Aqueous Suspension)
Etacrynic Acid, when dried, contains not less than 98.
Etacrynic Acid occurs as a white crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in methanol, freely soluble in ethanol (95), in acetic acid (100) and in diethyl ether, and very slightly soluble in water.
(1) Dissolve 0.2 g of Etacrynic Acid in 10 mL of acetic acid (100), and to 5 mL of this solution add 0.1 mL of bromine TS: the color of the test solution disappears. To the remaining 5 mL of the solution add 0.1 mL of potas-sium permanganate TS: the color of the test solution changes to light orange immediately. To 0.01 g of Etacrynic Acid add 1 mL of sodium hy-droxide TS, and heat in a water bath for 3 minutes. After cooling, add 1 mL of chromotropic acid TS, and heat in a water bath for 10 minutes: a deep purple color develops. Determine the absorption spectrum of a solution of Etacrynic Acid in methanol (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. JP XVIII Official Monographs / Etacrynic Acid Tablets 963 Perform the test with Etacrynic Acid as directed under Flame Coloration Test <1.04> (2): a green color ap-pears.
(1) Clarity and color of solution—Dissolve 1.0 g of Etacrynic Acid in 10 mL of methanol: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Eta- ‌crynic Acid according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Etacrynic Acid according to Method 3, and perform the test. Add 10 mL of a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and fire to burn (not more than 2 ppm). Related substances—Dissolve 0.20 g of Etacrynic Acid in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 3 mL of the sample solution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, ethyl acetate and acetic acid (100) (6:5:2) to a dis-tance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.25z (1 g, in vacu-um, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Etacrynic Acid, pre-viously dried, place in an iodine bottle, dissolve in 20 mL of acetic acid (100), and add exactly 20 mL of 0.05 mol/L bro-mine VS. To this solution add 3 mL of hydrochloric acid, stopper tightly at once, shake, and allow to stand in a dark place for 60 minutes. Add carefully 50 mL of water and 15 mL of potassium iodide TS, stopper tightly at once, shake well, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Per-form a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 15.16 mg of C13H12Cl2O4
Acrinol Hydrate occurs as a yellow crystalline powder. It is sparingly soluble in water, in methanol and in ethanol (99.5). Melting point: about 2459C (with decomposition). The pH of a solution of Acrinol Hydrate (1 in 100) is between 5.5 and 7.0.
(1) Determine the absorption spectrum of a solution of Acrinol Hydrate (3 in 250,000) as directed under ‌JP XVIII Official Monographs / Acrinol and Zinc Oxide Oil 415 Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Acrinol Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Acrinol Hydrate (1 in 100) add 5 mL of dilute sulfuric acid, shake well, allow to stand for about 10 minutes at room temperature, and filter: the fil-trate responds to Qualitative Tests <1.09> for lactate.
(1) Chloride <1.03>—Dissolve 1.0 g of Acrinol Hy-drate in 80 mL of water by warming on a water bath, cool, and add 10 mL of sodium hydroxide TS and water to make 100 mL. Shake well, allow to stand for 30 minutes, filter, to 40 mL of the filtrate add 7 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare 50 mL of the control solution with 4 mL of sodium hydroxide TS, 7 mL of dilute nitric acid, 0.30 mL of 0.01 mol/L hydrochloric acid VS and sufficient water (not more than 0.026z). Heavy metals <1.07>—Proceed with 1.0 g of Acrinol Hydrate according to Method 2, and perform the test. Pre- System suitability— Test for required detectability: Confirm that the peak area of acrinol obtained with 10 mL of the standard solution (2) is equivalent to 7 to 13z of that with 10 mL of the standard solution (1). System performance: When the procedure is run with 10 mL of the standard solution (1) under the above operating conditions, the number of theoretical plates and the symme-try factor of the peak of acrinol are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution (1) under the above operating conditions, the relative standard deviation of the peak area of acrinol is not more than 1.5z. Water <2.48> 4.5 – 5.5z (0.2 g, volumetric titration, direct titration) Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.27 g of Acrinol Hydrate, dissolve in 5 mL of formic acid, add 60 mL of a mixture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> immediately with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.34 mg of C H N O.C H O pare the control solution with 2.0 mL of Standard Lead So- 15 15 3 3 6 3 lution (not more than 20 ppm). Volatile fatty acids—Dissolve 0.5 g of Acrinol Hy-drate in a mixture of 20 mL of water and 5 mL of dilute sul-furic acid, shake well, filter, and heat the filtrate: no odor of volatile fatty acids is perceptible. Related substances—Dissolve 10 mg of Acrinol Hy-drate in 25 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solu-tion as the standard solution (1). Pipet 1 mL of the standard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 10 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than acrinol obtained from the sample solution is not larger than 3 times the peak area of acrinol from the standard solution (2), and the total area of the peaks other than acrinol from the sample solu-tion is not larger than the peak area of acrinol from the standard solution (1). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate in 900 mL of water, adjust to pH 2.8 with phosphoric acid, and add water to make 1000 mL. To 700 mL of this solution add 300 mL of acetonitrile for liquid chromatogra-phy, and add 1.0 g of sodium 1-octanesulfonate to dissolve. Flow rate: Adjust so that the retention time of acrinol is about 15 minutes. Time span of measurement: About 3 times as long as the retention time of acrinol, beginning after the solvent peak.
Ethambutol Hydrochloride, when dried, contains not less than 98.
Ethambutol Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a bitter taste. It is very soluble in water, soluble in methanol and in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution prepared by dissolving 1.0 g of Ethambutol Hydrochloride in 20 mL of water is between 3.4 and 4.0.
(1) To 10 mL of a solution of Ethambutol Hydrochloride (1 in 100) add 0.5 mL of copper (II) sulfate TS and 2 mL of sodium hydroxide TS: a deep blue color is produced. Dissolve 0.1 g of Ethambutol Hydrochloride in 40 mL of water, add 20 mL of 2,4,6-trinitrophenol TS, and allow to stand for 1 hour. Collect the precipitate, wash with solution, corresponding to that from the standard solution, has no more color than that from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Ethambutol Hydro-chloride, previously dried, dissolve in 20 mL of water, and add 1.8 mL of copper (II) sulfate TS. To the solution add 7 mL of sodium hydroxide TS with shaking, add water to make exactly 50 mL, and centrifuge. Pipet 10 mL of the su-pernatant liquid, add 10 mL of ammonia-ammonium chlo-ride buffer solution (pH 10.0) and 100 mL of water, and titrate <2.50> with 0.01 mol/L disodium dihydrogen ethyl-enediamine tetraacetate VS until the color of the solution changes from blue-purple through light red to light yellow (indicator: 0.15 mL of Cu-PAN TS). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.772 mg of C10H24N2O2.2HCl
Ethenzamide, when dried, contains not less than 98.
Ethenzamide occurs as white, crystals or crys-talline powder. It is soluble in methanol, in ethanol (95), and in acetone, and practically insoluble in water. It begins to sublime slightly at about 1059C.
(1) Determine the absorption spectrum of a solution of Ethenzamide in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ethenzamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ethenzamide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Ethenzamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.5 g of Ethen-zamide in 30 mL of acetone, add 6 mL of dilute nitric acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.7 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone and 6 mL of dilute nitric acid, and dilute with water to make 50 mL (not more than 0.050z). Sulfate <1.14>—Dissolve 0.5 g of Ethenzamide in 30 mL of acetone, add 1 mL of dilute hydrochloric acid, and dilute with water to 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.50 mL of 0.005 mol/L sulfuric acid VS add 30 mL of acetone and 1 mL of dilute hydrochloric acid, and dilute with water to 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Ethen- zamide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—To 0.40 g of Ethenzamide add 0.3 g of potassium nitrate and 0.5 g of anhydrous sodium carbon-ate, mix thoroughly, ignite the mixture gradually, and cool. Dissolve the residue in 10 mL of dilute sulfuric acid, and heat the solution until white fumes begin to evolve. After cooling, add water carefully to make 5 mL, use this solution as the test solution, and perform the test (not more than 5 ppm). Salicylamide—Dissolve 0.20 g of Ethenzamide in 15 mL of diluted ethanol (95) (2 in 3), and add 2 to 3 drops of dilute iron (III) chloride TS: no purple color develops. Loss on drying <2.41> Not more than 1.0z (1 g, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Ethenzamide and Ethenzamide RS, previously dried, and dissolve each in 70 mL of ethanol (95) by warming, and after cooling, add ethanol (95) to make exactly 100 mL. Pipet 5 mL each of these solutions, add ethanol (95) to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absorbances, AT and AS, of the sample solution and standard solution at 290 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using ethanol (95) as the blank. Amount (mg) of ethenzamide (C9H11NO2) ＝ MS × AT/AS MS: Amount (mg) of Ethenzamide RS taken
Ether contains not less than 96z and not more than 98z (by specific gravity) of ether (C4H10O).
Ether is a colorless, clear, mobile liquid, hav-ing a characteristic odor. It is miscible with ethanol (95). It is soluble in water. It is highly volatile and flammable. It is slowly oxidized by the action of air and light, with the formation of peroxides. Its vapor, when mixed with air and ignited, may explode violently. Boiling point: 35 – 379C Specific gravity <2.56> d 20: 0.718 – 0.721
(1) Foreign odor—Place 10 mL of Ether in an evaporating dish, and allow it to evaporate spontaneously to a volume of about 1 mL: no foreign odor is perceptible. Drop this residue onto a piece of clean, odorless filter paper to evaporate the ether: no foreign odor is perceptible. Acidity—Place 10 mL of diluted ethanol (95) (4 in 5) and 0.5 mL of phenolphthalein TS in a 50-mL glass-stoppered flask, and add 0.02 mol/L sodium hydroxide VS dropwise to produce a red color which persists after shaking for 30 seconds. Add 25 mL of Ether, stopper the flask, shake gently, and add 0.40 mL of 0.02 mol/L sodium hy-droxide VS with shaking: a red color develops. Aldehyde—Place 10 mL of Ether in a Nessler tube, add 1 mL of potassium hydroxide TS, and allow the mixture ‌968‌ to stand for 2 hours, protecting from light, with occasional shaking: no color is produced in the ether layer and the aqueous layer. Peroxide—Place 10 mL of Ether in a Nessler tube, add 1 mL of a freshly prepared solution of potassium iodide (1 in 10), shake for 1 minute, then add 1 mL of starch TS, and shake well: no color is produced in the ether layer and in the aqueous layer. Residue on evaporation—Evaporate 140 mL of Ether, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Ethinylestradiol, when dried, contains not less than 98.
Ethinylestradiol occurs as white to pale yellow, crystals or crystalline powder. It is odorless. It is freely soluble in pyridine and in tetrahydrofuran, soluble in ethanol (95) and in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Dissolve 2 mg of Ethinylestradiol in 1 mL of a mixture of sulfuric acid and ethanol (95) (1:1): a purplish red color develops with a yellow-green fluorescence. Add carefully 2 mL of water to this solution: the color of the solution changes to red-purple. (2) Transfer 0.02 g of Ethinylestradiol to a glass-stoppered test tube, dissolve in 10 mL of a solution of potas-sium hydroxide (1 in 20), add 0.1 g of benzoyl chloride, and shake. Collect the resulting precipitate, recrystallize from methanol, and dry in a desiccator (in vacuum, phosphorus (V) oxide): the precipitate melts <2.60> between 2009C and 2029C. D Optical rotation <2.49> [a]20: －26 – －319(after drying, 0.1 g, pyridine, 25 mL, 100 mm).
Estrone—Dissolve 5 mg of Ethinylestradiol in 0.5 mL of ethanol (95), and add 0.05 g of 1,3-dinitrobenzene. Add 0.5 mL of freshly prepared dilute potassium hydroxide-ethanol TS, allow to stand in a dark place for 1 hour, and add 10 mL of ethanol (95): the solution has no more color than the following control solution. Control solution: Proceed in the same manner as men-tioned above, omitting Ethinylestradiol. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). ‌JP XVIII Official Monographs / Ethinylestradiol Tablets 969 Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.2 g of Ethinylestradiol, previously dried, and dissolve in 40 mL of tetrahydrofuran. Add 10 mL of a solution of silver nitrate (1 in 20), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 29.64 mg of C20H24O2
Ethionamide, when dried, contains not less than 98.
Ethionamide occurs as yellow, crystals or crys-talline powder, having a characteristic odor. It is soluble in methanol and in acetic acid (100), sparingly soluble in ethanol (99.5) and in acetone, and practically in-soluble in water.
(1) Determine the absorption spectrum of a solution of Ethionamide in methanol (3 in 160,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ethionamide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Acidity—Dissolve 3.0 g of Ethionamide in 30 mL of methanol by warming, add 90 mL of water, allow to stand in ice water for 1 hour, and filter. To 80 mL of the fil-trate add 0.8 mL of cresol red TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Ethionamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ethionamide according to Method 3. Add 10 mL of a so-lution of magnesium nitrate hexahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and fire to burn (not more than 2 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.20 g of Ethionamide in 10 mL of acetone, and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solution (1). Separately, pipet exactly 0.2 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop with a mixture of ethyl acetate, hexane and metha-nol (6:2:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution (1), and number of the spot other than the principal spot from the sample solution which is more in-tense than the spot from the standard solution (2) is not more than one. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Ethionamide, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from orange-red to dark orange-brown (indicator: 2 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.62 mg of C8H10N2S
Ethosuximide occurs as a white, paraffin-like solid or powder. It is odorless or has a slight, characteristic odor. It is very soluble in methanol, in ethanol (95), in diethyl ether, and in N,N-dimethylformamide, and freely soluble in water. Melting point: about 489C
(1) To 0.2 g of Ethosuximide add 10 mL of sodium hydroxide TS, and boil: the gas evolved turns a moistened red litmus paper blue. Dissolve 0.05 g of Ethosuximide in 1 mL of ethanol (95), add 3 drops of a solution of copper (II) acetate mono-hydrate (1 in 100), warm slightly, and add 1 to 2 drops of so-dium hydroxide TS: a purple color is produced. Determine the absorption spectrum of a solution of Ethosuximide in ethanol (95) (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths.
(1) Clarity and color of solution—Dissolve 1.0 g of Ethosuximide in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—With 1.0 g of Ethosuximide, per- form the test. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). JP XVIII Official Monographs / Ethylcellulose 971 ‌Heavy metals <1.07>—Proceed with 1.0 g of Ethosux-imide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).‌ Arsenic <1.11>—Prepare the test solution with 1.0 g of Ethosuximide, according to Method 1, and perform the test (not more than 2 ppm). Acid anhydride—Dissolve 0.50 g of Ethosuximide in 1 mL of ethanol (95), add 1 mL of hydroxylammonium chlo-ride-iron (III) chloride TS, and allow to stand for 5 minutes. Add 3 mL of water, mix, and allow to stand for 5 minutes: the red to red-purple color of this solution is not more in-tense than that of the following control solution. Control solution: Dissolve 70 mg of succinic anhydride in ethanol (95) to make exactly 100 mL. To 1.0 mL of this solu-tion add 1 mL of hydroxylammonium chloride-iron (III) chloride TS, and proceed in the same manner. Cyanide—Dissolve 1.0 g of Ethosuximide in 10 mL of ethanol (95), and add 3 drops of iron (II) sulfate TS, 1 mL of sodium hydroxide TS and 2 to 3 drops of iron (III) chlo-ride TS. Warm gently, and acidify with dilute sulfuric acid: not a blue precipitate and a blue color are produced within 15 minutes. Water <2.48> Not more than 0.5z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Ethosuximide, dis-solve in 20 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 14.12 mg of C7H11NO2
Ethyl Icosapentate contains not less than 96.
Icosapentate is a colorless or pale yellow, clear liquid. It has a faint, characteristic odor. It is miscible with ethanol (99.5), with acetic acid (100) and with hexane. It is practically insoluble in water and in ethyl-ene glycol.
(1) To 20 mg of Ethyl Icosapentate add 3 mL of a solution of potassium hydroxide in ethylene glycol (21 in 100), stopper tightly while passing a current of nitro-gen, and heat at 1809C for 15 minutes. After cooling, add methanol to make 100 mL. To 4 mL of this solution add methanol to make 100 mL, and use this solution as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution, prepared in the same manner as the sample solution with 3 mL of the solution of potassium hydroxide in ethylene glycol (21 in 100), as a con-trol, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ethyl Icosapentate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ethyl Icosapentate as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Ethyl Icosapentate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.481 – 1.491 Specific gravity <2.56> d 20: 0.905 – 0.915 Acid value <1.13> Not more than 0.5. Saponification value <1.13> 165 – 175 Iodine value <1.13> 365 – 395 Perform the test with 20 mg of Ethyl Icosapentate.
(1) Heavy metals <1.07>—Mix 1.0 g of Ethyl Icosapentate with ethanol (99.5), and add 2 mL of dilute ace-tic acid and ethanol (99.5) to make 50 mL. Perform the test with this solution as the test solution. Control solution: To 1.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and ethanol (99.5) to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ethyl Icosapentate according to Method 3, and perform the test (not more than 2 ppm). Related substances—To 0.40 g of Ethyl Icosapentate add hexane to make 50 mL, and use this solution as the sam-ple solution. Perform the test with 1.5 mL of the sample solu- JP XVIII Official Monographs / Ethyl Icosapentate Capsules 975 ‌tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of these peaks by the area percentage method: the amount of the peak, having the relative retention time of about 0.53 to ethyl icosapentate, is not more than 0.5z, the amount of each peak, having the relative retention time of about 0.80 and 0.93, is not more than 1.0z, the amount of each peak other than the principal peak and the peak mentioned above is not more than 1.0z, and the total amount of these peaks other than the principal peak is not more than 3.5z. Operating conditions— Detector, column, column temperature, carrier gas and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of ethyl icosapentate, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the sample solution add hexane to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add hexane to make exactly 10 mL. Confirm that the peak area of ethyl icosapentate obtained with 1.5 mL of this solution is equivalent to 7 to 13z of that with 1.5 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 1.5 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ethyl icosapentate is not more than 2.0z. Peroxide—Weigh accurately about 1 g of Ethyl Icosapentate, put in a 200-mL glass-stoppered conical flask, add 25 mL of a mixture of acetic acid (100) and chloroform (3:2), and dissolve by gentle shaking. Add 1 mL of saturated potassium iodide solution TS, immediately stopper tightly, shake gently, and allow to stand in a dark place for 10 minutes. Then add 30 mL of water, shake vigorously for 5 to 10 seconds, and titrate <2.50> with 0.01 mol/L sodium thio-sulfate VS until the blue color of the solution disappears after addition of 1 mL of starch TS. Calculate the amount of peroxide by the following equation: not more than 2 mEq/kg. Amount (mEq/kg) of peroxide ＝ V/M × 10 V: Volume (mL) of 0.01 mol/L sodium thiosulfate VS consumed M: Amount (g) of Ethyl Icosapentate taken Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Ethyl Icosapentate, and add hexane to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of Ethyl Icosapentate RS, and add hexane to make exactly 10 mL. Pipet 2 mL of this solu-tion, add exactly 2 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 3 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethyl icosapentate to that of the internal standard. Amount (mg) of ethyl icosapentate (C22H34O2) ＝ MS × QT/QS × 5 MS: Amount (mg) of Ethyl Icosapentate RS taken Internal standard solution—A solution of methyl docosanate in hexane (1 in 125). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column in 4 mm inside diameter and 1.8 m in length, packed with siliceous earth for gas chroma-tography (175 to 246 mm in particle diameter), coated with diethylene glycol succinate polyester for gas chromatography in the ratio of 25z. Column temperature: A constant temperature of about 1909C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethyl icosapentate is about 30 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, the internal standard and ethyl icosapentate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ethyl icosapentate to that of the internal stand-ard is not more than 1.0z.
Containers—Tight containers. Storage—Being fully filled, or replacing the air with Nitro-
Ethyl Loflazepate, when dried, contains not less than 98.
Ethyl Loflazepate occurs as a white crystalline powder. It is freely soluble in dimethylsulfoxide, sparingly soluble in acetonitrile, slightly soluble in ethanol (99.5), and practi-cally insoluble in water. A solution of Ethyl Loflazepate in dimethylsulfoxide (1 in 50) shows no optical rotation. Melting point: about 1999C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ethyl Loflazepate in acetonitrile (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ethyl Loflazepate RS prepared in the same manner as the sample solution: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. (2) Determine the infrared absorption spectrum of Ethyl Loflazepate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of dried Ethyl Loflazepate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Soluble halides—To 1.0 g of Ethyl Loflazepate add 50 mL of water, allow to stand for 1 hour with occa-sional shaking, and filter. Discard the first 10 mL of the fil-trate, transfer 25 mL of the subsequent filtrate to a Nessler tube, add 6 mL of dilute nitric acid and water to make 50 mL, and use this solution as the test solution. Proceed as di-rected under Chloride Limit Test <1.03>. Prepare the control solution as follows: to 0.20 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL. Heavy metals <1.07>—Proceed with 1.0 g of Ethyl Loflazepate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ethyl Loflazepate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 20 mg of Ethyl Loflazepate in 20 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu- JP XVIII Official Monographs / Ethyl Loflazepate 977 tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with QS, of the peak area of ethyl loflazepate to that of the inter-nal standard. exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each Amount (mg) of ethyl loflazepate (C ＝ MS × QT/QS 18H14ClFN2O3) peak area by the automatic integration method: the peak area of the related substance A, having the relative retention time of about 1.15 to ethyl loflazepate, obtained from the sample solution is not larger than 1/5 times the peak area of ethyl loflazepate from the standard solution, the peak area of the related substance B, having the relative retention time of about 1.38, from the sample solution is not larger than 7/10 times the peak area of ethyl loflazepate from the stand-ard solution, and the area of the peak other than ethyl loflazepate and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of ethyl loflazepate from the standard solution. Furthermore, the total area of the peaks other than ethyl loflazepate from the sample solution is not larger than the peak area of ethyl loflazepate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.9 g of sodium dihydrogen phos-phate dihydrate in water to make 1000 mL, and adjust to pH 6.0 with a solution prepared by dissolving 9.0 g of disodium hydrogen phosphate dodecahydrate in water to make 1000 mL. To 500 mL of this solution add 500 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of ethyl loflazepate is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of ethyl loflazepate. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of ethyl loflazepate obtained with 5 mL of this solution is equivalent to 4 to 6z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of ethyl loflazepate are not less than 2500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ethyl loflazepate is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (0.2 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g, platinum crucible).
Weigh accurately about 10 mg each of Ethyl Loflazepate and Ethyl Loflazepate RS, both previously dried, add the internal standard solution to dissolve to make exactly 100 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and MS: Amount (mg) of Ethyl Loflazepate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile for liquid chromatography (1 in 3000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 229 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile for liquid chromatography and ethanol (95) (2:1:1). Flow rate: Adjust so that the retention time of ethyl loflazepate is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ethyl loflazepate are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ethyl loflazepate to that of the internal standard is not more than 1.0z.
Ethyl Parahydroxybenzoate contains not less than 98.
Ethyl Parahydroxybenzoate occurs as color-less crystals or a white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tone, and very slightly soluble in water.◆
Determine the infrared absorption spectrum of Ethyl Parahydroxybenzoate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Ethyl Parahydroxybenzoate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Ethyl Parahydroxybenzoate in ethanol (95) to make 10 mL: the solution is clear and not more intensely colored than the following control solution. Control solution: To 5.0 mL of Cobalt (II) Chloride CS, ‌12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity—To 2 mL of the solution of Ethyl Parahy-droxybenzoate obtained in (1) add 3 mL of ethanol (95), add 5 mL of freshly boiled and cooled water and 0.1 mL of bromocresol green-sodium hydroxide-ethanol TS, then add 0.1 mol/L sodium hydroxide VS until the solution shows a blue color: the volume of 0.1 mol/L sodium hydroxide VS
Weigh accurately about 50.0 mg each of Ethyl Par-ahydroxybenzoate and Ethyl Parahydroxybenzoate RS, dis-solve separately in 2.5 mL each of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ethyl parahydroxybenzoate in each solution. used does not exceed 0.1 mL. (3) Heavy metals <1.07>—Dissolve 1.0 g of Ethyl Par-ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute Amount (mg) of ethyl parahydroxybenzoate (C9H ＝ MS × AT/AS 10O3) acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm).◆ Related substances—Dissolve 50.0 mg of Ethyl Par-ahydroxybenzoate in 2.5 mL of methanol, and add the mo-bile phase to make exactly 50 mL. Pipet 10 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak area of parahydroxybenzoic acid having a relative retention time of about 0.5 to ethyl parahy-droxybenzoate obtained from the sample solution is not larger than the peak area of ethyl parahydroxybenzoate from the standard solution (0.5z). For the peak area of para-hydroxybenzoic acid, multiply its correction factor, 1.4. Fur-thermore, the area of the peak other than ethyl parahydroxy-benzoate and parahydroxybenzoic acid from the sample so-lution is not larger than the peak area of ethyl parahydroxy-benzoate from the standard solution (0.5z), and the total area of the peaks other than ethyl parahydroxybenzoate is not larger than 2 times the peak area of ethyl parahydroxy-benzoate from the standard solution (1.0z). For this calcu-lation the peak area not larger than 1/5 times the peak area of ethyl parahydroxybenzoate from the standard solution is excluded (0.1z). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of ethyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of ethyl parahydroxybenzo-ate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ethyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g). MS: Amount (mg) of Ethyl Parahydroxybenzoate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and potassium dihy-drogen phosphate solution (17 in 2500) (13:7). Flow rate: 1.3 mL per minute. System suitability— System performance: Dissolve 5 mg each of Ethyl Parahy-droxybenzoate, methyl parahydroxybenzoate and para-hydroxybenzoic acid in the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, and add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, parahydroxybenzoic acid, methyl parahydroxybenzoate and ethyl parahydroxybenzoate are eluted in this order, the rela-tive retention times of parahydroxybenzoic acid and methyl parahydroxybenzoate to ethyl parahydroxybenzoate are about 0.5 and about 0.8, respectively, and the resolution be-tween the peaks of methyl parahydroxybenzoate and ethyl parahydroxybenzoate is not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ethyl parahydroxybenzoate is not more than 0.85z.
Ethylcellulose is a partly O-ethylated cellulose. It contains not less than 44.
Ethylcellulose occurs as a white to yellowish white, amorphous powder or grains. It is soluble in dichloromethane. It forms a slightly white-turbid or white-turbid, viscous liquid upon addition of ethanol (95). To 1 g of Ethylcellulose add 100 mL of hot water, shake to become turbid, cool to room temperature, and add freshly boiled and cooled water to make 100 mL: the solution is neu-tral.◆ Identiˆcation Spread 2 drops of a solution of Ethylcellu-lose in dichloromethane (1 in 25) between sodium chloride plates, then remove one of the plates to evaporate the sol-vent, and determine the infrared absorption spectrum of the plate as directed in the ˆlm method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Viscosity <2.53> Weigh exactly a quantity of Ethylcellulose, equivalent to 5.00 g calculated on the dried basis, add 95 g of a mixture of 80 g of toluene and 20 g of ethanol (95), and shake to dissolve. Perform the test with this solution at 259C as directed in Method I: not less than 80.0z and not more than 120.0z of the labeled viscosity for a nominal viscosity more than 6 mPa･s, and not less than 75.0z and not more than 140.0z of the labeled viscosity for a nominal viscosity not more than 6 mPa･s.
(1) Acidity or alkalinity—To 0.5 g of Ethylcellu-lose add 25 mL of freshly boiled and cooled water, shake for 15 minutes, ˆlter through a glass ˆlter (G3), and use the ˆl-trate as the sample solution. To 10 mL of the sample solu-tion add 0.1 mL of dilute phenolphthalein TS and 0.5 mL of 0.01 mol/L sodium hydroxide VS: a light red color develops. To 10 mL of the sample solution add 0.1 mL of methyl red-sodium hydroxide TS and 0.5 mL of 0.01 mol/L hydrochlo-ric acid VS: a red color develops. Chloride—Disperse 0.250 g of Ethylcellulose in 50 mL of water, and boil with occasional shaking. Allow to cool, and ˆlter. Discard the ˆrst 10 mL of the ˆltrate, to 10 mL of the subsequent ˆltrate add water to make 15 mL, and use this solution as the sample solution. Separately, to 10 mL of Standard Chloride Solution add 5 mL of water, and use this solution as the control solution. To 15 mL each of the sample solution and control solution add 1 mL of 2 mol/L nitric acid TS, transfer to test tubes containing 1 mL of a so-lution of silver nitrate (17 in 1000), allow to stand for 5 minutes protecting from light, and compare the opalescence developed in the both solutions against a black background by viewing transversely: the opalescence developed in the sample solution is not more intense than that of the control solution (not more than 0.1z). (3) Heavy metal <1.07>—Proceed with 1.0 g of Ethyl- cellulose according to Method 2, and perform the test. Pre-pare the control solution with 4.0 mL of Standard Lead So-lution (not more than 40 ppm).◇ Acetaldehyde—Introduce 3.0 g of Ethylcellulose into a 250-mL glass-stoppered conical ‰ask, add 10 mL of water, and stir for 1 hour. Allow to stand for 24 hours, ˆlter, add water to the ˆltrate to make 100 mL, and use this solution as the sample solution. Separately, dissolve 1.0 g of acetalde-hyde for assay in water to make 100 mL. To 5 mL of this so-lution add water to make 500 mL. To 3 mL of this solution add water to make 100 mL, and use this solution as the con-trol solution. Transfer 5 mL each of the sample solution and control solution to 25-mL volumetric ‰asks, add 5 mL of a solution of 3-methyl-2-benzothiazolonehydrazone hydro-chloride monohydrate (1 in 2000), and heat in a water bath at 609C for 5 minutes. Add 2 mL of iron (III) chloride-amidosulfuric acid TS, and warm again at 609C for 5 minutes. After cooling, add water to make 25 mL, and com-pare the color of these solutions: the sample solution is not more intensely colored than the control solution (not more than 100 ppm). Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 2 hours). Residue on Ignition <2.44> Not more than 0.5z (1 g).
Weigh accurately about 30 mg of Ethylcellulose, transfer to a 5-mL pressure-tight serum vial, add exactly 60 mg of adipic acid, 2 mL of the internal standard solution and 1 mL of hydroiodic acid, seal the vial immediately with a septum coated with ‰uororesin and an aluminum cap or any other sealing system providing a su‹cient air-tightness, and weigh accurately the vial. Take care not to mix the con-tents in the vial before heating. Place the vial in an oven or heat in a suitable heater with continuous stirring, maintain-ing an internal temperature of about 115 ± 29C for 70 min. Allow to cool, and weigh accurately the vial. If the diŠerence of the mass between before heating and after heating is more than 10 mg, prepare a new sample solution. If the diŠerence of the mass between before heating and after heating is not more than 10 mg, after phase separation, pierce through the septum of the vial with a cooled syringe, and withdraw a su‹cient volume of the upper phase as the sample solution. Separately, place exactly 60 mg of adipic acid, 2 mL of the internal standard solution and 1 mL of hydroiodic acid in another serum vial, and seal immediately. Weigh accurately the vial, inject 25 mL of iodoethane for assay through the septum in the vial, and weigh again accurately. Shake thor-oughly, after phase separation, pierce through the septum of the vial with a cooled syringe, withdraw a su‹cient volume of the upper phase, and use this solution as the standard so-lution. Perform the test with 1 mL each of the sample solu-tion and standard solution as directed under Gas Chroma-tography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iodo-ethane to that of the internal standard. Amount (z) of ethoxy group (C2H5O) ＝ MS/MT × QT/QS × 28.89 MS: Amount (mg) of iodoethane for assay taken MT: Amount (mg) of Ethylcellulose taken, calculated on the dried basis Internal standard solution—A solution of n-octane in o-xylene (1 in 200). Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Column temperature: Maintain the temperature at 509C for 3 minutes, raise to 1009C at a rate of 109C per minute, then to 2509C at a rate of 359C per minute, and maintain at 2509C for 8 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2809C. ‌JP XVIII Official Monographs / Ethyl L-Cysteine Hydrochloride 973 Carrier gas: Helium. Flow rate: 4.2 mL per minute (the retention time of the in-ternal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, iodoethane and the internal standard are eluted in this order with the relative retention time of iodoethane to the in-ternal standard being about 0.6, and with the resolution be-tween these peaks being not less than 5.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of iodoethane to that of the internal standard is not more than 2.0z.
Ethylenediamine contains not less than 97.
Ethylenediamine is a clear, colorless to pale yellow liquid. It has an ammonia-like odor. It is miscible with water, with ethanol (95) and with diethyl ether. It has a caustic nature and an irritating property. It is gradually affected by air. Specific gravity d 20: about 0.898
(1) A solution of Ethylenediamine (1 in 500) is alkaline. To 2 mL of copper (II) sulfate TS add 2 drops of Ethylenediamine: a blue-purple color develops. To 0.04 g of Ethylenediamine add 6 drops of benzoyl chloride and 2 mL of a solution of sodium hydroxide (1 in 10), warm for 2 to 3 minutes with occasional shaking, collect the white precipitate formed, and wash with water. Dissolve the precipitate in 8 mL of ethanol (95) by warming, promptly add 8 mL of water, cool, filter the crystals, wash with water, and dry at 1059C for 1 hour: it melts <2.60> between 2479C and 2519C.
(1) Heavy metals <1.07>—Place 1.0 g of Ethylene-diamine in a porcelain crucible, evaporate to dryness on a water bath, cover loosely, ignite at a low temperature until charred, proceed according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Residue on evaporation—Pipet 5 mL of Ethylene-diamine, heat on a water bath to dryness, and dry to con-stant mass at 1059C: the mass of the residue does not exceed 3.0 mg. Distilling range <2.57> 114 – 1199C, not less than 95 volz.
Weigh accurately about 0.7 g of Ethylenediamine in a glass-stoppered conical flask, add 50 mL of water, and titrate <2.50> with 1 mol/L hydrochloric acid VS (indicator: 3 drops of bromophenol blue TS). Each mL of 1 mol/L hydrochloric acid VS ＝ 30.05 mg of C2H8N2
Containers—Tight containers. Storage—Light-resistant, and almost well-filled.
Ethylmorphine Hydrochloride Hydrate occurs as white to pale yellow, crystals or crystalline powder. It is very soluble in methanol and in acetic acid (100), freely soluble in water, soluble in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in diethyl ether. It is affected by light. Melting point: about 1239C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ethylmorphine Hydrochloride Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Ethyl-morphine Hydrochloride Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A solution of Ethylmorphine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －103 – －1069(0.4 g calcu-lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Ethylmorphine Hydrochlo-ride Hydrate in 10 mL of water: the pH of this solution is be-tween 4.0 and 6.0.
Related substances—Dissolve 0.20 g of Ethylmor-phine Hydrochloride Hydrate in 10 mL of diluted ethanol (1 in 2), and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add diluted ethanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of ethanol (99.5), toluene, acetone and ammonia solution (28) (14:14:7:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Water <2.48> 8.0 – 10.0z (0.25 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.5 g of Ethylmorphine Hy-drochloride Hydrate, and dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.99 mg of C19H23NO3.HCl
Etilefrine Hydrochloride occurs as white, crys-tals or crystalline powder. It is very soluble in water, freely soluble in ethanol (99.5), and sparingly soluble in acetic acid (100). It is gradually colored to yellow-brown by light. A solution of Etilefrine Hydrochloride (1 in 20) shows no optical rotation.
(1) Dissolve 5 mg of Etilefrine Hydrochlo-ride in 100 mL of diluted hydrochloric acid (1 in 1000). De-termine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Etilefrine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Etilefrine Hydrochloride (1 in 1000) re-sponds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 0.5 g of Etilefrine Hydrochloride in 10 mL of water: the solution is clear and colorless. Acidity or alkalinity—To 10 mL of a solution of Etilefrine Hydrochloride (1 in 50) add 0.1 mL of methyl red TS for acid or alkali test and 0.2 mL of 0.01 mol/L sodium hydroxide VS: a yellow color develops, and the necessary volume of 0.01 mol/L hydrochloric acid VS to change the color to red is not more than 0.4 mL. Sulfate <1.14>—Perform the test with 0.85 g of Etilefrine Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.020z). Heavy metals <1.07>—Dissolve 1.0 g of Etilefrine Hy- drochloride in 30 mL of water and 2 mL of acetic acid (100), adjust with sodium hydroxide TS to a pH of 3.3, add water to make 50 mL, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of Etilefrine Hydro-chloride, previously dried, dissolve in 20 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correctoin. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.77 mg of C10H15NO2.HCl
Etizolam, when dried, contains not less than 98.
Etizolam occurs as a white to pale yellow-white crystalline powder. It is soluble in ethanol (99.5), sparingly soluble in aceto-nitrile and in acetic anhydride, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Etizolam in ethanol (99.5) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Etizolam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Etizolam according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Etizolam in 50 mL of acetonitrile, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add aceto- area of etizolam is not more than 2z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Etizolam, previ-ously dried, dissolve in 70 mL of a mixture of acetic anhy-dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). The end point is the second equivalent point. Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 17.14 mg of C17H15ClN4S
Etodolac, when dried, contains not less than 98.
Etodolac occurs as white to pale yellow, crys-tals or crystalline powder. It is freely soluble in methanol and in ethanol (99.5), and practically insoluble in water. A solution of Etodolac in methanol (1 in 50) shows no op-tical rotation. Melting point: about 1479C (with decomposition).
(1) Determine the absorption spectrum of a solution of Etodolac in ethanol (99.5) (3 in 200,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Etodolac as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Etodolac according to Method 2, and perform the test. Pre- um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Etodolac, previ-ously dried, dissolve in 50 mL of ethanol (99.5), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 28.74 mg of C17H21NO3
Etoposide occurs as white, crystals or crystal-line powder. It is sparingly soluble in methanol, slightly soluble in ethanol (99.5), and very slightly soluble in water. Melting point: about 2609C (with decomposition).
(1) Determine the absorption spectrum of a solution of Etoposide in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Etoposide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Etoposide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Etoposide RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: －100 – －1059(0.1 g calcu-lated on the anhydrous basis, methanol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Etoposide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So- ‌988‌ lution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Etoposide in 10 mL of methanol, add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the area of the peak other than etoposide obtained from the sample solution is not larger than 1/5 times the peak area of etoposide from the standard solution, and the total area of the peaks other than etoposide is not larger than 1/2 times the peak area of etoposide from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of etoposide, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of etoposide ob-tained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of etoposide is not more than 2.0z. Water <2.48> Not more than 4.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 25 mg each of Etoposide and Etoposide RS (previously determined the water <2.48> in the same manner as Etoposide) dissolve separately in metha-nol to make exactly 25 mL. Pipet 10 mL each of these solu-tions, add exactly 5 mL of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of etoposide to that of the internal standard. Flow rate: Adjust so that the retention time of etoposide is about 20 minutes. System suitability— System performance: Dissolve 10 mg of Etoposide in 2 mL of methanol, add 8 mL of the mobile phase, and mix well. Add 0.1 mL of diluted acetic acid (100) (1 in 25) and 0.1 mL of phenolphthalein TS, and add sodium hydroxide TS until the color of the solution changes to faintly red. After allowing to stand for 15 minutes, add 0.1 mL of diluted ace-tic acid (100) (1 in 25). When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak of etoposide and the peak hav-ing the relative retention time of about 1.3 to etoposide is not less than 3. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of etoposide to that of the internal standard is not more than 1.0z.
Famotidine, when dried, contains not less than 98.
Famotidine occurs as white to yellowish white crystals. It is freely soluble in acetic acid (100), slightly soluble in ethanol (95), and very slightly soluble in water. It dissolves in 0.5 mol/L hydrochloric acid TS. It is gradually colored by light. Melting point: about 1649C (with decomposition).
(1) Determine the absorption spectrum of a solution of Famotidine in 0.05 mol/L potassium dihydrogen phosphate TS (1 in 50,000) as directed under Ultraviolet- Amount (mg) of etoposide (C29 H32O 13) ＝ MS × QT/QS visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in- MS: Amount (mg) of Etoposide RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 2,6-dichloro-phenol in methanol (3 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 290 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with phenylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 6.44 g of sodium sulfate decahy-drate in diluted acetic acid (100) (1 in 100) to make 1000 mL, and add 250 mL of acetonitrile. tensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Famotidine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Famotidine in 10 mL of 0.5 mol/L hydrochloric acid TS: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 2.0 g of Famoti- dine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of Famotidine in ‌JP XVIII Official Monographs / Famotidine Injection 989 10 mL of acetic acid (100), and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, and add ace-tic acid (100) to make exactly 100 mL. Pipet 1 mL, 2 mL and 3 mL of this solution, add acetic acid (100) to make exactly 10 mL, respectively, and use these solutions as the standard solution (1), the standard solution (2) and the standard solu-tion (3). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel (5 to 7 mm) with fluorescent indicator for thin-layer chromatography, and dry in a stream of nitrogen. Develop the plate with a mixture of ethyl acetate, methanol, toluene and ammonia solution (28) (40:25:20:2) to a distance of about 8 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 254 nm): the spots other than the principal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution (3). Total intensity of the spots other than the principal spot and the spot of the starting point from the sample solution is not more than 0.5z calcu-lated on the basis of intensities of the spots from the stand-ard solution (1) and the standard solution (2). Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Famotidine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.87 mg of C8H15N7O2S3
Indefinite small masses; externally yellow-brown to dark yellow-brown; easily broken; fractured sur-face has a glassy luster, and is not wet. Usually in a gall sac, occasionally taken out, the gall sac consists of a fibrous and strong membrane, 9 – 15 cm in length and 7 – 9 cm in width; externally dark brown and translucent. Odor, slight and characteristic; taste, extremely bitter.
To 0.1 g of pulverized Bear Bile, add 5 mL of methanol, warm in a water bath for 10 minutes. After cooling, filter, and use the filtrate as the sample solution. Separately, dissolve 10 mg of sodium tauroursodeoxycholate for thin-layer chromatography in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of acetic acid (100), toluene and water (10:10:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the stand-ard solution.
Other animal biles—Use the sample solution ob-tained in the Identification as the sample solution. Sepa-rately, dissolve 10 mg of sodium glycocholate for thin-layer chromatography and 20 mg of powdered porcine bile for thin-layer chromatography in 5 mL each of methanol, and use these solutions as the standard solution (1) and (2), re-spectively. Perform the test with these solutions as directed in the Identification: no spot obtained from the sample solu-tion appears at the position of the spot from the standard so-lution (1) and no grayish brown to black spot appears at the position of the spot at an Rf value of about 0.3 from the standard solution (2).
Felbinac Cataplasm contains not less than 90.
Weigh a quantity of Felbinac Cataplasm, equivalent to 10 mg of Felbinac, cut into minute pieces, add 20 mL of methanol, shake for 30 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of felbinac for assay in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (50:25:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the principal spot obtained from the sample solution and the spot from the standard solution is the same. pH Being specified separately when the drug is granted ap-proval based on the Law. Adhesiveness Being specified separately when the drug is granted approval based on the Law. Drug release Being specified separately when the drug is granted approval based on the Law.
Take exactly a quantity of Felbinac Cataplasm, equivalent to 70 mg of felbinac (C14H12O2), cut into minute pieces, add 150 mL of methanol, and heat under a reflux condenser. After cooling, separate the extraction liquid, add 20 mL of water to the residue, heat in a water bath at 759C for 10 minutes, then add 150 mL of methanol, and heat under a reflux condenser. After cooling, separate the ex-traction liquid, add 150 mL of methanol to the residue, and heat under a reflux condenser. After cooling, separate the extraction liquid, wash the residue and vessels with a small amount of methanol, combine the extraction liquids and washings, and add methanol to make exactly 500 mL. Pipet 6 mL of this solution, add exactly 3 mL of the internal stand-ard solution, add methanol to make 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 35 mg of felbinac for assay, previously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 250 mL. Pipet 6 mL of this solution, add exactly 3 mL of the in-ternal standard solution, add methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of felbinac to that of the internal stand-ard. water to make 500 mL. To this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of felbinac is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, felbinac and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of felbinac to that of the internal standard is not more than 1.0z.
Felbinac Tape contains not less than 90.
Cut up a quantity of Felbinac Tape, equiva-lent to 5 mg of Felbinac, add 30 mL of ethanol (95), and heat under a reflux condenser. After cooling, separate the ethanol extract, add ethanol (95) to make 50 mL, and filter. To 5 mL of the filtrate add ethanol (95) to make 100 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 251 nm and 255 nm. Adhesiveness Being specified separately when the drug is granted approval based on the Law. Drug release Being specified separately when the drug is granted approval based on the Law.
Take exactly a quantity of Felbinac Tape, equivalent to 35 mg of felbinac (C14H12O2), cut up them, add 60 mL of acetone, sonicate, and heat under a reflux condenser. After cooling, separate the acetone extract, and repeat the ex-traction twice more with 60 mL each of acetone by heating under a reflux condenser. After cooling, separate the extract, wash the residue and vessel with a small volume of acetone, combine the washings and the extracts, and add acetone to ‌998 make exactly 250 mL. Pipet 6 mL of this solution, add ex-actly 2 mL of the internal standard solution and the mobile phase to make 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 14 mg of felbinac for assay, previously dried at 1059C for 3 hours, and dis-solve in acetone to make exactly 100 mL. Pipet 6 mL of this solution, add exactly 2 mL of the internal standard solution and the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of felbinac to that of the internal standard. Amount (mg) of felbinac (C14H12O2) ＝ MS × QT/QS × 5/2 MS: Amount (mg) of felbinac for assay taken Internal standard solution—A solution of indomethacin in acetone (1 in 1250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (500:500:1). Flow rate: Adjust so that the retention time of felbinac is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, felbinac and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of felbinac to that of the internal standard is not more than 1.0z.
Felbinac, when dried, contains not less than 98.
Felbinac occurs as white to pale yellow-white, crystals or crystalline powder. It is soluble in methanol and in acetone, sparingly soluble in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Felbinac in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Fel-binac as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Felbinac in 40 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution by combining 0.30 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of acetone and 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.011z). Heavy metals <1.07>—Proceed with 1.0 g of Felbinac according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Felbinac in 10 mL of acetone, and use this solution as the sample solution. Amount [mg (potency)] of faropenem (C ＝ MS × QT/QS 12H15NO5S) Pipet 2 mL of the sample solution, and add acetone to make exactly 100 mL. Pipet 5 mL of the sample solution, add acetone to make exactly 50 mL, and use this solution as the MS: Amount [mg (potency)] of Faropenem Sodium RS taken Internal standard solution—Dissolve 0.5 g of m-hydorxya-cetophenone in 20 mL of acetonitrile, and add water to make 200 mL.
Weigh accurately the mass of not less than 5 Faro-penem Sodium Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg (potency) of faropenem (C12H15NO5S), add exactly 10 mL of the inter-nal standard solution, shake well, and add water to make 50 mL. Filter, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg (potency) of Faropenem Sodium RS, add exactly 10 mL of the internal standarad solution and water to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Faropenem Sodium Hydrate. Description Felbinac occurs as white to pale yellow-white, crystals or crystalline powder. It is soluble in methanol and in acetone, sparingly soluble in ethanol (95), and practically insoluble in water. Identification (1) Determine the absorption spectrum of a solution of Felbinac in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Fel-binac as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 163 – 1669C Purity (1) Chloride <1.03>—Dissolve 1.0 g of Felbinac in 40 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution by combining 0.30 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of acetone and 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.011z). Heavy metals <1.07>—Proceed with 1.0 g of Felbinac according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Felbinac in 10 mL of acetone, and use this solution as the sample solution. Amount [mg (potency)] of faropenem (C ＝ MS × QT/QS 12H15NO5S) Pipet 2 mL of the sample solution, and add acetone to make exactly 100 mL. Pipet 5 mL of the sample solution, add acetone to make exactly 50 mL, and use this solution as the MS: Amount [mg (potency)] of Faropenem Sodium RS taken Internal standard solution—Dissolve 0.5 g of m-hydorxya-cetophenone in 20 mL of acetonitrile, and add water to make 200 mL.
Felodipine occurs as pale yellow-white to light yellow-white, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (99.5), and practically insoluble in water. A solution of Felodipine in methanol (1 in 20) shows no optical rotation. Identiˆcation (1) Determine the absorption spectrum of a solution of Felodipine in methanol (1 in 62,500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Felodipine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals—Being speciˆed separately when the drug is granted approval based on the Law. (2) Related substances—Dissolve 25 mg of Felodipine in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this so-lution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than felodipine, related substance B having the relative retention time of about 0.7 to felodipine, and related substance C having the relative retention time of about 1.4 obtained from the sample solution is not larger than the peak area of felodipine from the standard solution. Furthermore, the total area of the peaks of related substances B and C from the sample solution is not larger than 10 times the peak area of felodipine from the standard solution, and the total area of the peaks other than felodipine and related sub-stances mentioned above from the sample solution is not larger than 3 times the peak area of felodipine from the standard solution. For this calculation the peak area less than 1/5 times the peak area of felodipine from the standard solution is excluded. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.2 g of sodium dihydrogen phos-phate dihydrate in 400 mL of water, adjust to pH 3.0 with phosphoric acid, and add 200 mL of methanol and 400 mL of acetonitrile. Flow rate: Adjust so that the retention time of felodipine is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of felodipine, beginning after the solvent peak. System suitability— Test for required detectability: When the procedure is run with 20 mL of the standard solution under the above operat-ing conditions, the SN ratio of the peak of felodipine is not less than 30. System performance: When the procedure is run with 20 JP XVIII Official Monographs / Felodipine Tablets 999 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of felodipine are not less than 5000 and not more than 1.5, respectively. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of felodipine is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.16 g of Felodipine, dis-solve in 25 mL of t-butyl alcohol and 25 mL of diluted per-chloric acid (17 in 200), and titrate <2.50> with 0.1 mol/L cerium (IV) sulfate VS (indicator: 50 mL of 1,10-phenanthro-line TS) until the color of the solution changes from orange to colorless. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L cerium (IV) sulfate VS ＝ 19.21 mg of C18H19Cl2NO4
Fenbufen, when dried, contains not less than 98.
Fenbufen occurs as a white crystalline powder. It has a bitter taste. It is sparingly soluble in acetone, slightly soluble in metha-nol, in ethanol (95) and in diethyl ether, and practically in-soluble in water. Melting point: about 1889C (with decomposition).
(1) Determine the absorption spectrum of a Amount (mg) of felodipine (C18H19Cl2NO4) ＝ M S × QT/QS solution of Fenbufen in ethanol (95) (1 in 200,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, MS: Amount (mg) of felodipine for assay taken, calcu-lated on the dried basis Internal standard solution—A solution of butyl para-hydroxybenzoate in methanol (1 in 6000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 264 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, water, a solution of sodium perchlorate monohydrate (281 in 2000) and diluted perchloric acid (17 in 200) (65:25:8:2). Flow rate: Adjust so that the retention time of felodipine is about 12 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and felodipine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of felodipine is not more than 1.0z.
(1) Heavy metals <1.07>—Take 2.0 g of Fenbufen, add 2 mL of sulfuric acid, and carbonize by gentle heating, proceed according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Fenbufen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.1 g of Fenbufen in 20 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, meth-anol and water (80:20:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Fenbufen, previ-ously dried, dissolve in 100 mL of ethanol (99.5), and titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS (potentiometric titration). Perform a blank determination in ‌JP XVIII Official Monographs / Fenofibrate 1001 the same manner, and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 25.43 mg of C16H14O3
Fenofibrate, when dried, contains not less than 98.
Fenofibrate occurs as a white to pale yellow-white crystalline powder. It is soluble in ethanol (99.5), and practically insoluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Fenofibrate in ethanol (99.5) (1 in 80,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Fenofibrate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Fenofibrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of dried Fenofibrate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Fenofibrate as directed under Flame Coloration Test <1.04>: a green color appears.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Fenofibrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.10 g of Fenofibrate in a mixture of acetonitrile and water (7:3) to make 100 mL. To 5 mL of this solution add a mixture of acetonitrile and water (7:3) to make 25 mL, and use this solution as the sample so-lution. Pipet 3 mL of the sample solution, add a mixture of acetonitrile and water (7:3) to make exactly 50 mL. Pipet 2.5 mL of this solution, add a mixture of acetonitrile and water (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of related sub-stance A having the relative retention time of about 1.4 to fenofibrate, obtained from the sample solution is not larger than 4/5 times the peak area of fenofibrate from the stand-ard solution, and the total area of the peaks other than fenofibrate from the sample solution is not larger than the peak area of fenofibrate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of fenofibrate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of acetonitrile and water (7:3) to make exactly 25 mL. Confirm that the peak area of fenofibrate obtained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: Dissolve 0.10 g each of Fenofibrate and 4-chlorobenzophenon in 100 mL of a mixture of aceto-nitrile and water (7:3). To 2 mL of this solution add a mix-ture of acetonitrile and water (7:3) to make 50 mL. To 1 mL of this solution add a mixture of acetonitrile and water (7:3) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-chlo-robenzophenon and fenofibrate are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fenofibrate is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 50 mg each of Fenofibrate and Fenofibrate RS, both previously dried, dissolve each in a mixture of acetonitrile and water (7:3) to make exactly 50 mL. Pipet 2 mL each of these solutions, add exactly 2 mL of the internal standard solution to each, add a mixture of acetonitrile and water (7:3) to make 50 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of fenofibrate to the internal standard. Amount (mg) of fenofibrate (C20H21ClO4) ＝ MS × QT/QS MS: Amount (mg) of Fenofibrate RS taken Internal standard solution—A solution of 4-chloroben-zophenon in a mixture of acetonitrile and water (7:3) (11 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 286 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and 0.02 mol/L phosphate buffer solution (pH 3.0) (7:3). Flow rate: Adjust so that the retention time of fenofibrate ‌1002 is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and fenofibrate are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of fenofibrate to that of the internal standard is not more than 1.0z.
Fentanyl Citrate contains not less than 98.
Fentanyl Citrate occurs as white, crystals or crystalline powder. It is freely soluble in methanol and in acetic acid (100), sparingly soluble in water and in ethanol (95), and very slightly soluble in diethyl ether.
(1) Dissolve 0.05 g of Fentanyl Citrate in 10 mL of 0.1 mol/L hydrochloric acid TS and ethanol (95) to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Fen-tanyl Citrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Fentanyl Citrate (1 in 100) responds to Qualitative Tests <1.09> (1) for citrate. pH <2.54> Dissolve 0.10 g of Fentanyl Citrate in 10 mL of water: the pH of this solution is between 3.0 and 5.0.
(1) Heavy metals <1.07>—Proceed with 0.5 g of Fentanyl Citrate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Fentanyl Citrate in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from ‌1004‌ the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.2 g, in vacu-um, silica gel, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 tablets of Fenofibrate Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 50 mg of fenofibrate (C20H21ClO4), add 30 mL of a mixture of acetonitrile and water (7:3), shake thoroughly, and add a mixture of aceto-nitrile and water (7:3) to make exactly 50 mL. Centrifuge this solution, pipet 2 mL of the supernatant liquid, add ex-actly 2 mL of the internal standard solution, add a mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Fenofibrate RS, previously dried in vacuum over phosphorus (V) oxide at 609C for 4 hours, and dissolve in a mixture of acetonitrile and water (7:3) to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, add a mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of fenofibrate to that of the internal standard. Amount (mg) of fenofibrate (C20H21ClO4) ＝ MS × QT/QS MS: Amount (mg) of Fenofibrate RS for assay taken Internal standard solution—A solution of 4-chloroben-zophenon in a mixture of acetonitrile and water (7:3) (11 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Fenofibrate. System suitability— Proceed as directed in the system suitability in the Assay under Fenofibrate.
Ferrous Sulfate Hydrate contains not less than 98.
Ferrous Sulfate Hydrate occurs as pale green, crystals or crystalline powder. It is odorless, and has an astringent taste. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It is efflorescent in dry air, and its surface becomes yellow-ish brown in moist air.
A solution of Ferrous Sulfate Hydrate (1 in 10) responds to Qualitative Tests <1.09> for ferrous salt and for sulfate.
(1) Clarity of solution—Dissolve 1.0 g of Ferrous Sulfate Hydrate in 20 mL of water and 1 mL of dilute sulfu-ric acid: the solution is clear. Acidity—To 5.0 g of powdered Ferrous Sulfate Hy-drate add 50 mL of ethanol (95), shake well for 2 minutes, and filter the mixture. To 25 mL of the filtrate add 50 mL of water, 3 drops of bromothymol blue TS and 0.5 mL of dilute sodium hydroxide TS: a blue color develops. Heavy metals <1.07>—Take 1.0 g of Ferrous Sulfate Hydrate in a porcelain dish, add 3 mL of aqua regia, and dissolve. Then evaporate on a water bath to dryness. To the residue add 5 mL of 6 mol/L hydrochloric acid TS, and dis-solve. Transfer this solution to a separator. Wash the por-celain dish with two 5-mL portions of 6 mol/L hydrochloric acid TS, and combine the washings and the solution in the separator. Pour two 40-mL portions and one 20-mL portion of diethyl ether in the separator, shaking each time to mix. Allow to stand, and discard each separated diethyl ether layer. To the aqueous layer add 0.05 g of hydroxylammo-nium chloride, dissolve, and heat on a water bath for 10 minutes. Cool, adjust the solution to a pH of 3 to 4 by drop-ping ammonia solution (28), add water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution as follows: take 2.5 mL of Stand-ard Lead Solution in a porcelain dish, add 3 mL of aqua re-gia, and proceed as directed for the preparation of the test solution (not more than 25 ppm).
Fexofenadine Hydrochloride contains not less than 98.
Fexofenadine Hydrochloride occurs as a white crystalline powder. It is very soluble in methanol, soluble in ethanol (99.5), and slightly soluble in water. A solution of Fexofenadine Hydrochloride in methanol (3 in 100) shows no optical rotation. Fexofenadine Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Fexofenadine Hydrochloride in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Fexofenadine Hy-drochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Fex-ofenadine Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Fexofenadine Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the RS according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals. A solution of Fexofenadine Hydrochloride in a mix-ture of water and methanol (1:1) (3 in 200) responds to Qualitative Tests <1.09> (2) for chloride. ‌JP XVIII Official Monographs / Fexofenadine Hydrochloride Tablets 1005
(1) Heavy metals <1.07>—Proceed with 2.0 g of Fexofenadine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 7.51 g of sodium dihydrogen phosphate dihydrate and 0.84 g of sodium perchlorate in 1000 mL of water, and adjust to pH 2.0 with phosphoric acid. In a mixture of this solution and aceto-nitrile for liquid chromatography (1:1) dissolve 25 mg of Fexofenadine Hydrochloride to make 25 mL, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the area of the peak other than fexo-fenadine obtained from the sample solution is not larger than the peak area of fexofenadine from the standard solu-tion. For the areas of the peaks, having the relative retention time of about 1.8 and about 3.3 to fexofenadine, multiply their correction factor, 1.5 and 0.9, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 6 times as long as the retention time of fexofenadine, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fexofenadine are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fexofenadine is not more than 2.0z. Water <2.48> Not more than 0.5z (0.25 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve 7.51 g of sodium dihydrogen phosphate di-hydrate and 0.84 g of sodium perchlorate in 1000 mL of water, and adjust to pH 2.0 with phosphoric acid. In a mix-ture of this solution and acetonitrile for liquid chromatogra-phy (1:1) dissolve accurately weighed about 25 mg each of Fexofenadine Hydrochloride and Fexofenadine Hydrochlo-ride RS (separately determine the water <2.48> in the same manner as Fexofenadine Hydrochloride), to make exactly 25 mL each. Pipet 3 mL each of these solutions, add the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of fexofenadine in each solution. Amount (mg) of fexofenadine hydrochloride (C32H39NO4.HCl) ＝ MS × AT/AS MS: Amount (mg) of Fexofenadine Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 650 mL of a solution, prepared by dis-solving 7.51 g of sodium dihydrogen phosphate dihydrate and 0.84 g of sodium perchlorate in 1000 mL of water and adjusting to pH 2.0 with phosphoric acid, add 350 mL of acetonitrile for liquid chromatography and 3 mL of triethylamine. Flow rate: Adjust so that the retention time of fexofena-dine is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fexofenadine are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fexofenadine is not more than 1.0z.
Filgrastim (Genetical Recombination) is an aqueous solution in which a desired product is a recombinant N-methionyl human granulocyte colony-stimulating factor consisting of 175 amino acid residues. It contains not less than 0.
Filgrastim (Genetical Recombination) occurs as a clear and colorless liquid.
(1) Take a volume of Filgrastim (Genetical Recombination), equivalent to 5 to 10 mg of protein depend-ing on the size of polyacrylamide gel for filgrastim, and add 10 mL of water. To 3 volumes of this solution add 1 volume of buffer solution for filgrastim sample, and use this solu-tion as the sample solution. Separately, take a volume of Filgrastim RS which contains equal amount of protein to Fil-grastim (Genetical Recombination) used above, proceed as directed for the sample solution, and use the solution so ob-tained as the standard solution. Set a polyacrylamide gel for filgrastim up to the electrophoresis apparatus, and put a nec-essary amount of buffer solution for SDS-polyacrylamide gel electrophoresis in the upper and lower reservoirs. Pipet the all amount of the sample solution and standard solution into each well of the gel, and start the electrophoresis setting the electrode of the lower reservoir as the anode. Stop the electrophoresis when the bromophenol blue band has been migrated to about the lower end of the gel. When stain the gel with a staining solution, which is prepared by dissolving 1.25 g of Coomassie brilliant blue R250 in a mixture of 450 mL of methanol, 100 mL of acetic acid (100) and water to make 1000 mL, stained bands obtained from the sample so-lution appear as similar migrating image at the same position as those from the standard solution. (2) Take a volume of Filgrastim (Genetical Recombina-tion) and Filgrastim RS, equivalent to about 80 mg of pro-tein, add 200 mL of the buffer solution for enzyme digestion, and add water to make 390 mL. To each of these solution add 10 mL of a solution containing 50 mg of V8 protease in 250 mL of water, incubate at 259C for 17 to 19 hours, then add 18 mL of a mixture of water and trifluoroacetic acid (19:1) to stop the reaction, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 70 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. When the chro- matograms obtained from these solutions are compared, both chromatograms show the similar peaks at the same retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 214 nm). Column: A stainless steel column 2.1 mm in inside diame-ter and 25 cm in length, packed with butylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water and trifluoroacetic acid (1000:1). Mobile phase B: A mixture of acetonitrile, water and trifluoroacetic acid (9000:1000:9). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 2 2 – 30 98 → 70 2 → 30 30 – 85 70 → 50 30 → 50 85 – 90 50 → 2 50 → 98 90 – 100 Flow rate: 0.20 mL per minute. System suitability— System performance: When the procedure is run with 70 mL of the standard solution under the above operating con-ditions, the resolutions between each adjacent peakpair of the major 8 peaks, which are eluted after the solvent peak appeared within 10 minutes, are not less than 1.5. pH <2.54> 3.7 – 4.3
(1) Multimers—Perform the test with 250 mL of Filgrastim (Genetical Recombination) as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method. Calculate their amounts of the peaks by the area percentage method; the total amount of the peaks other than filgrastim is not more than 2z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with hydrophilic silica gel for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.8 g of sodium chloride in 10 mL of dilute acetic acid and 900 mL of water, adjust to pH 5.5 with sodium hydroxide TS, then add 250 mg of sodium lauryl sulfate, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of filgrastim is about 17 minutes. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the time when the elution of filgrastim is com-pleted. System suitability— Test for required detectability: Measure exactly 10 mL of Filgrastim (Genetical Recombination), and add the mobile phase to make exactly 1000 mL. Confirm that the peak area of filgrastim obtained with 250 mL of this solution is 0.7 to 1.3z of that with 250 mL of Filgrastim (Genetical Recombi-nation). System performance: When the procedure is run with 10 mL of a solution containing 12.5 mg of egg albumin and 12.5 mg of myoglobin in 5 mL of water under the above operat- ing conditions, egg albumin and myoglobin are eluted in this order with the resolution between these peaks being not less than 1.7. System repeatability: When the test is repeated 6 times with 250 mL of Filgrastim (Genetical Recombination) under the above operating conditions, the relative standard devia-tion of the peak area of filgrastim is not more than 2.5z. Charge isomer—Perform the test with 100 mL of Fil-
(1) Protein content—Perform the test with exactly 200 mL each of Filgrastim (Genetical Recombination) and Filgrastim RS as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of filgrastim. Amount (mg) of protein in 1 mL of Filgrastim (Genetical Recombination) ＝ C × A /A grastim (Genetical Recombination) as directed under Liquid T S Chromatography <2.01> according to the following condi-tions, and determine the each peak area by the automatic in-tegration method. Calculate their amounts of the peaks by the area percentage method; the amount of charge isomer, having the relative retention time of about 0.87 to filgrastim, is not more than 3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 35 mm in length, packed with strongly acidic ion-exchange non-porous resin for liquid chromatography (2.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: To 900 mL of water add 1.14 mL of acetic acid (100), adjust to pH 5.4 with sodium hydroxide TS, and add water to make 1000 mL. Mobile phase B: Dissolve 5.84 g of sodium chloride in 1.14 mL of acetic acid (100) and 900 mL of water, adjust to pH 5.4 with sodium hydroxide TS, and add water to make 1000 mL. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. C: Protein concentration (mg/mL) of Filgrastim RS Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, 1-propanol and trifluoroacetic acid (699:300:1). Mobile phase B: A mixture of 1-propanol, water and trifluoroacetic acid (800:199:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 2 2 – 13 90 → 70 10 → 30 13 – 15 70 → 0 30 → 100 15 – 18 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Flow rate: Adjust so that the retention time of filgrastim is about 15 minutes. System suitability— 0 – 2 100 0 2 – 10 100 → 40 0 → 60 10 – 11 40 → 100 60 → 0 11 – 20 100 0 Flow rate: Adjust so that the retention time of filgrastim is about 14 minutes. Time span of measurement: From 6 minutes to 17 minutes. System suitability— Test for required detectability: Confirm that when per-form the test with 100 mL of the system suitability test solu-tion for filgrastim under the above operating conditions, the content of charge isomer is between 1.4 to 2.6z. System performance: When the procedure is run with 100 mL of the system suitability test solution for filgrastim under the above operating conditions, charge isomer peak and fil-grastim are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 100 mL of Filgrastim (Genetical Recombination) under the above operating conditions, the relative standard devia-tion of the peak area of filgrastim is not more than 2.5z. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 0.25 EU/mL. System performance: When the procedure is run with 200 mL of a solution prepared by dissolving 1 mg of uracil and 2 mg of diphenyl in 100 mL of a mixture of water, 1-propanol and trifluoroacetic acid (649:350:1) under the above operat-ing conditions, uracil and diphenyl are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 200 mL of Filgrastim RS under the above operating con-ditions, the relative standard deviation of the peak area of filgrastim is not more than 2.5z. Specific activity— Test cell: 32D clone3 cell. Sample dilution solution for assay: To Iscove's modified Dulbecco's fluid medium for filgrastim add 200 mmol/L L-glutamine solution and fetal calf serum to make 1 volz and 5 volz solution, respectively, and sterilize by filtration. Standard solutions Dilute Filgrastim RS by the sample dilution solution for assay to prepare not less than 5 serial dilutions started from any concentration SH so that all of their protein concentrations are within the range of 0.5 to 6 ng/mL, and use them as the standard solutions. Sample solutions Dilute Filgrastim (Genetical Recombination) by the sample dilution solution for assay to prepare not less than 5 serial dilutions in equal ratio started from any concentration UH so that all of their protein con-centrations are within the range of 0.5 to 6 ng/mL, and use them as the sample solutions. Procedure The procedure before stopping the incu-bation should be performed under aseptic condition. ‌JP XVIII Official Monographs / Filgrastim (Genetical Recombination) Injection 1009 Transfer exactly 100 mL of each concentration of the standard solutions and sample solutions to the wells of 96-well flat bottom microplates. Not less than three plates are F1: Value against m ＝ (nS ＋ nU)(r － 1) shown in the table. g ＝ VEF1/b2(SxxS ＋ SxxU) prepared for both standard solutions and sample solutions. Add exactly 100 mL of a test cell suspension containing 1 × 105 cells per mL in the sample dilution solution for assay to each well, and incubate under atmosphere of 5z carbon di-oxide at 37 ± 29C for 21 to 27 hours. After incubation, add 40 mL of fluorogenic substrate TS to each well, incubate under the same conditions as above for 21 to 51 hours, and measure fluorescence intensities at excitation wavelength 530 to 560 nm and at measurement wavelength 590 nm, using fluorescence microplate reader. Use the data from at least 3 plates and not less than 3 concentrations of the standard so-lution and sample solution for the calculation. Calculation Transform each concentration of the sample solutions and standard solutions to common logarithm, and name them as xU and xS, respectively, and their totals are named as XU and XS, respectively. The fluorescence intensities obtained from the sample solution Value of F1 against m m F1 m F1 m F1 161.45 4.667 25 4.242 18.51 4.600 26 4.225 10.129 4.543 27 4.210 7.709 4.494 28 4.196 6.608 4.451 29 4.183 5.987 4.414 30 4.171 5.591 4.381 40 4.085 5.318 4.351 60 4.001 5.117 4.325 120 3.920 4.965 4.301 / 3.841 4.844 4.279 4.747 4.260 and the standard solution are named as yU and yS, and their totals are named as YU and YS, respectively. The numbers of the concentrations of the sample solution and the standard solution are named as nU and nS, respectively, the number of the plate is r. Calculate the specific activity of Filgrastim (Genetical Recombination) by the following equation, using the protein content (mg/mL) obtained in (1). Specific activity (unit/mg) of Filgrastim (Genetical Recombi-nation) ＝ antilog M × biological activity of Filgrastim RS (unit/mL) × dilution factor for UH × UH dilution factor for SH SH × protein content (mg/mL) obtained in the Assay (1) M ＝ XS/nS － XU/nU － (SYS/nSr － SYU/nUr)/b b ＝ (SxyS ＋ SxyU)/(SxxS ＋ SxxU) SxyS ＝ SxSYS － XSSYS/nS SxyU ＝ SxUYU － XUSYU/nU SxxS ＝ rSxS2 － rXS2/nS SxxU ＝ rSxU2 － rXU2/nU The necessary requirements for validity of the test are fol-lowing three items: F?s is not less than F1 against m ＝ nS (r － 1) shown in the table below, and F?u is not less than F1 against m ＝ nU (r － 1) shown in the table. F?S ＝ VRS/VES VRS ＝ SxyS2/SxxS VES ＝ {SyS2 － S(YS2/r)}/{nS(r － 1)} F?U ＝ VRU/VEU VRU ＝ SxyU2/SxxU VEU ＝ {SyU2 － S(YU2/r)}/{nU(r － 1)} F? is smaller than F1 against m ＝ (nS ＋ nU)(r － 1) shown in the table below. F? ＝ VP/VE VP ＝ SxyS2/SxxS ＋ SxyU2/SxxU － (SxyS ＋ SxyU)2/ (SxxS ＋ SxxU) VE ＝ {SyS2 ＋ SyU2 － S(YS2/r) － S(YU2/r)}/{(nS ＋ nU)(r － 1)} L ≦ 0.3 VEF1{(1 － g)(1/nSr ＋ 1/nUr) L ＝ 2/b(1 － g) ＋ (SYS/nSr － SYU/nUr)2/b2(SxxS ＋ SxxU)}
Containers—Hermetic containers. Storage—Not exceeding 109C, avoiding freezing.
Flavoxate Hydrochloride occurs as white, crys-tals or crystalline powder. It is sparingly soluble in acetic acid (100) and in chlo-roform, slightly soluble in water and in ethanol (95), and practically insoluble in acetonitrile and in diethyl ether.
(1) Determine the absorption spectrum of a solution of Flavoxate Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flavoxate Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Flavoxate Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Flavoxate Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Flavoxate Hydrochloride according to Method 4, and per-form the test (not more than 1 ppm). Related substances—Dissolve 80 mg of Flavoxate Hydrochloride in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 20 mL, then pipet 1 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, reduced pressure, silica gel, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Flavoxate Hydro-chloride, previously dried, add 10 mL of acetic acid (100) and 40 mL of acetonitrile to dissolve, add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 42.79 mg of C24H25NO4.HCl
Flecainide Acetate occurs as a white crystalline powder, having slightly a characteristic or acetic acid like odor. It is freely soluble in methanol, in ethanol (95) and in acetic acid (100), and sparingly soluble in water. A solution of Flecainide Acetate in methanol (1 in 25) shows no optical rotation. Melting point: about 1509C (with decomposition).
(1) Dissolve 20 mg of Flecainide Acetate in 1 mL of water, add 1 mL of a solution of acetaldehyde (1 in 20), and shake. To this solution add dropwise at the same time 1 – 2 drops each of sodium pentacyanonitrosylferrate (III) dihydrate solution (1 in 10) and sodium hydrogen car-bonate TS: a blue precipitate is formed. Determine the absorption spectrum of a solution of Flecainide Acetate in ethanol (95) (13 in 100,000) as directed JP XVIII Official Monographs / Flecainide Acetate Tablets 1013 ‌under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flecainide Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Flecainide Acetate responds to Qualitative Tests <1.09> (1) for acetate. pH <2.54> The pH of a solution of 0.5 g of Flecainide Ace-tate in 20 mL of water is 6.7 to 7.1.
(1) Clarity and color of solution—Dissolve 0.25 g of Flecainide Acetate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Transfer 1.0 g of Flecainide Acetate in a porcelain crucible, and heat gentry to carbonize. After cooling, add 2 mL of sulfuric acid, heat carefully until white fumes are no longer evolved, then proceed according to Method 2 to prepare the test solution, and perform the test. Prepare the control solution as follows: Place 2 mL each of sulfuric acid and hydrochloric acid in a porcelain crucible, evaporate on a water bath, then evaporate to dry-ness on a sand bath, add to the residue 3 drops of hydrochlo-ric acid, then proceed in the same manner as for the test so-lution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). 2-Aminomethylpiperidine—Dissolve exactly 0.25 g of Flecainide Acetate in exactly 5 mL of methanol, and use this solution as the sample solution. Separately, dissolve exactly 50 mg of 2-aminomethylpiperidine in methanol to make ex-actly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone and ammonia solution (28) (20:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in methanol (1 in 500), and heat at 1059C for 2 to 5 minutes: the spot obtained from the sample solution, corresponding to the spot obtained from the stand-ard solution, is not more intense than the spot from the standard solution. Related substances—Dissolve 0.25 g of Flecainide Acetate in 25 mL of a mixture of water and acetonitrile (71:29), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (71:29) to make exactly 50 mL. Pipet 1 mL of this solution, add a mixture of water and acetonitrile (71:29) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the area of the peak other than flecainide obtained from the sample solution is not larger than the peak area of flecainide obtained from the standard solution, and the total area of the peaks other than flecainide from the sample solution is not larger than 2.5 times the peak area of flecainide from the standard solution. For the areas of the peaks, having the relative retention time of about 1.5 and about 2.9 to flecainide, multiply their cor-rection factors, 0.3 and 1.7, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile, acetic acid (100) and tetrabutylammonium hydroxide-methanol TS (142:58:2:1), adjusted to pH 5.8 with ammonia solution (28). Flow rate: Adjust so that the retention time of flecainide is about 4 minutes. Time span of measurement: About 5 times as long as the retention time of flecainide, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of water and acetonitrile (71:29) to make exactly 10 mL. Confirm that the peak area of flecainide obtained with 20 mL of this solution is equiva-lent to 7 – 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of flecainide are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flecainide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.67 kPa, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.6 g of Flecainide Acetate, previously dried, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform the blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 47.44 mg of C17H20F6N2O3.C2H4O2
Flopropione occurs as a white to pale yellow-brown crystalline powder. It is very soluble in N,N-dimethylformamide, freely solu-ble in methanol and in ethanol (99.5), and practically insolu-ble in water.
(1) Determine the absorption spectrum of a solution of Flopropione in ethanol (99.5) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of flopropione obtained with 20 mL of this solution is equivalent to 7 to 13z of that with from 20 mL of the standard solution. System performance: Dissolve 25 mg of ethyl parahy-droxybenzoate in 30 mL of acetonitrile, and add the mobile phase to make 50 mL. To 2.5 mL of this solution add 2 mL of the sample solution and the mobile phase to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, flopropione and ethyl par-ahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flopropione is not more than 1.0z. Water <2.48> Not more than 4.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Flopropione according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Flopropione in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of each peak other than flopropione obtained from the sample solu-tion is not larger than 1/10 times the peak area of flopropione from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 267 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (114:86:1). Flow rate: Adjust so that the retention time of flopropione is about 3 minutes. Time span of measurement: About 7 times as long as the retention time of flopropione.
Weigh accurately about 0.3 g of Flopropione, dis-solve in 30 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS Flopropione as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com- ＝ 18.22 mg of C9H 10O4 pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 177 – 1819C Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Flopropione according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Flopropione in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of each peak other than flopropione obtained from the sample solu-tion is not larger than 1/10 times the peak area of flopropione from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 267 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (114:86:1). Flow rate: Adjust so that the retention time of flopropione is about 3 minutes. Time span of measurement: About 7 times as long as the retention time of flopropione.
Fluconazole, when dried, contains not less than 99.
Fluconazole occurs as a white to pale yellow-white crystalline powder. It is soluble in ethanol (99.5), and slightly soluble in water. It dissolves in dilute hydrochloric acid.
(1) Dissolve 0.1 g of Fluconazole in 10 mL of dilute hydrochloric acid, and add 1 mL of Reinecke's salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Fluconazole in 0.01 mol/L hydrochloric acid-methanol TS (1 in 4000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Amount (mg) of flopropione (C9H10O4) ＝ M S × AT/AS Determine the infrared absorption spectrum of Fluconazole as directed in the potassium bromide disk MS: Amount (mg) of flopropione for assay taken, calcu-lated on the anhydrous basis method under Infrared Spectrophotometry <2.25>, and com- pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. ‌JP XVIII Official Monographs / Fluconazole Capsules 1019
(1) Clarity and color of solution—A solution ob-tained by dissolving 0.10 g of Fluconazole in 50 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Flucona- zole according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 30 mg of Fluconazole in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL. Pipet 1 mL of this so-lution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area of both solutions by the automatic integration method: the peak area of related substance I, having the relative retention time about 0.60 to fluconazole obtained from the sample solution is not larger than 6 times the peak area of fluconazole from the standard solution, the area of the peak other than fluconazole and the related substance I from the sample solution is not larger than the peak area of fluconazole from the standard solution, and the total area of the peaks other than fluconazole from the sample solution is not larger than 8 times the peak area of fluconazole from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of fluconazole is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of fluconazole, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of fluconazole obtained with 20 mL of this solution is equivalent to 35 to 65z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fluconazole are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fluconazole is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.25 g of Fluconazole, pre-viously dried, dissolve in 100 mL of a mixture of acetic anhy-dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form the blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 15.31 mg of C13H12F2N6O
Flucytosine, when dried, contains not less than 98.
Flucytosine occurs as a white crystalline pow-der. It is odorless. It is sparingly soluble in water, slightly soluble in metha-nol, in ethanol (95), in acetic anhydride and in acetic acid (100), and practically insoluble in diethyl ether. It dissolves in 0.1 mol/L hydrochloric acid TS. The pH of a solution of 1.0 g of Flucytosine in 100 mL of water is between 5.5 and 7.5. It is slightly hygroscopic. Melting point: about 2959C (with decomposition).
(1) Add 0.2 mL of bromine TS to 5 mL of a solution of Flucytosine (1 in 500): a yellow-brown color of bromine TS is immediately discharged. Further add 2 mL of barium hydroxide TS: a purple precipitate is formed. Proceed with 0.1 g of Flucytosine as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid. The solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Flucytosine in 0.1 mol/L hydrochloric acid TS (1 in 125,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 1.0 g of Flucytosine in 100 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Flucytosine in 80 mL of water by heating on a water bath. After cooling, to 40 mL of this solution add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Fluoride—Dissolve 0.10 g of Flucytosine in 10.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20). Trans-fer 5.0 mL of this solution to a 20-mL volumetric flask, add 10 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerrous ni-trate TS (1:1:1), and add water to make 20 mL. Allow the mixture to stand for 1 hour, and use this solution as the sam-ple solution. Separately, transfer 4.0 mL of Standard Fluo-rine Solution to a 20-mL volumetric flask, add 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20), add 10 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerrous ni-trate TS (1:1:1). Proceed in the same manner as directed in the preparation of the sample solution, and use this solution as the standard solution. Transfer 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20) to a 20-mL volumetric flask, proceed in the same manner as directed in the prepara-tion of the standard solution, and use this solution as the blank solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 600 nm, using the blank solution as the control as directed under Ultravio-let-visible Spectrophotometry <2.24>: AT is not larger than AS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Flucyto- sine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Flucytosine according to Method 2, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Flucytosine in 5 mL of diluted methanol (1 in 2), and use this solution as the sample solution. Measure accurately 1 mL of the sample solution, add diluted methanol (1 in 2) to make exactly 25 mL. Measure accurately 1 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (5:3:2) to a distance of about 12 cm, air-dry the plate, and observe the spots under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Flucytosine—Weigh accurately about 0.2 g of Flucytosine, previously dried, dissolve in 40 mL of acetic acid (100), add 100 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 12.91 mg of C4H4FN3O (2) Fluorine—Weigh accurately about 10 mg of Flucyto-sine, previously dried, and proceed as directed in the deter-mination of fluorine under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide VS and 20 mL of water as the absorbing liquid.
Fludiazepam, when dried, contains not less than 99.
Fludiazepam occurs as white to light yellow, crystals or crystalline powder. It is very soluble in chloroform, freely soluble in metha-nol, in ethanol (95), in acetic acid (100) and in diethyl ether, and practically insoluble in water. roform to make exactly 50 mL. Pipet 2 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethyl acetate (10:7) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
(1) Prepare the test solution with 0.01 g of Fludiazepam as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Fludiazepam in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wave-lengths. Separately, determine the absorption spectrum of a solution of Fludiazepam in methanol (1 in 20,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Fludiazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Fludiazepam as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—Dissolve 1.0 g of Fludiazep-am in 50 mL of diethyl ether, add 50 mL of water, and shake. Separate the water layer, wash it with two 20-mL por-tions of diethyl ether, and filter the water layer. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 2.0 g of Fludia- zepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Fludiazepam in 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chlo-
Weigh accurately about 0.5 g of Fludiazepam, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.28 mg of C H ClFN O 16 12 2 Identification (1) Prepare the test solution with 0.01 g of Fludiazepam as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Fludiazepam in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wave-lengths. Separately, determine the absorption spectrum of a solution of Fludiazepam in methanol (1 in 20,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Fludiazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Fludiazepam as directed under Flame Coloration Test <1.04> (2): a green color appears. Melting point <2.60> 91 – 949C Purity (1) Chloride <1.03>—Dissolve 1.0 g of Fludiazep-am in 50 mL of diethyl ether, add 50 mL of water, and shake. Separate the water layer, wash it with two 20-mL por-tions of diethyl ether, and filter the water layer. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 2.0 g of Fludia- zepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Fludiazepam in 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chlo-
フルドロコルチゾン酢酸エステル
Fludrocortisone Acetate, when dried, contains not less than 97.
Fludrocortisone Acetate occurs as a white to pale yellow, crystals or crystalline powder. It is soluble in acetone, sparingly soluble in ethanol (95), and practically insoluble in water. Melting point: about 2209C (with decomposition).
(1) Prepare the test solution by proceeding with 10 mg of Fludrocortisone Acetate according to the Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide VS and 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Fludrocortisone Acetate in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Fludrocortisone Acetate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same ‌1024 wavelengths. Determine the infrared absorption spectrum of Fludrocortisone Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Fludrocortisone Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋131 – ＋1389(after drying, 0.1 g, acetone, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 0.5 g of Fludrocortisone Acetate according to Method 2, and per-form the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 30 ppm). (2) Related substances—Dissolve 20 mg of Fludrocorti-sone Acetate in 10 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than fludrocortisone acetate ob-tained from the sample solution is not larger than 1/4 times the peak area of fludrocortisone acetate from the standard solution, and the total area of the peaks other than fludrocortisone acetate from the sample solution is not larger than 1/2 times the peak area of fludrocortisone ace-tate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and tetrahydrofuran (13:7). Flow rate: Adjust so that the retention time of fludrocorti-sone acetate is about 10 minutes. Time span of measurement: About 2 times as long as the retention time of fludrocortisone acetate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of fludrocortisone acetate ob-tained with 20 mL of this solution is equivalent to 4.0 to 6.0z of that with 20 mL of the standard solution. System performance: Dissolve 2 mg each of Fludrocor-tisone Acetate and hydrocortisone acetate in 50 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrocor-tisone acetate and fludrocortisone acetate are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fludrocortisone acetate is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 1009C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati- num crucible).
Weigh accurately about 25 mg each of Fludrocor-tisone Acetate and Fludrocortisone Acetate RS, previously dried, and dissolve separately in ethanol (95) to make exactly 100 mL. Pipet 4 mL each of these solutions, add ethanol (95) to make exactly 100 mL, and use these solutions as the sam-ple solution and the standard solution, respectively. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 238 nm. Amount (mg) of fludrocortisone acetate (C23H31FO6) ＝ MS × AT/AS MS: Amount (mg) of Fludrocortisone Acetate RS taken
Flunitrazepam, when dried, contains not less than 99.
Flunitrazepam occurs as a white to pale yellow crystalline powder. It is freely soluble in acetic acid (100), soluble in acetic an-hydride and in acetone, slightly soluble in ethanol (99.5) and in diethyl ether, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Flunitrazepam in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Flunitrazepam, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Chloride <1.03>—To 1.0 g of Flunitrazepam add 50 mL of water, allow to stand for 1 hour with occa-sional stirring, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test with this solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.022z). Heavy metals <1.07>—Proceed with 2.0 g of Flunitrazepam according to Method 4 using a platinum cru- ‌JP XVIII Official Monographs / Fluocinolone Acetonide 1025 cible, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Flunitra-zepam in 10 mL of acetone, and use this solution as the sam-ple solution. Pipet 2 mL of the sample solution, and add ace-tone to make exactly 20 mL. Pipet 1 mL of this solution, add acetone to make exactly 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1,2-dichloroethane, diethyl ether and ammonia solution (28) (200:100:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): number of the spots other than the principal spot ob-tained from the sample solution is not more than 2, and they are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.5 g of Flunitrazepam, previously dried, dissolve in 20 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.33 mg of C16H12FN3O3
Fluocinolone Acetonide occurs as white, crys-tals or crystalline powder. It is freely soluble in acetic acid (100) and in acetone, solu-ble in ethanol (99.5), sparingly soluble in methanol, and practically insoluble in water. Melting point: 266 – 2749C (with decomposition). It shows crystal polymorphism.
(1) To 2 mg of Fluocinolone Acetonide add 2 mL of sulfuric acid: a yellow color is produced. Dissolve 0.01 g of Fluocinolone Acetonide in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: a red precipi-tate is produced. Proceed with 0.01 g of Fluocinolone Acetonide as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solu-tion responds to Qualitative Tests <1.09> for fluoride. Determine the infrared absorption spectrum of Fluocinolone Acetonide, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of previously dried Fluocino-lone Acetonide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Fluocinolone Aceto-nide and Fluocinolone Acetonide RS in acetone, respec-tively, then evaporate the acetone to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋98 – ＋1089(after drying, 0.1 g, methanol, 10 mL, 100 mm).
Related substances—Dissolve 15 mg of Fluocino-lone Acetonide in 25 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the total area of the peaks other than fluocinolone acetonide obtained from the sample solution is not larger than the peak area of fluocinolone acetonide from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water-saturated chloroform, methanol and acetic acid (100) (200:3:2). Flow rate: Adjust so that the retention time of fluocino-lone acetonide is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of fluocinolone acetonide, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of fluocinolone acetonide obtained with 20 mL of this solution is equivalent to 4 to 6z of that with 20 mL of the standard solution. System performance: Dissolve 15 mg each of Fluocinolone Acetonide and triamcinolone acetonide in 25 mL of the mo-bile phase. To 5 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, triamcino-lone acetonide and fluocinolone acetonide are eluted in this order with the resolution between these peaks being not less than 1.9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌1026 areas of fluocinolone acetonide is not more than 1.0z. Loss on drying <2.41> Not more than 1.0z (0.2 g, in vacu-um, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.2 g, platinum crucible).
Weigh accurately about 20 mg each of Fluocinolone Acetonide and Fluocinolone Acetonide RS, previously dried, and dissolve in 40 mL each of methanol, add exactly 10 mL each of the internal standard solution, then add water to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of fluocinolone acetonide to that of the internal stand-ard. Fluocinonide
Fluocinonide, when dried, contains not less than 97.
Fluocinonide occurs as white, crystals or crys-talline powder. It is sparingly soluble in chloroform, slightly soluble in acetonitrile, in methanol, in ethanol (95) and in ethyl ace-tate, and practically insoluble in water. It shows crystal polymorphism.
(1) To 0.01 g of Fluocinonide add 4 mL of water and 1 mL of Fehling's TS, and heat: a red precipitate is formed. Prepare the test solution with 0.01 g of Fluocinonide as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as an absorbing liquid: the test solu-tion responds to Qualitative Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Fluocinonide in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Fluocinonide RS prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectra of Fluocinonide and Fluocinonide RS, previously dried, as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare both spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears in the absorption spectra, dissolve the sample and the RS in ethyl acetate, re-spectively, evaporate the ethyl acetate, and perform the test with the residue in the same manner. D Optical rotation <2.49> [a]20: ＋81 – ＋899(after drying, 0.2 g, chloroform, 20 mL, 100 mm).
Related substances—Dissolve 10 mg of Fluocinonide in 2 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and methanol (97:3) ‌JP XVIII Official Monographs / Fluorescein Sodium 1027 to a distance of about 12 cm, and air-dry the plate. Spray evenly alkaline blue tetrazolium TS on the plate: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g, platinum crucible).
Weigh accurately about 20 mg each of Fluocinonide and Fluocinonide RS, previously dried, dissolve each in 50 mL of acetonitrile, to each add exactly 8 mL of the internal standard solution and water to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of fluocinonide to that of the internal standard, respectively. Amount (mg) of fluocinonide (C26H32F2O7) ＝ MS × QT/QS MS: Amount (mg) of Fluocinonide RS taken Internal standard solution—A solution of propyl benzoate in acetonitrile (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of fluocinonide is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, fluocinonide and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of fluocinonide to that of the internal stand-ard is not more than 1.0z.
Fluorometholone, when dried, contains not less than 97.
Fluorometholone occurs as a white to light yellow-white crystalline powder. It is odorless. It is freely soluble in pyridine, slightly soluble in metha-nol, in ethanol (99.5) and in tetrahydrofuran, and practically insoluble in water and in diethyl ether.
(1) Proceed with 7 mg of Fluorometholone as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as an absorbing liquid: the liquid re-sponds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Fluorometholone in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Fluorometholone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Fluorometholone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Fluorometho-lone RS: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: ＋52 – ＋609(after drying, 0.1 g, pyridine, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Fluorometholone according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 20 mg of Fluorometho-lone in 10 mL of tetrahydrofuran, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add tetrahydrofuran to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer chromatography <2.03>. Spot 25 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, acetone and methanol (45:5:1) to a dis-tance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 1.0z (0.2 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2 z (0.2 g, platinum crucible).
Weigh accurately about 0.1 g each of Fluorometho-lone and Fluorometholone RS, previously dried, and dis-solve each in methanol to make exactly 100 mL. Pipet 5 mL each of these solutions, and add diluted methanol (7 in 10) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution and diluted methanol (7 in 10) to make 100 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions and calculate the ratios, QT and QS, of the peak area of fluorometholone to that of the internal standard. Amount (mg) of fluorometholone (C22H29FO4) ＝ MS × QT/QS MS: Amount (mg) of Fluorometholone RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 25 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Diluted methanol (7 in 10). Flow rate: Adjust so that the retention time of fluorometholone is about 8 minutes. Selection of column: Proceed with 20 mL of the standard solution under the above operating conditions, and calculate the resolution. Use a column giving elution of fluorometho-lone and the internal standard in this order with the resolu-tion between these peaks being not less than 4.
Fluorouracil, when dried, contains not less than 98.
Fluorouracil occurs as white, crystals or crys-talline powder. It is odorless. It is freely soluble in N,N-dimethylformamide, sparingly soluble in water, slightly soluble in ethanol (95), and practi-cally insoluble in diethyl ether. Melting point: about 2829C (with decomposition).
(1) Add 0.2 mL of bromine TS to 5 mL of a solution of Fluorouracil (1 in 500): the color of bromine TS is discharged. Further add 2 mL of barium hydroxide TS: a purple precipitate is formed. Proceed with 0.01 g of Fluorouracil as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Fluorouracil in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.
(1) Clarity and color of solution—Add 20 mL of water to 0.20 g of Fluorouracil, and dissolve by warming: the solution is clear and colorless. Fluoride—Dissolve 0.10 g of Fluorouracil in 10.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20). Trans-fer 5.0 mL of this solution to a 20-mL volumetric flask, add 10 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1), and add water to make 20 mL. Allow to stand for 1 hour, and use this solution as the sample solu-tion. Separately, transfer 1.0 mL of Standard Fluorine Solu-tion to a 20-mL volumetric flask, add 5.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20), and add 10 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1). Proceed in the same manner as directed for the preparation of the sample solution, and use this solution as the standard solution. Perform the test as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using a solution, prepared with 5.0 mL of diluted 0.01 mol/L sodium hydrox-ide TS (1 in 20) in the same manner, as the blank: the absor-bance of the sample solution at 600 nm is not larger than that of the standard solution (not more than 0.012z). Heavy metals <1.07>—Proceed with 1.0 g of Fluoro-uracil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Fluorouracil in a crucible add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), ignite the ethanol to burn, and incinerate by strong heating at 7509C to 8509C. If a car-bonized substance remains in this method, moisten with a small amount of nitric acid, and incinerate by strong heat-ing. Cool, add 10 mL of dilute hydrochloric acid to the residue, dissolve it by warming on a water bath, use this so-lution as the test solution, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Fluorouracil in 10 mL of water, and use this solution as the sample solu-tion. Measure exactly 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetone and water (7:4:1) to a distance of about 12 cm, air-dry the plate, and examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Fluorouracil—Weigh accurately about 0.2 g of Fluorouracil, previously dried, dissolve in 20 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hydroxide VS until the color of the solution changes from yellow through blue-green to blue (indicator: 3 drops of thymol blue-dimethylformamide TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 13.01 mg of C4H3FN2O2 (2) Fluorine—Weigh accurately about 4 mg of Fluoro-uracil, previously dried, and proceed as directed in the deter-mination of fluorine under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid.
フルフェナジンエナント酸エステル
Fluphenazine Enanthate, when dried, contains not less than 98.
Fluphenazine Enanthate is a light yellow to yellowish orange viscous liquid. It is generally clear, and can be opaque by producing crystals. It is freely soluble in methanol and in diethyl ether, soluble in ethanol (95) and in acetic acid (100), and practically in-soluble in water.
(1) Prepare the test solution with 0.01 g of Fluphenazine Enanthate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> for fluoride. Dissolve 2 mg of Fluphenazine Enanthate in 200 mL of a solution of hydrochloric acid in methanol (17 in 2000). Determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the ard solution. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Fluphenazine Enan-thate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.49 mg of C29H38F3N3O2S
Flurazepam Hydrochloride, when dried, contains not less than 99.
Flurazepam Hydrochloride occurs as white to yellowish white, crystals or crystalline powder. It is freely soluble in water, in ethanol (95), in ethanol (99.5) and in acetic acid (100). Melting point: about 1979C (with decomposition).
(1) Determine the absorption spectrum of a solution of Flurazepam Hydrochloride in sulfuric acid-ethanol TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flurazepam Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Flurazepam Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Flurazepam Hydrochloride in 20 mL of water: the pH of this solution is between 5.0 and 6.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Fluphenazine Enanthate according to Method 2, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). (2) Related substances—Dissolve 0.25 g of Fluphenazine Enanthate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ace-tone, hexane and ammonia solution (28) (16:6:1) to a dis-tance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Then spray evenly diluted sulfuric acid (1 in 2) on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand- Description Flurazepam Hydrochloride occurs as white to yellowish white, crystals or crystalline powder. It is freely soluble in water, in ethanol (95), in ethanol (99.5) and in acetic acid (100). Melting point: about 1979C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Flurazepam Hydrochloride in sulfuric acid-ethanol TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flurazepam Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Flurazepam Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Flurazepam Hydrochloride in 20 mL of water: the pH of this solution is between 5.0 and 6.0. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Flurazepam Hydrochloride in 10 mL of water: the solu-tion is clear and colorless to pale yellow. JP XVIII Official Monographs / Flurbiprofen 1031 Sulfate <1.14>—Perform the test with 1.5 g of Flurazepam Hydrochloride. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.011z). Heavy metals <1.07>—Proceed with 1.0 g of Flura- ‌zepam Hydrochloride in a platinum crucible according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.05 g of Flurazepam Hydrochloride in 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 50 mL. Pipet 1 mL of this solution, add ethanol (95) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Place the plate in a chamber filled with ammonia vapor, allow to stand for about 15 minutes, and immediately develop the plate with a mixture of diethyl ether and diethylamine (39:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): not more than 3 spots other than the principal spot and the spot on the start-ing point obtained from the sample solution appear, and are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Flurazepam Hydro-chloride, previously dried, dissolve in 10 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.22 mg of C21H23ClFN3O.HCl
Flurbiprofen, when dried, contains not less than Identification (1) Determine the absorption spectrum of a solution of Flurbiprofen in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.
Flurbiprofen occurs as a white crystalline pow-der. It has a slightly irritating odor. It is freely soluble in methanol, in ethanol (95), in acetone and in diethyl ether, soluble in acetonitrile, and practically insoluble in water. A solution of Flurbiprofen in ethanol (95) (1 in 50) shows no optical rotation. standard solution add a mixture of water and acetonitrile (11:9) to make exactly 25 mL. Confirm that the peak area of flurbiprofen obtained with 20 mL of this solution is equiva-lent to 16 to 24z of that with 20 mL of the standard solu-tion. System performance: Dissolve 0.04 g of flurbiprofen and 0.02 g of butyl parahydroxybenzoate in 100 mL of a mixture ‌1032 of water and acetonitrile (11:9). To 5 mL of this solution add a mixture of water and acetonitrile (11:9) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, butyl parahydroxybenzoate and flurbiprofen are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flurbiprofen is not more than 2.0z. Loss on drying <2.41> Not more than 0.10z (1 g, in vacu-um at a pressure not exceeding 0.67 kPa, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
(1) Determine the absorption spectrum of a solution of Flurbiprofen in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Flurbiprofen, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.6 g of Flurbi-profen in 40 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.015z). Heavy metals <1.07>—Dissolve 2.0 g of Flurbiprofen in 30 mL of acetone, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 30 mL of acetone, 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Related substances—Dissolve 20 mg of Flurbiprofen in 10 mL of a mixture of water and acetonitrile (11:9), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (11:9) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of the peak other than flurbiprofen obtained from the sample solu-tion is not larger than the peak area of flurbiprofen from the standard solution, and the total area of these peaks is not larger than 2 times the peak area of flurbiprofen from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (12:7:1). Flow rate: Adjust so that the retention time of flur-biprofen is about 20 minutes. Time span of measurement: About twice as long as the retention time of flurbiprofen, beginning after the solvent peak. 98.0z of flurbiprofen (C15 H13 FO2). System suitability— Test for required detectability: To exactly 5 mL of the Description Flurbiprofen occurs as a white crystalline pow-der. It has a slightly irritating odor. It is freely soluble in methanol, in ethanol (95), in acetone and in diethyl ether, soluble in acetonitrile, and practically insoluble in water. A solution of Flurbiprofen in ethanol (95) (1 in 50) shows no optical rotation. standard solution add a mixture of water and acetonitrile (11:9) to make exactly 25 mL. Confirm that the peak area of flurbiprofen obtained with 20 mL of this solution is equiva-lent to 16 to 24z of that with 20 mL of the standard solu-tion. System performance: Dissolve 0.04 g of flurbiprofen and 0.02 g of butyl parahydroxybenzoate in 100 mL of a mixture ‌1032 of water and acetonitrile (11:9). To 5 mL of this solution add a mixture of water and acetonitrile (11:9) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, butyl parahydroxybenzoate and flurbiprofen are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flurbiprofen is not more than 2.0z. Loss on drying <2.41> Not more than 0.10z (1 g, in vacu-um at a pressure not exceeding 0.67 kPa, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.6 g of Flurbiprofen, previ-ously dried, dissolve in 50 mL of ethanol (95), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 24.43 mg of C15H13FO2
Flutamide, when dried, contains not less than 98.
Flutamide occurs as a light yellow crystalline powder. It is freely soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Flutamide in ethanol (95) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and Flutamide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Dissolve 40 mg of Flutamide in 50 mL of methanol, and use this solution as the sample solu-tion. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of the sample solution by the automatic integration method and calculate the amounts of them by the area percentage method: the amount of each peak other than flutamide is not more than 0.3z, and the total amount of the peaks other than flutamide is not more than 0.5z. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Time span of measurement: About 2 times as long as the retention time of flutamide, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the sample so-lution, add methanol to make 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add methanol to make exactly 20 mL. Confirm that the peak area of flutamide obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of flutamide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
(1) Heavy metals <1.07>—Proceed with 2.0 g of MS: Amount (mg) of Flutamide RS taken Internal standard solution—A solution of testosterone in methanol (9 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about ‌JP XVIII Official Monographs / Flutoprazepam Tablets 1033‌ 259C. Mobile phase: A mixture of methanol and 0.05 mol/L potassium dihydrogen phosphate TS (7:4). Flow rate: Adjust so that the retention time of flutamide is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, flutamide and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flutamide is not more than 1.0z.
Weigh accurately about 40 mg each of Flutamide and Flutamide RS, previously dried, and dissolve separately in methanol to make exactly 25 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard so-lution, add methanol to make 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of flutamide to that of the internal standard. compare the spectrum with the Reference Spectrum or the Amount (mg) of flutamide (C H F N O ) spectrum of a solution of Flutamide RS prepared in the same manner as the sample solution: both spectra exhibit similar ＝ MS × QT/QS 11 11 3 2 3 intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Flu-tamide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Flutamide RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Melting point <2.60> 109 – 1139C Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of MS: Amount (mg) of Flutamide RS taken Internal standard solution—A solution of testosterone in methanol (9 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about ‌JP XVIII Official Monographs / Flutoprazepam Tablets 1033‌ 259C. Mobile phase: A mixture of methanol and 0.05 mol/L potassium dihydrogen phosphate TS (7:4). Flow rate: Adjust so that the retention time of flutamide is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, flutamide and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flutamide is not more than 1.0z.
Flutoprazepam, when dried, contains not less than 99.
Flutoprazepam occurs as a white to light yel-low, crystals or crystalline powder. It is freely soluble in ethyl acetate, soluble in ethanol (99.5) and in acetic anhydride, and practically insoluble in water.
(1) Dissolve 2 mg of Flutoprazepam in 200 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flutoprazepam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Flutoprazepam as directed un-der Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—To 1.0 g of Flutoprazepam add 50 mL of water, allow to stand for 1 hour with occa-sional shaking, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con- trol solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 1.0 g of Fluto- prazepam according to Method 2, and perform the test. Pre-pare the control solution with 1.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Flutopra-zepam in 20 mL of ethyl acetate, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add ethyl acetate to make exactly 50 mL. Pipet 1 mL of this solu-tion, add ethyl acetate to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (3:2) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.5 g of Flutoprazepam, previously dried, dissolve in 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.28 mg of C19H16ClFN2O
Fluvoxamine Maleate contains not less than 98.
Fluvoxamine Maleate occurs as a white crystal-line powder. It is freely soluble in ethanol (99.5), and sparingly soluble in water. 15 minutes, and add the mobile phase to make exactly 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of fluto-prazepam for assay, previously dried at 1059C for 2 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution.
(1) Dissolve 10 mg of Fluvoxamine Male-ate in 5 mL of water, neutralize with dilute sodium hydrox-ide TS, then add 1 mL of ninhydrin TS, and heat in a water bath at 60 – 709C for 5 minutes: a blue-purple color de-velops. Determine the absorption spectrum of a solution of Fluvoxamine Maleate (1 in 50,000) as directed under Ultravi-olet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a JP XVIII Official Monographs / Fluvoxamine Maleate 1035 solution of Fluvoxamine Maleate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Fluvoxamine Maleate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Fluvoxamine Maleate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Fluvoxamine Maleate (1 in 500) add 1 drop of potassium permanganate TS: the red color of the TS disappears immediately.
(1) Clarity and color of solution—A solution ob-tained by dissolving 0.5 g of Fluvoxamine Maleate in 50 mL of water is clear and colorless. Chloride <1.03>—Perform the test with 1.0 g of Fluvoxamine Maleate. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.009z). Sulfate <1.14>—Perform the test with 1.0 g of Fluvoxamine Maleate. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.017z). Heavy metals <1.07>—Proceed with 1.0 g of Fluvoxa- mine Maleate according to Method 2, using alumina ceramic crucible, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Fluvoxamine Maleate in 20 mL of a mixture of methanol for liquid chro-matography and water (7:3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol for liquid chromatography and water (7:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the areas of the peaks, having the relative retention time of about 0.76, about 0.82, about 0.89, about 1.58 and about 1.66 to fluvoxamine, obtained from the sample solution are not larger than 1/5 times, 3/10 times, 7/10 times, 1/10 times and 1/10 times the peak area of fluvoxamine from the standard solution, respectively, and the total area of the peaks other than fluvoxamine from the sample solution is not larger than 1.5 times the peak area of fluvoxamine from the standard solution. For the areas of the peaks, having the relative retention times of about 0.76, about 0.89, about 1.58 and about 1.66 to fluvoxamine, mul-tiply their correction factors, 0.87, 2.00, 0.67 and 2.76, re-spectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 12.67 g of diammonium hydrogen phosphate and 0.85 g of sodium 1-heptanesulfonate in 900 mL of water, adjust to pH 2.0 with phosphoric acid, and add water to make 1000 mL. To 300 mL of this solution add 700 mL of methanol for liquid chromatography. Flow rate: Adjust so that the retention time of fluvoxa- mine is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of fluvoxamine, beginning after the peak of maleic acid. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add a mixture of methanol for liquid chromatogra-phy and water (7:3) to make exactly 20 mL. Confirm that the peak area of fluvoxamine obtained with 10 mL of this solu-tion is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fluvoxamine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fluvoxamine is not more than 2.0z. Loss on drying <2.41> Not more than 0.1z (1 g, in vacu-um, 509C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 20 mg each of Fluvoxamine Maleate and Fluvoxamine Maleate RS (separately determine the loss on drying <2.41> under the same condition as Fluvoxamine Maleate), dissolve each in 10 mL of the mobile phase, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of fluvoxamine to that of the internal standard. Amount (mg) of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) ＝ MS × QT/QS MS: Amount (mg) of Fluvoxamine Maleate RS taken, cal-culated on the dried basis Internal standard solution—A solution of diphenylamine in methanol (7 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.8 g of diammonium hydrogen phosphate and 0.8 g of sodium 1-heptanesulfonate in water to make 300 mL, add 700 mL of methanol, and adjust to pH 3.5 with phosphoric acid. Flow rate: Adjust so that the retention time of fluvoxa-mine is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, fluvoxamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times ‌1036 with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of fluvoxamine to that of the internal standard is not more than 1.0z.
Foeniculated Ammonia Spirit is a colorless to yellow liquid, having a characteristic odor. It has a slightly sweet, pungent taste. Specific gravity d 20: about 0.85 Alcohol number <1.01> Not less than 7.8 (Method 2).
Formalin Water contains not less than 0.
Formalin Water is a clear, colorless liquid. It has a slight odor of formaldehyde. It is almost neutral.
Transfer 20 mL of Formalin Water, measured exactly, to a 100-mL volumetric flask containing 2.5 mL of 1 mol/L sodium hydroxide VS, and add water to make 100 mL. Pipet 10 mL of this solution, and proceed as directed in the Assay under Formalin. Each mL of 0.05 mol/L iodine VS ＝ 1.501 mg of CH2O
Formalin is a clear, colorless liquid. Its vapor is irritating to the mucous membrane. It is miscible with water and with ethanol (95). When stored for a long time, especially in a cold place, it may become cloudy.
(1) Dilute 2 mL of Formalin with 10 mL of water in a test tube, and add 1 mL of silver nitrate-ammonia TS: a gray precipitate is produced, or a silver mirror is formed on the wall of the test tube. (2) To 5 mL of sulfuric acid in which 0.1 g of salicylic acid has been dissolved add 2 drops of Formalin, and warm the solution: a persistent, dark red color develops.
Acidity—Dilute 20 mL of Formalin with 20 mL of water, and add 5.0 mL of 0.1 mol/L sodium hydroxide VS and 2 drops of bromothymol blue TS: a blue color develops. Residue on ignition <2.44> Not more than 0.06 w/vz (5 mL, after evaporation).
Weigh accurately a weighing bottle containing 5 mL of water, add about 1 g of Formalin, and weigh accurately again. Add water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 50 mL of 0.05 mol/L iodine VS and 20 mL of potassium hydroxide TS, and allow to stand for 15 minutes at an ordinary temperature. To this mixture add 15 mL of dilute sulfuric acid, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indica-tor: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 1.501 mg of CH2O
ホルモテロールフマル酸塩水和物
Formoterol Fumarate Hydrate contains not less than 98.
Formoterol Fumarate Hydrate occurs as a white to yellowish white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, very slightly soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. A solution of Formoterol Fumarate Hydrate in methanol (1 in 100) shows no optical rotation. Melting point: about 1389C (with decomposition).
(1) Dissolve 0.5 g of Formoterol Fumarate Hydrate in 20 mL of 0.5 mol/L sulfuric acid TS, and extract with three 25-mL portions of diethyl ether. Wash the com-bined diethyl ether extracts with 10 mL of 0.5 mol/L sulfuric acid TS, and evaporate the ether layer under reduced pres-sure, and dry the residue at 1059C for 3 hours: the residue melts <2.60> at about 2909C (with decomposition, in a sealed tube). Determine the absorption spectrum of a solution of Formoterol Fumarate Hydrate in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of For-moterol Fumarate Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Formoterol Fumarate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related Substances—Dissolve 0.20 g of Formoterol Fumarate Hydrate in 10 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 200 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform, 1,4-dioxane, ethanol (99.5) and ammonia solution (28) (20:20:10:3) to a distance of about 12 cm, and air-dry the tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.24 mg of (C19H24N2O4)2.C4H4O4
Weigh accurately about 0.7 g of Formoterol Fumarate Hydrate, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten- (C19H24N2O4)2.C4H4O4.2H2O: 840.91 N-(2-Hydroxy-5-{(1RS)-1-hydroxy- 2-[(1RS)-2-(4-methoxyphenyl)- methylethylamino]ethyl}phenyl)formamide hemifumarate monohydrate [43229-80-7, anhydride] Formoterol Fumarate Hydrate contains not less than 98.5z of formoterol fumarate [(C19H24N2O4)2. C4H4O4: 804.88], calculated on the anhydrous basis. Description Formoterol Fumarate Hydrate occurs as a white to yellowish white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, very slightly soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. A solution of Formoterol Fumarate Hydrate in methanol (1 in 100) shows no optical rotation. Melting point: about 1389C (with decomposition). Identification (1) Dissolve 0.5 g of Formoterol Fumarate Hydrate in 20 mL of 0.5 mol/L sulfuric acid TS, and extract with three 25-mL portions of diethyl ether. Wash the com-bined diethyl ether extracts with 10 mL of 0.5 mol/L sulfuric acid TS, and evaporate the ether layer under reduced pres-sure, and dry the residue at 1059C for 3 hours: the residue melts <2.60> at about 2909C (with decomposition, in a sealed tube). Determine the absorption spectrum of a solution of Formoterol Fumarate Hydrate in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of For-moterol Fumarate Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Formoterol Fumarate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related Substances—Dissolve 0.20 g of Formoterol Fumarate Hydrate in 10 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 200 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform, 1,4-dioxane, ethanol (99.5) and ammonia solution (28) (20:20:10:3) to a distance of about 12 cm, and air-dry the tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.24 mg of (C19H24N2O4)2.C4H4O4
Powdered Longgu occurs as a light grayish white to light grayish brown. It is odorless and tasteless.
(1) Dissolve 0.5 g of Powdered Longgu in 10 mL of dilute hydrochloric acid: it evolves a gas, and forms a slightly brownish and turbid solution. Pass the gas evolved through calcium hydroxide TS: a white precipitate is produced. The turbid solution obtained in (1) has a characteris-tic odor. Filter this solution, and neutralize filtrate with ammonia TS: the solution responds to Qualitative test <1.09> (1), (2) and (3) for calcium salt. Dissolve 0.1 g of Powdered Longgu in 5 mL of nitric acid by warming, and add hexaammonium heptamolybdate TS: a yellow precipitate is produced.
(1) Heavy metals <1.07>—To 2.0 g of Powdered Longgu add 5 mL of water, shake to mix, add gradually 6 mL of hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 50 mL of water, and filter. To 25 mL of the filtrate add 2 mL of dilute acetic acid, 1 drop of ammonia TS and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con- ‌2066‌ Lonicera Leaf and Stem / Crude Drugs and Related Drugs trol solution as follows: evaporate 3 mL of hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid and 2.0 mL of Standard Lead Solution, and add water to make 50 mL (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.20 g of Powdered Longgu according to Method 2, and perform the test (not more than 10 ppm).
Irregular masses or fragments, occasionally cylindrical masses; externally light grayish white, sometimes with grayish black or yellow-brown spots here and there; the outer part consists of a layer 2 – 10 mm in thickness, and is minute in texture, surrounding the light brown, porous por-tion; heavy and hard, but somewhat fragile in texture; when crushed, it changes into pieces and powder. Odorless, tasteless, and strongly adhesive to the tongue on licking.
(1) Dissolve 0.5 g of pulverized Longgu in 10 mL of dilute hydrochloric acid: it evolves a gas, and forms a slightly brownish and turbid solution. Pass the gas evolved through calcium hydroxide TS: a white precipitate is produced. The turbid solution obtained in (1) has a characteris-tic odor. Filter this solution and neutralize filtrate with am-monia TS: this solution responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. Dissolve 0.1 g of pulverized Longgu in 5 mL of nitric acid by warming, and add hexaammonium heptamolybdate TS: a yellow precipitate is produced.
(1) Heavy metals <1.07>—To 2.0 g of pulverized Longgu, add 5 mL of water, shake, add gradually 6 mL of hydrochloric acid, evaporate on a water bath to dryness, dis-solve the residue in 50 mL of water, and filter. To 25 mL of the filtrate, add 2 mL of dilute acetic acid, 1 drop of ammo-nia TS and water to make 50 mL. Perform the test with this solution as the test solution. Prepare the control solution as follows: evaporate 3 mL of hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL, and use this solution as the control solution (not more than 20 ppm). When being shown as extracts, infusions and decoctions on the label, the procedure and the limit are as follows. To 20.0 g of pulverized Longgu, add 80 mL of water, shake occasionally in a water bath, heat to make about 40 mL, allow to cool, and filter. Proceed with this solution ac-cording to Method 3, and perform the test. To the control solution, add 1.0 mL of Standard Lead Solution (not more than 0.5 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.20 g of pulverized Longgu according to Method 2, and perform the test (not more than 10 ppm). When being shown the restricted utilization forms as ``ex-tracts, infusions and decoctions only'', the procedure and the limit are as follows. Put 4.0 g of pulverized Longgu in a centrifuge tube, add 30 mL of water, and heat in a water bath with occasional shaking to make about 15 mL. After cooling, centrifuge, and perform the test using the supernatant liquid as the test solution (not more than 0.5 ppm).
Fructose, when dried, contains not less than 98.
Fructose occurs as colorless to white, crystals or crystalline powder. It is odorless and has a sweet taste. It is very soluble in water, sparingly soluble in ethanol (95) and practically insoluble in diethyl ether. It is hygroscopic.
(1) Add 2 to 3 drops of a solution of Fruc-tose (1 in 20) to 5 mL of boiling Fehling's TS: a red precipi-tate is produced. (2) Determine the infrared absorption spectrum of Fruc-tose as directed in the paste method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 4.0 g of Fructose in 20 mL of water: the pH of the solution is between 4.0 and 6.5.
(1) Clarity and color of solution—Dissolve 25.0 g of Fructose in 50 mL of water: the solution is clear and has no more color than the following control solution. Control solution: To a mixture of 1.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS, and add water to make 10.0 mL. To 3.0 mL of the solution add water to make 50 mL. Acidity—Dissolve 5.0 g of Fructose in 50 mL of freshly boiled and cooled water, and add 3 drops of phenol-phthalein TS and 0.60 mL of 0.01 mol/L sodium hydroxide VS: a red color develops. Chloride <1.03>—Perform the test with 2.0 g of Fruc- tose. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Sulfate <1.14>—Perform the test with 2.0 g of Fruc- tose. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Sulfite—Dissolve 0.5 g of Fructose in 5 mL of water, and add 0.25 mL of 0.02 mol/L iodine: the color of the solu-tion is yellow. Heavy metals <1.07>—Proceed with 5.0 g of Fructose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 4 ppm). Calcium—Dissolve 0.5 g of Fructose in 5 mL of water, add 2 to 3 drops of ammonia TS and 1 mL of ammo-nium oxalate TS, and allow to stand for 1 minute: the solu- tion is clear. Arsenic <1.11>—Dissolve 1.5 g of Fructose in 5 mL of water, heat with 5 mL of dilute sulfuric acid and 1 mL of bromine TS on a water bath for 5 minutes, concentrate to 5 mL, and cool. Perform the test with this solution as the test solution (not more than 1.3 ppm). 5-Hydroxymethylfurfurals—Dissolve 5.0 g of Fruc-tose in 100 mL of water, and read the absorbance at 284 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.32. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 4 g of Fructose, previously dried, dissolve in 0.2 mL of ammonia TS and 80 mL of water, and after standing for 30 minutes add water to make exactly 100 mL, and determine the optical rotation, aD, in a 100-mm cell at 20 ± 19C as directed under Optical Rotation Determination <2.49>. Amount (mg) of fructose (C6H12O6) ＝ |aD| × 1087.0
Fudosteine, when dried, contains not less than 99.
Fudosteine occurs as white, crystals or crystal-line powder. It is freely soluble in water, slightly soluble in acetic acid (100), and practically insoluble in ethanol (99.5). It dissolves in 6 mol/L hydrochloric acid TS. Melting point: about 2009C (with decomposition).
(1) To 5 mL of a solution of fudosteine (1 in 1000) add 2 mL of sodium hydroxide TS, shake well, add 0.3 mL of sodium pentacyanonitrosylferrate (III) TS, and shake well again. After allowing to stand at 409C for 10 minutes, cool the solution in an ice bath for 2 minutes, add 2 mL of dilute hydrochloric acid, and shake: a red-orange color develops. (2) Determine the infrared absorption spectrum of Fudosteine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －7.4 – －8.99(after drying, 1 g, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm).
(1) Chloride <1.03>—Dissolve 0.20 g of Fudosteine in 10 mL of water and 20 mL of nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of nitric acid and water to make 50 mL (not more than 0.044z). Heavy metals <1.07>—Proceed with 2.0 g of Fudoste- ine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Fudosteine according to Method 3, and perform the test (not more than 1 ppm). L-Cystine—Dissolve exactly 0.25 g of Fudosteine in the mobile phase to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, dissolve exactly 25 mg of L-cystine in 2 mL of 1 mol/L hydrochloric acid TS, then add the mobile phase to make exactly 50 mL. Pipet 2.5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the peak area of L-cystine obtained from the sample solution is not larger than the peak area of L-cystine from the standard so-lution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A solution of sodium 1-hexanesulfonate in diluted phosphoric acid (1 in 1000) (1 in 1250). Flow rate: Adjust so that the retention time of fudosteine is about 8 minutes. System suitability— System performance: Dissolve 25 mg of L-cystine in 2 mL of 1 mol/L hydrochloric acid TS, add 25 mg of Fudosteine, and add the mobile phase to make 50 mL. Take 2.5 mL of this solution, add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, L-cystine and fudosteine are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of L-cystine is not more than 2.0z. Related substances—Dissolve 0.25 g of Fudosteine in the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 100 mL, pipet 2.5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than fudosteine obtained from the sample solu-tion is not larger than the peak area of fudosteine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 559C. Mobile phase: Diluted phosphoric acid (1 in 1000). ‌1048 Flow rate: Adjust so that the retention time of fudosteine is about 3 minutes. Time span of measurement: About 10 times as long as the retention time of fudosteine, beginning after the peak of fudosteine. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fudosteine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fudosteine is not more than 2.0z. Loss on drying <2.41>—Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44>—Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Fudosteine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 17.92 mg of C6H13NO3S
Furosemide, when dried, contains not less than 98.
Furosemide occurs as white, crystals or crystal-line powder. It is freely soluble in N,N-dimethylformamide, soluble in methanol, sparingly soluble in ethanol (99.5), slightly soluble in acetonitrile and in acetic acid (100), and practically insolu-ble in water. It dissolves in dilute sodium hydroxide TS. It is gradually colored by light. Melting point: about 2059C (with decomposition).
(1) Dissolve 25 mg of Furosemide in 10 mL of methanol. To 1 mL of this solution add 10 mL of 2 mol/L hydrochloric acid TS. Heat the solution under a reflux con-denser on a water bath for 15 minutes, cool, and add 18 mL of sodium hydroxide TS to make weakly acidic: the solution responds to Qualitative Tests <1.09> for primary aromatic amines, producing a red to red-purple color. Determine the absorption spectrum of a solution of Furosemide in dilute sodium hydroxide TS (1 in 125,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Furosemide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Furosemide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Furosemide RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Furosemide in 10 mL of a solution of sodium hydroxide (1 in 50): the solution is clear and colorless. Chloride <1.03>—Dissolve 2.6 g of Furosemide in 90 mL of dilute sodium hydroxide TS, add 2 mL of nitric acid, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.40 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.020z). Sulfate <1.14>—To 20 mL of the filtrate obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution as follows: To 0.35 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydro-chloric acid and water to make 50 mL (not more than 0.030z). Heavy metals <1.07>—Proceed with 2.0 g of Furosemide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 25 mg of Furosemide in 25 mL of the dissolving solution, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the dissolving solution to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of each peak appeared ahead of the peak of furosemide ob-tained from sample solution is not larger than 2/5 times the peak area of furosemide from the standard solution, the area of each peak appeared behind the peak of furosemide is not larger than 1/4 times the peak area of furosemide from the standard solution, and the total area of these peaks is not larger than 2 times the peak area of furosemide from the standard solution. Dissolving solution—To 22 mL of acetic acid (100) add a mixture of water and acetonitrile (1:1) to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, tetrahydrofuran and acetic acid (100) (70:30:1). Flow rate: Adjust so that the retention time of furosemide is about 18 minutes. Time span of measurement: About 2.5 times as long as the retention time of furosemide, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 2 mL of the standard solution, and add the dissolving solution to make exactly 50 mL. Confirm that the peak area of furosemide obtained with 20 mL of this solution is equivalent to 3.2 to 4.8z of that with 20 mL of the standard solution. ‌1050 System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of furosemide is not less than 7000 and not more than 1.5, respectively. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of furosemide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Furosemide, previ-ously dried, dissolve in 50 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color of the solution changes from yellow to blue (indica-tor: 3 drops of bromothymol blue TS). Perform a blank de-termination with a mixture of 50 mL of N,N-dimethylfor-mamide and 15 mL of water in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 33.07 mg of C12H11ClN2O5S
Fursultiamine Hydrochloride contains not less than 98.
Fursultiamine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless or has a charac-teristic odor, and has a bitter taste. It is freely soluble in water, in methanol and in ethanol (95). It shows crystal polymorphism.
(1) Dissolve 5 mg of Fursultiamine Hydro-chloride in 6 mL of 0.1 mol/L hydrochloric acid TS, add 0.1 g of zinc powder, allow to stand for several minutes, and filter. To 3 mL of the filtrate, add 3 mL of sodium hy-droxide TS and 0.5 mL of potassium hexacyanoferrate (III) TS, then add 5 mL of 2-methyl-1-propanol, shake vigorously for 2 minutes, allow to stand to separate the 2-methyl-1-propanol layer, and examine under ultraviolet light (main wavelength: 365 nm): the 2-methyl-1-propanol layer shows a blue-purple fluorescence. The fluorescence disappears by acidifying, and appears again by alkalifying. Determine the infrared absorption spectrum of Fur-sultiamine Hydrochloride, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 24 hours, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Fursultiamine Hy- ‌1052‌ drochloride RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 24 hours: both spectra exhibit similar intensities of absorption at the same wave numbers. If any differences appear, dissolve the Fursultiamine Hydro-chloride in water, evaporate the water, and dry the residue in a desiccator (in vacuum, phosphorus (V) oxide) for 24 hours, and repeat the test. A solution of Fursultiamine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Fursultiamine Hydrochloride in 20 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Proceed with 1.5 g of Fursultiamine Hydrochloride, and perform the test. Prepare the control so-lution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.011z). Heavy metals <1.07>—Proceed with 1.0 g of Fursul- tiamine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Fursultiamine Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condition. Determine each peak area of each solution by the automatic integration method: the total area of the peaks other than fursultiamine obtained from the sample solution is not larger than the peak area of fursultiamine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in the operating conditions in the Assay. Detection sensitivity: Adjust the detection sensitivity so that the peak height of fursultiamine from 10 mL of the standard solution is between 20 mm and 30 mm. Time span of measurement: About 3 times as long as the retention time of fursultiamine. aminobenzoate in ethanol (95) (3 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 1.01 g of sodium 1-heptane sul-fonate in 1000 mL of diluted acetic acid (100) (1 in 100). To 675 mL of this solution add 325 mL of a mixture of metha-nol and acetonitrile (3:2). Flow rate: Adjust so that the retention time of Fursultia-mine is about 9 minutes. Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions and calculate the resolution. Use a column giving elution of fursultiamine and the internal standard in this order with the resolution be-tween these peaks being not less than 10.
Gabexate Mesilate, when dried, contains not less than 98.
Gabexate Mesilate occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in ethanol (95).
(1) To 4 mL of a solution of Gabexate Mesilate (1 in 2000) add 2 mL of 1-naphthol TS and 1 mL of diacetyl TS, and allow to stand for 10 minutes: a red color develops. Dissolve 1 g of Gabexate Mesilate in 5 mL of water, add 2 mL of sodium hydroxide TS, and heat in a water bath for 5 minutes. After cooling, add 2 mL of dilute nitric acid and 5 mL of ethanol (95), shake, add 5 drops of iron (III) chloride TS, and shake: a purple color develops. Determine the absorption spectrum of a solution of Gabexate Mesilate (1 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Gabexate Mesilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. A 0.1 g portion of Gabexate Mesilate responds to Qualitative Tests <1.09> (1) for mesilate. pH <2.54> Dissolve 1.0 g of Gabexate Mesilate in 10 mL of water: the pH of the solution is between 4.7 and 5.7. JP XVIII Official Monographs / b-Galactosidase (Aspergillus) 1053 ‌
(1) Clarity and color of solution—Dissolve 1.0 g of Gabexate Mesilate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Gabexate Mesilate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Dissolve 2.0 g of Gabexate Mesilate in 20 mL of 1 mol/L hydrochloric acid TS by heating in a water bath, and continue the heating for 20 minutes. After cooling, centrifuge, and use 10 mL of the supernatant liquid as the test solution. Perform the test (not more than 2 ppm). Ethyl parahydroxybenzoate—Weigh 50 mg of Gabexate Mesilate, previously dried, and dissolve in dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, dissolve 5.0 mg of ethyl parahydroxybenzoate in dilute ethanol to make exactly 100 mL. Pipet 1 mL of this solution, and add dilute ethanol to make exactly 20 mL. To exactly 5 mL of this solu-tion add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethyl parahydroxybenzoate to that of the internal standard: QT is not larger than QS. Internal standard solution—A solution of butyl parahy-droxybenzoate in dilute ethanol (1 in 5000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay. Related substances—Dissolve 0.20 g of Gabexate Mesilate in 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate until it has no acetic odor. Spray evenly a solution of 8-quinolinol in acetone (1 in 1000) on the plate, and after air-drying, spray evenly bromine-sodium hydroxide TS: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 55 mg each of Fursultiamine Hydrochloride and Fursultiamine Hydrochloride RS (previ-ously determined the water <2.48> in the same manner as Fursultiamine Hydrochiloride) and dissolve each in 50 mL of water, and add exactly 10 mL each of the internal standard solution, then add water to make exactly 100 mL. To 8 mL each of the solution add water to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of fursultiamine to that of the internal standard. Amount (mg) of fursultiamine hydrochloride (C17H26N4O3S2.HCl) ＝ MS × QT/QS MS: Amount (mg) of Fursultiamine Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl 4- Description Gabexate Mesilate occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in ethanol (95). Identification (1) To 4 mL of a solution of Gabexate Mesilate (1 in 2000) add 2 mL of 1-naphthol TS and 1 mL of diacetyl TS, and allow to stand for 10 minutes: a red color develops. Dissolve 1 g of Gabexate Mesilate in 5 mL of water, add 2 mL of sodium hydroxide TS, and heat in a water bath for 5 minutes. After cooling, add 2 mL of dilute nitric acid and 5 mL of ethanol (95), shake, add 5 drops of iron (III) chloride TS, and shake: a purple color develops. Determine the absorption spectrum of a solution of Gabexate Mesilate (1 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Gabexate Mesilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. A 0.1 g portion of Gabexate Mesilate responds to Qualitative Tests <1.09> (1) for mesilate. pH <2.54> Dissolve 1.0 g of Gabexate Mesilate in 10 mL of water: the pH of the solution is between 4.7 and 5.7. JP XVIII Official Monographs / b-Galactosidase (Aspergillus) 1053 ‌Melting point <2.60> 90 – 939C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Gabexate Mesilate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Gabexate Mesilate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Dissolve 2.0 g of Gabexate Mesilate in 20 mL of 1 mol/L hydrochloric acid TS by heating in a water bath, and continue the heating for 20 minutes. After cooling, centrifuge, and use 10 mL of the supernatant liquid as the test solution. Perform the test (not more than 2 ppm). Ethyl parahydroxybenzoate—Weigh 50 mg of Gabexate Mesilate, previously dried, and dissolve in dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, dissolve 5.0 mg of ethyl parahydroxybenzoate in dilute ethanol to make exactly 100 mL. Pipet 1 mL of this solution, and add dilute ethanol to make exactly 20 mL. To exactly 5 mL of this solu-tion add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethyl parahydroxybenzoate to that of the internal standard: QT is not larger than QS. Internal standard solution—A solution of butyl parahy-droxybenzoate in dilute ethanol (1 in 5000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay. Related substances—Dissolve 0.20 g of Gabexate Mesilate in 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate until it has no acetic odor. Spray evenly a solution of 8-quinolinol in acetone (1 in 1000) on the plate, and after air-drying, spray evenly bromine-sodium hydroxide TS: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 50 mg each of Gabexate Mesilate and Gabexate Mesilate RS, previously dried, and dissolve each in dilute ethanol to make exactly 100 mL. Pipet 5 mL each of these solutions, add exactly 5 mL each of the internal standard solution, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of gabexate to that of the internal standard. Amount (mg) of gabexate mesilate (C16H23N3O4.CH4O3S) ＝ MS × QT/QS MS: Amount (mg) of Gabexate Mesilate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in dilute ethanol (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, a solution of so-dium lauryl sulfate (1 in 1000), a solution of sodium 1-hep-tane sulfonate (1 in 200) and acetic acid (100) (540:200:20:1). Flow rate: Adjust so that the retention time of gabexate is about 13 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, the internal standard and gabexate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of gabexate to that of the internal standard is not more than 1.0z.
Powdered Gambir occurs as a red-brown to dark brown powder. It has a slight odor, and an extremely astringent and bitter taste. Under a microscope <5.01>, Powdered Gambir, immersed in olive oil or liquid paraffin, consists of masses of needle crystals or yellow-brown to red-brown angular fragments, and reveals epidermal tissue and thick-walled hairs.
(1) To 0.2 g of Powdered Gambir add 10 mL of water, warm in a water bath for 5 minutes with occa-sional shaking, and filter. Cool the filtrate, and add 2 to 3 drops of gelatin TS: a white turbidity or precipitate is pro-duced. (2) Shake 0.1 g of Powdered Gambir with 20 mL of dilute ethanol for 2 minutes, and filter. Mix 1 mL of the fil-trate with 9 mL of dilute ethanol, and to the solution add 1 mL of vanillin-hydrochloric acid TS: a light red to red-brown color develops. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 70.0z.
Brown to dark brown, brittle mass; inside light brown. Odor, slight; taste, extremely astringent and bitter.
(1) To 0.2 g of pulverized Gambir add 10 mL of water, warm in a water bath for 5 minutes with occa-sional shaking, and filter. Cool the filtrate, and add 2 to 3 drops of gelatin TS: a white turbidity or precipitate is pro-duced. (2) Shake 0.1 g of pulverized Gambir with 20 mL of dilute ethanol for 2 minutes, and filter. Mix 1 mL of the fil-trate with 9 mL of dilute ethanol, and to the solution add 1 mL of vanillin-hydrochloric acid TS: a light red to red-brown color develops. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 70.0z.
Gati‰oxacin Hydrate occurs as white to pale yellow, crystals or crystalline powder. It is slightly soluble in methanol and in ethanol (99.5), and very slightly soluble in water. It dissolves in sodium hydroxide TS. It is gradually colored to pale yellow by light. A solution of Gati‰oxacin Hydrate in dilute sodium hy-droxide TS (1 in 100) shows no optical rotation. Identiˆcation (1) Determine the absorption spectrum of a solution of Gati‰oxacin Hydrate in dilute sodium hydroxide TS (1 in 100,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Gati‰oxacin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Gati‰oxacin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Gati‰oxacin RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Gati‰oxacin Hydrate in 10 mL of sodium hydroxide TS: the solution is clear. Perform the test with the solution as di-rected under Methods for Color Matching <2.65>: the solu-tion is not more colored than diluted Matching Fluid O (1 in 5). Heavy metals <1.07>—Proceed with 1.0 g of Gati‰ox-acin Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Gati‰oxacin Hydrate in 50 mL of the dissolving solution, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, and add the dissolving solution to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolving solution to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con- ditions. Determine each peak area by the automatic integra-tion method: the peak area of the related substance A, hav-ing the relative retention time of about 1.2 to gati‰oxacin, obtained from the sample solution is not larger than 2 times the peak area of gati‰oxacin from the standard solution, and the area of the peak other than gati‰oxacin and the peak mentioned above from the sample solution is not larger than the peak area of gati‰oxacin from the standard solution. Furthermore, the total area of the peaks other than gati‰ox-acin from the sample solution is not larger than 3 times the <2.48> in the same manner as Gati‰oxacin Hydrate), and dis-solve each in the dissolving solution to make exactly 100 mL. Pipet 2 mL each of these solution, add exactly 2 mL of the internal standard solution to them, add the dissolving solu-tion to make 25 mL, and use these solutions as the sample solution and standard solutions. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of gati‰oxacin to that of the internal standard. peak area of gati‰oxacin from the standard solution. Dissolving solution: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (4:1). Amount (mg) of gati‰oxacin (C ＝ MS × QT/QS 19H22FN3O4) Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 325 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: A mixture of diluted triethylamine (1 in 100), adjusted to pH 4.3 with phosphoric acid, and aceto-nitrile (22:3). Mobile phase B: A mixture of diluted triethylamine (1 in 100), adjusted to pH 4.3 with phosphoric acid, and aceto-nitrile (1:1). ‌Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: Amount (mg) of Gati‰oxacin RS taken, calculated on the anhydrous basis Internal standard solution—A solution of methyl 4-aminobenzoate in the dissolving solution (1 in 4000). Dissolving solution: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (4:1). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted triethylamine (1 in 100), adjusted to pH 4.5 with phosphoric acid, and aceto-nitrile (87:13). Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Flow rate: Adjust so that the retention time of gati‰oxacin is about 5 minutes. 0 – 15 100 0 15 – 30 100 → 0 0 → 100 30 – 40 0 100 Flow rate: 1.0 mL per minute (the retention time of gati‰oxacin is about 16 minutes). Time span of measurement: About 2.5 times as long as the retention time of gati‰oxacin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the dissolving solution to make exactly 10 mL. Conˆrm that the peak area of gati‰oxacin obtained with 20 mL of this solution is equivalent to 40 to 60z of that with 20 mL of the standard solution. System performance: Dissolve 20 mg of methyl 4-aminobenzoate in 50 mL of the dissolving solution. To 5 mL of this solution add 1 mL of the sample solution and the dis-solving solution to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating condi-tions, gati‰oxacin and methyl 4-aminobenzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of gati‰oxacin is not more than 3.0z. Water <2.48> 6.0 – 9.0z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg of Gati‰oxacin Hy-drate and Gati‰oxacin RS (separately determine the water System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, gati‰oxacin and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of gati‰oxacin to that of the internal standard is not more than 1.0z.
Gefarnate is a mixture of 4E geometrical isomer. It contains not less than 98.
Gefarnate is a light yellow to yellow, clear oily liquid. It is miscible with acetonitrile, with ethanol (99.5) and with cyclohexane. It is practically insoluble in water.
Determine the infrared absorption spectrum of Gefarnate as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Gefarnate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 20: 0.906 – 0.914
(1) Acidity—To 1.0 g of Gefarnate add 30 mL of neutralized ethanol. To this solution add 1 drop of phenol-phthalein TS and 0.40 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Proceed with 2.0 g of Gefar- nate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Use a solution of Gefarnate in acetonitrile (1 in 500) as the sample solution. Pipet 2 mL of the sample solution, add acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than the peak of gefar-nate obtained from the sample solution is not larger than 1/2 times the peak area of gefarnate from the standard solution, and the total area of the peaks other than the peak of gefar-nate from the sample solution is not larger than the peak area of gefarnate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of gefarnate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, add acetonitrile to make exactly 20 mL. Confirm that the peak area of gefarnate obtained with 2 mL of this so-lution is equivalent to 7 to 13z of that of gefarnate with 2 mL of the standard solution. System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of gefarnate are not less than 4000, and be-tween 0.9 and 1.2, respectively. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of gefarnate is not more than 1.0z. Isomer ratio To 1 mL of Gefarnate add 100 mL of ethanol (99.5), and use this solution as the sample solution. Perform the test with 4 mL of the sample solution as directed under Gas Chromatography <2.02> according to the following con-ditions. Determine the areas of two adjacent peaks, Aa and Ab, having the retention time of about 37 minutes, where Aa is the peak area of shorter retention time and Ab is the peak area of longer retention time: Aa/(Aa ＋ Ab) is between 0.2 and 0.3. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 160 cm in length, packed with polyethylene glycol 20M for gas chromatography coated at the ratio of 5z on acid-treated and silanized siliceous earth for gas chromatography (149 to 177 mm in particle diameter). Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen. Flow rate: Adjust so that the reaction time of the peak showing earlier elution of the two peaks of gefarnate is about 35 minutes. System suitability— System performance: When the procedure is run with 4 mL of the sample solution under the above conditions: the reso-lution between the two peaks of gefarnate is not less than 1.0. System repeatability: When the test is repeated 6 times with 4 mL of the sample solution under the above operating ‌JP XVIII Official Monographs / Gefitinib 1059 conditions: the relative standard deviation of the peak area of gefarnate with the shorter retention time of the two peaks of gefarnate is not more than 2.0z.
Weigh accurately about 50 mg each of Gefarnate and Gefarnate RS, add exactly 5 mL of the internal standard solution and 20 mL of acetonitrile, and use these solutions as the sample solution and standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of gefarnate to that of the internal standard. Amount (mg) of gefarnate (C27H44O2) ＝ MS × QT/QS MS: Amount (mg) of Gefarnate RS taken Internal standard solution—A solution of tris (4-t-butyl-phenyl) phosphate in acetonitrile (1 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with phenylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile for liquid chroma-tography, water and phosphoric acid (700:300:1). Flow rate: Adjust so that the retention time of gefarnate is about 19 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the internal standard and gefarnate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of gefarnate to that of the internal standard is not more than 1.0z.
Gefitinib occurs as a white powder. It is slightly soluble in ethanol (99.5), and practically in-soluble in water.
(1) Determine the absorption spectrum of a solution of Gefitinib in a mixture of trifluoroacetic acid so-lution (1 in 500) and acetonitrile (3:2) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Gefitinib RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Gefitinib as directed in the potassium bromide disk method under Infrared Spectrometry <2.25>, and compare the spec-trum with the Reference Spectrum or the spectrum of Gefitinib RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. Or, perform the test by the diffuse reflectance method, and compare the spectrum with the spectrum of Gefitinib RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers.
(1) Heavy metals—Being specified separately when the drug is granted approval based on the Law. (2) Related substances－Use the sample solution ob- tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of trifluoroacetic acid solu-tion (1 in 500) and acetonitrile (3:2) to make exactly 100 mL. Pipet 1 mL of this solution, and add a mixture of trifluoroa-cetic acid solution (1 in 500) and acetonitrile (3:2) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of related substance A having the rel-ative retention time of about 0.13 to gefitinib obtained from the sample solution is not larger than the peak area of gefitinib from the standard solution, the peak area of related substance B having the relative retention time of about 1.3 from the sample solution is not larger than 2 times the peak area of gefitinib from the standard solution, and the area of the peak other than gefitinib and the peaks mentioned above from the sample solution is not larger than the peak area of gefitinib from the standard solution. Furthermore, the total area of the peaks other than gefitinib from the sample solu-tion is not larger than 4 times the peak area of gefitinib from ‌the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of gefitinib, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of trifluoroacetic acid solution (1 in 500) and acetonitrile (3:2) to make exactly 10 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, the SN ratio of the peak of gefitinib is not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of gefitinib is not more than 2.0z. Water <2.48> Not more than 0.4z (0.1 g, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (1.0 g, platinum crucible).
Weigh accurately about 35 mg each of Gefitinib and Gefitinib RS (separately determine the water <2.48> in the same manner as Gefitinib), add 85 mL of a mixture of trifluoroacetic acid solution (1 in 500) and acetonitrile (3:2) to each, sonicate to dissolve, add a mixture of trifluoroacetic acid solution (1 in 500) and acetonitrile (3:2) to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of gefitinib in each solution. Amount (mg) of gefitinib (C22H24ClFN4O3) ＝ MS × AT/AS MS: Amount (mg) of Gefitinib RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 247 nm). Column: A stainless steel column 3 mm in inside diameter and 10 cm in length, packed with octadesylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of ammonium acetate solution (3 in 310) and acetonitrile (31:19). Flow rate: 0.9 mL per minute (the retention time of gefitinib is about 5.5 minutes). System suitability— System performance: Dissolve 15 mg of 3,4-dichloroani-line in 60 mL of the standard solution. When the procedure is run with 5 mL of this solution under the above operating conditions, 3,4-dichloroaniline and gefitinib are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of gefitinib is not more than 1.0z.
Gelatin occurs as colorless or white to light yellow-brown sheets, shreds, granules or powder. It is freely soluble in hot water, and practically insoluble in ethanol (95). The gelling grade does not dissolve in water, but slowly swells and softens when immersed in it, gradually absorbing water 5 to 10 times its own mass. The gelling grade derived from an acid-treated collagen ex-hibits an isoelectric point between pH 7.0 and 9.0, and the gelling grade derived from an alkali-treated collagen exhibits an isoelectric point between pH 4.5 and 5.0. The non-gelling grade is freely soluble in water.◆
(1) Dissolve 1.00 g of Gelatin in freshly boiled and cooled water at about 559C to make 100 mL, and use this solution as the sample solution. To 2 mL of the sam-ple solution keeping at about 559C add 0.05 mL of copper (II) sulfate TS. Mix and add 0.5 mL of 2 mol/L sodium hy-droxide TS: a violet color is produced. In a test tube about 15 mm in internal diameter, place JP XVIII Official Monographs / Gelatin 1061 0.5 g of Gelatin, add 10 mL of water, and allow to stand for 10 minutes. Heat at 609C for 15 minutes, then keep the tube upright at 2 to 89C for 6 hours, and invert the tube: the con-tents do not flow out immediately for the gelling grade. In case of the non-gelling grade the contents flow out immedi-ately. Apply to the non-gelling grade. Place 0.5 g of Gelatin in a 250-mL flask and add 10 mL of water and 5 mL of sul-furic acid. Cover with a watch glass or other instrument, avoiding complete closure, and heat at 1059C for 4 hours. Cool, add 200 mL of water, and then adjust to pH 6.0 to 8.0 with a sodium hydroxide solution (1 in 5). In a test tube, place 2 mL of this solution, add 2 mL of the oxidizing rea-gent, shake, and allow to stand for 20 minutes. Add 2 mL of the colored solution, shake, and warm in a water bath of 609C for about 15 minutes: a red to purple color develops. Oxidizing reagent: Dissolve 5.53 g of disodium hydrogen phosphate dodecahydrate and 0.48 g of citric acid monohy-drate in water to make 100 mL. Dissolve 1.4 g of sodium toluenesulfonchloramide trihydrate in this solution. Prepare before use. Colored solution: Dissolve 1.0 g of 4-dimethylaminoben-zaldehyde in 3.5 mL of a solution of perchloric acid (1 in 2) and add slowly 6.5 mL of 2-propanol. Prepare before use. Gel strength (Bloom value) Apply to the gelling grade. De-termine the mass (g) necessary to produce the force which, applied to a plunger 12.7 mm in diameter, makes a depres-sion 4 mm deep in a gel having a concentration of 6.67z and matured at 109C. Apparatus: Texture analyzer or gelometer with a cylindrical piston 12.7 ± 0.1 mm in diameter with a plane pressure surface and a sharp bottom edge, and with a bottle 59 ± 1 mm in internal diameter and 85 mm high (jelly cup). Procedure: Place 7.5 g of Gelatin in a jelly cup, add 105 mL of water, close the cup, and allow to stand for 1 to 4 hours. Heat in a water bath at 65 ± 29C for 15 minutes. While heating, stir gently with a glass rod. Ensure that the solution is uniform and any condensed water on the inner walls of the cup is incorporated. Allow to cool at room tem-perature for 15 minutes and transfer the cup to a thermostat-ically controlled bath at 10.0 ± 0.19C, and fitted with a device to ensure that the platform on which the cup stands is perfectly horizontal. Close the cup, and allow to stand for 17 ± 1 hours. Remove the sample cup from the bath and quickly wipe the water from the exterior of the cup. Center the cup on the platform of the apparatus so that the plunger contacts the sample as nearly at its midpoint as possible, and start the measurement with 4 mm depression distance and 0.5 mm per second test speed: 80 to 120z of the labeled nominal value. pH <2.54> pH at 559C of the sample solution obtained in Identification (1) is 3.8 – 7.6.
◇(1) Heavy metals <1.07>—Proceed with 0.5 g of Gelatin according to Method 2, and perform the test. Pre-pare the control solution with 2.5 mL of Standard Lead So-lution (not more than 50 ppm).◇ Iron—To 5.00 g of Gelatin, in a glass-stoppered flask, add 10 mL of hydrochloric acid, close the flask, and heat in a water bath at 75 – 809C for 2 hours. If necessary for proper solubilization, the gelatin may be allowed to swell after addition of the acid and before heating, the heating time may be prolonged and a higher temperature may be used. After cooling, adjust the content of the flask to 100.0 g with water, and use this solution as the sample solution. Separately, place 5.00 g each of Gelatin in three glass-stop- pered flasks, proceed with them in the same manner as the sample solution, then add 10 mL, 20 mL and 30 mL of Stan-dard Iron Solution (2) for Atomic Absorption Spectrophoto-metry exactly to each flask separately. Adjust the content of these flasks to 100.0 g each with water, and use these solu-tions as the standard solutions. The amount of the standard solution to be added may be adjusted according to the sen-sitivity of the instrument to be used. Perform the test with the sample solution and standard solutions as directed in the standard addition method under Atomic Absorption Spec-trophotometry <2.23> according to the following conditions, and determine the content of iron: not more than 30 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Iron hollow cathode lamp. Wavelength: 248.3 nm. Chromium—Use the sample solution obtained in (2) as the sample solution. Separately, place 5.00 g each of Gela-tin in three glass-stoppered flasks, proceed with them in the same manner as the sample solution, then add 0.25 mL, 0.50 mL and 0.75 mL of Standard Chromium Solution for Atomic Absorption Spectrophotometry exactly to each flask separately. Adjust the content of these flasks to 100.0 g each with water, and use these solutions as the standard solutions. The amount of the standard solution to be added may be adjusted according to the sensitivity of the instrument to be used. Perform the test with the sample solution and standard solutions as directed in the standard addition method under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and determine the content of chro-mium: not more than 10 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Chromium hollow cathode lamp. Wavelength: 357.9 nm. Zinc—Use the sample solution obtained in (2) as the sample solution. Separately, place 5.00 g each of Gelatin in three glass-stoppered flasks, proceed with them in the same manner as the sample solution, then add 7.5 mL, 15 mL and 22.5 mL of Standard Zinc Solution for Atomic Absorption Spectrophotometry exactly to each flask separately. Adjust the content of these flasks to 100.0 g each with water, and use these solutions as the standard solutions. The amount of the standard solution to be added may be adjusted according to the sensitivity of the instrument to be used. Perform the test with the sample solution and standard solutions as directed in the standard addition method under Atomic Absorption Spectrophotometry <2.23> according to the fol-lowing conditions, and determine the content of zinc: not more than 30 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm. (5) Arsenic <1.11>—Take 15.0 g of Gelatin in a flask, add 60 mL of diluted hydrochloric acid (1 in 5), and dissolve by heating. Add 15 mL of bromine TS, heat until the excess of bromine is expelled, neutralize with ammonia TS, add 1.5 g of disodium hydrogen phosphate dodecahydrate, and allow to cool. To this solution add 30 mL of magnesia TS, allow to stand for 1 hour, and collect the precipitates. Wash the precipitates with five 10-mL portions of diluted ammonia TS (1 in 4), and dissolve in diluted hydrochloric acid (1 in 4) to make exactly 50 mL. Perform the test with 5 mL of this solution: the solution has no more color than the following color standard. Color standard: Proceed with 15 mL of Standard Arsenic ‌1062 Solution, instead of Gelatin, in the same manner (not more than 1 ppm).◇ Peroxides— Enzyme reaction: Peroxidase transfers oxygen from peroxides to an organic redox indicator which is converted to a blue oxidation product. The intensity of the color obtained is proportional to the quantity of peroxide and can be com-pared with a color scale provided with the test strips, to de-termine the peroxide concentration. Procedure: Weigh 20.0 ± 0.1 g of Gelatin in a beaker, add 80.0 ± 0.2 mL of water, and stir to moisten all the gelatin. Allow to stand at room temperature for 1 – 3 hours. Cover the beaker with a watch-glass, and heat the beaker for 20 ± 5 minutes in a water bath at 65 ± 29C for dissolving the sample. Stir the contents of the beaker with a glass rod to achieve a homogeneous solution, and use this as the sample solution. Dip a peroxide test strip for 1 second into the sample solution, such that the reaction zone is prop-erly wetted. Remove the test strip, shake off excess liquid, and compare the reaction zone after 15 seconds with the color scale provided. Multiply the concentration read from the color scale by a factor of 5 to calculate the concentration of peroxide in the test substance: not more than 10 ppm. Suitability test: To exactly 10 mL of Standard Hydrogen Peroxide Solution add water to make exactly 300 mL. Pipet 2 mL of this solution, add water to make exactly 1000 mL (2 ppm). Dip a peroxide test strip for 1 second into this solution, such that the reaction zone is properly wetted. Remove the test strip, shake off excess liquid and compare the color of the reaction zone after 15 seconds with the color scale: the color of the zone is equivalent to 2 ppm of the color scale. Sulfur dioxide— Apparatus: Use as shown in the figure. A: Three-necked round-bottomed flask (500 mL) B: Cylindrical dropping funnel (100 mL) C: Condenser D: Test tube E: Tap Procedure: Introduce 150 mL of water into the three-necked round-bottomed flask and pass carbon dioxide through the whole system at a rate of 100 mL per minute. Place 10 mL of hydrogen peroxide-sodium hydroxide TS in the test tube. After 15 minutes, remove the cylindrical drop-ping funnel without interrupting the stream of carbon di-oxide, and introduce through the opening into the three-necked round-bottomed flask about 25.0 g of Gelatin with the aid of 100 mL of water. Pour 80 mL of 2 mol/L hydrochloric acid TS into the funnel, open the tap to in-troduce the hydrochloric acid into the three-necked round-bottomed flask ◇and close the tap while several mL of the hydrochloric acid remains, in order to avoid losing sulfur di-oxide◇, and boil the mixture for 1 hour. Transfer the con-tents of the test tube with the aid of a little water to a 200 mL wide-necked conical flask. Heat the flask in a water bath for 15 minutes and cool. Add 0.1 mL of bromophenol blue TS and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the amount of sulfur dioxide from the following expression: it is not more than 50 ppm. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 M: Amount (g) of Gelatin taken V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed Conductivity <2.51> Perform the test at 30 ± 1.09C with the sample solution obtained in Identification (1), without temperature compensation: not more than 1 mS･cm－1. Loss on drying <2.41> Not more than 15.0z (5 g, 1059C, 16 hours). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 103 CFU/g and 102 CFU/g, respectively. Escherichia coli and Salmonella are not observed.
◇Containers—Tight containers.◇ Storage—Protect from heat and moisture.
Weigh accurately a suitable amount of Insu-lin Human (Genetical Recombination), and dissolve in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains 2.0 mg. Transfer 500 mL of this solution into a clean test tube, add 2.0 mL of HEPES buffer solution (pH 7.5) and 400 mL of V8-protease TS, incubate at 259C for 6 hours, then add 2.9 mL of ammonium sulfate buffer solu-tion to stop the reaction, and use this solution as the sample solution. Separately, proceed with Insulin Human RS in the same manner as above, and use this solution as the standard solution. Perform the test with 50 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and compare the chromatograms obtained from these solu-tions: a similar peak is observed at the same retention time in the both chromatograms. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, ammonium sulfate buffer solution and acetonitrile (7:2:1). Mobile phase B: A mixture of water, acetonitrile and am-monium sulfate buffer solution (2:2:1). Flowing of mobile phase: Change the mixing ratio of the mobile phase A and B linearly from 9:1 to 3:7 in 60 minutes after sample injection, further change to 0:10 linearly in 5 minutes, and then flow the mobile phase B only for 5 minutes. Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con- Insulin Human (Genetical Recombination) / Official Monographs ditions, the symmetry factor of the two larger peaks which appear next to the first peak just after the solvent peak are not more than 1.5 respectively, and the resolution between these peaks is not less than 3.4.
(1) Related substances—Perform this procedure rapidly. Dissolve 7.5 mg of Insulin Human (Genetical Recombination) in 2 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak area of human insulin, A1, the peak area of the desamido substance having the relative retention time of about 1.3 to human insulin, AD, and the total area of the peaks other than the solvent peak, AT: the amounts of the desamido substance and related substances other than the desamido substance are not more than 2.0z, respectively. Previously, perform the test with 0.01 mol/L hydrochloric acid TS in the same manner to confirm the sol-vent peak. Amount (z) of the desamido substance ＝ AD/AT × 100 Amount (z) of related substances other than the desamido substance ＝ [{AT － (A1 ＋ AD)}/AT] × 100 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of phosphoric acid-sodium sulfate buffer solution (pH 2.3) and acetonitrile for liquid chromatography (41:9). Mobile phase B: A mixture of phosphoric acid-sodium sul-fate buffer solution (pH 2.3) and acetonitrile for liquid chro-matography (1:1). Flowing of mobile phase: Flow a mixture of the mobile phase A and B (78:22) for 36 minutes before and after the sample injection, then change the mixing ratio to 33:67 linearly in 25 minutes, and maintain this ratio for 6 minutes. Then flow the first mixture (78:22) for the next 15 minutes. Adjust the mixing ratio of the first mixture so that the reten-tion time of human insulin is about 25 minutes. Flow rate: 1.0 mL per minute. Time span of measurement: For about 75 minutes after the sample is injected. System suitability— Test for required detectability: Confirm that the peak height of the desamido substance obtained with 20 mL of human insulin desamido substance-containing TS is between 30z and 70z of the full scale. System performance: When the procedure is run with 20 mL of human insulin desamido substance-containing TS under the above operating conditions, human insulin and human insulin desamido substance are eluted in this order with the resolution between these peaks being not less than 2.0, and the symmetry factor of the peak of human insulin is not more than 1.8. High-molecular mass proteins—Dissolve 4 mg of In-sulin Human (Genetical Recombination) in 1 mL of 0.01 mol/L hydrochloric acid TS. Perform the test with 100 mL of this solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate each peak area: the total of areas of the peaks having smaller retention time than human insulin is not more than 1.0z of the total area of all peaks. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 276 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of L-arginine (1 in 1000), acetonitrile and acetic acid (100) (13:4:3). Flow rate: Adjust so that the retention time of human in-sulin is about 20 minutes. Time span of measurement: Until the peak of human insu-lin monomer has appeared. System suitability— Test for required detectability: Confirm that the peak height of the dimer obtained with 100 mL of human insulin dimer-containing TS is between 10z and 50z of the full scale. System performance: When the procedure is run with 100 mL of human insulin dimer-containing TS under the above operating conditions, polymer, dimer and monomer are eluted in this order, and the ratio, H1/H2, of the peak height of the dimer H1 to the height of the bottom between the peaks of the dimer and the monomer H2 is not less than 2.0. Product related impurities—Being specified sepa-rately when the drug is granted approval based on the Law. Process related impurities—Being specified sepa-rately when the drug is granted approval based on the Law.
Containers—Well-closed contain-ers. Light-resistant, and in a cold place
Gentamicin Sulfate is the sulfate of a mixture of aminoglycoside substances having antibacterial ac-tivity produced by the growth of Micromonospora purpurea or Micromonospora echinospora. It contains not less than 590 mg (potency) and not more than 775 mg (potency) per mg, calculated on the dried basis.
Gentamicin Sulfate occurs as a white to light yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic.
(1) Dissolve 50 mg each of Gentamicin Sul- JP XVIII Official Monographs / Gentamicin Sulfate 1065 fate and Gentamicin Sulfate RS in 10 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and standard solution on a plate of silica gel for thin-layer chromatography. Separately, shake a mixture of chlo-roform, ammonia solution (28) and methanol (2:1:1) in a separator, and allow the mixture to stand for more than 1 hour. To 20 mL of the lower layer so obtained add 0.5 mL of methanol, and use this as the developing solvent. Develop the plate with the developing solvent to a distance of about 17 cm in a developing container with a cover, having an opening of about 20 mm2, and without putting a filter paper in the container, and air-dry the plate. Allow the plate to stand in iodine vapors: three principal spots obtained from the sample solution are the same with the corresponding spots from the standard solution in color tone and the Rf value, respectively. (2) Dissolve 50 mg of Gentamicin Sulfate in 5 mL of water, and add 0.5 mL of barium chloride TS: a white pre-cipitate is formed. D Optical rotation <2.49> [a]25: ＋107 – ＋1219(0.25 g calcu-lated on the dried basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.20 g of Gentamicin Sulfate in 5 mL of water is between 3.5 and 5.5. Content ratio of the active principle Dissolve 50 mg of Gentamicin Sulfate in water to make 10 mL, and use this so-lution as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Separately, shake a mix-ture of chloroform, ammonia solution (28) and methanol (2:1:1) in a separator, and allow the mixture to stand for more than 1 hour. To 20 mL of the lower layer so obtained add 0.5 mL of methanol, and use this as the developing sol-vent. Develop the plate with the developing solvent to a dis-tance of about 17 cm in a developing container with a cover, having an opening of about 20 mm2, without putting a filter paper in the container, and air-dry the plate. Allow the plate to stand in iodine vapor. Determine the integral absor-bances, Aa, Ab and Ac, of the colored spots of gentamicin C1 (Rf value: about 0.3), gentamicin C2 (Rf value: about 0.2) and gentamicin C1a (Rf value: about 0.1), respectively, by a densitometer (wavelength: 450 nm) while covering the plate with a glass plate, and calculate these amounts by the follow-ing formulae: gentamicin C1 is between 25z and 55z, gen-tamicin C2 is between 25z and 50z, and gentamicin C1a is between 5z and 30z. Amount (z) of gentamicin C1 ＝ Aa/(Aa ＋ 1.35Ab ＋ Ac) × 100 Amount (z) of gentamicin C2 ＝ 1.35Ab/(Aa ＋ 1.35Ab ＋ Ac) × 100 Amount (z) of gentamicin C1a ＝ Ac/(Aa ＋ 1.35Ab ＋ Ac) × 100
(1) Clarity and color of solution—Dissolve 1.0 g of Gentamicin Sulfate in 10 mL of water: the solution is clear and its absorbance at 400 nm determined as directed under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.08. Heavy metals <1.07>—Proceed with 2.0 g of Gen- tamicin Sulfate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Gentamicin Sulfate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Separately, shake a mixture of chloroform, ammonia solution (28) and methanol (2:1:1) in a separator, and allow the mixture to stand for more than 1 hour. To 20 mL of the lower layer so obtained add 0.5 mL of methanol, and use this as the developing solvent. Develop the plate with the developing solvent to a distance of about 17 cm in a developing container with a cover, having an opening of about 20 mm2, without putting a filter paper in the container, and air-dry the plate. Allow the plate to stand in iodine vapor, and compare the colored spots while cover-ing with a glass plate: the spots other than the spots of gen-tamicin C1 (Rf value: about 0.3), gentamicin C2 (Rf value: about 0.2) and gentamicin C1a (Rf value: about 0.1) obtained from the sample solution are not more intense than the spot of gentamicin C2 from the standard solution. Loss on drying <2.41> Not more than 18.0z (0.15 g, reduced pressure not exceeding 0.67 kPa, 1109C, 3 hours). Handle the sample avoiding absorption of moisture. Residue on ignition <2.44> Not more than 1.0z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus epidermidis ATCC 12228 Agar media for seed and base layer— Glucose 1.0 g Peptone 6.0 g Meat extract 1.5 g Yeast extract 3.0 g Sodium chloride 10.0 g Agar 15.0 g Water 1000 mL Mix all the ingredients, and sterilize. Adjust the pH of the solution so that it will be 7.8 to 8.0 after sterilization. Agar medium for transferring test organisms—Use the medium ii in 2) Medium for other organisms under (2) Agar media for transferring test organisms. Standard solutions—Weigh accurately an amount of Gentamicin Sulfate RS, equivalent to about 25 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 159C or lower, and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, re-spectively. Sample solutions—Weigh accurately an amount of Gentamicin Sulfate, equivalent to about 25 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and the low concentra- ‌1066‌ tion sample solution, respectively.
Glacial Acetic Acid contains not less than 99.
Glacial Acetic Acid is a clear, colorless, vola-tile liquid, or colorless or white, crystalline masses. It has a pungent, characteristic odor. It is miscible with water, with ethanol (95) and with diethyl ether. Boiling point: about 1189C Specific gravity d 20: about 1.049
A solution of Glacial Acetic Acid (1 in 3) changes blue litmus paper to red, and responds to Qualitative Tests <1.09> for acetate. Congealing point <2.42> Not less than 14.59C.
(1) Chloride—To 10 mL of Glacial Acetic Acid add water to make 100 mL, and use this solution as the sam-ple solution. To 10 mL of the sample solution add 5 drops of silver nitrate TS: no opalescence is produced. Sulfate—To 10 mL of the sample solution obtained in (1) add 1 mL of barium chloride TS: no turbidity is pro-duced. Heavy metals <1.07>—Evaporate 2.0 mL of Glacial Acetic Acid on a water bath to dryness. Dissolve the residue in 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution by adding 2.0 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Potassium permanganate-reducing substances—To 20 mL of the sample solution obtained in (1) add 0.10 mL of 0.02 mol/L potassium permanganate VS: the red color does not disappear within 30 minutes. Non-volatile residue—Evaporate 10 mL of Glacial Acetic Acid on a water bath to dryness, and dry at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg.
Place 10 mL of water in a glass-stoppered flask, and weigh accurately. Add about 1.5 g of Glacial Acetic Acid, weigh accurately again, then add 30 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 drops of phenolphthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 60.05 mg of C2H4O2
Glibenclamide, when dried, contains not less than 98.
Glibenclamide occurs as white to pale yellow-ish white, crystals or crystalline powder. It is freely soluble in dimethylformamide, sparingly solu-ble in chloroform, slightly soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Glinbenclamide in methanol (1 in 10,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Gli-benclamide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Glibenclamide as directed under Flame Coloration Test <1.04> (2): a green color ap-pears.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Glibenclamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.20 g of Glibencla-mide in 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 20 mL. Pipet 1 mL of this solu-tion, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, chloroform and diluted ammonia TS (4 in 5) (11:7:2) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours).
Weigh accurately about 0.9 g of Glibenclamide, pre-viously dried, dissolve in 50 mL of N,N-dimethylformamide, ‌1068 and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (in-dicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner with a solution prepared by adding 18 mL of water to 50 mL of N,N-dimethylfor-mamide, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 49.40 mg of C23H28ClN3O5S
Gliclazide, when dried, contains not less than 98.
Gliclazide is a white crystalline powder. It is sparingly soluble in acetonitrile and in methanol, slightly soluble in ethanol (99.5) and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Gliclazide in methanol (1 in 62,500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Gliclazide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Gliclazide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Conduct this procedure within 2 hours after preparation of the sample solution. Dissolve 50 mg of Gliclazide in 23 mL of acetonitrile, add water to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (11:9) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of water and acetonitrile (11:9) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than gliclazide obtained from the sample solution is not larger than the peak area of gliclazide from the standard so-lution, and the total area of the peaks other than the peak of
Weigh accurately about 0.3 g of Gliclazide, previ-ously dried, dissolve in 30 mL of a mixture of acetic anhy-dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.34 mg of C H N O S 15 21 3 3 Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Gliclazide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Conduct this procedure within 2 hours after preparation of the sample solution. Dissolve 50 mg of Gliclazide in 23 mL of acetonitrile, add water to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (11:9) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of water and acetonitrile (11:9) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than gliclazide obtained from the sample solution is not larger than the peak area of gliclazide from the standard so-lution, and the total area of the peaks other than the peak of
Glimepiride occurs as a white crystalline pow-der. It is slightly soluble in dichloromethane, very slightly solu-ble in methanol and in ethanol (99.5), and practically insolu-ble in water. Melting point: about 2029C (with decomposition).
(1) Determine the absorption spectrum of a solution of Glimepiride in methanol (1 in 125,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Glimepiride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Glimepiride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Glimepiride RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Glimepiride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). cis-Isomer—Dissolve 10 mg of Glimepiride in 5 mL of dichloromethane, add the mobile phase to make 20 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area of both solutions by the automatic in-tegration method: the area of the peak, having the relative retention time of about 0.9 to glimepiride, obtained from the sample solution is not larger than 3/4 times the peak area of glimepiride from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 3 mm in inside diameter and 15 cm in length, packed with diol silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of heptane for liquid chromatog-raphy, 2-propanol for liquid chromatography, and acetic acid (100) (900:100:1). Flow rate: Adjust so that the retention time of glimepiride is about 14 minutes. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of glimepiride obtained with 10 mL of this solution is equivalent to 35 to 65z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z. Related substances—Keep the sample solution and standard solution below 49C after preparing. Dissolve 20 mg of Glimepiride in 100 mL of a mixture of acetonitrile for liq-uid chromatography and water (4:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.25 to glimepiride, obtained from the sample solution is not larger than 4 times the peak area of glimepi-ride from the standard solution, the area of the peak, having the relative retention time of about 1.1, is not larger than 2 times the peak area of glimepiride from the standard solu-tion, the area of the peak, having the relative retention time of about 0.32, is not larger than 1.5 times the peak area of glimepiride from the standard solution, the area of peak other than glimepiride and above mentioned peak from the sample solution is not larger than the peak of glimepiride from the standard solution, and the total area of the peaks other than glimepiride and the peak, having the relative retention time of about 0.25, from the sample solution is not larger than 5 times the peak area of glimepiride from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of glimepiride, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of glimepiride obtained with 20 mL of this solution is equivalent to 35 to 65z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 9000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times ‌1070 with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z. Water <2.48> Not more than 0.5z (0.25 g, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 20 mg each of Glimepiride and Glimepiride RS (separately determine the water <2.48> in the same manner as Glimepiride), dissolve each substance in a mixture of acetonitrile for liquid chromatography and water (4:1) to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 20 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of glimepi-ride in each solution. sure, suspend the residue with 1 mL of water, and filter under reduced pressure. Wash the residue with 1 mL of water, dry at 1059C for 1 hour. Determine the infrared ab-sorption spectrum as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 3370 cm－1, 3290 cm－1, 2930 cm－1, 1708 cm－1, 1674 cm－1, 1347 cm－1, 1156 cm－1 and 618 cm－1. Purity Related substances—Keep the sample solution and standard solution below 49C after preparation. To a quan-tity of powdered Glimepiride Tablets, equivalent to 9 mg of Glimepiride, wet with 0.5 mL of water, add a mixture of acetonitrile for liquid chromatography and water (4:1) to make 50 mL, shake, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of acetonitrile for liquid chromatogra-phy and water (4:1) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly Amount (mg) of glimepiride (C ＝ MS × AT/AS 24H34N4O5S) 5 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.5 g of sodium dihydrogen phos-phate dihydrate in 500 mL of water, adjust to pH 2.5 with phosphoric acid, and add 500 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of glimepiride is about 17 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 9000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glimepiride is not more than 1.0z.
Glucagon (Genetical Recombination) is a recom-binant human glucagon, and is a peptide consisting of 29 amino acid residues. It contains not less than 92.
Glucagon (Genetical Recombination) occurs as a white lyophilized powder. It is practically insoluble in water and in ethanol (99.5). It is hygroscopic.
(1) Dissolve 5 mg of Glucagon (Genetical Recombination) in 1 mL of 0.01 mol/L hydrochloric acid TS. To 200 mL of this solution add 800 mL of 0.1 mol/L am-monium hydrogen carbonate TS and 25 mL of enzyme TS for glucagon, react at 379C for 2 hours, add 120 mL of acetic acid (100) to stop the reaction, and use this solution as the sample solution. Separately, dissolve a suitable amount of Glucagon RS in 0.1 mol/L ammonium hydrogen carbonate TS so that each mL contains 1 mg of glucagon. To 1000 mL of this solution add 25 mL of enzyme TS for glucagon, react at 379C for 2 hours, add 120 mL of acetic acid (100) to stop the reaction, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and compare the chromatograms obtained from these solutions: both chromatograms show the similar peaks at the same retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4 mm in inside diameter and 50 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 229C. Mobile phase A: To 0.5 mL of trifluoroacetic acid add 1000 mL of water. Mobile phase B: To 0.5 mL of trifluoroacetic acid add 600 mL of ethanol (99.5) and 400 mL of water. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the peaks 1, 2, 3, 4 and 5 are eluted in this order, and the resolution between the peak 2 and the peak 3 is not less than 1.5. (2) Perform the test with 15 mL each of the sample solu-tion and standard solution obtained in the Assay as directed under Liquid chromatography <2.01> according to the condi-tions described in the Assay: the retention times of the prin-cipal peaks obtained from the sample solution and the stand-ard solution are the same.
Related substances and desamido substances—Conduct this procedure at a temperature between 29C and 89C. Dissolve 50 mg of Glucagon (Genetical Recombination) in 100 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Perform the test with 15 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the total amount of desamido substance 1 having the relative retention time of about 1.1 to glucagon, desamido substance 2 having the relative retention time of about 1.2, desamido substance 3 having the relative retention time of about 1.3 and desamido substance 4 having the rela-tive retention time of about 1.4 is not more than 0.8z, and the total amount of peaks other than glucagon is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 37 minutes after injec-tion, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: When the procedure is run with 15 mL of the standard solution obtained in the Assay under the above operating conditions, the peak corre-sponding to the desamido substance 2 is detected. Water <2.48> Not more than 10z (50 mg, coulometric titration).
Conduct this procedure at a temperature between 29C and 89C. Weigh accurately about 50 mg of Glucagon (Genetical Recombination), dissolve in 100 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Separately dissolve Glucagon RS in 0.01 mol/L hydrochloric acid TS so that each mL contains about 0.5 mg of glucagon, and use this solution as the standard so-lution. Perform the test with exactly 15 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of glucagon in each solution. Amount (z) of glucagon ＝ AT/AS × CS/CT × 100 ‌JP XVIII Official Monographs / Glucose 1073 CS: Concentration (mg/mL) of the standard solution CT: Concentration (mg/mL) of the sample solution The calculated amount (z) of glucagon is corrected by the water content to obtain the amount (z) of glucagon on the anhydrous basis. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 3 mm in inside diameter and 150 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase A: Dissolve 16.3 g of potassium dihydrogen phosphate in 750 mL of water, adjust to pH 2.7 with phos-phoric acid, add water to make 800 mL, and add 200 mL of acetonitrile for liquid chromatography. Mobile phase B: A mixture of water and acetonitrile (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Glucose
Glucose Hydrate occurs as white, crystals or crystalline powder, and has a sweet taste. It is freely soluble in water, sparingly soluble in methanol, and slightly soluble in ethanol (95).◆
◇(1) Add 2 to 3 drops of a solution of Glu-cose Hydrate (1 in 20) to 5 mL of boiling Fehling's TS: a red precipitate is produced.◇ Perform the test with 20 mL each of the sample solu-tion and standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions: the principal peak in the chromatogram obtained from the sample solution is similar in retention time and size to the principal peak in the chromatogram from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay.
(1) Clarity and color of solution—Dissolve 10.0 g of Glucose Hydrate in 15 mL of water, and use this solution as the test solution. Perform the test with the test solution as directed under Turbidity Measurement <2.61>: the solution is clear. Perform the test with the test solution according to Method 2 under Methods for Color Matching <2.65>: the so-lution is not more colored than Matching Fluid BY7. (2) Heavy metals <1.07>—Proceed with 5.0 g of Glu-cose Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 4 ppm).◆ Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 250 mL, and use this solution as the standard solution (1). Pipet 25 mL of the standard solution (1), add water to make exactly 200 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution, the standard solution (1) and the standard solution (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the total area of the peaks of maltose and isomaltose, having the relative retention time of about 0.8 to glucose, obtained from the sample solution, is not larger than the peak area of glucose from the standard solution (1) (not more than 0.4z), and the peak area of mal-totriose, having the relative retention time of about 0.7 from the sample solution, is not larger than 1/2 times the peak area of glucose from the standard solution (1) (not more than 0.2z), and the peak area of fructose, having the rela-tive retention time of about 1.3 from the sample solution, is not larger than 3 times the peak area of glucose from the standard solution (2) (not more than 0.15z), and the area of the peak other than glucose and the peaks mentioned above from the sample solution is not larger than 2 times the peak area of glucose from the standard solution (2) (not more than 0.10z). Furthermore, the total area of the peaks other than glucose from the sample solution is not larger than 1.25 times the peak area of glucose from the standard solution (1) (not more than 0.5z). For these calculations the peak areas not larger than the peak area of glucose from the standard solution (2) are excluded (disregard limit: 0.05z). Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of glucose. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Confirm that the peak area of glucose obtained with 20 mL of the standard solution (2) is equivalent to 8.8 to 16.3z of that with 20 mL of the standard solution (1). System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviation of the peak area of glucose is not more than 1.0z.◇ Dextrin—To 1.0 g of powdered Glucose Hydrate add 20 mL of ethanol (95), and boil under a reflux condenser: the solution is clear. Soluble starch and sulfite—To 7.4 g of Glucose Hy-drate add 15 mL of water, dissolve by heating on a water bath, cool, and add 25 mL of 0.05 mol/L iodine VS: a yellow color develops (not more than 15 ppm as SO3). ‌JP XVIII Official Monographs / Purified Glucose 1075 Conductivity <2.51> Dissolve 20.0 g of Glucose Hydrate in a fleshly boiled and cooled distilled water to make 100 mL, and use this solution as the sample solution. Measure the conductivity of the sample solution at 25 ± 0.19C while gently stirring with a magnetic stirrer: not more than 20 mS･ cm－1. Water <2.48> 7.5 – 9.5z (0.25 g, volumetric titration, direct titration).
Weigh accurately about 0.33 g of Glucose Hydrate and 0.3 g of ◆Glucose RS◆ (separately determine the water <2.48> in the same manner as Purified Glucose), dissolve separately in water to make exactly 10 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of glucose in each solution. Purified Glucose
Glucose is a-D-glucopyranose, b-D-glucopyranose, or a mixture of them. It, when dried, contains not less than 99.
Glucose occurs as white, crystals or crystalline powder. It is odorless, and has a sweet taste. 0 – 25* 61 39 25 – 29 61 → 12 39 → 88 29 – 30 12 88 30 – 31 12 → 61 88 → 39 31 – 37 61 39 * Adjust the time for the isocratic condition so that the gradient starts after the desamido substance 4 is eluted. Flow rate: 0.5 mL per minute. System suitability— System performance: Dissolve Glucagon RS in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains 0.5 mg of glucagon. Warm this solution at 509C for 48 hours, and use this solution as the solution for system suitability test. When the procedure is run with 15 mL of the solution for system suitability test under the above operating conditions, four peaks corresponding to the desamido sub-stances 1, 2, 3 and 4 eluted after the principal peak are clear-ly detected, the total amount of these peaks is not less than 7z, and the resolution between glucagon and the desamido substance 1 is not less than 1.5. Furthermore, when the procedure is run with 15 mL of the standard solution under the above operating conditions, the symmetry factor of the principal peak is not more than 1.8. System repeatability: When the test is repeated 5 times with the standard solution under the above operating condi-tions, the relative standard deviation of the peak area of glucagon is not more than 2.0z.
Add 2 to 3 drops of a solution of Glucose (1 in 20) to 5 mL of boiling Fehling's TS: a red precipitate is produced.
(1) Clarity and color of solution—Add 25 g of Glucose to 30 mL of water in a Nessler tube, warm at 609C in a water bath until solution is effected, cool, and add water to make 50 mL: the solution is clear and has no more color than the following control solution. Control solution: To a mixture of 1.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS, and 2.0 mL of Copper (II) Sulfate CS, add water to make 10.0 mL. To 3.0 mL of this solution add water to make 50 mL. Acidity—Dissolve 5.0 g of Glucose in 50 mL of freshly boiled and cooled water, and add 3 drops of phenol-phthalein TS and 0.60 mL of 0.01 mol/L sodium hydroxide VS: a red color develops. Chloride <1.03>—Perform the test with 2.0 g of Glu- cose. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Sulfate <1.14>—Perform the test with 2.0 g of Glu- cose. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 5.0 g of Glucose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 4 ppm). Arsenic <1.11>—Dissolve 1.5 g of Glucose in 5 mL of water, add 5 mL of dilute sulfuric acid and 1 mL of bromine TS, heat on a water bath for 5 minutes, and concentrate to 5 mL. After cooling, perform the test with this solution as the test solution (not more than 1.3 ppm). Dextrin—To 1.0 g of Glucose add 20 mL of ethanol (95), and boil under a reflux condenser: the solution is clear. Soluble starch and sulfite—Dissolve 1.0 g of Glucose in 10 mL of water, and add 1 drop of iodine TS: a yellow color develops. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (2 g). ‌1074
Weigh accurately about 10 g of Glucose, previously dried, dissolve in 0.2 mL of ammonia TS and water to make exactly 100 mL, allow to stand for 30 minutes, and deter-mine the optical rotation, aD, of this solution at 20 ± 19C in a 100-mm cell as directed under Optical Rotation Determina- tion <2.49>. Amount (mg) of glucose (C6H12O6) ＝ aD × 1895.4
Containers—Tight containers. Storage—Light-resistant, and not exceeding – 159C.
Glutathione, when dried, contains not less than 98.
Glutathione occurs as a white crystalline pow-der. It is freely soluble in water, and practically insoluble in ethanol (99.5). Melting point: about 1859C (with decomposition).
Determine the infrared absorption spectrum of Glutathione, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. D Optical rotation <2.49> [a]20: －15.5 – －17.59(after dry-ing, 2 g, water, 50 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Glutathione in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Glutathione according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Glutathione according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Glutathione in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak having the relative retention time of about 4 to glutathione obtained from the sample solution is not larger than 3/4 times the peak area of glutathione from the standard solution, and the total area of the peaks other than glutathione is not larger than the peak area of glutathione from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate and 2.02 g of sodium 1-heptane sulfonate in 1000 mL of water, and adjust the pH to 3.0 with phosphoric acid. To 970 mL of this solution add 30 mL of methanol. Flow rate: Adjust so that the retention time of glutathione is about 5 minutes. Time span of measurement: About 6 times as long as the retention time of glutathione, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of glutathione obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of glutathione, 10 mg of D-phenylglycine and 50 mg of ascorbic acid in 100 mL of water. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, ascorbic acid, glutathione and D-phenylglycine are eluted in this order, and the resolutions between the peaks of ascorbic acid and glutathione and between the peaks of glutathione and D-phenylglycine are not less than 5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glutathione is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Glutathione, previ-ously dried, dissolve in 50 mL of a solution of metaphos- ‌1080 phoric acid (1 in 50), and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L iodine VS ＝ 30.73 mg of C10H17N3O6S
Glycerin is a clear, colorless, viscous liquid. It has a sweet taste. It is miscible with water and with ethanol (99.5). It is hygroscopic.
Determine the infrared absorption spectrum of Glycerin as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.449 – 1.454 Fatty acids and esters—Mix 50 g of Glycerin with 50 mL of freshly boiled and cooled water, add exactly 10 mL of 0.1 mol/L sodium hydroxide VS, boil the mixture for 15 minutes, cool, and titrate <2.50> the excess sodium hydroxide with 0.1 mol/L hydrochloric acid VS: 0.1 mol/L sodium hydroxide VS consumed is not more than 3.0 mL (indicator: 3 drops of phenolphthalein TS). Perform a blank determina-tion in the same manner. Ethylene glycol, diethylene glycol and related sub-stances—Weigh accurately about 5.88 g of Glycerin, mix with methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g each of ethylene glycol and diethylene glycol, mix with methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion and transfer into a 100-mL volumetric flask. Separately, weigh 5.0 g of glycerin for gas chromatography, mix with a suitable amount of methanol and put in the volumetric flask, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and determine the peak areas, AT1 and AS1, of ethylene glycol and, AT2 and AS2, of diethylene glycol by the automatic integration method. The amounts of ethylene glycol and diethylene glycol, calculated by the following equations, are not more than 0.1z, respectively. The amount of the peak other than glycerin, ethylene glycol and diethylene glycol obtained from the sample solution, calcu-lated by the area percentage method, is not more than 0.1z, and the total amount of the peaks other than glycerin is not more than 1.0z. Amount (z) of ethylene glycol Specific gravity <2.56> d 20: 1.221 – 1.230 ＝ MS1/MT × AT1/AS1 × 5
(1) Color—Place 50 mL of Glycerin in a Nessler tube, and observe downward: the solution has no more color than the following control solution. Control solution: Place 0.40 mL of Iron (III) Chloride CS in a Nessler tube, and add water to make 50 mL. Acidity or alkalinity—To 2 mL of Glycerin add 8 mL of water and mix: the solution is neutral. Chloride <1.03>—Take 10.0 g of Glycerin, and per- form the test: Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.001z). Sulfate <1.14>—Take 10.0 g of Glycerin, and perform the test. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.002z). Ammonium—To 5 mL of Glycerin add 5 mL of a so-lution of sodium hydroxide (1 in 10), and boil: the gas evolved does not change moistened red litmus paper to blue. Heavy metals <1.07>—Proceed with 5.0 g of Glycerin according to Method 1, and perform the test: Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Calcium—To 5 mL of the solution obtained in (2) add 3 drops of ammonium oxalate TS: the solution remains unchanged. Arsenic <1.11>—Prepare the test solution with 1.0 g of Glycerin according to Method 1, and perform the test (not more than 2 ppm). Acrolein, glucose, and other reducing sub-stances—To 1.0 g of Glycerin add 1 mL of ammonia TS, mix, and warm in a water bath at 609C for 5 minutes: no yel-low color is produced. Take the solution out of the water bath, add 3 drops of silver nitrate TS immediately, and allow to stand in a dark place for 5 minutes: the color of the solu-tion does not change, and no turbidity is produced. Amount (z) of diethylene glycol ＝ MS2/MT × AT2/AS2 × 5 MS1: Amount (g) of ethylene glycol taken MS2: Amount (g) of diethylene glycol taken MT: Amount (g) of Glycerin taken Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused-silica column 0.32 mm in inside diame- ter and 30 m in length, coated the inner surface with 14z cyanopropylphenyl-86z dimethyl silicone polymer for gas chromatography 1 mm in thickness. Column temperature: Inject at a constant temperature of about 1009C, raise the temperature at the rate of 7.59C per minute to 2209C, and maintain at a constant temperature of about 2209C. Injection port temperature: A constant temperature of about 2209C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: about 38 cm per second. Split ratio: 1:20. Time span of measurement: About 3 times as long as the retention time of glycerin, beginning after the solvent peak. System suitability— System performance: Mix 50 mg each of ethylene glycol, diethylene glycol and glycerin for gas chromatography with 100 mL of methanol. When the procedure is run with 1 mL of this solution under the above operating conditions, ethyl-ene glycol, diethylene glycol and glycerin are eluted in this order, and the resolution between the peaks of ethylene glycol and diethylene glycol is not less than 40, and between ‌JP XVIII Official Monographs / Concentrated Glycerin 1081 the peaks of diethylene glycol and glycerin is not less than 10. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviations of the peak area of ethylene glycol and diethylene glycol are not more than 10z, respectively. Readily carbonizable substances—To 5 mL of Glycerin add carefully 5 mL of sulfuric acid for readily car-bonizable substances, mix gently at a temperature between 189C and 209C, and allow to stand for 1 hour between 159C and 259C: the solution has not more color than Matching Fluid H. Water <2.48> 13 – 17z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Weigh accurately about 10 g of Glycerin in a tared crucible, heat to boiling, and fire to burn immediately. After cooling, moisten the residue with 1 to 2 drops of sulfuric acid, and ignite cautiously to constant mass: the mass of the residue is not more than 0.01z.
Weigh accurately about 0.2 g of Glycerin, transfer into a glass-stoppered flask, add 50 mL of water, mix, add exactly 50 mL of sodium periodate TS, shake, and allow to stand in a dark place at a room temperature for about 30 minutes. Add 10 mL of a mixture of water and ethylene glycol (1:1), allow to stand for about 20 minutes, add 100 mL of water, and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (indicator: 2 drops of phenolphthalein TS). Per-form a blank determination in the same manner, and make the necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 9.209 mg of C3H8O3
Glyceryl Monostearate occurs as white to light yellow, waxy masses, thin flakes, or granules. It has a char-acteristic odor and taste. It is very soluble in hot ethanol (95), soluble in chlo-roform, sparingly soluble in diethyl ether, and practically in-soluble in water and in ethanol (95). It is slowly affected by light.
(1) Heat 0.2 g of Glyceryl Monostearate with 0.5 g of potassium hydrogen sulfate until thoroughly charred: the irritative odor of acrolein is perceptible. (2) Dissolve 0.1 g of Glyceryl Monostearate in 2 mL of ethanol (95) by warming, heat with 5 mL of dilute sulfuric acid in a water bath for 30 minutes, and cool: a white to yel-low solid is produced. This separated solid dissolves when shaken with 3 mL of diethyl ether. ‌JP XVIII Official Monographs / Gonadorelin Acetate 1083‌ Melting point <1.13> Not below 559C. Acid value <1.13> Not more than 15. Saponification value <1.13> 157 – 170 Iodine value <1.13> Not more than 3.0. Use chloroform in-stead of cyclohexane.
<1.13> Acidity or alkalinity—To 1.0 g of Glyceryl Monostearate add 20 mL of boiling water, and cool with swirling: the solution is neutral. Residue on ignition <2.44> Not more than 0.1z (1 g).
Gonadorelin Acetate contains not less than 96.
Gonadorelin Acetate occurs as a white to pale yellow powder. It is odorless or has a slight, acetic odor. It is freely soluble in water, in methanol and in acetic acid (100), and sparingly soluble in ethanol (95). It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Gonadorelin Acetate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Gonadorelin Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Gonadorelin Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 20 mg of Gonadorelin Acetate in 0.5 mL of ethanol (99.5), add 1 mL of sulfuric acid, and heat: the odor of ethyl acetate is perceptible. D Optical rotation <2.49> [a]20: －53.0 – －57.09(0.1 g calcu- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) Mobile phase D (volz) lated on the anhydrous basis, diluted acetic acid (100) (1 in 100), 10 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Gonadorelin Acetate in 10 mL of water: the pH of this solution is between 4.8 and 5.8. Constituent amino acids Put 10 mg of Gonadorelin Ace-tate in a test tube for hydrolysis, add 0.5 mL of hydrochloric acid and 0.5 mL of a solution of mercaptoacetic acid (2 in 25), seal the tube under reduced pressure, and heat at 1109C for 5 hours. After cooling, open the tube, transfer the hydrolyzate into a beaker, and evaporate to dryness on a water bath. Add exactly 100 mL of 0.02 mol/L hydrochloric acid TS to dissolve the residue, and use this solution as the sample solution. Separately, weigh exactly 0.105 g of L-serine, 0.147 g of L-glutamic acid, 0.115 g of L-proline, 75 mg of glycine, 0.131 g of L-leucine, 0.181 g of L-tyrosine, 0.210 g of L-histidine hydrochloride monohydrate, 0.204 g of L-tryptophan and 0.211 g of L-arginine hydrochloride, which are all previously dried at 1059C for 3 hours, add 50 mL of 1 mol/L hydrochloric acid TS to dissolve them, and add water to make exactly 1000 mL. Pipet 10 mL of this so-lution, add 0.02 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: the peaks of nine constituent amino acids are observed on the chromato-gram obtained from the sample solution, and their respective molar ratios to arginine are 0.7 – 1.0 for serine and trypto-phan, 0.8 – 1.2 for proline, 0.9 – 1.1 for glutamic acid, leu-cine, tyrosine and histidine, respectively, and 1.8 – 2.2 for glycine. Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 nm for proline and 570 nm for others). Column: A stainless steel column 4 mm in inside diameter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with a sulfonated polystyrene copolymer (5 mm in particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Mobile phase: Prepare the mobile phases A, B, C and D according to the following table. Mobile phase A B C D 0 ¿ 9 100 0 0 0 9 ¿ 25 0 100 0 0 25 ¿ 61 0 100 → 0 0 → 100 0 61 ¿ 76 0 0 100 0 76 ¿ 96 0 0 0 100 Reaction reagent: Dissolve 204 g of lithium acetate dihy-drate in 336 mL of water, add 123 mL of acetic acid (100) and 401 mL of 1-methoxy-2-propanol, and use as Solution A. Separately, dissolve 39 g of ninhydrin and 81 mg of so-dium borohydride in 979 mL of 1-methoxy-2-propanol, and use as Solution B. Mix the same volume of Solution A and Solution B before use. Flow rate of mobile phase: 0.25 mL per minute. Flow rate of reaction reagent: 0.3 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, serine, glutamic acid, proline, glycine, leucine, tyro-sine, histidine, tryptophan and arginine are eluted in this order with enough separation between these peaks.
(1) Clarity and color of solution—Dissolve 1.0 g of Glycine in 10 mL of water: the solution is clear and color-less. Chloride <1.03>—Perform the test with 0.5 g of Gly- cine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of Gly- cine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium<1.02>—Perform the test using 0.25 g of Glycine. Prepare the control solution with 5.0 mL of Stand-ard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of Glycine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Glycine according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Glycine in 25 mL of water and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex- 5-Oxo-L-prolyl-L-histidyl-L-tryptophyl-L-seryl-L-tyrosyl-glycyl-L-leucyl-L-arginyl-L-prolyl-glycinamide diacetate [34973-08-5] Gonadorelin Acetate contains not less than 96.0z and not more than 102.0z of gonadorelin acetate (C55H75N17O13.2C2H4O2), calculated on the anhydrous basis. Description Gonadorelin Acetate occurs as a white to pale yellow powder. It is odorless or has a slight, acetic odor. It is freely soluble in water, in methanol and in acetic acid (100), and sparingly soluble in ethanol (95). It is hygroscopic. Identification (1) Determine the absorption spectrum of a solution of Gonadorelin Acetate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Gonadorelin Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Gonadorelin Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 20 mg of Gonadorelin Acetate in 0.5 mL of ethanol (99.5), add 1 mL of sulfuric acid, and heat: the odor of ethyl acetate is perceptible. D Optical rotation <2.49> [a]20: －53.0 – －57.09(0.1 g calcu- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) Mobile phase D (volz) lated on the anhydrous basis, diluted acetic acid (100) (1 in 100), 10 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Gonadorelin Acetate in 10 mL of water: the pH of this solution is between 4.8 and 5.8. Constituent amino acids Put 10 mg of Gonadorelin Ace-tate in a test tube for hydrolysis, add 0.5 mL of hydrochloric acid and 0.5 mL of a solution of mercaptoacetic acid (2 in 25), seal the tube under reduced pressure, and heat at 1109C for 5 hours. After cooling, open the tube, transfer the hydrolyzate into a beaker, and evaporate to dryness on a water bath. Add exactly 100 mL of 0.02 mol/L hydrochloric acid TS to dissolve the residue, and use this solution as the sample solution. Separately, weigh exactly 0.105 g of L-serine, 0.147 g of L-glutamic acid, 0.115 g of L-proline, 75 mg of glycine, 0.131 g of L-leucine, 0.181 g of L-tyrosine, 0.210 g of L-histidine hydrochloride monohydrate, 0.204 g of L-tryptophan and 0.211 g of L-arginine hydrochloride, which are all previously dried at 1059C for 3 hours, add 50 mL of 1 mol/L hydrochloric acid TS to dissolve them, and add water to make exactly 1000 mL. Pipet 10 mL of this so-lution, add 0.02 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: the peaks of nine constituent amino acids are observed on the chromato-gram obtained from the sample solution, and their respective molar ratios to arginine are 0.7 – 1.0 for serine and trypto-phan, 0.8 – 1.2 for proline, 0.9 – 1.1 for glutamic acid, leu-cine, tyrosine and histidine, respectively, and 1.8 – 2.2 for glycine. Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 nm for proline and 570 nm for others). Column: A stainless steel column 4 mm in inside diameter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with a sulfonated polystyrene copolymer (5 mm in particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Mobile phase: Prepare the mobile phases A, B, C and D according to the following table. Mobile phase A B C D 0 ¿ 9 100 0 0 0 9 ¿ 25 0 100 0 0 25 ¿ 61 0 100 → 0 0 → 100 0 61 ¿ 76 0 0 100 0 76 ¿ 96 0 0 0 100 Reaction reagent: Dissolve 204 g of lithium acetate dihy-drate in 336 mL of water, add 123 mL of acetic acid (100) and 401 mL of 1-methoxy-2-propanol, and use as Solution A. Separately, dissolve 39 g of ninhydrin and 81 mg of so-dium borohydride in 979 mL of 1-methoxy-2-propanol, and use as Solution B. Mix the same volume of Solution A and Solution B before use. Flow rate of mobile phase: 0.25 mL per minute. Flow rate of reaction reagent: 0.3 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, serine, glutamic acid, proline, glycine, leucine, tyro-sine, histidine, tryptophan and arginine are eluted in this order with enough separation between these peaks. Purity (1) Clarity and color of solution—A solution ob-tained by dissolving 0.10 g of Gonadorelin Acetate in 10 mL of water is clear, and the absorbance of this solution at 350 nm determined as directed under Ultraviolet-visible Spectro-photometry <2.24> is not more than 0.10. (2) Related substances—Dissolve 50 mg of Gonadorelin Acetate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than gonadorelin from the sample solution is not larger than 1/5 times the peak area of gonadorelin from the standard so-lution, and the total area of the peaks other than gonadore-lin from the sample solution is not larger than 3/5 times the peak area of gonadorelin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of gonadorelin, beginning after the solvent peak. Trisodium citrate dihy- drate 6.19 g 7.74 g 26.67 g — System suitability— Sodium hydroxide — — — 8.00 g Sodium chloride 5.66 g 7.07 g 54.35 g — Citric acid monohydrate 19.80 g 22.00 g 6.10 g — Ethanol (99.5) 130 mL 20 mL — 100 mL Benzyl alcohol — — 5 mL — Thiodiglycol 5 mL 5 mL — — Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of gonadorelin obtained with 10 mL of this solution is equivalent to 1 to 3z of that with 10 mL of the standard solution. Lauromacrogol solution in diethyl ether (1 in 4) 4 mL 4 mL 4 mL 4 mL System performance: Dissolve 4 mg of Gonadorelin Ace- Caprylic acid 0.1 mL 0.1 mL 0.1 mL 0.1 mL tate in a suitable amount of the mobile phase, add 5 mL of a Water a sufficient amount a sufficient amount a sufficient amount a sufficient amount solution of phenacetin in acetonitrile (1 in 1000) and the mo-bile phase to make 50 mL. When the procedure is run with Total volume 1000 mL 1000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B, C and D as directed in the following table. 10 mL of this solution under the above operating conditions, gonadorelin and phenacetin are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- JP XVIII Official Monographs / Human Chorionic Gonadotrophin 1085 ing conditions, the relative standard deviation of the peak area of gonadorelin is not more than 5z. Water <2.48> Not more than 8.0z (0.15 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (0.1 g).
Weigh accurately about 20 mg of Gonadorelin Ace-tate and Gonadorelin Acetate RS (separately determine the water <2.48> in the same manner as Gonadorelin Acetate) and dissolve in diluted acetic acid (100) (1 in 1000) to make gonadotrophin Units per mg, and contains not less than 3000 chorionic gonadotrophin Units per mg pro-tein. It contains not less than 80z and not more than 125z of the labeled human chorionic gonadotrophin Units. Description Human Chorionic Gonadotrophin occurs as a white to light yellow-brown powder. It is freely soluble in water. Identification Calculate b by the following equation, using exactly 25 mL each. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution and add water to make 25 mL, and use these solutions as the sample solu-tion and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed Y3 and Y4 b ＝ E/I E ＝ (Y3 obtained in the Assay: b is not more than 120. － Y4)/f under Liquid Chromatography <2.01> according to the fol- lowing conditions, and calculate the ratios, QT and QS, of f: Number of test animals per group I ＝ log (T /T ) the peak area of gonadorelin to that of the internal standard. H L Amount (mg) of gonadorelin acetate (C H N O .2C H O ) Purity (1) Clarity and color of solution—Dissolve 0.05 g of Human Chorionic Gonadotrophin in 5 mL of isotonic so- 55 75 17 13 2 4 2 dium chloride solution: the solution is clear and colorless or ＝ MS × QT/QS MS: Amount (mg) of Gonadorelin Acetate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of phenacetin in a mixture of water and acetonitrile (3:2) (1 in 1000). Operating conditions— ‌Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0) and acetonitrile (90:17). Flow rate: Adjust so that the retention time of gonadore-lin is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, gonadorelin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of gonadorelin to that of the internal standard is not more than 1.5z.
Goreisan Extract contains not less than 0.
Goreisan Extract occurs as a light red-brown to light brown powder, or a black-brown viscous extract. It has a characteristic odor, and a slightly sweet first, bitter, then acrid taste.
(1) Weigh exactly 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 20 mL of water and 2 mL of ammonia solution (28), and shake. Add 20 mL of a mixture of hexane and ethyl acetate (20:1), shake, centri-fuge, and separate the upper layer. Add 20 mL of a mixture of hexane and ethyl acetate (20:1) to the aqueous layer, shake, centrifuge, and separate the upper layer. Combine these extracts, evaporate the solvent under low pressure (in vacuo), add exactly 2 mL of methanol to the residue, and use this solution as the sample solution. Separately, weigh ex-actly 10 mg of alisol A for thin-layer chromatography, and dissolve in exactly 10 mL of methanol. Pipet 1 mL of this so-lution, add methanol to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl formate, water and formic acid (30:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid-acetic acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and examine under ultra-violet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution, and it is larger and more intense than the spot from the standard solution (Alisma Tuber). For preparation prescribed Atractylodes Rhizome—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red to red-purple spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 25 mL of hexane, and shake. Separate the hexane layer, and evaporate the solvent under low pressure (in vacuo), add 0.5 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a green-brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhizome). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 50 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, diethyl ether and methanol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydradine TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 5 mL of hexane, and shake. Centrifuge this solution, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-2-methoxycinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance ‌2018 Goshajinkigan Extract / Crude Drugs and Related Drugs of about 7 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
Conduct this procedure using light-resistant vessels. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of (E )-cinnamic acid for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of (E )-cinnamic acid in each solution. Amount (mg) of (E )-cinnamic acid ＝ MS × AT/AS × 1/20 MS: Amount (mg) of (E )-cinnamic acid for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (750:250:1). Flow rate: 1.0 mL per minute (the retention time of (E )-cinnamic acid is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of (E )-cinnamic acid are not less than 5000 and not more than1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of (E )-cinnamic acid is not more than 1.5z.
Goshajinkigan Extract contains not less than 4 mg and not more than 16 mg of loganin, not less than 6 mg and not more than 18 mg of paeoniflorin (C23H28O11: 480.
Goshajinkigan Extract occurs as a brown to dark brown powder or black-brown viscous extract. It has slightly a characteristic odor and an acid taste.
(1) To 1.0 g of the dry extract (or 3.0 g of the viscous extract), add 10 mL of water, shake, then add 30 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of water, methanol and 1-butanol (1:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool; a dark-green spot is observed at an Rf value of about 0.6 (Rehmannia Root). To 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of loganin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer JP XVIII Crude Drugs and Related Drugs / Goshajinkigan Extract 2019 chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (6:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-methoxybezal-dehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Cornus Fruit). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract), add 10 mL of sodium carbonate TS, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of alisol A for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid-acetic acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yel-low fluorescent spot from the standard solution (Alisma Tuber). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract), add 10 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and diethyl ether (5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange spot from the standard solution (Moutan Bark). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate 1 mL of the hexane layer, add 0.5 mL of sodium hydroxide TS, shake, centrifuge, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, diethyl ether and methanol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. To 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 10 mL of water, shake, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-2-methoxycinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution. To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract), add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in va-cuo), add 1 mL of acetonitrile to the residue, and use this so-lution as the sample solution. Separately, dissolve 1 mg of benzoylmesaconine hydrochloride for thin-layer chromatog-raphy in 10 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Pow-dered Processed Aconite Root). To 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 0.3 g of pulverized plantago seed for thin-layer chromatography, add 1 mL of methanol, warm on a water bath for 3 minutes, centrifuge after cool-ing, and use the supernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, ethyl acetate, water and acetic acid (100) (10:10:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the deep blue spot (Rf value: about 0.3) from the standard solution (Plantago Seed). To 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 2 g of achyranthes root for thin-layer chromatography, add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-propanol, ethyl acetate and water (4:4:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sul- Goshajinkigan Extract / Crude Drugs and Related Drugs furic acid on the plate and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value (around 0.4) with the dark red spot from the standard solution (Achyran-thes Root).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under the Extracts (4), and perform the test (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Aconitum diester alkaloids (aconitine, jesaconitine, hypaconitine and mesaconitine)—Weigh accurately 1.0 g of the dry extract (or an amount of the viscous extract, equiva-lent to 1.0 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS and shake for 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ing conditions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5z. Loss on drying <2.41> The dry extract: Not more than 9.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Loganin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of loganin for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of loganin in each solution. ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 Amount (mg) of loganin ＝ MS × AT/AS × 1/2 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, and evaporate to dryness under low pressure (in vacuo). Dissolve the residue with exactly 10 mL of a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1). Centrifuge this solution, and use the supernatant liquid as the sample solution. Sepa-rately, pipet 1 mL of aconitum diester alkaloids standard so-lution for purity, add a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make ex-actly 10 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions: the heights of the peaks corresponding to aconitine, jesaconi-tine, hypaconitine and mesaconitine from the sample solu-tion are not higher than the respective heights corresponding to aconitine, jesaconitine, hypaconitine and mesaconitine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of mesaconitine is about 31 minutes). System suitability— System performance: When the procedure is run with 20 mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between their peaks is not less than 1.5 respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- MS: Amount (mg) of loganin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, acetonitrile and metha-nol (55:4:1). Flow rate: 1.2 mL per minute (the retention time of loga-nin is about 25 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of loganin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of loganin is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis JP XVIII Crude Drugs and Related Drugs / Goshajinkigan Extract 2021 ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Total alkaloids—Weigh accurately about 1 g of the dry extract (or an amount of the viscous extract, equivalent to about 1 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, and evaporate to dryness under low pressure (in vacuo). Dissolve the residue with a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make exactly 10 mL. Centrifuge this so-lution, and use the supernatant liquid as the sample solution. Perform the test with exactly 20 mL each of the sample solu-tion and the aconitum monoester alkaloids standard solution TS for assay as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas of benzoylmesaconine, benzoylhypaconine and 14-anisoylaconine, ATM and ASM, ATH and ASH, as well as ATA and ASA, in each solution, respectively. Amount (mg) of benzoylmesaconine hydrochloride ＝ CSM × ATM/ASM × 10 Amount (mg) of benzoylhypaconine hydrochloride ＝ CSH × ATH/ASH × 10 Amount (mg) of 14-anisoylaconine hydrochloride ＝ CSA × ATA/ASA × 10 CSM: Concentration (mg/mL) of benzoylmesaconine hy-drochloride for assay in aconitum monoester alkaloids standard solution TS for assay CSH: Concentration (mg/mL) of benzoylhypaconine hy- drochloride for assay in aconitum monoester alkaloids standard solution TS for assay length: 231 nm for benzoylmesaconine and benzoylhypaco-nine; 254 nm for 14-anisoylaconine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of ben-zoylmesaconine is about 15 minutes). System suitability— System performance: When the procedure is run with 20 mL of the aconitum monoester alkaloids standard solution TS for assay under the above operating conditions, the num-ber of theoretical plates and the symmetry factor of the peak of benzoylmesaconine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the aconitum monoester alkaloids standard solution TS for assay under the above operating conditions, the relative standard deviation of the peak areas of ben-zoylmesaconine, benzoylhypaconine and 14-anisoylaconine is not more than 1.5z.
Goshuyuto Extract contains not less than 0.
Goshuyuto Extract occurs as a light brown to light red-yellow powder, or a black-brown viscous extract. It has a slight odor and a hot and bitter taste. Identiˆcation (1) To 1.0 g of the dry extract (or 3.0 g of CSA : Concentration (mg/mL) of 14-anisoylaconine hydro-chloride for assay in aconitum monoester alkaloids standard solution TS for assay the viscous extract) add 10 mL of sodium hydroxide TS, shake, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, to 1 g of pulverized euodia fruit add 10 mL of methanol, shake, cen- Operating conditions— Detector: An ultraviolet absorption photometer (wave- trifuge, and use the supernatant liquid as the standard solu-tion. Perform the test with these solutions as directed under Goshajinkigan Extract / Crude Drugs and Related Drugs Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, 2-propanol, water and formic acid (7:7:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot (Rf value: about 0.5) from the standard so-lution (Euodia Fruit). lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 30 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, weigh accurately about 10 mg of evodiamine for assay, and dissolve in methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of evodiamine in each solution. To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, add 25 mL of diethyl Amount (mg) of evodiamine ＝ MS × AT/AS × 1/4 ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzal-dehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of sodium hydroxide TS, shake, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginseno-side Rb1 RS or ginsenoside Rb1 for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solu-tion (Ginseng).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed MS: Amount (mg) of evodiamine for assay taken, calculat-ed on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 282 nm). Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (620:380:1). Flow rate: 1.0 mL per minute (the retention time of evodiamine is about 18 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of evodiamine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of evodiamine is not more than 1.5z. (2) [6]-Gingerol—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 30 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, weigh accurately about 10 mg of [6]-gingerol for assay, dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of [6]-gingerol in each solution. under Extracts (4), and perform the test (not more than 30 ppm). Amount (mg) of [6]-gingerol ＝ MS × AT/AS × 1/20 Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 11.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Evodiamine—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva- MS: Amount (mg) of [6]-gingerol for assay taken Operating conditions— Detector, column, column temperature and mobile phase: Proceed as directed in the operating conditions in (1). Flow rate: 1.0 mL per minute (the retention time of [6]-gingerol is about 14 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-gingerol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌JP XVIII Crude Drugs and Related Drugs / Exsiccated Gypsum 2023‌ ing conditions, the relative standard deviation of the peak area of [6]-gingerol is not more than 1.5z. Ginsenoside Rb1—Weigh accurately about 2 g of the dry extract (or an amount of the viscous extract, equivalent to about 2 g of the dried substance), add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 15 mL of diluted methanol (3 in 5), and repeat the same procedure. Combine all the supernatant liquids, and add diluted metha- Gypsum Gypsum Fibrosum
Guaifenesin, when dried, contains not less then 98.
Guaifenesin occurs as white, crystals or crys-talline powder. It is freely soluble in ethanol (95), and sparingly soluble in water. A solution of ethanol (95) (1 in 20) shows no optical rota-tion.
(1) Determine the absorption spectrum of a solution of Guaifenesin (1 in 50,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Guaifenesin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Guaifenesin, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Guaifenesin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Guaifenesin in 100 mL of water: the pH of the solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 0.20 g of Guaifenesin in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.7 g of Guaifenesin in 25 mL of water by warming. Cool, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using ‌JP XVIII Official Monographs / Guanabenz Acetate 1089 this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.020z). Heavy metals <1.07>—Dissolve 2.0 g of Guaifenesin in 25 mL of water by warming. Cool, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Guaifenesin according to Method 3, and perform the test (not more than 2 ppm). Free guaiacol—To 1.0 g of Guaifenesin add exactly 25 mL of water, dissolve by warming, cool, and use this so-lution as the sample solution. Separately, dissolve 0.100 g of guaiacol in water to make exactly 1000 mL. Pipet 3 mL of this solution, add exactly 22 mL of water, and use this solu-tion as the standard solution. To each of the sample solution and standard solution add 1.0 mL of potassium hexacyano-ferrate (III) TS and 5.0 mL of a solution of 4-aminoantipy-rine (1 in 200), and immediately after shaking for exactly 5 seconds add a solution of sodium hydrogen carbonate (1 in 1200) to make exactly 100 mL. Determine the absorbances of these solutions at 500 nm exactly 15 minutes after the addi-tion of the 4-aminoantipyrine solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using a solution, prepared in the same manner with 25 mL of water, as the blank: the absorbance of the solution obtained from the sample solution is not more than that from the standard so-lution. Related substances—Dissolve 1.0 g of Guaifenesin in 100 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, ethanol (95), and ammonia solution (28) (40:10:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1109C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 60 mg of Guaifenesin and Guaifenesin RS, previously dried, and dissolve each then in water to make exactly 100 mL. Pipet 5 mL of these solu-tions, and add water to make exactly 100 mL, and use these solutions as the sample solution and standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 273 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Guanabenz Acetate
Guanabenz Acetate, when dried, contains not less than 98.
Guanabenz Acetate occurs as white, crystals or crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol and in ethanol (95), slightly soluble in water, and practically insoluble in diethyl ether. It is gradually affected by light. Melting point: about 1909C (with decomposition).
(1) To 5 mL of a solution of Guanabenz Acetate (1 in 1000) add 0.5 mL of a diluted ethanol (95) (5 in 6) which contains 16 g of urea and 0.2 g of 1-naphthol in 100 mL, and add 1 mL of N-bromosuccinimide TS: a purple color develops. Determine the absorption spectrum of a solution of Guanabenz Acetate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Guanabenz Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. To 0.1 g of Guanabenz Acetate add 5 mL of water and 1 mL of ammonia TS, shake, filter, and neutralize the filtrate with dilute hydrochloric acid: the solution responds to Qualitative Tests <1.09> (3) for acetate.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Guanabenz Acetate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.05 g of Guanabenz Acetate in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add methanol to make exactly 10 mL, then pipet 1 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Amount (mg) of guaifenesin (C10H14O4) ＝ M S × AT/AS Chromatography <2.03>. Spot 5 mL each of the sample solu- tion and standard solution on a plate of silica gel with fluo- MS: Amount (mg) of Guaifenesin RS taken
Weigh accurately about 0.25 g of Guanabenz Ace-tate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.11 mg of C H Cl N .C H O Heavy metals <1.07>—Proceed with 2.0 g of Guanethidine Sulfate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 0.5 g of Guanethidine Sul-fate, previously dried, dissolve in 2 mL of formic acid, add 70 mL of a mixture of acetic anhydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. 8 8 2 4 2 4 2
Guanethidine Sulfate, when dried, contains not less than 98.
Guanethidine Sulfate occurs as white, crystals or crystalline powder. It is odorless or has a slight, character-istic odor and a bitter taste. It is very soluble in formic acid, freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. Melting point: 251 – 2569C (an evacuated sealed capillary tube, with decomposition).
(1) To 4 mL of a solution of Guanethidine Sulfate (1 in 4000) add 2 mL of 1-naphthol TS, 1 mL of dia-cetyl TS and 15 mL of water, and allow to stand for 30 minutes: a red color develops. Determine the infrared absorption spectrum of Guanethidine Sulfate, previously dried, as directed in the po-tassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Guanethidine Sulfate (1 in 10) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Guanethidine Sulfate in 50 mL of water: the pH of the solution is between 4.7 and 5.7.
(1) Clarity and color of solution—Dissolve 1.0 g of Guanethidine Sulfate in 50 mL of water: the solution is clear and colorless. (2) Methylisothiourea sulfate—Dissolve 2.0 g of Guanethidine Sulfate in 80 mL of sodium hydroxide TS, and allow to stand for 10 minutes. Add 60 mL of hydrochloric acid, 2 g of sodium bromide and water to make 200 mL. Then, to this solution add 0.70 mL of 1/60 mol/L potassium bromate VS and 2 mL of zinc iodide-starch TS: a blue color develops. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.64 mg of C10H22N4.H2SO4
Exsiccated Gypsum occurs as a white to grayish white powder. It is odorless and tasteless. It is slightly soluble in water, and practically insoluble in ethanol (95). It absorbs moisture slowly on standing in air to lose its solidifying property. When it is heated to yield an anhydrous compound at a temperature above 2009C, it loses its solidifying property.
Shake 1 g of Exsiccated Gypsum with 20 mL of water for 5 minutes, and filter: the filtrate responds to the Qualitative Tests <1.09> (2) and (3) for calcium salt and to the Qualitative Tests <1.09> for sulfate.
Alkalinity—Take 3.0 g of Exsiccated Gypsum in a glass-stoppered test tube, add 10 mL of water and 1 drop of phenolphthalein TS, and shake vigorously: no red color de-velops. Solidification To 10.0 g of Exsiccated Gypsum add 10 mL of water, stir immediately for 3 minutes, and allow to stand: the period until water no longer separates, when the material is pressed with a finger, is not more than 10 minutes from the time when the water was added. Hachimijiogan Extract / Crude Drugs and Related Drugs
Gypsum occurs as lustrous, white, heavy, fibrous, crystalline masses, which easily split into needles or very fine crystalline powder. It is odorless and tasteless. It is slightly soluble in water.
To 1 g of pulverized Gypsum add 20 mL of water, allow to stand with occasional shaking for 30 minutes, and filter: the filtrate responds to the Qualitative Tests <1.09> (2) and (3) for calcium salt and to the Qualita-tive Tests <1.09> for sulfate.
(1) Heavy metals <1.07>—Boil 4.0 g of pulverized Gypsum with 4 mL of acetic acid (100) and 96 mL of water for 10 minutes, cool, add water to make exactly 100 mL, and filter. Perform the test using 50 mL of the filtrate as the test solution. Prepare the control solution as follows: to 4.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Gypsum according to Method 2, and perform the test (not more than 5 ppm).
Hachimijiogan Extract contains not less than 4 mg and not more than 16 mg of loganin, not less than 6 mg and not more than 18 mg (for preparation prescribed 3 g of Moutan Bark) or not less than 5 mg and not more than 15 mg (for preparation prescribed gel for thin-layer chromatography.
Hachimijiogan Extract occurs as a grayish brown to brown powder or black-brown viscous extract. It has a characteristic odor and a slightly bitter and acid taste.
(1) To 1.0 g of the dry extract (or 3.0 g of the viscous extract), add 10 mL of water, shake, then add 30 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica phy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (6:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-methoxybezal-dehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Cornus Fruit). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract), add 10 mL of sodium carbonate TS, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of alisol A for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid-acetic acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Alisma Tuber). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract), add 10 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and diethyl ether (5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange spot from the standard solution (Moutan Bark). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, JP XVIII Crude Drugs and Related Drugs / Hachimijiogan Extract 2025 separate 1 mL of the hexane layer, add 0.5 mL of sodium hy-droxide TS, shake, centrifuge, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, diethyl ether and methanol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. To 2.0 g of the dry extract (or 6.0 g of the viscous extract), add 10 mL of water, shake, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-2-methox-ycinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the stand-ard solution. To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract), add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes and centrifuge. Separate the diethyl ether layer, evaporate the diethyl ether layer under low pres-sure (in vacuo), add 1 mL of acetonitrile to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of benzoylmesaconine hydrochloride for thin-layer chromatography in 10 mL of ethanol (99.5), and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Processed Aconite Root or Powdered Processed Aconite Root).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under the Extracts (4), and perform the test (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Aconitum diester alkaloids (aconitine, jesaconitine, hypaconitine and mesaconitine)—Weigh accurately 1.0 g of the dry extract (or an amount of the viscous extract, equiva-lent to 1.0 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS and shake for 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue with exactly 10 mL of a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1), centrifuge this solution, and use the supernatant liquid as the sample solution. Sepa-rately, pipet exactly 1 mL of aconitum diester alkaloids standard solution for purity, add a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions: the heights of the peaks corresponding to aconitine, jesaconitine, hypaconitine and mesaconitine from the sample solution are not higher than the respective heights corresponding to aconitine, jesaconitine, hypaconitine and mesaconitine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of mesaconitine is about 31 minutes). System suitability— System performance: When the procedure is run with 20 mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between their peaks is not less than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5 z. Loss on drying <2.41> The dry extract: Not more than 8.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Loganin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of loganin for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac- Hachimijiogan Extract / Crude Drugs and Related Drugs cording to the following conditions, and determine the peak areas, AT and AS, of loganin in each solution. ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Amount (mg) of loganin ＝ MS × AT/AS × 1/2 Total alkaloids—Weigh accurately about 1 g of the dry extract (or an amount of the viscous extract, equivalent MS: Amount (mg) of loganin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, acetonitrile and metha-nol (55:4:1). Flow rate: 1.2 mL per minute (the retention time of loga-nin is about 25 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of loganin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of loganin is not more than 1.5z. (2) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine to about 1 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue with a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make exactly 10 mL. Centrifuge this so-lution, and use the supernatant liquid as the sample solution. Perform the test with exactly 20 mL each of the sample solu-tion and the aconitum monoester alkaloids standard solution TS for assay as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas of benzoylmesaconine, benzoylhypaconine and 14-anisoylaconine, ATM and ASM, ATH and ASH, as well as ATA and ASA, in each solution, respectively. Amount (mg) of benzoylmesaconine hydrochloride ＝ CSM × ATM/ASM × 10 Amount (mg) of benzoylhypaconine hydrochloride ＝ CSH × ATH/ASH × 10 Amount (mg) of 14-anisoylaconine hydrochloride ＝ CSA × ATA/ASA × 10 CSM: Concentration (mg/mL) of benzoylmesaconine hy-drochloride for assay in aconitum monoester alkaloids standard solution TS for assay CSH: Concentration (mg/mL) of benzoylhypaconine hy- drochloride for assay in aconitum monoester alkaloids standard solution TS for assay the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 CSA : Concentration (mg/mL) of 14-anisoylaconine hydro-chloride for assay in aconitum monoester alkaloids standard solution TS for assay MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for benzoylmesaconine and benzoylhypaco-nine; 254 nm for 14-anisoylaconine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of ben-zoylmesaconine is about 15 minutes). System suitability— System performance: When the procedure is run with 20 mL of the aconitum monoester alkaloids standard solution TS for assay under the above operating conditions, the num-ber of theoretical plates and the symmetry factor of the peak of benzoylmesaconine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the aconitum monoester alkaloids standard so-lution TS for assay under the above operating conditions, ‌JP XVIII Crude Drugs and Related Drugs / Hangekobokuto Extract 2027 the relative standard deviation of the peak areas of ben-zoylmesaconine, benzoylhypaconine and 14-anisoylaconine is not more than 1.5z.
Haloperidol, when dried, contains not less than 99.
Haloperidol occurs as white to pale yellow, crystals or powder. It is freely soluble in acetic acid (100), sparingly soluble in methanol, slightly soluble in 2-propanol and in ethanol (99.5), and practically insoluble in water.
(1) Dissolve 30 mg of Haloperidol in 100 mL of 2-propanol. To 5 mL of the solution add 10 mL of 0.1 mol/L hydrochloric acid TS and 2-propanol to make 100 ‌JP XVIII Official Monographs / Haloperidol Fine Granules 1091 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Halo-peridol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—To 1.0 g of Haloperidol add 50 mL of water, shake, and filter. To 25 mL of the filtrate add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Halo- peridol according to method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Related substances—Dissolve 25 mg of Haloperidol in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than haloperidol obtained from the sample solution is not larger than the peak area of haroperidol from the standard solu-tion, and the total area of these peaks is not larger than 2 times the peak area of haloperidol from the standard solu-tion. For the areas of the peaks, having the relative retention time of about 0.5, about 1.2 and about 2.6 to haloperidol, multiply their correction factors, 0.75, 1.47 and 0.76, respec-tively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.95 g of trisodium citrate dihy-drate in 900 mL of water, adjust to pH 3.5 with dilute hy-drochloric acid, and add water to make 1000 mL. To 300 mL of this solution add 700 mL of methanol and 1.0 g of sodium lauryl sulfate. Flow rate: Adjust so that the retention time of haloperidol is about 9 minutes. Time span of measurement: About 3 times as long as the retention time of haloperidol, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of haloperidol obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of haloperidol are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of haloperidol is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of haloperidol, previ-ously dried, and dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 1 drop of crystal violet TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.59 mg of C21H23ClFNO2
Halothane is a clear, colorless, and mobile liq-uid. It is miscible with ethanol (95), with diethyl ether and with isooctane. It is slightly soluble in water. It is a volatile, nonflammable liquid, and setting fire to its heated vapor does not support combustion. It is affected by light. D Refractive index n20: 1.369 – 1.371
Transfer about 3 mL of Halothane to a gas cell having light path 10 cm in length, and determine the in-frared absorption spectrum as directed in the gas sampling method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 20: 1.872 – 1.877
(1) Acidity or alkalinity—Shake 60 mL of Halo-thane with 60 mL of freshly boiled and cooled water vigor-ously for 3 minutes. Separate the water layer, and use this as the sample solution. To 20 mL of the sample solution add 1 drop of bromocresol purple TS and 0.10 mL of 0.01 mol/L sodium hydroxide VS: a red-purple color develops. To 20 mL of the sample solution add 1 drop of bromocresol purple TS and 0.6 mL of 0.01 mol/L hydrochloric acid VS: a yellow color is produced. (2) Halide and halogen—To 5 mL of the sample solution obtained in (1) add 1 drop of nitric acid and 0.20 mL of sil-ver nitrate TS: no turbidity is produced. To 10 mL of the sample solution obtained in (1) add 1 mL of potassium 509C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of the internal standard is 2 to 3 minutes. Selection of column: Mix 3 mL of Halothane and 1 mL of the internal standard. Proceed with 1 mL of this solution under the above operating conditions, and calculate the reso-lution. Use a column giving elution of the internal standard and halothane in this order with the resolution between these peaks being not less than 10. Detection sensitivity: Adjust the detection sensitivity so that the peak height of the internal standard obtained with 5 mL of the sample solution composes 30 to 70z of the full scale. Time span of measurement: About 3 times as long as the retention time of halothane. Distilling range <2.57> Not less than 95 volz distils within a 19C range between 499C and 519C. Thymol To 0.50 mL of Halothane add 5.0 mL of isooctane and 5.0 mL of titanium (IV) oxide TS, shake vigorously for 30 seconds, and allow to stand: the separated upper layer has more color than the following control solution A, and has no more color than the following control solution B. Control solution: Dissolve 0.225 g of thymol for assay in isooctane to make exactly 100 mL. To 10 mL each of this so-lution, accurately measured, add isooctane to make exactly 150 mL and 100 mL, respectively. Proceed with 0.50 mL each of these solutions in the same manner as Halothane, and use the separated upper layers so obtained as the control solution A and B, respectively.
Containers—Tight containers. Storage—Light-resistant, and not exceeding 309C.
Haloxazolam, when dried, contains not less than 99.
Haloxazolam occurs as white, crystals or crys-talline powder. It is odorless and tasteless. It is freely soluble in acetic acid (100), sparingly soluble in acetonitrile, in methanol and in ethanol (99.5), slightly solu-ble in diethyl ether, and practically insoluble in water. Melting point: about 1839C (with decomposition).
(1) Dissolve 10 mg of Haloxazolam in 10 mL of methanol, add 1 drop of hydrochloric acid: the solu-tion shows a yellow-green fluorescence under ultraviolet light (main wavelength: 365 nm). To this solution add 1 mL of sodium hydroxide TS: the fluorescence disappears imme-diately. Prepare the test solution with 50 mg of Haloxazolam as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 20 mL of dilute sodium hydroxide TS and 1 mL of hydrogen peroxide (30) as an absorbing liquid: the test solution responds to Qualitative Tests <1.09> for bro-mide and for fluoride. Determine the absorption spectrum of a solution of Haloxazolam in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Haloxazolam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Arsenic <1.11>—To 1.0 g of Haloxazolam in a de-composition flask add 5 mL of nitric acid and 2 mL of sulfu-ric acid, place a small funnel on the mouth of the flask, and heat carefully until white fumes are evolved. After cooling, add 2 mL of nitric acid, heat, repeat this procedure twice, add several 2-mL portions of hydrogen peroxide (30), and heat until the solution is colorless to pale yellow. After cool-ing, add 2 mL of a saturated solution of ammonium oxalate monohydrate, and heat until white fumes are evolved. After cooling, add water to make 5 mL, and perform the test with this solution: the solution has no more color than the follow-ing control solution (not more than 2 ppm). Control solution: Proceed in the same manner as above without using Haloxazolam, add 2.0 mL of Standard Arsenic Solution and water to make 5 mL, and proceed in the same manner as the test solution. Related substances—Dissolve 0.10 g of Haloxazolam in 100 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area of both solutions by the automatic integration method: the total area of all peaks other than haloxazolam from the sample solution is not larger than the peak area of the haloxazolam from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.2 g of boric acid and 7.5 g of potassium chloride in 900 mL of water, adjust the pH with triethylamine to 8.5, and add water to make 1000 mL. To 300 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of haloxazol-am is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of haloxazolam, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add acetonitrile to make exactly 50 mL. Confirm that the peak area of haloxazolam obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 1 cm Absorbance <2.24> E1z anol, 1000 mL). (247 nm): 390 – 410 (10 mg, meth- mL of the standard solution. System performance: Dissolve 10 mg each of Haloxa-zolam and cloxazolam in 200 mL of acetonitrile. When the
(1) Clarity and color of solution—Dissolve 0.10 g of Haloxazolam in 20 mL of ethanol (99.5): the solution is clear and colorless. Soluble halides—To 1.0 g of Haloxazolam add 50 mL of water, allow to stand for 1 hour with occasional shak-ing, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test with this solution as directed under Chloride Limit Test <1.03>. Prepare the control solution with 0.10 mL of 0.01 mol/L hy-drochloric acid VS. Heavy metals <1.07>—Proceed with 1.0 g of Haloxa- zolam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). procedure is run with 10 mL of this solution under the above operating conditions, haloxazolam and cloxazolam are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of haloxazolam is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible). ‌1096
Weigh accurately about 0.5 g of Haloxazolam, pre-viously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.72 mg of C17H14BrFN2O2
Hangekobokuto Extract contains not less than 2 mg and not more than 6 mg of magnolol, not less than 4 mg (for preparation prescribed 2 g of Perilla Herb) or not less than 6 mg (for preparation prescribed 3 g of Perilla Herb) of rosmarinic acid, and not less than 0.
Hangekobokuto Extract is a light brown to dark brown powder or black-brown viscous extract. It has a characteristic odor and has a bitter and astringent taste first then pungent later.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of magnolol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solu-tion (Magnolia Bark). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of 0.1 mol/L hydrochloric acid TS, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of rosmarinic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (60:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solution (Perilla Herb). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane and acetone (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 11.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 14.0z, calculated on the dried basis.
(1) Magnolol—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of magnolol for assay, and dissolve in diluted methanol (7 in 10) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of magnolol in each solution. Amount (mg) of magnolol ＝ MS × AT/AS × 1/8 MS: Amount (mg) of magnolol for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌2028 Hangeshashinto Extract / Crude Drugs and Related Drugs length: 289 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of [6]-gingerol in each solution. acid (100) (50:50:1). Flow rate: 1.0 mL per minute (the retention time of mag- Amount (mg) of [6]-gingerol ＝ MS × AT/AS × 1/20 nolol is about 15 minutes). System suitability— System performance: Dissolve 1 mg each of magnolol for assay and honokiol in diluted methanol (7 in 10) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, honokiol and mag-nolol are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of magnolol is not more than 1.5z. Rosmarinic acid—Conduct this procedure using light-resistant vessels. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of rosmarinic acid for assay, dissolve in diluted methanol (7 in 10) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of rosmarinic acid in each solution. Amount (mg) of rosmarinic acid ＝ MS × AT/AS × 1/4 MS: Amount (mg) of rosmarinic acid for assay taken, cal-culated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 330 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (800:200:1). Flow rate: 1.0 mL per minute (the retention time of ros-marinic acid is about 11 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rosmarinic acid are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosmarinic acid is not more than 1.5z. [6]-Gingerol—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of [6]-gingerol for assay, dissolve in MS: Amount (mg) of [6]-gingerol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (620:380:1). Flow rate: 1.0 mL per minute (the retention time of [6]-gingerol is about 15 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-gingerol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-gingerol is not more than 1.5z.
Hangeshashinto Extract contains not less than 70 mg and not more than 210 mg (for preparation prescribed 2.
Hangeshashinto Extract is a light yellow to yel-low-brown powder or black-brown viscous extract. It has a slightly odor and a hotter, bitter and slightly sweet taste.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Take the diethyl ether layer, evapo-rate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Scutellaria Root). For preparation prescribed Processed Ginger—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of [6]-shogaol for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 1 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of cyclohexane and ethyl ace-tate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Processed Ginger). For preparation prescribed Ginger—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1- butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rg1 RS or ginsenoside Rg1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Ginseng). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solu-tion. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 0.5 g of the dry extract (or 1.5 g of the viscous extract) with 10 mL of methanol, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, dissolve 1 mg of coptisine chloride for thin-layer chromatography in 5 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28) and metha-nol (15:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Coptis Rhizome).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract—Not more than 9.5z (1 g, 1059C, 5 hours). The viscous extract—Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Baicalin—Weigh accurately about 0.1 g of the Hangeshashinto Extract / Crude Drugs and Related Drugs dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of baicalein for resolution check in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and bai-calein is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Berberine—Weigh accurately about 0.2 g of the dry JP XVIII Crude Drugs and Related Drugs / Hemp Fruit 2031 ‌extract (or an amount of the viscous extract, equivalent to about 0.2 g of the dried substance), add exactly 50 mL of the mobile phase, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography‌ <2.01> according to the following conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine chloride (C20H18ClNO4) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: 1.0 mL per minute (the retention time of ber-berine is about 8 minutes). System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, palmatine and berberine are eluted in this order with the resolution be-tween these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Hochuekkito Extract contains not less than 16 mg and not more than 64 mg of hesperidin, not less than 0.
Hochuekkito Extract occurs as a light brown to brown powder or black-brown viscous extract. It has a slight odor, and a sweet and bitter taste.
(1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, and shake. Centrifuge this solution, and use the 1-butanol layer as the sample solu-tion. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). For preparation prescribed Atractylodes Rhizome—To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 30 mL of water, shake, then add 50 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the red spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, shake, and separate the hexane layer. Evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying, heated the plate at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhizome). To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract) add 40 mL of a solution of potassium hydroxide in methanol (1 in 50), shake for 15 minutes, centrifuge, and evaporate the solvent under low pressure (in vacuo). Add 30 mL of water and 20 mL of diethyl ether to the residue, shake, remove the diethyl ether layer, and separate the aque-ous layer. To the aqueous layer add 20 mL of 1-butanol, shake, and separate the 1-butanol layer. To the 1-butanol JP XVIII Crude Drugs and Related Drugs / Hochuekkito Extract 2033 layer add 20 mL of water, shake, separate the 1-butanol layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of astragalo-side IV for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of octadecylsilanized silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, water, 1-butanol and acetic acid (100) (60:30:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-brown spot from the standard solution (Astragalus Root). To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract) add 30 mL of water, shake, then add 50 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Japanese Angelica Root). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 30 mL of water, shake, then add 50 mL of 1-butanol, shake, and separate the 1-butanol layer. Evaporate the solvent under low pressure (in vacuo), add 3 mL of meth-anol to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of hesperidin for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL of the sample solution and 20 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, ace-tone, water and acetic acid (100) (10:6:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and expose to ammonia vapor: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (Citrus Unshiu Peel). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, and shake. Centrifuge this solution, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatogra-phy in 1 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 30 mL of water, shake, then add 50 mL of 1-butanol, shake, and separate the 1-butanol layer. Evaporate the solvent under low pressure (in vacuo), add 3 mL of meth-anol to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). For preparation prescribed Ginger—To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 30 mL of water, shake, then add 50 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of diethyl ether to the residue, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of [6]-gingerol for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). For preparation prescribed Processed Ginger—Put 10 g of the dry extract (or 30 g of the viscous extract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determi-nation, and heat to boil under a reflux condenser. The grad-uated tube of the apparatus is previously filled with water to the standard line, and 2 mL of hexane is added to the grad-uated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample solution. Separately, dissolve 1 mg of [6]-shogaol for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 60 mL of the sample solution and 10 mL of the standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of cyclohexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Processed Ginger). Hochuekkito Extract / Crude Drugs and Related Drugs To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 30 mL of water, shake, then add 50 mL of 1-butanol, shake, and separate the 1-butanol layer. Evaporate the solvent under low pressure (in vacuo), add 3 mL of meth-anol to the residue, and use this solution as the sample solu-tion. Use (E )-isoferulic acid-(E )-ferulic acid TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetone and water (20:12:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and exa-mine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the light yellow-white fluorescent spot from the standard solution (Cimicifuga Rhizome).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed in the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 11.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Hesperidin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.1 g of the dried substance), add exactly 50 mL of diluted tetrahydrofuran (1 in 4), shake for 30 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 10 mg of hesperidin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add diluted tetra-hydrofuran (1 in 4) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of hesperidin in each solution. Amount (mg) of hesperidin ＝ MS × AT/AS × 1/20 MS: Amount (mg) of hesperidin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (82:18:1). Flow rate: 1.0 mL per minute (the retention time of hesperidin is about 15 minutes). System suitability— System performance: Dissolve 1 mg each of hesperidin for assay and naringin for thin-layer chromatography in diluted methanol (1 in 2) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, naringin and hesperidin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hesperidin is not more than 1.5z. Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of sai-kosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 CS: Concentration (mg/mL) of saikosaponin b2 in saiko-saponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of saiko-saponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as de-scribed above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the ‌JP XVIII Crude Drugs and Related Drugs / Houttuynia Herb 2035‌ standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. diately. Resorcinol-coloring substances—Mix well 5 g of Honey with 15 mL of diethyl ether, filter, and evaporate the diethyl ether solution at ordinary temperature. To the residue add 1 to 2 drops of resorcinol TS: a yellow-red color Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) may develop in the solution of resorcinol and in the residue, and a red to red-purple color which does not persist more than 1 hour. MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Homatropine Hydrobromide contains not less than lution with 0.
Homatropine Hydrobromide occurs as white, crystals or crystalline powder. It is odorless. It is freely soluble in water, sparingly soluble in ethanol (95), slightly soluble in acetic acid (100), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. It is affected by light. Melting point: about 2149C (with decomposition).
(1) To 5 mL of a solution of Homatropine Hydrobromide (1 in 20) add 2 to 3 drops of iodine TS: a brown precipitate is produced. Dissolve 0.05 g of Homatropine Hydrobromide in 5 mL of water, and add 3 mL of 2,4,6-trinitrophenol TS: a yellow precipitate is produced. Filter the precipitate, wash with five 10-mL portions of water, and dry at 1059C for 2 hours: it melts <2.60> between 1849C and 1879C. A solution of Homatropine Hydrobromide (1 in 20) responds to Qualitative Tests <1.09> for bromide.
(1) Acidity—Dissolve 1.0 g of Homatropine ‌Hydrobromide in 20 mL of water, and add 0.40 mL of 0.01 mol/L sodium hydroxide VS and 1 drop of methyl red-methylene blue TS: a green color develops. Atropine, hyoscyamine and scopolamine—To 10 mg of Homatropine Hydrobromide add 5 drops of nitric acid, evaporate on a water bath to dryness, and cool. Dissolve the residue in 1 mL of N,N-dimethylformamide, and add 5 to 6 drops of tetraethylammonium hydroxide TS: no red-purple color is produced. Related substances—Dissolve 0.15 g of Homatropine Hydrobromide in 3 mL of water, and use this solution as the sample solution. To 1 mL of the sample solution add 2 to 3 drops of tannic acid TS: no precipitate is produced. To 1 mL of the sample solution add 2 to 3 drops each of dilute hydrochloric acid and platinic chloride TS: no pre-cipitate is produced. Loss on drying <2.41> Not more than 1.5z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (0.2 g).
Dissolve by warming about 0.4 g of Homatropine Hydrobromide in 60 mL of a mixture of acetic anhydride and acetic acid (100) (7:3). Cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.63 mg of C16H21NO3.HBr
Homochlorcyclizine Hydrochloride, when dried, contains not less than 98.
Homochlorcyclizine Hydrochloride occurs as white to pale brown, crystals or powder. It is very soluble in water, freely soluble in acetic acid (100), slightly soluble in ethanol (99.5), and very slightly soluble in acetonitrile and in acetic anhydride. It dissolves in 0.1 mol/L hydrochloric acid TS. It is hygroscopic. It is colored slightly by light. A solution of Homochlorcyclizine Hydrochloride (1 in 10) shows no optical rotation. Melting point: about 2279C (with decomposition).
(1) Determine the absorption spectrum of a solution of Homochlorcyclizine Hydrochloride in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Homochlorcyclizine Hydrochloride, previously dried, as di-rected in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. A solution of Homochlorcyclizine Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Homochlorcyclizine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Homochlor-cyclizine Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Measure exactly 1 mL of the sample solution, add the mobile phase to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas by the automatic integration method: the areas of the peaks other than homochlorcycli-zine obtained from the sample solution are not larger than 1/2 times the peak area of homochlorcyclizine from the standard solution, and the total area of the peaks other than homochlorcyclizine from the sample solution is not larger than the peak area of homochlorcyclizine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 223 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and per-chloric acid (134:66:1). Flow rate: Adjust so that the retention time of homochlor-cyclizine is about 10 minutes. Time span of measurement: About 2 times as long as the retention time of homochlorcyclizine. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 50 mL. Confirm that the peak area of homochlorcyclizine ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 5 mg each of Homochlor-cyclizine Hydrochloride and methyl parahydroxybenzoic acid in 100 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, methyl parahydroxybenzoic acid and homo-chlorcyclizine are eluted in this order with the resolution be-tween these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of homochlorcyclizine is not more than 1.0z. Loss on drying <2.41> Not more than 2.0z (1 g, 1109C, 4 hours). ‌JP XVIII Official Monographs / Hydralazine Hydrochloride for Injection 1109‌‌ Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Homochlorcyclizine Hydrochloride, previously dried, dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.39 mg of C H ClN .2HCl 50 mL of water: the pH of the solution is between 3.5 and 4.5. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Hydralazine Hydrochloride in 50 mL of water: the solu-tion is clear, and colorless or pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Hydrala- zine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). 19 23 2
Human Menopausal Gonadotrophin is a dried preparation of gonad-stimulating hormone obtained from the urine of postmenopausal healthy women, after processing for virus removal or inactivation. It has follicle-stimulating hormonal action and luteiniz-ing hormonal action. It contains not less than 40 follicle-stimulating hor-mone Units per mg.
Human Menopausal Gonadotrophin occurs as a white to pale yellow powder. It is soluble in water. Ratio of interstitial cell-stimulating hormone to follicle-stimulating hormone Perform the test according to the fol-lowing method: the ratio of the unit of interstitial cell-stimulating hormone (luteinizing hormone) to that of fol-licle-stimulating hormone is not more than 1. Test animals—Select healthy male albino rats weigh-ing about 45 to 65 g. Standard solutions—Dissolve Menopausal Gonadotrophin RS in bovine serum albumin-sodium chlo-ride-phosphate buffer solution (pH 7.2) to prepare three kinds of solutions, containing 10, 20 and 40 interstitial cell-stimulating hormone (luteinizing hormone) units per 1.0 mL, respectively. Inject these solutions into three groups consist-ing of five test animals each, and weigh their seminal vesicles as directed in (iv). According to the result of the test, desig-nate the concentration of the reference standard, which will make the mass of the seminal vesicle 20 to 35 mg, as the high-dose standard solution, SH. Dilute the SH to 1.5 to 2.0 times the initial volume with the bovine serum albumin-sodium chloride-phosphate buffer solution (pH 7.2) and designate this solution as the low-dose standard solution, SL. Sample solutions—Weigh accurately a suitable amount of Human Menopausal Gonadotrophin, and dis-solve in the bovine serum albumin-sodium chloride-phosphate buffer solution (pH 7.2) to prepare the high-dose sample solution, TH and the low-dose sample solution, TL, so that their concentrations are similar to those of the corre-sponding standard solutions, respectively. Store these solu-tions at 2 – 89C. Procedure—Divide the test animals at random into 4 groups, A, B, C and D, with not less than 10 animals and equal numbers in each group. Inject subcutaneously once every day 0.2 mL each of SH, SL, TH and TL to each animal in the respective groups for five days. On the sixth day, ex-cise the seminal vesicles, remove extraneous tissue, remove fluid adhering to the vesicles and the contents of the vesicles by lightly pressing between filter papers, and weigh the vesi-cles. Calculation—Proceed as directed in (v) in the Assay by changing the mass of ovaries to the mass of seminal vesi-cles and the unit of follicle-stimulating hormone to the unit of interstitial cell-stimulating hormone to read. Water <2.48> Not more than 5.0z (0.2 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Dissolve Human Menopausal Gonadotrophin in water for bacterial endotoxins test to pre-pare a solution containing 75 follicle-stimulating hormone Units per mL, and perform the test: less than 0.66 EU/ follicle-stimulating hormone Unit. Specific activity Perform the test with Human Menopausal Gonadotrophin according to the following method, and cal-culate the specific activity using the amount (Unit) obtained in the Assay: it is not less than 50 follicle-stimulating hor-mone Units per 1 mg of protein. Sample solution—Weigh accurately about 10 mg of Human Menopausal Gonadotrophin, dissolve in water so that each mL contains exactly 200 mg, and use this solution as the sample solution. Standard solutions—Weigh accurately about 10 mg of bovine serum albumin, and dissolve in water to make ex-actly 20 mL. To this solution add water to make four solu-tions containing exactly 300 mg, 200 mg, 100 mg and 50 mg of the albumin per mL, respectively, and use these solutions as the standard solutions. Procedure—To glass test tubes, about 18 mm in in-side diameter and about 130 mm in height, add separately exactly 0.5 mL each of the sample solution and the standard solutions. To these tubes add exactly 5 mL of alkaline cop-per TS, warm in a water bath at 309C for 10 minutes, then add exactly 0.5 mL of diluted Folin's TS (1 in 2), and warm in a water bath at 309C for 20 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-metry <2.24>, and determine the absorbances at 750 nm, using a liquid obtained with 0.5 mL of water in the same manner as above as a blank. Prepare a calibration curve from the absorbances of the standard solutions, with absorbance on the vertical axis and concentration on the horizontal axis. Calculate the amount of protein in the sample solution from the absorbance of the sample solution using the curve, and calculate the protein content of the sample.
Test animals—Select healthy female albino rats weighing about 45 to 65 g. Standard solutions—Dissolve Human Menopausal Gonadotrophin RS in human chorionic gonadotrophin TS to prepare three solutions which contain 0.75, 1.5 and 3.0 fol-licle-stimulating hormone Units per 1.0 mL, respectively. In-ject these solutions into three groups consisting of five test animals each, and weigh their ovaries, as directed in (iv). Ac-cording to the result of the test, designate the concentration of the reference standard, which will make the mass of the o-vary about 120 to 160 mg, as the high-dose standard solu-tion, SH. Dilute the SH to 1.5 to 2.0 times the initial volume ‌1088 with the human chorionic gonadotrophin TS, and designate the solution as the low-dose standard solution, SL. Sample solutions—Weigh accurately a suitable amount of Human Menopausal Gonadotrophin, dissolve in human chorionic gonadotrophin TS, and prepare the high-dose sample solution, TH, and the low-dose sample solution, TL, which have similar numbers of units to those of corre-sponding standard solutions in equal volume, respectively. Procedure—Divide the test animals at random into 4 groups, A, B, C and D, with not less than 10 animals and equal numbers in each group. Inject subcutaneously 0.2 mL each of SH, SL, TH and TL into the animals in each group, once in the afternoon on the first day, three times in the morning, noon and afternoon on the second day, and two times in the morning and afternoon on the third day. On the fifth day, excise the ovaries, remove the fat and extraneous tissue, remove fluid adhering to the ovaries by lightly press-ing between filter papers, and immediately weigh the ovaries. Calculation—Designate the mass of ovaries by SH, SL, TH and TL as y1, y2, y3 and y4, respectively. Sum up y1, y2, y3 and y4 on each set to obtain Y1, Y2, Y3 and Y4. Units of follicle-stimulating hormone per mg of Human Menopausal Gonadotrophin ＝ antilog M × (units per mL of SH) × b/a M ＝ IYa/Yb I ＝ log (SH/SL) ＝ log (TH/TL) Ya ＝ －Y1 － Y2 ＋ Y3 ＋ Y4 Yb ＝ Y1 － Y2 ＋ Y3 － Y4 a: Mass (mg) of Human Menopausal Gonadotrophin taken b: Total volume (mL) of the high dose of the test solution prepared by diluting with human chorionic gonadotro-phin TS F? computed by the following equation should be smaller than F1 against n when s2 is calculated. And compute L (P ＝ 0.95) by the following equation: L should be not more than 0.3. If F? exceeds F1, or if L exceeds 0.3, repeat the test in-creasing the number of the test animals or arranging the assay method in a better way until F? is smaller than F1 or L is not more than 0.3. F? ＝ (Y1 － Y2 － Y3 ＋ Y4)2/(4fs2) f: Number of test animals per group s2 ＝ {Sy2 － (Y/f)}/n Sy2: The sum of the squares of each y1, y2, y3 and y4 Y ＝ Y12 ＋ Y22 ＋ Y32 ＋ Y42 (C － 1) (CM 2 ＋ I 2) n ＝ 4(f － 1) L ＝ 2 C ＝ Yb2/(Yb2 － 4fs2t2) t2: Value shown in the following table against n used to calculate s2 n t2 ＝ F1 n t2 ＝ F1 n t2 ＝ F1 161.45 4.667 4.242 18.51 4.600 4.225 10.129 4.543 4.210 7.709 4.494 4.196 6.608 4.451 4.183 5.987 4.414 4.171 5.591 4.381 4.085 5.318 4.351 4.001 5.117 4.325 3.920 4.965 4.301 ∞ 3.841 4.844 4.279 4.747 4.260
Hydralazine Hydrochloride, when dried, contains not less than 98.
Hydralazine Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is soluble in water, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 2759C (with decomposition).
(1) Determine the absorption spectrum of a solution of Hydralazine Hydrochloride (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Hydralazine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Hydralazine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Hydralazine Hydrochloride in
Hydrochloric Acid Lemonade is a clear, color-less liquid. It has a sweet, cool, acid taste.
Hydrochloric Acid contains not less than 35.
Hydrochloric Acid is a colorless liquid having a pungent odor. It is fuming but ceases to fume when it is diluted with 2 volumes of water. Specific gravity d 20: about 1.18.
(1) Allow a glass stick wet with ammonia TS to come near the surface of Hydrochloric Acid: a remarkable white smoke evolves. (2) A solution of Hydrochloric Acid (1 in 100) changes blue litmus paper to red, and responds to Qualitative Tests <1.09> for chloride.
(1) Sulfate <1.14>—To 15 mL of Hydrochloric Acid add water to make 50 mL, and use this solution as the sample solution. To 3.0 mL of the sample solution add 5 mL of water and 5 drops of barium chloride TS, and allow to stand for 1 hour: no turbidity is produced. Sulfite—To 3.0 mL of the sample solution obtained in (1) add 5 ml of water and 1 drop of iodine TS: the color of iodine TS does not disappear. Bromide or iodide—Place 10 mL of the sample solu-tion obtained in (1) in a glass-stoppered test tube, add 1 mL of chloroform and 1 drop of 0.002 mol/L potassium per-manganate VS, and shake well: the chloroform layer remains colorless. Bromine or chlorine—Place 10 mL of the sample so- lution obtained in (1) in a glass-stoppered test tube, add 5 drops of potassium iodide TS and 1 mL of chloroform, and shake for 1 minute: the chloroform layer remains free from a purple color. Heavy metals <1.07>—Evaporate 5 mL of Hydro- chloric Acid on a water bath to dryness, and add 2 mL of dilute acetic acid and water to the residue to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution as follows: to 3.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.7 mL of Hydrochloric Acid according to Method 1, and per-form the test (not more than 1 ppm). Mercury—Dilute 20 mL of Hydrochloric Acid with water to make exactly 100 mL, and use the solution as the sample solution. Perform the test with the sample solution as directed under Atomic Absorption Spectrophotometry <2.23> (cold vapor type). Place the sample solution in a sam- ple bottle of the atomic absorption spectrophotometer, add 10 mL of tin (II) chloride-sulfuric acid TS, connect the bottle immediately to the spectrophotometer, circulate air, and de-termine the absorbance AT of the sample solution after the recorder reading has risen rapidly, and becomes constant at a wavelength of 253.7 nm. On the other hand, to 8 mL of Standard Mercury Solution add water to make exactly 100 mL, and determine the absorbance AS of the solution ob-tained by the same procedure as used for the sample solu-tion: AT is smaller than AS (not more than 0.04 ppm). Residue on ignition <2.44> Pipet 10 mL of Hydrochloric Acid, add 2 drops of sulfuric acid, evaporate to dryness, and ignite: not more than 1.0 mg of residue remains.
Weigh accurately a glass-stoppered flask containing 20 mL of water, add about 3 mL of Hydrochloric Acid, and weigh accurately again. Dilute with 25 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 to 3 drops of methyl red TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 36.46 mg of HCl
Hydrochlorothiazide, when dried, contains not less than 99.
Hydrochlorothiazide occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in acetone, sparingly soluble in aceto-nitrile, very slightly soluble in water and in ethanol (95), and practically insoluble in diethyl ether. It dissolves in sodium hydroxide TS. Melting point: about 2679C (with decomposition).
(1) To 5 mg of Hydrochlorothiazide add 5 mL of chromotropic acid TS, and allow to stand for 5 minutes: a purple color develops. Fuse a mixture of 0.1 g of Hydrochlorothiazide and 0.5 g of sodium carbonate decahydrate cautiously: the gas evolved changes moistened red litmus paper to blue. After cooling, crush with a glass rod, add 10 mL of water, stir, and filter. To 4 mL of the filtrate add 2 drops of hydrogen peroxide (30), 5 mL of diluted hydrochloric acid (1 in 5) and 2 to 3 drops of barium chloride TS: a white precipitate is produced. To 4 mL of the filtrate obtained in (2) add 5 mL of dilute nitric acid and 3 drops of silver nitrate TS: a white precipitate is produced. Dissolve 12 mg of Hydrochlorothiazide in 100 mL of sodium hydroxide TS. Dilute 10 mL of the solution with water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of a solution of Hydro-chlorothiazide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Chloride <1.03>—Dissolve 1.0 g of Hydrochlo-rothiazide in 30 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.036z). Sulfate <1.14>—Dissolve 1.0 g of Hydrochloro- thiazide in 30 mL of acetone, add 1 mL of dilute hydrochlo-ric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.005 mol/L sulfuric acid VS add 30 mL of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Hydro- chlorothiazide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). ‌JP XVIII Official Monographs / Hydrocortisone 1113 Primary aromatic amines—Dissolve 80 mg of Hydro-chlorothiazide in acetone to make exactly 100 mL. Measure exactly 1 mL of the solution, add 3.0 mL of dilute hydro-chloric acid, 3.0 mL of water and 0.15 mL of sodium nitrite TS, shake, and allow to stand for 1 minute. Shake this solu-tion with 1.0 mL of ammonium amidosulfate TS, allow to stand for 3 minutes, then add 1.0 mL of N,N-diethyl-N?-1-naphthylenediamine oxalate TS, shake, and allow to stand for 5 minutes. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 1.0 mL of acetone in the same man-ner as the blank: the absorbance at 525 nm is not more than 0.10. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 30 mg each of Hydrochloro-thiazide and Hydrochlorothiazide RS, previously dried, and dissolve in 150 mL of the mobile phase, add exactly 10 mL each of the internal standard solution, then add the mobile phase to make 200 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of hydrochlorothiazide to that of the internal standard. Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × QT/QS MS: Amount (mg) of Hydrochlorothiazide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in acetonitrile (9 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.1 mol/L sodium dihydrogen phosphate TS (pH 3.0) and acetonitrile (9:1). Flow rate: Adjust so that the retention time of hydrochlo-rothiazide is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, hydrochlorothiazide and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of hydrochlorothiazide to that of the inter-nal standard is not more than 1.0z.
Hydrocortisone Acetate, when dried, contains not less than 97.
Hydrocortisone Acetate occurs as white, crys-tals or crystalline powder. It is freely soluble in dimethylsulfoxide, slightly soluble in methanol and in ethanol (95), and practically insoluble in water. Melting point: about 2209C (with decomposition). It shows crystal polymorphism.
(1) Add 2 mL of sulfuric acid to 2 mg of Hydrocortisone Acetate: the solution shows a yellowish green fluorescence immediately, and the color of the solution gradually changes through orange-yellow to dark red. This solution shows a strong light green fluorescence under ultra-violet light. Add carefully 10 mL of water to this solution: the color changes from yellow to orange-yellow with a light green fluorescence, and a yellow-brown, flocculent precipi-tate is formed. Dissolve 0.01 g of Hydrocortisone Acetate in 1 mL of methanol by warming, add 1 mL of Fehling's TS, and heat: an orange to red precipitate is formed. To 0.05 g of Hydrocortisone Acetate add 2 mL of po-tassium hydroxide-ethanol TS, and heat on a water bath for 5 minutes. Cool, add 2 mL of diluted sulfuric acid (2 in 7), and boil gently for 1 minute: the odor of ethyl acetate is per-ceptible. Determine the infrared absorption spectra of Hydro-cortisone Acetate and Hydrocortisone Acetate RS, previ-ously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: both the sample and the RS exhibit similar intensities of absorption at the same wave numbers. If any difference appears, dissolve the sample and the Reference Standard in ethanol (95), respectively, evaporate to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]25: ＋154 – ＋1649(after drying, 50 mg, dimethylsulfoxide, 10 mL, 100 mm).
Related substances—Dissolve 40 mg of Hydrocorti-sone Acetate in 25 mL of a mixture of chloroform and meth-anol (9:1), and use this solution as the sample solution. Pipet 2 mL of the sample solution, add a mixture of chloroform and methanol (9:1) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of dichloromethane, diethyl ether, methanol and water (160:30:8:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly alkaline blue tetrazolium TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Dissolve about 20 mg each of Hydrocortisone Ace-tate and Hydrocortisone Acetate RS, previously dried and accurately weighed, in methanol, add exactly 10 mL each of the internal standard solution, then add methanol to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of hydrocortisone acetate to that of the internal standard, re-spectively. Amount (mg) of hydrocortisone acetate (C23H32O6) ＝ MS × QT/QS MS: Amount (mg) of Hydrocortisone Acetate RS taken ‌JP XVIII Official Monographs / Hydrocortisone Butyrate 1115‌ Internal standard solution—A solution of benzyl parahy-droxybenzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (13:7). Flow rate: Adjust so that the retention time of hydrocorti- sone acetate is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, hydrocortisone acetate and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of hydrocortisone acetate to that of the in-ternal standard is not more than 1.0z.
Hydrocortisone Butyrate, when dried, contains not less than 96.
Hydrocortisone Butyrate occurs as a white powder. It is odorless. It is freely soluble in chloroform, soluble in methanol, sparingly soluble in ethanol (99.5) and practically insoluble in water. Melting point: about 2009C (with decomposition).
(1) Add 2 mL of sulfuric acid to 2 mg of Hydrocortisone Butyrate: the solution shows a yellowish green fluorescence immediately, and the color of the solution gradually changes through orange-yellow to dark red. This solution shows a strong light green fluorescence under ultra-violet light (main wavelength: 254 nm). Add carefully 10 mL of water to this solution: the color changes from yellow to orange-yellow with a light green fluorescence, and a yellow-brown, flocculent precipitate is formed. Dissolve 0.01 g of Hydrocortisone Butyrate in 1 mL of methanol by warming, add 1 mL of Fehling's TS, and heat: an orange to red precipitate is formed. To 50 mg of Hydrocortisone Butyrate add 2 mL of potassium hydrox-ide-ethanol TS, and heat on a water bath for 5 minutes. Cool, add 2 mL of diluted sulfuric acid (2 in 7), and boil gently for 1 minute: the odor of ethyl butyrate is perceptible. Determine the infrared absorption spectrum of Hy-drocortisone Butyrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋48 – ＋529(after drying, 0.1 g, chloroform, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of ‌1116 Hydrocortisone Butyrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Hydrocorti- ethanol (99.5) to make exactly 50 mL. Determine the absor-bance A of this solution at the wavelength of maximum ab-sorption at about 241 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. sone Butyrate in 50 mL of a mixture of acetonitrile and the mobile phase A (4:1), and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add a mixture of Amount (mg) of hydrocortisone butyrate (C ＝ A/375 × 25,000 25H 36O6) acetonitrile and the mobile phase A (4:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than hydrocortisone butyrate obtained from the sample solution is not larger than the peak area of hydro-cortisone butyrate from the standard solution, and the total area of the peaks other than hydrocortisone butyrate from the sample solution is not larger than 2 times the peak area of hydrocortisone butyrate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 1 g of potassium dihydrogen
ヒドロコルチゾンコハク酸エステル
Hydrocortisone Succinate, when dried, contains not less than 97.
Hydrocortisone Succinate occurs as a white crystalline powder. It is very soluble in methanol, freely soluble in ethanol (99.5), and practically insoluble in water. It shows crystal polymorphism.
(1) To 3 mg of Hydrocortisone Succinate add 2 mL of sulfuric acid: the solution shows a yellowish green fluorescence immediately, and the color of the solution gradually changes through orange-yellow to dark red. This solution shows a strong light green fluorescence under ultra-violet light. Add carefully 10 mL of water to this solution: the color changes from yellow to orange-yellow with a light green fluorescence, and a yellow-brown flocculent precipi-tate is formed. (2) Determine the infrared absorption spectrum of Hy-drocortisone Succinate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer- JP XVIII Official Monographs / Hydrocotarnine Hydrochloride Hydrate 1119 ‌ence Spectrum or the spectrum of previously dried Hydro-cortisone Succinate RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. If any differ-ence appears between the spectra, dissolve Hydrocortisone Succinate and Hydrocortisone Succinate RS in methanol, respectively, then evaporate the methanol to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋147 – ＋1539(after drying, 0.1 g, ethanol (99.5), 10 mL, 100 nm).
Related substances—Dissolve 25 mg of Hydrocorti-sone Succinate in exactly 10 mL of methanol, and use this solution as the sample solution. Separately, dissolve 25 mg of hydrocortisone in exactly 10 mL of methanol. Pipet 1 mL of this solution, dilute with methanol to exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 3 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, ethanol (99.5) and formic acid (150:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 50 mg each of Hydrocorti-sone Succinate and Hydrocortisone Succinate RS, previously dried, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL each of the internal standard solution, then add methanol to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following oper-ating conditions, and calculate the ratios, QT and QS, of the peak area of hydrocortisone succinate to that of the internal standard, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of hydrocortisone succinate to that of the internal standard is not more than 1.0z.
Hydrocortisone, when dried, contains not less than 97.
Hydrocortisone occurs as a white crystalline powder. It is sparingly soluble in methanol and in ethanol (99.5), and very slightly soluble in water. Melting point: 212 – 2209C (with decomposition). It shows crystal polymorphism.
(1) Add 2 mL of sulfuric acid to 2 mg of Hydrocortisone: the solution shows a yellow-green fluores-cence immediately, and the color of the solution changes gradually from orange to dark red. Dilute carefully the solu-tion with 10 mL of water: the color changes through yellow to orange-yellow with green fluorescence, and a small amount of a flocculent precipitate is formed. Dissolve 0.01 g of Hydrocortisone in 1 mL of metha-nol, add 1 mL of Fehling's TS, and heat: a red precipitate is formed. Determine the infrared absorption spectrum of Hy-drocortisone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Hydrocortisone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Hydrocortisone and Hydrocortisone RS in ethanol (95), respectively, then evaporate the ethanol to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]25: ＋160 – ＋1709(after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm).
Related substances—Dissolve 20 mg of Hydrocorti-sone in 10 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (9:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (17:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g). ‌Assay Dissolve about 20 mg each of Hydrocortisone and Hydrocortisone RS, previously dried and accurately weighed, in 20 mL each of a mixture of chloroform and methanol (9:1), add 10 mL each of the internal standard so-lution, then add a mixture of chloroform and methanol (9:1) to make 50 mL, and use these solutions as the sample solu-tion and standard solution. Perform the test with 5 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of hydrocortisone to that of the internal standard, respectively. Amount (mg) of hydrocortisone (C21H30O5) ＝ MS × QT/QS MS: Amount (mg) of Hydrocortisone RS taken Internal standard solution—A solution of prednisone in a mixture of chloroform and methanol (9:1) (9 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of chloroform, methanol and acetic acid (100) (1000:20:1). Flow rate: Adjust so that the retention time of hydrocorti-sone is about 15 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and hydrocortisone are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of hydrocortisone to that of the internal stand-ard is not more than 1.0z.
Hydrocotarnine Hydrochloride Hydrate, when dried, contains not less than 98.
Hydrocotarnine Hydrochloride Hydrate oc-curs as white to pale yellow, crystals or crystalline powder. It is freely soluble in water, sparingly soluble in ethanol (95) and in acetic acid (100), and slightly soluble in acetic an-hydride.
(1) Determine the absorption spectrum of a solution of Hydrocotarnine Hydrochloride Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Hydrocotarnine Hydrochloride Hydrate as directed in the Amount (mg) of hydrocortisone succinate (C ＝ MS × QT/QS 25H 34O8) potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab- MS: Amount (mg) of Hydrocortisone Succinate RS taken Internal standard solution—A solution of butyl parahydroxy benzoate in methanol (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetic acid-sodium acetate buffer solution (pH 4.0) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of hydrocorti-sone succinate is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, hydrocortisone succinate and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. sorption at the same wave numbers. A solution of Hydrocotarnine Hydrochloride Hy-drate (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Hydrocotarnine Hydrochloride Hydrate in 20 mL of water: the pH of the solution is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Hydrocotarnine Hydrochloride Hydrate in 10 mL of water: the solution is clear, and when perform the test with this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, using water as the blank, the absorbance at 400 nm is not more than 0.17. Heavy metals <1.07>—Proceeds with 1.0 g of Hydrocotarnine Hydrochloride Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Hydrocotar-nine Hydrochloride Hydrate in 10 mL of diluted ethanol ‌1120‌ (99.5) (1 in 2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ethanol (99.5) (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ace-tone, toluene, ethanol (99.5) and ammonia water (28) (20:20:3:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Hydrocotarnine Hydrochloride Hydrate, previously dried. Dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3) by warming. Cool, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 25.77 mg of C12H15NO3.HCl
Hydrogenated Oil occurs as a white mass or powder and has a characteristic odor and a mild taste. It is freely soluble in diethyl ether, very slightly soluble in ethanol (95), and practically insoluble in water. The oil obtained by hydrogenation of castor oil is slightly soluble in diethyl ether, very slightly soluble in ethanol (95), and practically insoluble in water. Acid value <1.13> Not more than 2.0.
(1) Moisture and coloration—Hydrogenated Oil (5.0 g), melted by heating on a water bath, forms a clear liq-uid, from which no water separates. In a 10-mm thick layer of the liquid, it is colorless or slightly yellow. Alkalinity—To 2.0 g of Hydrogenated Oil add 10 mL of water, melt by heating on a water bath, and shake vigor-ously. After cooling, add 1 drop of phenolphthalein TS to the separated water layer: no color develops. Chloride—To 1.5 g of Hydrogenated Oil add 30 mL of ethanol (95), boil for 10 minutes under a reflux con-denser, and filter after cooling. To 20 mL of the filtrate add 5 drops of a solution of silver nitrate in ethanol (95) (1 in 50): the turbidity of the solution does not exceed that of the following control solution. Control solution: To 1.0 mL of 0.01 mol/L hydrochloric acid VS add ethanol (95) to make 20 mL, then add 5 drops of a solution of silver nitrate in ethanol (95) (1 in 50). Heavy metals—Heat 2.0 g of Hydrogenated Oil with 5 mL of dilute hydrochloric acid and 10 mL of water on a water bath for 5 minutes with occasional shaking. After cooling, filter, and make 5 mL of the filtrate weakly alkaline with ammonia TS, then add 3 drops of sodium sulfide TS: the solution remains unchanged. Nickel—Place 5.0 g of Hydrogenated Oil in a quartz or porcelain crucible, heat slightly with caution at the begin-ning, and, after carbonization, incinerate by strong heating (500 ± 209C). Cool, add 1 mL of hydrochloric acid, evapo-rate on a water bath to dryness, dissolve the residue in 3 mL of dilute hydrochloric acid, and add 7 mL of water. Then add 1 mL of bromine TS and 1 mL of a solution of citric acid monohydrate (1 in 5), make alkaline with 5 mL of am-monia TS, and cool in running water. To this solution add 1 mL of dimethylglyoxime TS, add water to make 20 mL, and use this solution as the test solution. Allow to stand for 5 minutes: the solution has no more color than the following control solution. Control solution: Evaporate 1 mL of hydrochloric acid on a water bath to dryness, add 1 mL of Standard Nickel Solu-tion and 3 mL of dilute hydrochloric acid, and add 6 mL of water. Then proceed as directed in the test solution, add water to make 20 mL, and allow to stand for 5 minutes. Residue on ignition <2.44> Not more than 0.1z (5 g).
Hydrophilic Petrolatum is white in color. It has a slight, characteristic odor. When mixed with an equal volume of water, it retains the consistency of ointment.
Hydrous Lanolin is Purified Lanolin to which water is added. It contains not less than 70z and not more than 75z of Purified Lanolin (as determined by the test for Residue on evaporation).
Hydrous Lanolin is a yellow-white, ointment-like substance, and has a slight, characteristic odor, which is not rancid. It is soluble in diethyl ether and in cyclohexane, with the separation of water. When melted by heating on a water bath, it separates into a clear oily layer and a clear aqueous layer. Melting point: about 399C.
Dissolve 1 g of Hydrous Lanolin in 50 mL of cyclohexane, and remove the separated water. Superimpose carefully 1 mL of the cyclohexane solution on 2 mL of sulfu-ric acid: a red-brown color develops at the zone of contact, and sulfuric acid layer shows a green fluorescence. Acid value <1.13> Not more than 1.0. Iodine value 18 – 36 Heat a suitable amount of Hydrous Lanolin on a water bath to remove its almost moisture, then weigh accurately about 0.8 g of the treated Hydrous Lanolin in a glass-stoppered 500-mL flask, and add 10 mL of cyclo-hexane to dissolve, and add exactly 25 mL of Hanus's TS, and mix well. If a clear solution is not obtained, add more cyclohexane to make clear, and allow the mixture to stand for 1 hour between 209C and 309C in a light-resistant, well-closed container while occasional shaking. Add 20 mL of a solution of potassium iodide (1 in 10) and 100 mL of water, shake, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Per-form a blank determination in the same manner. Iodine value ＝ (a － b) × 1.269/M M: amount (g) of Hydrous Lanolin taken a: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con-sumed in the blank determination b: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con-sumed in the titration
(1) Acidity or alkalinity—To 5 g of Hydrous Lanolin add 25 mL of water, boil for 10 minutes, and cool. Add water to restore the previous mass, and separate the aqueous layer: the aqueous layer is neutral. Chloride <1.03>—To 2.0 g of Hydrous Lanolin add 40 mL of water, boil for 10 minutes, and cool. Add water to restore the previous mass, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Use this solution as the test solution, and perform the test. Pre-pare the control solution with 1.0 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.036z). Ammonia—To 10 mL of the aqueous layer obtained in (1) add 1 mL of sodium hydroxide TS, and boil: the gas evolved does not turn moistened red litmus paper to blue. Water-soluble organic substances—To 5 mL of the aqueous layer obtained in (1) add 0.25 mL of 0.002 mol/L potassium permanganate VS, and allow to stand for 5 minutes: the red color of the solution does not disappear. Petrolatum—Dissolve 1.0 g of the dried residue ob-tained in the Residue on evaporation in 10 mL of a mixture of tetrahydrofuran and isooctane (1:1), and use this solution as the sample solution. Add dissolve 20 mg of vaseline in 10 mL of a mixture of tetrahydrofuran and isooctane (1:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with isooctane to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 2) on the plate, heat the plate at 809C for 5 minutes, cool, and examine under ultraviolet light (main wavelength: 365 nm): no fluorescent spot is observed in the same level with the spot of standard solution. For this test use a thin-layer plate previously developed with isooctane to the upper end, dried in air, and heated at 1109C for 60 minutes. Residue on evaporation Weigh accurately about 12.5 g of Hydrous Lanolin, dissolve in 50 mL of diethyl ether, put it in a separator, transfer the separated aqueous layer to another separator, add 10 mL of diethyl ether, shake, and combine the diethyl ether layer and diethyl ether in the first separator. Shake the diethyl ether layer with 3 g of anhy-drous sodium sulfate, and filter through dry filter paper. Wash the separator and the filter paper with two 20-mL por-tions of diethyl ether, combine the washings with the filtrate, evaporate on a water bath until the odor of diethyl ether is no longer perceptible, and dry in a desiccator (in vacuum, silica gel) for 24 hours: the content is not less than 70z and not more than 75z.
Hydroxocobalamin Acetate contains not less than 96.
Hydroxocobalamin Acetate occurs as dark red, crystals or powder. It is odorless. It is freely soluble in water and in methanol, slightly solu-ble in ethanol (95), and practically insoluble in diethyl ether. It is hygroscopic. ‌JP XVIII Official Monographs / Hydroxyethylcellulose 1121
(1) Determine the absorption spectrum of a solution of Hydroxocobalamin Acetate in acetic acid-sodium acetate buffer solution (pH 4.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Mix 1 mg of Hydroxocobalamin Acetate with 50 mg of potassium hydrogen sulfate, and fuse by igniting. Cool, break up the mass with a glass rod, add 3 mL of water, and dissolve by boiling. Add 1 drop of phenolphthalein TS, and add dropwise sodium hydroxide TS until the solution de-velops a light red. Then add 0.5 g of sodium acetate trihy-drate, 0.5 mL of dilute acetic acid and 0.5 mL of a solution of disodium 1-nitroso-2-naphthol-3,6-disulfonate (1 in 500): a red to orange-red color develops immediately. Then add 0.5 mL of hydrochloric acid, and boil for 1 minute: the red color does not disappear. Add 0.5 mL of ethanol (99.5) and 1 mL of sulfuric acid to 20 mg of Hydroxocobalamin Acetate, and heat the mixture: the odor of ethyl acetate is perceptible.
Conduct this procedure using light-resistant vessels. Dissolve 75 mg of Hydroxocobalamin Acetate in 100 mL of the dissolving solution, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the dissolv-ing solution to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the total area of the peaks other than hydroxocobalamin obtained from the sample solution is not larger than the peak area of hydroxocobalamin from the standard solution. Dissolving solution: A mixture of water, mobile phase C and methanol (41:5:4). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 351 nm). Column: Connect two columns which are 4.6 mm in inside diameter and 10 cm in length, composed of octadecyl-silanized monolithic silica for liquid chromatography, hav-ing a bimodal pore structure with 2 mm macropore and 13 nm mesopore, coated with polyether ether ketone. Column temperature: A constant temperature of about 309C. Mobile phase A: Water. Mobile phase B: Methanol. Mobile phase C: Dissolve 15.6 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust to pH 3 with diluted phosphoric acid (1 in 100). Flowing of mobile phase: Control the gradient by mixing the mobile phase A, B and C as directed in the following table. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add the dissolving solution to make exactly 50 mL. Confirm that the peak area of hydroxocobalamin obtained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of hydroxocobalamin are not less than 4000 and not more than 2.4, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydroxocobalamin is not more than 2.0z. Water <2.48> 8.0 – 12.0z (50 mg, volumetric titration, direct titration).
Weigh accurately about 0.1 g of Hydroxocobalamin Acetate, and dissolve in water to make exactly 500 mL. Pipet 5 mL of this solution, and add acetic acid-sodium acetate buffer solution (pH 4.5) to make exactly 25 mL. Determine the absorbance, A, of this solution at 351 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of hydroxocobalamin acetate (C62H89CoN13O15P.C2H4O2) ＝ A/187 × 25,000
Hydroxyethylcellulose is partly O-(2-hydrox-yethylated) cellulose. It contains not less than 30.
Hydroxyethylcellulose occurs as a white to yellowish white, powder or grains. It is practically insoluble in ethanol (95). Flow rate: 2 mL per minute. Time span of measurement: For 40 minutes after injection of the sample solution. It forms a viscous liquid upon addition of water. It is hygroscopic.◆ Identiˆcation (1) Determine the infrared absorption spec- trum of Hydroxyethylcellulose as directed in the ATR method under Infrared Spectrophotometry <2.25> and com-pare the spectrum with the spectrum of Hydroxyethylcellu-lose for Identiˆcation RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers. (2) Disperse 1.0 g of Hydroxyethylcellulose, calculated on the dried basis, in 50 mL of freshly boiled and cooled water. After 10 minutes, add freshly boiled and cooled water to make 100 mL, stir to dissolve completely, and use this so-lution as the sample solution. Boil 10 mL of the sample solu-tion: the solution is clear. Viscosity <2.53> Weigh exactly a quantity of Hydrox-yethylcellulose, equivalent to 10.00 g calculated on the dried basis, add 400 mL of water, stir to dissolve, and add water to make exactly 500.0 g. Remove air bubbles, and use this solu-tion as the sample solution. Perform the test with the sample solution at 20 ± 0.19C as directed in Method II under Vis-cosity Determination, using a beaker with an inside diameter of not less than 70 mm and a single cylinder-type rotational viscometer, according to the following operating conditions: not less than 75z and not more than 140z of the labeled viscosity. Operating conditions— Apparatus: Brookˆeld type viscometer LV or RV model. Rotor No., rotation frequency and calculation multiplier: According to the following table, depending on the labeled viscosity. 100 mL, and use these solutions as the standard solutions. If the viscosity of Hydroxyethylcellulose is more than 1000 mPa･s, pipet 1 mL, 2 mL and 4 mL of the standard nitrate stock solution, add the diluting solution to each to make exactly 100 mL, and use these solutions as the standard solu-tions. Perform the test with the sample solution and stand-ard solutions using a nitrate-selective electrode as an indica-tor electrode, a silver-silver chloride electrode as a reference electrode and diluted ammonium sulfate TS (1 in 30) as ref-erence electrolyte. Calculate the concentration of nitrates in the sample solution using a calibration curve obtained from the potential diŠerences of the standard solutions: not more than 3.0z, calculated on the dried basis, if Hydroxyethylcel-lulose has a viscosity of not more than 1000 mPa･s, and not more than 0.2z, calculated on the dried basis, if Hydrox-yethylcellulose has a viscosity of more than 1000 mPa･s. Diluting solution: To a mixture of 50 mL of 1 mol/L sul-furic acid TS and 800 mL of water add 135 g of potassium dihydrogen phosphate, and add water to make 1000 mL. To this solution add water to make exactly 25 times the initial volume. In order to determine the applicable limit, determine the viscosity using the following procedure. Introduce a quantity of Hydroxyethylcellulose, equivalent to 2.00 g calculated on the dried basis, into 50 g of water, stir, add water to make 100 g, and stir to dissolve com-pletely. Determine the viscosity using a rotating viscometer at 259C and at a shear rate of 100 s－1 for substances with an expected viscosity less than 100 mPa･s, at a shear rate of Labeled viscosity (mPa･s) Model Rotor No. Rotation frequency /min Calculation multiplier 10 s－1 for substances with an expected viscosity not less than less than 200 LV Not less than 200 and less than 4000 LV 〃 〃 10,000 LV 〃 10,000 〃 50,000 RV 〃 50,000 RV 100 mPa･s and not more than 20,000 mPa･s, and at a shear rate of 1 s－1 for substances with an expected viscosity more than 20,000 mPa･s. If it is impossible to obtain a shear rate of exactly 10 s－1 or 100 s－1 respectively, use a rate slightly higher and a rate slightly lower and interpolate. (3) Heavy metal <1.07>—Proceed with 1.0 g of Hydrox-yethylcellulose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Procedure of apparatus: Read a value after 2 minutes of rotation, and stop the rotation for at least 2 minutes. Repeat this procedure more two times, and average the three ob-served values.◆ pH <2.54> The pH of the sample solution obtained in the Identiˆcation (2) is between 5.5 and 8.5.
(1) Chloride—To 1 mL of the sample solution ob-tained in the Identiˆcation (2) add water to make 30 mL, and use this solution as the sample solution. Separately, to 10 mL of Standard Chloride Solution add 5 mL of water, and use this solution as the control solution. To 15 mL each of the sample solution and control solution add 1 mL of diluted nitric acid (1 in 5), transfer to test tubes containing 1 mL of a solution of silver nitrate (17 in 1000), allow to stand for 5 minutes protecting from light, and compare the opalescence developed in the both solutions against a black background by viewing transversely: the opalescence developed in the sample solution is not more intense than that of the control solution (not more than 1.0z). (2) Nitrate—Prepare the solutions before use. Dissolve 0.50 g of Hydroxyethylcellulose in the diluting solution to make exactly 100 mL, and use this solution as the sample so-lution. Separately, dissolve 0.8154 g of potassium nitrate in the diluting solution to make 1000 mL, and use this solution as the standard nitrate stock solution. If the viscosity of Hydroxyethylcellulose is not more than 1000 mPa･s, pipet 10 mL, 20 mL and 40 mL of the standard nitrate stock solution, add the diluting solution to each to make exactly Solution (not more than 20 ppm).◇ (4) Aldehydes—Introduce 1.0 g of Hydroxyethylcellu-lose into a glass-stoppered test tube, add 10 mL of ethanol (99.5), stopper the tube tightly, and stir for 30 minutes. Cen-trifuge, and use the supernatant liquid as the sample solu-tion. Use Standard Glyoxal Solution as the control solution. Pipet 2 mL each of the sample solution and control solution, to each add 5 mL of a solution prepared by dissolving 4 g of 3-methyl-2-benzothiazolonehydrazone hydrochloride mono-hydrate in diluted acetic acid (100) (4 in 5) to make 1000 mL, shake to homogenize, and allow to stand for 2 hours. Com-pare the color of these solutions: the sample solution is not more intensely colored than the control solution (not more than 20 ppm). Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 3 hours). Residue on Ignition <2.44> Not more than 4.0z if the viscosity of Hydroxyethylcellulose is not more than 1000 mPa･s, and not more than 1.0z if the viscosity of Hydrox-yethylcellulose is more than 1000 mPa･s (1 g). In order to determine the applicable limit, determine the viscosity ac-cording to the method in the Purity (2).
Weigh accurately about 30 mg of Hydroxyethylcel-lulose, transfer to a 5-mL pressure-tight serum vial, add ex-actly 60 mg of adipic acid, 2 mL of the internal standard so-lution and 1 mL of hydroiodic acid, seal the vial immediately with a septum coated with ‰uororesin and an aluminum cap ‌JP XVIII Official Monographs / Hydroxypropylcellulose 1123 or any other sealing system providing a su‹cient air-tight-ness, and weigh accurately the vial. Take care not to mix the content of the vial before heating. Place the vial in an oven or heat in a suitable heater with continuous stirring, main-taining the internal temperature of about 165 ± 29C for 2.5 hours. Allow to cool and weigh accurately the vial. If the diŠerence of the mass between before heating and after heat-ing is more than 10 mg, prepare a new sample solution. If the diŠerence of the mass between before heating and after heating is not more than 10 mg, after phase separation, pierce through the septum of the vial with a cooled syringe, and withdraw a su‹cient volume of the upper phase as the sample solution. Separately, place exactly 60 mg of adipic acid, 2 mL of the internal standard solution and 1 mL of hydroiodic acid in another serum vial, and seal immediately. Weigh accurately the vial, inject 55 mL of iodoethane for assay through the septum in the vial, and weigh again accu-rately. Shake thoroughly, after phase separation, pierce through the septum of the vial with a cooled syringe, and withdraw a su‹cient volume of the upper phase as the stand-ard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chro-matography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iodo-ethane to that of the internal standard. Amount (z) of hydroethoxy group (C2H5O2) ＝ MS/MT × QT/QS × 39.15 MS: Amount (mg) of iodoethane for assay taken MT: Amount (mg) of Hydroxyethylcellulose taken, calcu-lated on the dried basis Internal standard solution—A solution of n-octane in o-xy-lene (1 in 200). Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Column temperature: Maintain the temperature at 509C for 3 minutes, raise to 1009C at a rate of 109C per minute, then raise to 2509C at a rate of 359C per minute, and main-tain at 2509C for 8 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2809C. Carrier gas: Helium. Flow rate: 4.2 mL per minutes (the retention time of the internal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, iodoethane and the internal standard are eluted in this order with the relative retention time of iodoethane to the in-ternal standard being about 0.6 and the resolution between these peaks being not less than 5.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of iodoethane to that of the internal standard is not more than 2.0z. ◆Containers and storage Containers—Tight containers.◆ Hydroxypropylcellulose
Hydroxypropylcellulose is partially O-(2-hydrox-ypropylated) cellulose. It contains not less than 53.
Hydroxypropylcellulose occurs as a white to yellowish white powder. It forms a viscous liquid upon addition of water or ethanol (95).◆ Identiˆcation (1) Dissolve 1 g of Hydroxypropylcellulose in 100 mL of water, transfer 1 mL of the solution to a glass plate, and allow the water to evaporate: a thin ˆlm is formed. Determine the infrared absorption spectrum of Hydroxypropylcellulose as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If there are an absorption at about 1719 cm－1, disregard the absorption. pH <2.54> Disperse evenly 1.0 g of Hydroxypropylcellulose in 100 mL of freshly boiled water, and allow to cool the mix-ture while stirring with a magnetic stirrer: the pH of the solu-tion is between 5.0 and 8.0.
(1) Heavy metals<1.07>—Proceed with 1.0 g of Hydroxypropylcellulose according to Method 2 and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm).◇ Silicon dioxide—Apply to Hydroxypropylcellulose, if the addition of silicon dioxide is stated on the label and if more than 0.2z residue is found in the Residue on ignition test. Weigh accurately the crucible containing the residue tested in the Residue on ignition of Hydroxypropylcellulose (a (g)). Moisten the residue with water, and add 5 mL of hydro‰uoric acid, in small portions. Evaporate it on a steam bath to dryness and cool. Add 5 mL of hydro‰uoric acid and 0.5 mL of sulfuric acid, and evaporate to dryness. Slowly in-crease the temperature until all the acids have been volati-lized, and ignite at 1000 ± 259C. Cool the crucible in a desiccator, and weigh (b (g)). Calculate the amount of silicon dioxide by the following equation: not more than 0.6z. ‌1124 Amount (z) of silicon dioxide (SiO2) ＝ (a － b)/M × 100 M: Amount (g) of Hydroxypropylcellulose used for residue on ignition test Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.8z (1 g, plati-num crucible).
Weigh accurately about 30 mg of Hydroxypropylcel-lulose, transfer to a reaction vial, add exactly 60 mg of adip-ic acid, 2 mL of the internal standard solution and 1 mL of hydriodic acid, stopper the vial tightly, and weigh accu-rately. Place the vial in an oven or heat by a suitable heater with continuous stirring, maintaining the internal tempera-ture of 115 ± 29C for 70 minutes. Allow the vial to cool and weigh accurately. If the diŠerence of the mass between be-fore heating and after heating is more than 10 mg, prepare a new test solution. If the diŠerence of the mass between be-fore heating and after heating is not more than 10 mg, after phase separation by allowing the vial to stand, pierce through the septum of the vial with a cooled syringe, and withdraw a su‹cient volume of the upper phase as the sam-ple solution. Separately, place exactly 60 mg of adipic acid, 2 mL of internal standard solution and 1 mL of hydriodic acid in an another reaction vial, stopper tightly, and weigh accu-rately. Inject 25 mL of isopropyl iodide for assay through the septum, and again weigh accurately. Shake the vial thor-oughly, and after phase separation by allowing the vial to stand, pierce through the septum of the vial with a cooled syringe, and withdraw a su‹cient volume of the upper phase as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of isopropyl iodide to that of the internal standard. Amount (z) of hydroxypropoxy group (C3H7O2) ＝ MS/MT × QT/QS × 1.15 × 44.17 MS: Amount (mg) of isopropyl iodide for assay taken MT: Amount (mg) of Hydroxypropylcellulose taken, cal-culated on the dried basis 1.15: Correction factor Internal standard solution—A solution of methylcyclohexan in o-xylene (1 in 50). Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Column: A fused silica tube 0.53 mm in diameter and 30 m in length, coated with methylsilicone polymer for gas chromatography in 3 mm thickness. Column Temperature: Maintain the temperature at 409C for 3 minutes, raise to 1009C at a rate of 109C per minute, then raise to 2509C at a rate of 509C per minute, and main-tain at 2509C for 3 minutes. Injection port temperature: A constant temperature of about 1809C. Detector temperature: A constant temperature of about 2809C. Carrier gas: Helium. Flow rate: 52 cm per second (the retention time of the in-ternal standard is about 8 minutes). Split ratio: 1:50. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi- tions, isopropyl iodide and the internal standard are eluted in this order with the relative retention time of isopropyl iodide to the internal standard being about 0.8, and with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of isopropyl iodide to that of the internal standard is not more than 2.0z.
Hydroxyzine Hydrochloride, when dried, contains not less than 98.
Hydroxyzine Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is very soluble in water, freely soluble in methanol, in ethanol (95) and in acetic acid (100), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: about 2009C (with decomposition).
(1) To 5 mL of a solution of Hydroxyzine Hydrochloride (1 in 100) add 2 to 3 drops of ammonium thiocyanate-cobalt (II) nitrate TS: a blue precipitate is ＝ MS/MT × QT/QS × 44.17 3 7 2 formed. (2) Determine the absorption spectrum of a solution of MS: Amount (mg) of isopropyl iodide for assay taken MT: Amount (mg) of Low Substituted Hydroxypropylcel-lulose taken, calculated on the dried basis Internal standard solution—A solution of n-octane in o-xy-lene (3 in 100). Operating conditions— Detector: A thermal conductivity detector or hydrogen flame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Use a guard column if necessary. Column temperature: Maintain the temperature at 509C for 3 minutes after injection, raise to 1009C at a rate of 109C per minute, then to 2509C at a rate of 359C per minute and maintain at 2509C for 8 minutes. Injection port temperature: 2509C. Detector temperature: 2809C. Hydroxyzine Hydrochloride in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Hydroxyzine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Hydroxyzine Hydrochloride in 20 mL of water: the pH of this solution is between 1.3 and 2.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Hydroxyzine Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Hydrox- yzine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). ‌1126 Related substances—Dissolve 0.20 g of Hydroxyzine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ethanol (95) and ammonia solution (28) (150:95:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot obtained from the standard solution. Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.1 g of Hydroxyzine Hy-drochloride, previously dried, dissolve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 22.39 mg of C21H27ClN2O2.2HCl
Hydroxyzine Pamoate contains not less than 98.
Hydroxyzine Pamoate occurs as a light yellow crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in N,N-dimethylformamide, slightly soluble in acetone, and practically insoluble in water, in methanol, in ethanol (95) and in diethyl ether.
(1) To 0.1 g of Hydroxyzine Pamoate add 25 mL of sodium hydroxide TS, and shake well. Extract with 20 mL of chloroform, and use the chloroform layer as the sample solution. Use the water layer for test (4). To 5 mL of the sample solution add 2 mL of ammonium thiocyanate-cobalt (II) nitrate TS, shake well, and allow to stand: a blue color is produced in the chloroform layer. Evaporate 2 mL of the sample solution obtained in on a water bath to dryness, and dissolve the residue in 0.1 mol/L hydrochloric acid TS to make 500 mL. Determine the absorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Perform the test with Hydroxyzine Pamoate as di-rected under Flame Coloration Test <1.04> (2): a green color appears. To 1 mL of the water layer obtained in (1), add 2 mL of 1 mol/L hydrochloric acid TS: a yellow precipitate is pro-duced. Collect the precipitate, dissolve the precipitate in 5 mL of methanol, and add 1 drop of iron (III) chloride TS: a green color is produced.
(1) Clarity and color of solution—Dissolve 1.0 g of Hydroxyzine Pamoate in 10 mL of N,N-dimethylfor-mamide: the solution is clear, and shows a slightly greenish, light yellow-brown color. Chloride <1.03>—To 0.3 g of Hydroxyzine Pamoate add 6 mL of dilute nitric acid and 10 mL of water, shake for 5 minutes, and filter. Wash the residue with two 10-mL por-tions of water, combine the washings with the filtrate, and add water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.80 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.095z). Heavy metals <1.07>—Proceed with 1.0 g of Hydro- xyzine Pamoate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Hydroxyzine Pamoate according to Method 3, and per-form the test (not more than 1 ppm). Related substances—Dissolve 0.40 g of Hydroxyzine Pamoate in 10 mL of a mixture of sodium hydroxide TS and acetone (1:1), and use the solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of sodium hydroxide TS and acetone (1:1) to make exactly 20 mL. Pipet 5 mL of this solution, add a mixture of sodium hy-droxide TS and acetone (1:1) to make exactly 50 mL, and use the solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, ethanol (95) and ammonia TS (150:95:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate: the spots other than hydroxyzine and pamoic acid obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.5z (1 g).
Weigh accurately about 0.6 g of Hydroxyzine Pamo-ate, add 25 mL of sodium hydroxide TS, shake well, and ex-tract with six 25-mL portions of chloroform. Filter each ex-tract through 5 g of anhydrous sodium sulfate on a pledget of absorbent cotton. Combine the chloroform extracts, and evaporate the combined chloroform extracts on a water bath to about 30 mL. Add 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank de- ‌JP XVIII Official Monographs / Hypromellose 1127 ‌termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.16 mg of C H ClN O .C H O water (2:1) to make 50 mL (not more than 0.024z). (3) Heavy metals <1.07>—Proceed with 2.0 g of Hymecromone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead 21 27 2 2 23 16 6 Solution (not more than 10 ppm).
Hymecromone, when dried, contains not less than 98.
Hymecromone occurs as white, crystals or crystalline powder. It is odorless and tasteless. It is freely soluble in N,N-dimethylformamide, sparingly soluble in ethanol (95), in ethanol (99.5) and in acetone, slightly soluble in diethyl ether, and practically insoluble in water.
(1) Dissolve 2 mg of Hymecromone in 5 mL of ammonia-ammonium chloride buffer solution (pH 11.0): the solution shows an intense blue-purple fluores-cence. Dissolve 25 mg of Hymecromone in 5 mL of diluted ethanol (95) (1 in 2), and add 1 drop of iron (III) chloride TS: initially a blackish brown color develops, and when allowed to stand the color changes to yellow-brown. Determine the absorption spectrum of a solution of Himecromone in ethanol (99.5) (1 in 250,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Himecromone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.8 g of Hymecro-mone in 40 mL of a mixture of acetone and water (2:1), and add 6 mL of dilute nitric acid and a mixture of acetone and water (2:1) to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and a mixture of acetone and water (2:1) to make 50 mL (not more than 0.011z). (2) Sulfate <1.14>—Dissolve 0.8 g of Hymecromone in 40 mL of a mixture of acetone and water (2:1), and add 1 mL of dilute hydrochloric acid and a mixture of acetone and water (2:1) to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid and a mixture of acetone and Arsenic <1.11>—Prepare the test solution with 1.0 g of Hymecromone according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 80 mg of Hymecro-mone in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add ethanol (95) to make exactly 50 mL. Pipet 1 mL of this solu-tion, add ethanol (95) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform and ethanol (95) (10:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 5 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Hymecromone, previously dried, dissolve in 90 mL of N,N-dimethylfor-mamide, and titrate <2.50> with 0.1 mol/L tetramethylam-monium hydroxide VS (potentiometric titration). Separately, perform a blank determination in the same manner with a solution prepared by adding 14 mL of water to 90 mL of N,N-dimethylformamide, and make any necessary correc-tion. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 17.62 mg of C10H8O3
ヒプロメロース酢酸エステルコハク酸エステル
Hypromellose Acetate Succinate is an acetic acid and monosuccinic acid mixed ester of hypromellose. It contains not less than 12.
Hypromellose Acetate Succinate occurs as a white to yellowish white, powder or granules. It is practically insoluble in water and in ethanol (99.5). It dissolves in sodium hydroxide TS. It is hygroscopic.
Determine the infrared absorption spectrum of Hypromellose Acetate Succinate as directed in the ATR method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 2840 cm－1, 1737 cm－1, 1371 cm－1, 1231 cm－1 and 1049 cm－1. Viscosity <2.53> To 2.00 g of Hypromellose Acetate Suc-cinate, previously dried, add dilute sodium hydroxide TS to make 100.0 g, stopper tightly, and dissolve by shaking for 30 minutes. Perform the test with this solution at 209C accord-ing to Method 1: 80 – 120z of the labeled viscosity.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Hypromellose Acetate Succinate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Free acetic acid and free succinic acid—Weigh accu-rately about 0.1 g of Hypromellose Acetate Succinate, add exactly 4 mL of 0.02 mol/L phosphate buffer solution (pH 7.5), stopper tightly, and stir for 2 hours. Then add exactly 4 mL of diluted phosphoric acid (1 in 500), turn the test tube upside down several times, centrifuge, and use the superna-tant liquid as the sample solution. Separately, place 20 mL of water in a 100-mL volumetric flask, weigh the mass of the flask accurately, then add 2.0 mL of acetic acid (100), weigh the mass of the flask to calculate the accurate mass of added acetic acid, and dilute with water to volume. Pipet 6 mL of this solution, add water to make exactly 100 mL, and use this solution as the acetic acid stock solution. Separately, weigh accurately about 0.13 g of succinic acid, dissolve in cinic acid, dissolve in water to make exactly 100 mL, and use this solution as the succinic acid stock solution. Pipet 4 mL each of the acetic acid stock solution and the succinic acid stock solution, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, ATA, ATS and ASA, ASS, of acetic acid and succinic acid in each solution. Amount (z) of acetyl group (C H O) water to make exactly 100 mL, and use this solution as the 2 3 succinic acid stock solution. Pipet exactly 4 mL each of the acetic acid stock solution and the succinic acid stock solu-tion, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas of acetic acid and succinic acid of both solutions, ATA, ATS and ASA, ASS, and calculate the amount of free acetic acid and free succinic acid by the following expressions: the total amount is not more than 1.0z. Amount (z) of free acetic acid (C2H4O2) ＝ MSA/MT × ATA/ASA × 48/625 Amount (z) of free succinic acid (C4H6O4) ＝ MSS/MT × ATS/ASS × 32/25 MSA: Amount (mg) of acetic acid (100) taken MSS: Amount (mg) of succinic acid taken MT: Amount (mg) of Hypromellose Acetate Succinate taken, calculated on the dried basis Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay (1). Test for required detectability: To 3 mL of the standard solution add the mobile phase to make 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak areas of acetic acid and succinic acid obtained with 10 mL of this solution are equivalent to 7 to 13z of corresponding those with 10 mL of the solution for system suitability test. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.2z (1 g).
(1) Acetyl group and succinyl group—Weigh accu-rately about 30 mg of Hypromellose Acetate Succinate, add exactly 10 mL of sodium hydroxide TS, stopper tightly, and stir for 4 hours. Add exactly 10 mL of diluted phosphoric acid (17 in 200), turn the test tube upside down several times, and filter the solution through a membrane filter with a pore size 0.22 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, place 20 mL of water in a 100-mL volumetric flask, weigh the mass of the flask accurately, then add 2.0 mL of acetic acid (100), weigh the mass of the flask to calculate the ac-curate mass of added acetic acid, and dilute with water to volume. Pipet 6 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the acetic acid stock solution. Separately, weigh accurately about 0.13 g of suc- ＝ (MSA/MT × ATA/ASA × 24/125 － Afree) × 0.717 Amount (z) of succinyl group (C4H5O3) ＝ (MSS/MT × ATS/ASS × 16/5 －Sfree) × 0.856 MSA: Amount (mg) of acetic acid (100) taken MSS: Amount (mg) of succinic acid taken MT: Amount (mg) of Hypromellose Acetate Succinate taken, calculated on the dried basis Afree: Amount (z) of free acetic acid obtained in the Puri-ty (2) Sfree: Amount (z) of free succinic acid obtained in the Purity (2) Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: 0.02 mol/L potassium dihydrogen phos-phate TS, adjusted to pH 2.8 with phosphoric acid. Flow rate: Adjust so that the retention time of succinic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, acetic acid and succinic acid are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of acetic acid and succinic acid is not more than 2.0z. Methoxy group and hydroxypropoxy group Apparatus—Reaction bottle: A 5 mL pressure-tight glass vial, having 20 mm in outside diameter, 50 mm in height, the neck 20 mm in outside diameter and 13 mm in in-side diameter, equipped with a septum of butyl rubber proc-essed the surface with fluoroplastics, which can be fixed tightly to vial with aluminum cap, or equivalent. Heater: A square-shaped aluminum block, having holes 20 mm in inside diameter and 32 mm in depth, adopted to the reaction bottle. Capable of stirring the content of the bottle by means of magnetic stirrer or reciprocal shaker about 100 times per minute. Procedure—Weigh accurately about 65 mg of Hypromellose Acetate Succinate, place in the reaction bot-tle, add 0.06 to 0.10 g of adipic acid, 2.0 mL of the internal standard solution and 2.0 mL of hydriodic acid, stopper the bottle immediately, and weigh accurately. Stir or shake for 60 minutes while heating so that the temperature of the bottle content is 130 ± 29C. In a case where the magnetic stirrer or shaker is not available, heat for 30 minutes with repeated shaking at 5 minute intervals by hand, and continue heating for additional 30 minutes. Allow the bottle to cool, ‌JP XVIII Official Monographs / Hypromellose Phthalate 1131 and again weigh accurately. If the mass loss is less than 0.50z or there is no evidence of a leak, use the upper layer of the mixture as the sample solution. Separately, put 0.06 to 0.10 g of adipic acid, 2.0 mL of the internal standard solu-tion and 2.0 mL of hydriodic acid in a reaction bottle, stop-per the bottle immediately, and weigh accurately. Add 45 mL of iodomethane for assay and 15 to 22 mL of isopropyl iodide for assay through the septum using a micro-syringe with weighing accurately every time, stir thoroughly, and use the upper layer of the mixture as the standard solution. Per-form the test with 1 to 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios of the peak areas of iodomethane and isopropyl iodide to the peak area of the internal standard, QTa, QTb and QSa, QSb. Amount (z) of methoxy group (CH3O) ＝ MSa/MT × QTa/QSa × 21.86 Amount (z) of hydroxypropoxy group (C3H7O2) ＝ MSb/MT × QTb/QSb × 44.17 MSa: Amount (mg) of iodomethane for assay taken MSb: Amount (mg) of isopropyl iodide for assay taken MT: Amount (mg) of Hypromellose Acetate Succinate taken, calculated on the dried basis Internal standard solution—A solution of n-octane in o-xylene (3 in 100). Operating conditions— Column: A glass tube 3 – 4 mm in inside diameter and 200731 27.0 35.0 1.8 – 3 m in length, packed with siliceous earth for gas chro- 220824 21.0 27.0 matography, 120 to 150 mm in diameter coated with methyl Detector: A thermal conductivity detector or hydrogen flame-ionization detector. Hypromellose Phthalate
ヒプロメロースフタル酸エステル
Hypromellose Phthalate is a monophthalic acid ester of hypromellose. It contains methoxy group (－OCH3: 31.
Hypromellose Phthalate occurs as white, powder or granules. It is practically insoluble in water, in acetonitrile and in ethanol (99.5). It becomes a viscous liquid when a mixture of methanol and dichloromethane (1:1) or a mixture of ethanol (99.5) and acetone (1:1) is added. It dissolves in sodium hydroxide TS.◆
Determine the infrared absorption spec-trum of Hypromellose Phthalate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ Viscosity <2.53> To 10 g of Hypromellose Phthalate, previ-ously dried at 1059C for 1 hour, add 90 g of a mixture of methanol and dichloromethane in equal mass ratio, and stir to dissolve. Determine the viscosity at 20 ± 0.19C as di-rected in Method 1 under Viscosity Determination: the vis-cosity is not less than 80z and not more than 120z of the labeled unit.
(1) Chloride <1.03>—Dissolve 1.0 g of Hypromel-lose Phthalate in 40 mL of 0.2 mol/L sodium hydroxide VS, add 1 drop of phenolphthalein TS, and add dilute nitric acid dropwise with vigorous stirring until the red color is dis-charged. Further add 20 mL of dilute nitric acid with stir-ring. Heat on a water bath with stirring until the gelatinous precipitate formed turns to granular particles. After cooling, centrifuge, and take off the supernatant liquid. Wash the precipitate with three 20-mL portions of water by centrifug- ‌1132‌ ing each time, combine the supernatant liquid and the wash-ings, add water to make 200 mL, and filter. Perform the test with 50 mL of the filtrate. Control solution: To 0.50 mL of 0.01 mol/L hydrochloric acid VS add 10 mL of 0.2 mol/L sodium hydroxide VS and 7 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.07z). (2) Heavy metals <1.07>—Proceed with 2.0 g of Hypromellose Phthalate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Phthalic acid—Weigh accurately about 0.2 g of Hypromellose Phthalate, add about 50 mL of acetonitrile to dissolve partially with the aid of ultrasonic waves, add 10 mL of water, and dissolve further with the ultrasonic waves. After cooling, add acetonitrile to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 12.5 mg of phthalic acid, dissolve in about 125 mL of acetonitrile by mixing, add 25 mL of water, then add acetonitrile to make exactly 250 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of phthalic acid in each solution: amount of phthalic acid (C8H6O4: 166.13) is not more than 1.0z. Amount (z) of phthalic acid ＝ MS/MT × AT/AS × 40 MS: Amount (mg) of phthalic acid taken MT: Amount (mg) of Hypromellose Phthalate taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A stainless steel column about 4.6 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of 0.1z trifluoroacetic acid and acetonitrile (9:1). Flow rate: About 2.0 mL per minute. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of phthalic acid are not less than 2500 and not more than 1.5, respectively.◆ System repeatability: When repeat the test 6 times with 10 mL of the standard solution under the above operating con-ditions, the relative standard deviation of the peak area of phthalic acid is not more than 1.0z. Water <2.48> Not more than 5.0z (1 g, volumetric titra-tion, direct titration, using a mixture of ethanol (99.5) and dichloromethane (3:2) instead of methanol for water deter-mination). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 1 g of Hypromellose Phtha-late, dissolve in 50 mL of a mixture of ethanol (95), acetone and water (2:2:1), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 2 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Amount (z) of carboxybenzoyl group (C8H5O3) ＝ {(0.01 × 149.1 × V )/M}－ {(2 × 149.1 × P)/166.1} P: Amount (z) of phthalic acid obtained in the Purity (3) V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed M: Amount (g) of Hypromellose Phthalate taken, calcu-lated on the anhydrous basis
Hypromellose occurs as a white to yellowish Not less than 600 and less than 1400 3 60 20 tion lier white, powder or grains. It is practically insoluble in ethanol (99.5). It swells with water and becomes a clear or slightly turbid, viscous solution.◆ Identiˆcation (1) Disperse evenly 1.0 g of Hypromellose over the surface of 100 mL of water in a beaker, while gently tapping the top of the beaker, if necessary, and allow the beaker to stand: it aggregates on the surface of water. Add 1.0 g of Hypromellose to 100 mL of hot water, and stir: it becomes a suspension. Cool the suspension to 109C, and stir: the resulting liquid is a clear or a slightly cloudy, viscous ‰uid. To 0.1 mL of the viscous ‰uid obtained in (2) add 9 mL of diluted sulfuric acid (9 in 10), shake, heat in a water bath for exactly 3 minutes, and immediately cool in ice water. Add carefully 0.6 mL of ninhydrin TS, shake, and allow to stand at 259C: the solution shows a red color ˆrst, then changes to purple color within 100 minutes. Pour and spread out 2 to 3 mL of the viscous ‰uid obtained in (2) onto a glass plate, and allow the water to evaporate: a transparent ˆlm results. Pipet 50 mL of water, add exactly 50 mL of the vis-cous ‰uid obtained in (2), and warm to raise the temperature at a rate of 2 to 59C per minute while stirring: the tempera-ture, when a white turbidity of the solution starts to increase, is not less than 509C. Viscosity <2.53> (i) Method I: Apply to Hypromellose having a labeled viscosity of less than 600 mPa･s. Put an ex-act amount of Hypromellose, equivalent to 4.000 g calcu-lated on the dried basis, in a tared, wide-mouth bottle, add water (between 909C and 999C) to make 200 g, stopper the bottle, stir by mechanical means at 350 to 450 revolutions per minute for 10 to 20 minutes to get a homogeneous dis-persion. If necessary, take oŠ the sample attached on the walls of the bottle, put them in the dispersed solution, and dissolve by continuing the stirring in a water bath at not exceeding 109C for 20 to 40 minutes. Add cold water, if nec-essary, to make 200 g, and use this solution as the sample so-lution. Centrifuge the solution if necessary to expel any en-trapped air bubbles. Perform the test with the sample solu-tion at 20 ± 0.19C as directed in Method I under Viscosity Determination: not less than 80z and not more than 120z of the labeled viscosity. (ii) Method II: Apply to Hypromellose having a labeled viscosity of not less than 600 mPa･s. Put an exact amount of Hypromellose, equivalent to 10.00 g calculated on the dried basis, in a tared, wide-mouth bottle, add water (between 909C and 999C) to make 500 g, and prepare the sample solu-tion in the same manner as directed in Method I. Perform 〃 〃 〃 〃 〃 〃 99,500 〃 99,500 Procedure of apparatus: Read the value after 2 minutes of rotation, and stop the rotation for at least 2 minutes. Repeat this procedure more two times, and average the three ob-served values. pH <2.54> The pH of the sample solution obtained in the Viscosity, measured after 5 minutes immersing the electrode in the sample solution, is between 5.0 and 8.0.
Heavy metals—Put 1.0 g of Hypromellose in a 100-mL Kjeldahl ‰ask, add a su‹cient amount of a mixture of nitric acid and sulfuric acid (5:4) to wet the sample, and heat gently. Repeat this procedure until to use totally 18 mL of the mixture of nitric acid and sulfuric acid (5:4). Then boil gently until the solution changes to black. After cooling, add 2 mL of nitric acid, and heat until the solution changes to black. Repeat this procedure until the solution no longer changes to black, and heat strongly until dense white fumes are evolved. After cooling, add 5 mL of water, boil gently until dense white fumes are evolved, then heat until the volume of the solution becomes to 2 to 3 mL. After cooling, if the solution reveals yellow color by addition of 5 mL of water, add 1 mL of hydrogen peroxide (30), and heat until the volume of the solution becomes to 2 to 3 mL. After cool-ing, dilute the solution with 2 to 3 mL of water, transfer to a Nessler tube, add water to make 25 mL, and use this solution as the sample solution. Separately, put 2.0 mL of Standard Lead Solution in a 100-mL Kjeldahl ‰ask, add 18 mL of the mixture of nitric acid and sulfuric acid (5:4) and an amount of nitric acid equal to that used for preparation of the sam-ple solution, and heat until white fumes are evolved. After cooling, add 10 mL of water. In the case where hydrogen peroxide (30) is added for the preparation of the sample so-lution, add the same amount of hydrogen peroxide (30), then proceed in the same manner for preparation of the sample solution, and use so obtained solution as the control solu-tion. Adjust the sample solution and the control solution to pH 3.0 to 4.0 with ammonia solution (28), and add water to make 40 mL, respectively. To these solutions add 1.2 mL of thioacetamide-alkaline glycerin TS, 2 mL of acetate buŠer solution (pH 3.5) and water to make 50 mL, separately. After allowing to stand for 5 minutes, observe vertically both tubes on a white background: the color obtained with the sample solution is not more intense than that with the control solution (not more than 20 ppm).◇ Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, ‌JP XVIII Official Monographs / Hypromellose Acetate Succinate 1129 hour). Residue on ignition <2.44> Not more than 1.5z (1 g).
(i) Apparatus—Reaction vial: A 5-mL pressure-tight serum vial, having 20 mm in outside diameter and 50 mm in height, the neck 20 mm in outside diameter and 13 mm in inside diameter, equipped with a septum of butyl-rubber processed the surface with ‰uoroplastics, which can be ˆxed tightly to vial with aluminum cap, or equivalent. Heater: A square-shaped aluminum block, having holes 20 mm in diameter and 32 mm in depth, adopted to the reaction vial. Capable of stirring the content of the reaction vial by means of magnetic stirrer or of reciprocal shaker about 100 times per minute. (ii) Procedure—Weigh accurately about 65 mg of Hypromellose, transfer to a reaction vial, add 60 to 100 mg of adipic acid, 2.0 mL of the internal standard solution and .0 mL of hydroiodic acid, immediately stopper the vial tightly, and weigh accurately. Using a magnetic stirrer equipped in the heating module, or using a shaker, stir for 60 minutes while heating so that the temperature of the vial content is 130 ± 29C. In the case when a magnetic stirrer or shaker is not available, heat for 30 minutes with repeated shaking at 5-minute intervals by hand, and continue heating for an additional 30 minutes. Allow the vial to cool, and again weigh accurately. If the mass loss is less than 26 mg and there is no evidence of a leak of the content, use the up-per layer of the mixture as the sample solution. Separately, put 60 to 100 mg of adipic acid, 2.0 mL of the internal stand-ard solution and 2.0 mL of hydroiodic acid in a reaction vial, immediately stopper the vial tightly, and weigh accu-rately. Add 45 mL of iodomethane for assay and 15 to 22 mL of isopropyl iodide for assay through the septum using a micro-syringe with weighing accurately every time, shake thoroughly, and use the upper layer of the content as the standard solution. Perform the test with 1 to 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con-ditions, and calculate the ratios, QTa and QTb, of the peak area of iodomethane and isopropyl iodide to that of the in-ternal standard obtained from the sample solution, and QSa and QSb, of the peak area of iodomethane and isopropyl iodide to that of the internal standard from the standard so-lution. Content (z) of methoxy group (CH3O) ＝ MSa/M × QTa/QSa × 21.86 Content (z) of hydroxypropoxy group (C3H7O2) ＝ MSb/M × QTb/QSb × 44.17 MSa: Amount (mg) of iodomethane for assay taken MSb: Amount (mg) of isopropyl iodide for assay taken M: Amount (mg) of Hypromellose taken, calculated on the dried basis Internal standard solution—A solution of n-octane in o-xy-lene (3 in 100). Operating conditions— Detector: A thermal conductivity detector or hydrogen ‰ame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Use a guard column, if necessary. Column temperature: Maintain the temperature at 509C for 3 minutes, raise to 1009C at a rate of 109C per minute, then raise to 2509C at a rate of 359C per minute, and main-tain at 2509C for 8 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2809C. Carrier gas: Helium. Flow rate: 4.3 mL per minute (the retention time of the in-ternal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 – 2 mL of the standard solution under the above operating con-ditions, iodomethane, isopropyl iodide and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 1 – 2 mL of the standard solution under the above oper-ating conditions, the relative standard deviations of the ratio of the peak area of iodomethane and isopropyl iodide to that of the internal standard are not more than 2.0z, respec-tively.
Ibudilast, when dried, contains not less than 98.
Ibudilast occurs as a white crystalline powder. It is very soluble in methanol, freely soluble in ethanol (99.5) and in acetic anhydride, and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Ibudilast in methanol (1 in 250,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ibudilast as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ibudilast according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Ibudilast in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this so-lution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: each peak area other than ibudilast obtained from the sample solution is not larger than the peak area of ibudilast from the stand-ard solution, and the total area of the peaks other than ibudilast is not larger than 3 times the peak area of ibudilast from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌JP XVIII Official Monographs / Ibuprofen Piconol 1133‌ length: 292 nm). Column: A stainless steel column 2.6 mm in inside diame-ter and 15 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of hexane and ethyl acetate (50:1). Flow rate: Adjust so that the retention time of ibudilast is about 9 minutes. Time span of measurement: About 4 times as long as the retention time of ibudilast, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of ibudilast obtained with 10 mL of this solution is equivalent to 40 to 60z of that with 10 mL of the standard solution. System performance: To 5 mL of the sample solution add the mobile phase to make 50 mL. To 2 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the number of theoretical plates and the symme-try factor of the peak of ibudilast are not less than 3500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ibudilast is not more than 3.0z. Loss on drying <2.41> Not more than 0.3z (1 g, in vacu-um, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Ibudilast, previ-ously dried, dissolve in 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 23.03 mg of C14H18N2O
Ibuprofen Piconol contains not less than 98.
Ibuprofen Piconol occurs as a clear, colorless to pale yellow liquid. It is odorless or has a slight characteris-tic odor. It is miscible with methanol, with ethanol (95), with ace-tone and with acetic acid (100). It is practically insoluble in water. It decomposes on exposure to light. It shows no optical rotation.
(1) Dissolve 10 mg of Ibuprofen Piconol in 250 mL of ethanol (95). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ibuprofen Piconol as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.529 – 1.532 Specific gravity <2.56> d 20: 1.046 – 1.050
(1) Chloride <1.03>—Dissolve 0.5 g of Ibuprofen Piconol in 20 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate <1.14>—Dissolve 0.5 g of Ibuprofen Piconol in 20 mL of acetone, add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 20 mL of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.038z). Heavy metals <1.07>—Proceed with 4.0 g of Ibuprofen Piconol according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 5 ppm). Related substances—Dissolve 0.10 g of Ibuprofen Piconol in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, ethyl acetate, acetic acid (100) and methanol (30:10:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of phos-phomolybdic acid n-hydrate in ethanol (95) (1 in 10) on the plate, and heat the plate at 1709C for 10 minutes: the num-ber of spots other than the dark brown principal spot ob-tained from the sample solution is two or less, and they are not more intense than the dark brown spot from the stand-ard solution. Water <2.48> Not more than 0.1z (5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Ibuprofen Piconol, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.74 mg of C19H23NO2
Ibuprofen, when dried, contains not less than 98.
Ibuprofen occurs as a white crystalline pow-der. It is freely soluble in ethanol (95) and in acetone, and practically insoluble in water. It dissolves in dilute sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Ibuprofen in dilute sodium hydroxide TS (3 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ibuprofen, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Ibuprofen according to Method 2, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ibuprofen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Ibuprofen in 5 mL of acetone , and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane, ethyl ace-tate and acetic acid (100) (15:5:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Ibuprofen, previ-ously dried, dissolve in 50 mL of ethanol (95), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 20.63 mg of C13H18O2
Ichthammol, calculated on the dried basis, contains not less than 2.
Ichthammol is a red-brown to black-brown, viscous fluid. It has a characteristic odor. It is miscible with water. It is partially soluble in ethanol (95) and in diethyl ether.
(1) To 4 mL of a solution of Ichthammol (3 in 10) add 8 mL of hydrochloric acid: a yellow-brown to blackish brown, oily or resinous mass is produced. Cool the mass with ice to solidify, and discard the water layer. Wash the residue with diethyl ether: a part of the mass dissolves but it does not dissolve completely even when it is washed until almost no color develops in the washing. Perform the following tests with this residue. To 0.1 g of the residue add 1 mL of a mixture of ethanol (95) and diethyl ether (1:1): it dissolves. To 0.1 g of the residue add 2 mL of water: it dis-solves. To 1 mL of this solution add 0.4 mL of hydrochloric acid: a yellow-brown to blackish brown oily or resinous sub-stance is produced. ‌1136 To 1 mL of the solution obtained in (ii) add 0.3 g of sodium chloride: a yellow-brown or blackish brown oily or resinous substance is produced. (2) Boil 2 mL of a solution of Ichthammol (1 in 10) with 2 mL of sodium hydroxide TS: the gas evolved changes moistened red litmus paper to blue. Loss on drying <2.41> Not more than 50z (0.5 g, 1059C, 6 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
(1) Ammonia—Weigh accurately about 5 g of Ichthammol, transfer to a Kjeldahl flask, and add 60 mL of water, 1 mL of 1-octanol and 4.5 mL of a solution of sodium hydroxide (2 in 5). Connect the flask to a distilling tube with a spray trap and a condenser, and immerse the lower outlet of the condenser in the receiver containing exactly 30 mL of 0.25 mol/L sulfuric acid VS. Distil slowly, collect about 50 mL of the distillate, and titrate <2.50> the excess sulfuric acid with 0.5 mol/L sodium hydroxide VS (indicator: 3 drops of methyl red TS). Perform a blank determination in the same manner. Each mL of 0.25 mol/L sulfuric acid VS ＝ 8.515 mg of NH3 Ammonium sulfate—Weigh accurately about 1 g of Ichthammol, add 25 mL of ethanol (95), stir thoroughly, and filter. Wash with a mixture of ethanol (95) and diethyl ether (1:1) until the washings are clear and colorless. Dry the filter paper and the residue in air, dissolve the residue in 200 mL of hot water acidified slightly with hydrochloric acid, and filter. Boil the filtrate, add 30 mL of barium chloride TS slowly, heat for 30 minutes on a water bath, and filter. Wash the precipitate with water, dry, and ignite to constant mass. Weigh the residue as barium sulfate (BaSO4: 233.39). Amount (mg) of ammonium sulfate [(NH4)2SO4] ＝ amount (mg) of barium sulfate (BaSO4) × 0.566 Total sulfur—Weigh accurately about 0.6 g of Ichthammol, transfer to a 200-mL Kjeldahl flask, and add 30 mL of water and 5 g of potassium chlorate, then add slowly 30 mL of nitric acid, and evaporate the mixture to about 5 mL. Transfer the residue to a 300-mL beaker with the aid of 25 mL of hydrochloric acid, and evaporate again to 5 mL. Add 100 mL of water, boil, filter, and wash with water. Heat the combined filtrate and washings to boil, add gradually 30 mL of barium chloride TS, heat the mixture on a water bath for 30 minutes, and filter. Wash the precipitate with water, dry, and ignite to constant mass. Weigh the residue as barium sulfate (BaSO4). Amount (mg) of total sulfur (S) ＝ amount (mg) of barium sulfate (BaSO4) × 0.13739
Idarubicin Hydrochloride contains not less than 960 mg (potency) and not more than 1030 mg (potency) per mg, calculated on the anhydrous basis.
Idarubicin Hydrochloride occurs as a yellow-red powder. It is sparingly soluble in methanol, slightly soluble in water and in ethanol (95), and practically insoluble in aceto-nitrile and in diethyl ether.
(1) Determine the absorption spectra of a solution of Idarubicin Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Idarubicin Hydrochloride RS prepared in the same manner as the sam-ple solution: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectra of Idarubi-cin Hydrochloride and Idarubicin Hydrochloride RS as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare these spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 2 mg of Idarubicin Hydrochloride in 3 mL of water, and add 1 mL of dilute nitric acid and 3 drops of silver nitrate TS: a white turbidity is produced. D Optical rotation <2.49> [a]20: ＋188 – ＋2019(20 mg calcu-lated on the anhydrous basis, methanol, 20 mL, 100 mm). pH <2.54> Dissolve 10 mg of Idarubicin Hydrochloride in 10 mL of water: the pH of the solution is between 5.0 and 6.5.
(1) Clarity and color of solution—Dissolve 10 mg of Idarubicin Hydrochloride in 10 mL of water: the solution is clear and yellow-red in color. Silver— Dissolve exactly 0.10 g of Idarubicin Hydro-chloride in diluted nitric acid (1 in 200) to make exactly 20 mL, and use this solution as the sample solution. Separately, to exactly 5 mL of Standard Silver Solution for Atomic Ab-sorption Spectrophotometry add diluted nitric acid (1 in 200) to make exactly 50 mL. Pipet a suitable amount of this solu-tion, dilute exactly it with diluted nitric acid (1 in 200) so that each mL contains 0.05 mg, 0.075 mg, 0.1 mg and 0.2 mg of sil- ‌JP XVIII Official Monographs / Idarubicin Hydrochloride for Injection 1137 ver (Ag: 107.87), and use this solution as the standard solu-tion. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectropho-tometry <2.23> according to the following conditions, and calculate the amount of silver in the sample solution using the calibration curve obtained with the absorbances of the standard solution: not more than 20 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Silver hollow-cathode lamp. Wavelength: 328.1 nm. Related substances—Conduct this procedure using light-resistant vessels. Perform the test with 20 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic in-tegration method, and calculate their amounts by the area percentage method: the amount of the peak other than idarubicin is not more than 1.0z, and the total amount of the peaks other than idarubicin is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3.3 times as long as the retention time of idarubicin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase without sodium lauryl sulfate to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for sys-tem suitability test, and add the mobile phase without so-dium lauryl sulfate to make exactly 20 mL. Confirm that the peak area of idarubicin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 10.2 g of potassium dihydrogen-phosphate in a suitable amount of water, add 1 mL of phos-phoric acid and water to make 750 mL, and add 250 mL of tetrahydrofuran. To 500 mL of this solution add 0.72 g of sodium lauryl sulfate and 0.5 mL of N,N-dimethyl-n-octyla-mine, and adjust to pH 4 with 2 mol/L sodium hydroxide TS. Flow rate: Adjust so that the retention time of idarubicin is about 15 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating conditions, the number of theoretical plates of the peak of idarubicin is not less than 3000. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of idarubicin is not more than 2.0z.
Idoxuridine, when dried, contains not less than 98.
Idoxuridine occurs as colorless, crystals or a white, crystalline powder. It is odorless. It is freely soluble in dimethylamide, slightly soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. It dissolves in sodium hydroxide TS. Melting point: about 1769C (with decomposition).
(1) Dissolve 0.01 g of Idoxuridine in 5 mL of water by warming, add 5 mL of diphenylamine-acetic acid TS, and heat for 5 minutes: a blue color develops. Heat 0.1 g of Idoxuridine: a purple gas evolves. Dissolve 2 mg of Idoxuridine in 50 mL of 0.01 mol/L sodium hydroxide. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Idoxuridine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]20: ＋28 – ＋319(after drying, 0.2 g, sodium hydroxide TS, 20 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.20 g of Idoxuridine in 5 mL of a solution of sodium hydroxide (1 in 200): the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Idoxuri- dine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Idoxuridine in exactly 10 mL of a mixture of dilute ethanol and ammonia solution (28) (99:1), and use this solution as the sample solu-tion. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 50 mL of the sample solution on a plate of silica gel with fluorescent indi-cator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and diluted 2-propanol (2 in 3) (4:1) to a distance of about 10 cm, and air-dry the plate. Then develop two-dimensionally at right angles to the first, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): any spot other than the principal spot does not appear. Iodine and iodide—Dissolve 0.10 g of Idoxuridine in 20 mL of water and 5 mL of sodium hydroxide TS, and add immediately 5 mL of dilute sulfuric acid under ice-cooling. Allow to stand for 10 minutes with occasional shaking, and ‌JP XVIII Official Monographs / Idoxuridine Ophthalmic Solution 1139 filter. Transfer the filtrate into a Nessler tube, add 10 mL of chloroform and 3 drops of a solution of potassium iodate (1 in 100), shake for 30 seconds, and allow to stand: the chloroform layer has no more color than the following control solution. Control solution: Weigh accurately 0.111 g of potassium iodide, and dissolve in water to make 1000 mL. To exactly 1 mL of this solution add 19 mL of water, 5 mL of sodium hy-droxide TS and 5 mL of dilute sulfuric acid, mix, and filter. Transfer the filtrate to a Nessler tube, and proceed in the same manner. Loss on drying <2.41> Not more than 0.5z (2 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.7 g of Idoxuridine, previ-ously dried, dissolve in 80 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hydroxide VS until the color of the solution changes from yellow through yellow-green to blue (indicator: 5 drops of thymol blue-dimethylformamide TS). Perform a blank de-termination, and make any necessary correction in the same manner. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 35.41 mg of C9H11IN2O5
Ifenprodil Tartrate contains not less than 98.
Ifenprodil Tartrate occurs as a white crystal-line powder. It is odorless. It is freely soluble in acetic acid (100), soluble in ethanol (95), slightly soluble in water and in methanol, and practi-cally insoluble in diethyl ether. roform, and collect the water layer. Evaporate 30 mL of the water layer on a water bath to dryness, and after cooling, dissolve the residue in 6 mL of water: the solution responds to Qualitative Tests <1.09> for tartrate.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ifenprodil Tartrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.30 g of Ifenprodil Tartrate in 10 mL of diluted ethanol (95) (3 in 4), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add diluted ethanol (95) (3 in 4) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thinlayer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, 1-butanol and ammonia solution (28) (140:40:20:1) to a distance of about 10 cm, and air-dry the plate. Spray hydrogen hexachloroplatinate (IV)-potas-sium iodide TS evenly on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> Not more than 4.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Ifenprodil Tartrate, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.05 mg of (C21H27NO2)2.C4H6O6
Imidapril Hydrochloride occurs as white crys-tals. It is freely soluble in methanol, soluble in water, and spar-ingly soluble in ethanol (99.5). Dissolve 1.0 g of Imidapril Hydrochloride in 100 mL of water: the pH of the solution is about 2. Melting point: about 2039C (with decomposition).
(1) To 3 mL of a solution of Imidapril Hy-drochloride (1 in 50) add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the infrared absorption spectrum of Imi-dapril Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Imidapril Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: －65.0 – －69.09(after dry- ing, 0.1 g, methanol, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Imidapril Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Imidapril Hy-drochloride in 50 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak having the relative retention time of about 0.45 to imidapril, obtained from the sample solution, is not larger than 2/5 times the peak area of imidapril from the standard solution, and the area of each peak other than imidapril and the peak mentioned above from the sample so-lution is not larger than 1/5 times the peak area of imidapril from the standard solution. Furthermore, the total area of the peaks other than imidapril from the sample solution is not larger than 1/2 times the peak area of imidapril from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 2.7 with phosphoric acid. To 600 mL of this solution add 400 mL of methanol. Flow rate: Adjust so that the retention time of imidapril is about 8 minutes. Time span of measurement: About 2 times as long as the retention time of imidapril, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of imidapril obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of imidapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imidapril is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Imidapril Hydro-chloride, previously dried, dissolve in 70 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS from the first equivalent point to the second equivalent point (poten-tiometric titration). ‌JP XVIII Official Monographs / Imidapril Hydrochloride Tablets 1143 Each mL of 0.1 mol/L sodium hydroxide VS ＝ 44.19 mg of C20H27N3O6.HCl
Imipenem Hydrate contains not less than 980 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the anhydrous basis.
Imipenem Hydrate occurs as a white to light yellow crystalline powder. It is sparingly soluble in water, and practically insoluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Imipenem Hydrate in 0.1 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 7.0) (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- Amount (mg) of imidapril hydrochloride (C ＝ MS × QT/QS 20H27N3O6.HCl) ence Spectrum or the spectrum of a solution of Imipenem RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same MS: Amount (mg) of imidapril hydrochloride for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in diluted methanol (2 in 5) (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about wavelengths. (2) Determine the infrared absorption spectrum of Imi-penem Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Imipenem RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋89 – ＋949(50 mg calcu-lated on the anhydrous basis, 0.1 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 7.0), 10 mL, 100 mm). ‌JP XVIII Official Monographs / Imipenem and Cilastatin Sodium for Injection 1145 pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Imipenem Hydrate in 200 mL of water is between 4.5 and 7.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Imipenem Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Put 2.0 g of Imipenem Hydrate in a crucible, add 5 mL of nitric acid and 1 mL of sulfuric acid, and heat carefully until white fumes evolve. After cooling, add 2 mL of nitric acid, heat, and repeat this procedure once more. Then add 2 mL of hydrogen peroxide (30), heat, and repeat this procedure several times until the color of the solu-tion changes to colorless to pale yellow. After cooling, heat again until white fumes evolve. After cooling, add water to make 5 mL, and perform the test with this solution as the test solution (not more than 1 ppm). Related substances—Dissolve 50 mg of Imipenem Hydrate in 50 mL of 0.1 mol/L 3-(N-morpholino)pro-panesulfonic acid buffer solution (pH 7.0) and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add 0.1 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak area of thienamycin, having the relative retention time of about 0.8 to imipenem, obtained from the sample solution is not larger than 1.4 times the peak area of imipenem from the standard solution, the area of the peak other than imipenem and thienamycin from the sample solution is not larger than 1/3 times the peak area of imipenem from the standard solu-tion, and the total area of the peaks other than imipenem and thienamycin from the sample solution is not larger than the peak area of imipenem from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of imipenem. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 5 mL of the standard solution, add 0.1 mol/L 3-(N-morpholino)-propanesulfonic acid buffer solution (pH 7.0) to make ex-actly 50 mL. Confirm that the peak area of imipenem ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imipenem is not more than 2.0z. Water <2.48> Not less than 5.0z and not more than 8.0z (20 mg, coulometric titration, water evaporation tempera-ture: 1409C). Residue on ignition <2.44> Not more than 0.2z (1 g).
Perform the procedure within 30 minutes after preparation of the sample solution and standard solution. Weigh accurately an amount of Imipenem Hydrate and Imi-penem RS, equivalent to about 50 mg (potency), dissolve each in 0.1 mol/L 3-(N-morpholino)-propanesulfonic acid buffer solution (pH 7.0) to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution, within 30 minutes after prepara-tion of these solutions, as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and determine the peak areas, AT and AS, of imipenem in each solution. Amount [ mg (potency)] of imipenem (C12H17N3O4S) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Imipenem RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.1 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 7.0) and acetonitrile (100:1). Flow rate: Adjust so that the retention time of imipenem is about 6 minutes. System suitability— System performance: Dissolve 50 mg of Imipenem Hy-drate and 75 mg of resorcinol in 50 mL of 0.1 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0). When the procedure is run with 10 mL of this solution under the above operating conditions, imipenem and resorcinol are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imipenem is not more than 0.80z.
Imipramine Hydrochloride, when dried, contains not less than 98.
Imipramine Hydrochloride occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. The pH of a solution of 0.1 g of Imipramine Hydrochlo-ride in 10 mL of water is between 4.2 and 5.2. It is gradually colored by light.
(1) Dissolve 5 mg of Imipramine Hydro-chloride in 2 mL of nitric acid: a deep blue color develops. Dissolve 5 mg of Imipramine Hydrochloride in 250 mL of 0.01 mol/L hydrochloric acid TS. Determine the absorption spectrum of the solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Imipramine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Dissolve 0.05 g of Imipramine Hydrochloride in 5 mL of water, add 1 mL of ammonia TS, allow to stand for 5 minutes, filter, and acidify the filtrate with dilute nitric acid: it responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Imipramine Hydrochloride in 10 mL of water: the solu-tion is clear, and has no more color than the following con-trol solution. Control solution: Take exactly 1.0 mL of Cobalt (II) Chloride CS, 2.4 mL of Iron (III) Chloride CS, 0.4 mL of Copper (II) Sulfate CS and 6.2 mL of diluted hydrochloric acid (1 in 40), and mix them. Pipet 0.5 mL of this solution, and add exactly 9.5 mL of water. Iminodibenzyl—Dissolve 50 mg of Imipramine Hy-drochloride in 10 mL of a mixture of hydrochloric acid and ethanol (95) (1:1) in a 25-mL brown volumetric flask. Cool the flask in ice water, add 5 mL of an ethanol (95) solution of furfural (1 in 250) and 5 mL of hydrochloric acid, and allow to stand at 259C for 3 hours. Add a mixture of hydro-chloric acid and ethanol (95) (1:1) to make 25 mL, and deter-mine the absorbance of this solution at 565 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.16. Related substances—Dissolve 0.20 g of Imipramine Hydrochloride in 10 mL of ethanol (95), and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add ethanol (95) to make exactly 50 mL. Pipet 5 mL of this solution, add ethanol (95) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetic acid (100), hydrochloric acid and water (11:7:1:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly potassium dichromate-sulfuric acid TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Imipramine Hydro-chloride, previously dried, and dissolve in 20 mL of water. Add 5 mL of sodium hydroxide TS, and extract with three 20-mL portions of chloroform. Filter each extract through a pledget of absorbent cotton on which a small quantity of an-hydrous sodium sulfate is placed. Combine the chloroform extracts, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the yellow solution changes to red-purple (indicator: 10 drops of metanil yellow TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.69 mg of C19H24N2.HCl
Indapamide occurs as a white crystalline pow-der. It is freely soluble in ethanol (99.5), and practically insolu-ble in water. A solution of Indapamide in ethanol (99.5) (1 in 10) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Indapamide in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Indapamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Inda-pamide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Indapamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Indapamide as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—To 1.5 g of Indapamide add 50 mL of water, shake for 15 minutes, allow to stand in an ice bath for 30 minutes, and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochlo-ric acid VS (not more than 0.01z). Heavy metals <1.07>—Proceed with 2.0 g of Indapa- mide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.10 g of Indapamide in 5 mL of ethanol (99.5), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add ethanol (99.5) to make exactly 200 mL, and use this solution as the stand-ard solution (1). Pipet 5 mL of the standard solution (1), add ethanol (99.5) to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tions (1) and (2) on a plate of silica gel with fluorescent indi-cator for thin-layer chromatography. Immediately develop ‌JP XVIII Official Monographs / Indapamide Tablets 1149 the plate with a mixture of ethyl acetate, cyclohexane and acetic acid (100) (100:80:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1), and the total amount of these related substances, calculated by comparison with the spots from the standard solutions (1) and (2), is not more than 2.0z. Loss on drying <2.41> Not more than 3.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 1109C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Take 20 Imipramine Hydrochloride Tablets, add ex-actly 200 mL of 0.01 mol/L hydrochloric acid TS, and shake well until the tablets are completely disintegrated. After cen-trifuging the solution, pipet a volume of the supernatant liq-uid, equivalent to about 25 mg of imipramine hydrochloride (C19H24N2.HCl), add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 25 mg of Imipra-mine Hydrochloride RS, previously dried at 1059C for 2 hours, dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solu-tion. Pipet 3 mL each of these solutions into separators which contain 15 mL of potassium hydrogen phthalate buffer solution (pH 5.6), 8 mL of bromocresol green-sodium hydroxide TS and 30 mL of chloroform, and shake. Filter the chloroform layer through a pledget of absorbent cotton into a 100-mL volumetric flask. Repeat the extraction with two 30-mL portions of chloroform, combine the chloroform layers in the 100-mL volumetric flask, and add chloroform to make exactly 100 mL. Perform the test with these solu-tions as directed under Ultraviolet-visible Spectrophotome-try <2.24>, using a solution obtained by proceeding with 3 mL of 0.01 mol/L hydrochloric acid TS in the same manner as the blank. Determine the absorbances, AT and AS, of these solutions at 416 nm. Amount (mg) of imipramine hydrochloride (C19H24N2.HCl) ＝ MS × AT/AS MS: Amount (mg) of Imipramine Hydrochloride RS taken
Indenolol Hydrochloride is a mixture of (2RS )-1-(3H-Inden-4-yloxy)-3-(1-methylethyl)aminopropan-2-ol monohydrochloride and (2RS )-1-(3H-Inden-7-yloxy)-3-(1-methylethyl)aminopropan-2-ol monohy-drochloride. When dried, it contains not less than 98.
Indenolol Hydrochloride occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in water and in acetic acid (100), soluble in ethanol (95) and in chloroform, slightly soluble in acetic anhydride, very slightly soluble in ethyl acetate, and practi-cally insoluble in diethyl ether. The pH of a solution of 1.0 g of Indenolol Hydrochloride in 10 mL of water is between 3.5 and 5.5. It is colored by light.
(1) Dissolve 0.1 g of Indenolol Hydrochlo-ride in 1 to 2 drops of dilute hydrochloric acid and 5 mL of water, and add 1 mL of Reinecke salt TS: a red-purple pre-cipitate is formed. Determine the absorption spectrum of a solution of Indenolol Hydrochloride (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wave-lengths. Separately, determine the absorption spectrum of a solution of Indenolol Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of In-denolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Indenolol Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. 1 cm Absorbance <2.24> E1z 10 mg, water, 1000 mL). (250 nm): 330 – 340 (after drying,
(1) Clarity and color of solution—Dissolve 1.0 g of Indenolol Hydrochloride in 10 mL of water: the solution is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of In- ‌JP XVIII Official Monographs / Indigocarmine 1151 denolol Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Indenolol Hydrochloride according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Indenolol Hy-drochloride in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1,2-dichloroethane, ethanol (99.5) and ammonia solution (28) (70:15:2) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 5 mg of Indenolol Hydrochloride in 1.0 mL of a mixture of ethyl acetate and trifluoroacetic an-hydride for gas chromatography (9:1), and use this solution as the sample solution. Perform the test with 2 mL of the sample solution as directed under Gas Chromatography <2.02> according to the following conditions. Determine the areas of two adjacent peaks, Aa and Ab, having the retention times of about 16 minutes, where Aa is the peak area of shorter retention time and Ab is the peak area of longer retention time: the ratio Aa/(Aa ＋ Ab) is between 0.6 and 0.7. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column about 2 mm in inside diameter and about 2 m in length, packed with siliceous earth for gas chromatography (150 to 180 mm in particle diameter) coated with 65z phenyl-methyl silicon polymer for gas chromatog-raphy at the ratio of 2z. Column temperature: A constant temperature between 1509C and 1709C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of the peak showing earlier elution of the two peaks of indenolol hydro-chloride is about 16 minutes. Selection of column: Proceed with 2 mL of the sample so-lution under the above operating conditions, and calculate the resolution. Use a column with the resolution between the two peaks being not less than 1.1.
Weigh accurately about 0.5 g of Indenolol Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (4:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solu-tion changes from purple through blue to green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.38 mg of C15H21NO2.HCl
Indigocarmine, when dried, contains not less than 95.
Indigocarmine occurs as a blue to dark blue, powder or granules. It is odorless. It is sparingly soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic. When compressed, it has a coppery luster.
(1) A solution of Indigocarmine (1 in 100) is dark blue in color. Perform the following tests with this solution as the sample solution: the dark blue color of each solution disappears. ( i ) Add 1 mL of nitric acid to 2 mL of the sample solu-tion; ( ii ) Add 1 mL of bromine TS to 2 mL of the sample so-lution; Add 1 mL of chlorine TS to 2 mL of the sample so-lution; Add 2 mL of sodium hydroxide TS and 0.2 g of zinc powder to 2 mL of the sample solution, and warm. Dissolve 0.1 g of Indigocarmine in 100 mL of a solu-tion of ammonium acetate (1 in 650). To 1 mL of the solu-tion add a solution of ammonium acetate (1 in 650) to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Ignite 1 g of Indigocarmine to carbonize. After cool-ing, add 20 mL of water to the residue, shake, and filter the mixture: the filtrate responds to Qualitative Tests <1.09> for sodium salt and for sulfate. pH <2.54> Dissolve 0.10 g of Indigocarmine in 20 mL of water: the pH of the solution is between 5.0 and 6.0.
(1) Water-insoluble substances—To 1.00 g of In-digocarmine add 200 mL of water, shake, and filter through a tared glass filter (G4). Wash the residue with water until the blue color of the filtrate becomes practically colorless, and dry the residue at 1059C for 4 hours: the mass of the residue does not exceed 5.0 mg. (2) Arsenic <1.11>—Place 0.8 g of Indigocarmine in a Kjeldahl flask, add 5 mL of sulfuric acid and 5 mL of nitric acid, and ignite gently. Repeat the addition of 2 to 3 mL of nitric acid occasionally, and continue to heat until a colorless to light yellow solution is obtained. After cooling, add 15 mL of a saturated ammonium oxalate solution, heat the so-lution until dense white fumes are evolved, and concentrate to 2 to 3 mL. After cooling, dilute with water to 10 mL, and perform the test with 5 mL of this solution as the test solu-tion (not more than 5 ppm). ‌1152‌‌ Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not less than 28z and not more than 38z (after drying, 1 g).
Weigh accurately about 0.5 g of Indigocarmine, pre-viously dried, add 15 g of sodium hydrogen tartrate mono-hydrate, and dissolve in 200 mL of water, boil with bubbling of a stream of carbon dioxide, and titrate <2.50>, while being hot, with 0.1 mol/L titanium (III) chloride VS until the color of the solution changes from blue through yellow to orange. Each mL of 0.1 mol/L titanium (III) chloride VS ＝ 23.32 mg of C16H8N2Na2O8S2
Dissolve a quantity of Indometacin Supposi-tories, equivalent to 50 mg of Indometacin, in 20 mL of methanol by warming, add methanol to make 50 mL, and filter if necessary. To 2 mL of this solution add methanol to make 100 mL, and use this solution as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 317 nm and 321 nm. Amount (mg) of indometacin (C ＝ MS × QT/QS 19H 16ClNO4) Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. MS: Amount (mg) of Indometacin RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and diluted phos-phoric acid (1 in 1000) (7:3). Flow rate: Adjust so that the retention time of indometa-cin is about 8 minutes. System suitability— System performance: Dissolve 50 mg of 4-chlorobenzoic acid, 30 mg of butyl parahydroxybenzoate and 50 mg of in-dometacin in 50 mL of methanol. To 5 mL of this solution add the mobile phase to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-chlorobenzoic acid, butyl parahydroxybenzo-ate and indometacin are eluted in this order, with the resolu-tion between the peaks of 4-chlorobenzoic acid and butyl parahydroxybenzoate being not less than 2.0, and between the peaks of butyl parahydroxybenzoate and indometacin being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of indometacin to that of the internal stand-ard is not more than 1.0z.
Weigh accurately not less than 20 Indometacin Suppositories, cut into small pieces carefully, and mix well. Weigh accurately a portion of the mass, equivalent to about 50 mg of indometacin (C19H16ClNO4), add 40 mL of tetra-hydrofuran, warm at 409C, dissolve by shaking, cool, and add tetrahydrofuran to make exactly 50 mL. Filter the solu-tion, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 3 mL of the internal stand-ard solution, and add the mobile phase to make 100 mL. Allow the solution to stand for 30 minutes, filter through a membrane filter (0.5 mm pore size), discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of In-dometacin RS, previously dried at 1059C for 4 hours, and dissolve in tetrahydrofuran to make exactly 50 mL. Pipet 5 mL of the solution, proceed in the same manner as the sam-ple solution, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of indometacin to that of the internal standard, respectively. ‌JP XVIII Official Monographs / Insulin Human (Genetical Recombination) 1155‌ Amount (mg) of indometacin (C19H16ClNO4) ＝ MS × QT/QS MS: Amount (mg) of Indometacin RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica Insulin Human (Genetical Recombination)
Indometacin, when dried, contains not less than dark blue color disappears, and a yellow-brown color de-velops.
Indometacin occurs as a white to light yellow, fine, crystalline powder. It is sparingly soluble in methanol, in ethanol (95) and in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS. It is colored by light. Melting point: 155 – 1629C It shows crystal polymorphism.
(1) Dissolve 2 mg of Indometacin in 100 mL of methanol. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Indometacin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Indo-metacin, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Indometacin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. ‌JP XVIII Official Monographs / Indometacin Capsules 1153 If any difference appears between the spectra, recrystallize the sample and the RS with diethyl ether, filter and dry the crystals, and perform the test with the crystals. Perform the test with Indometacin as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Acidity—To 1.0 g of Indometacin add 50 mL of water, shake for 5 minutes, and filter. To the filtrate add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenolphthalein TS: a red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Indo- metacin according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Indometacin according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Indometacin in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dehy-drated diethyl ether and acetic acid (100) (100:3) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Measure exactly a volume of Indigocarmine Injection, equivalent to about 0.2 g of indigocarmine (C16H8N2Na2O8S2), add 6 g of sodium hydrogen tartrate monohydrate, and dissolve in water to make 200 mL. Then boil under a carbon dioxide stream, and proceed as directed in the Assay under Indigocarmine. Description Indometacin occurs as a white to light yellow, fine, crystalline powder. It is sparingly soluble in methanol, in ethanol (95) and in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS. It is colored by light. Melting point: 155 – 1629C It shows crystal polymorphism. Identification (1) Dissolve 2 mg of Indometacin in 100 mL of methanol. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Indometacin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Indo-metacin, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Indometacin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. ‌JP XVIII Official Monographs / Indometacin Capsules 1153 If any difference appears between the spectra, recrystallize the sample and the RS with diethyl ether, filter and dry the crystals, and perform the test with the crystals. Perform the test with Indometacin as directed under Flame Coloration Test <1.04> (2): a green color appears. Purity (1) Acidity—To 1.0 g of Indometacin add 50 mL of water, shake for 5 minutes, and filter. To the filtrate add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenolphthalein TS: a red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Indo- metacin according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Indometacin according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Indometacin in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dehy-drated diethyl ether and acetic acid (100) (100:3) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.7 g of Indometacin, previ-ously dried, dissolve in 60 mL of methanol, add 30 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS make 50 mL, then to 2 mL of this solution add methanol to make 100 mL, and use this solution as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 317 nm and 321 nm. Purity Related substances—Powder the content of Indo-metacin Capsules. To a quantity of the powder, equivalent to 0.10 g of Indometacin, add exactly 10 mL of methanol, shake well, filter, and use the filtrate as the sample solution. Dissolve 25 mg of Indometacin RS in methanol to make ex-actly 50 mL. Pipet 1 mL of the solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Proceed as directed in the Purity (4) under In-dometacin. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the content of 1 capsule of Indometacin Capsules, and dissolve in methanol to make exactly V mL so that each mL contains about 1 mg of indometacin (C19H16ClNO4). Filter the solution, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 3 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Indo-metacin RS, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solu-tion, add exactly 3 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this so-lution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of indometacin (C19H16ClNO4) ＝ MS × QT/QS × V/25 MS: Amount (mg) of Indometacin RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 1000). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 900 mL of a mixture of water and phosphate buffer solution (pH 7.2) (4:1) as the dissolution medium, the dissolution rate in 20 minutes of Indometacin Capsules is not less than 75z. ＝ 35.78 mg of C 19H 16ClNO4 Start the test with 1 capsule of Indometacin Capsules, withdraw not less than 20 mL of the medium at the specified
Iodine, Salicylic Acid and Phenol Spirit
ヨード・サリチル酸・フェノール精
Iodine, Salicylic Acid and Phenol Spirit contains not less than 1.
Iodine, Salicylic Acid and Phenol Spirit is a dark red-brown liquid, having the odor of phenol.
(1) To a mixture of 1 mL of starch TS and 9 mL of water add 1 drop of Iodine, Salicylic Acid and Phenol Spirit: a dark blue-purple color develops (iodine). To 1 mL of Iodine, Salicylic Acid and Phenol Spirit add 5 mL of ethanol (95) and water to make 50 mL. To 1 mL of this solution add hydrochloric acid-potassium chloride buffer solution (pH 2.0) to make 50 mL, and to 15 mL of this solution add 5 mL of a solution of iron (III) nitrate enneahydrate (1 in 200): a red-purple color is produced (sali-cylic acid). Shake 1 mL of Iodine, Salicylic Acid and Phenol Spirit with 1 mL of sodium thiosulfate TS, add 20 mL of water and 5 mL of dilute hydrochloric acid, and extract with 25 mL of diethyl ether. Wash the diethyl ether extract with two 25-mL portions of sodium hydrogen carbonate TS, and extract with 10 mL of dilute sodium hydroxide TS. Shake 1 mL of the extract with 1 mL of sodium nitrite TS and 1 mL of dilute hydrochloric acid, and add 3 mL of sodium hydrox-ide TS: a yellow color is developed (phenol). Shake 1 mL of Iodine, Salicylic Acid and Phenol Spirit with 1 mL of sodium thiosulfate TS, add 20 mL of water and 5 mL of dilute hydrochloric acid, extract with 10 mL of diethyl ether, and use the diethyl ether extract as the sample solution. Dissolve 25 mg of salicylic acid, 10 mg of phenol and 40 mg of benzoic acid in 5 mL each of diethyl ether, respectively, and use these solutions as the standard solutions (1), (2) and (3). Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel with fluorescent indica-tor for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the 3 ‌1174 spots from the sample solution show the same Rf value as the corresponding spots of the standard solutions (1), (2) and (3). Spray evenly iron (III) chloride TS on the plate: the spot from standard solution (1) and the corresponding spot from the sample solution acquires a purple color.
(1) Iodine—Pipet 4 mL of Iodine, Salicylic Acid and Phenol Spirit, add ethanol (95) to make exactly 50 ml, and use this solution as the sample solution. On the other hand, weigh accurately about 1.2 g of iodine for assay and about 0.8 g of potassium iodide for assay, previously dried at 1059C for 4 hours, and dissolve in ethanol (95) to make exactly 100 ml. Pipet 4 ml of this solution, add ethanol (95) to make exactly 50 ml, and use this solution as the standard solution. Pipet 3 ml each of the sample solution and stand-ard solution, to each add exactly 25 ml of a mixture of chlo-roform and hexane (2:1), and shake. Further add exactly 10 ml of water, shake and separate the chloroform-hexane layers [use the water layers in (2)]. Filter through a pledget of absorbent cotton, and determine the absorbances of the fil-trates from the sample solution and standard solution, re-spectively, AT and AS, at 512 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>, using a mixture of chloroform and hexane (2:1) as the blank. Amount (mg) of iodine (I) ＝ MS × AT/AS × 1/25 MS: Amount (mg) of iodine for assay taken Potassium iodide—Separate the water layers of the sample solution and standard solution obtained in the Assay (1), pipet 8 ml each of the water layers, and add 1 ml of diluted dilute hydrochloric acid (1 in 2) and 1 ml of sodium nitrite TS. Immediately after shaking, add exactly 10 ml of a mixture of chloroform and hexane (2:1), shake, and proceed in the same manner as for the Assay (1). Amount (mg) of potassium iodide (KI) ＝ MS × AT/AS × 1/25 MS: Amount (mg) of potassium iodide for assay taken Salicylic acid, phenol and benzoic acid—Pipet 2 mL of Iodine, Salicylic Acid and Phenol Spirit, add 20 mL of diluted methanol (1 in 2) and 0.1 mol/L sodium thiosulfate VS until the color of iodine disappears, add exactly 20 mL of the internal standard solution, then add diluted methanol (1 in 2) to make 200 mL, and use this solution as the sample solution. Weigh accurately about 0.2 g of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours, about 80 mg of phenol for assay, and 0.32 g of benzoic acid, previously dried in a desiccator (silica gel) for 3 hours, dis-solve in diluted methanol (1 in 2) to make exactly 50 mL. Pipet 25 mL of this solution, add exactly 20 mL of the inter-nal standard solution and diluted methanol (1 in 2) to make 200 mL, and use this solution as the standard solution. Per-form the test with 3 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QTa, QTb and QTc, of the peak areas of salicylic acid, phenol and benzoic acid to those of the internal standard of the sample solution, and the ratios, QSa, QSb and QSc, of the peak areas of salicylic acid, phenol and benzoic acid to those of the internal standard of the standard solution. Amount (mg) of salicylic acid (C7H6O3) ＝ MSa × QTa/QSa × 1/2 Amount (mg) of phenol (C6H6O) ＝ MSb × QTb/QSb × 1/2 Amount (mg) of benzoic acid (C7H6O2) ＝ MSc × QTc/QSc × 1/2 MSa: Amount (mg) of salicylic acid for assay taken MSb: Amount (mg) of phenol for assay taken MSc: Amount (mg) of benzoic acid taken Internal standard solution—A solution of theophylline in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 25 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: Room temperature. Mobile phase: A mixture of 0.1 mol/L phosohate buffer solution (pH 7.0) and methanol (3:1). Flow rate: Adjust so that the retention time of salicylic acid is about 6 minutes. Selection of column: Dissolve 0.2 g of benzoic acid, 0.2 g of salicylic acid and 50 mg of theophylline in 100 mL of diluted ethanol (1 in 2). To 10 mL of this solution add 90 mL of diluted methanol (1 in 2). Proceed with 10 mL of this solu-tion under the above operating conditions. Use a column giving elution of benzoic acid, salicylic acid and theophylline in this order, and clearly dividing each peak.
Iodine Tincture contains not less than 5.
Iodine Tincture is a dark red-brown liquid, and has a characteristic odor. Specific gravity d 20: about 0.97
(1) To a mixture of 1 mL of starch TS and 9 mL of water add 1 drop of Iodine Tincture: a dark blue-purple color develops. (2) Evaporate 3 mL of Iodine Tincture to dryness on a water bath, and heat gently over a free flame: a white residue is formed which responds to Qualitative Tests <1.09> for po-tassium salt and iodide. Alcohol number <1.01> Not less than 6.6 (Method 2). Per-form the pretreatment (ii) in the Method 1.
(1) Iodine—Pipet 5 mL of Iodine Tincture, add 0.5 g of potassium iodide, 20 mL of water and 1 mL of dilute hydrochloric acid, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 2 mL of starch TS). Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 12.69 mg of I (2) Potassium iodide—Pipet 5 mL of Iodine Tincture into an iodine flask, add 20 mL of water, 50 mL of hydro-chloric acid and 5 mL of chloroform. Cool to room tempera-ture, and titrate <2.50> with 0.05 mol/L potassium iodate VS until the red-purple color disappears from the chloroform layer, with agitating the mixture vigorously and continuous-ly. After the chloroform layer has been decolorized, allow the mixture to stand for 5 minutes. If the color reappears, the mixture should be titrated <2.50> further with 0.05 mol/L potassium iodate VS. Calculate the amount (mg) of potas-sium iodide from the number of mL (a) of 0.05 mol/L potas-sium iodate VS used as above and the number of mL (b) of 0.1 mol/L sodium thiosulfate VS used in the titration under the Assay (1). Amount (mg) of potassium iodide (KI) ＝ 16.60 ×{a － (b/2)}
Iodine occurs as grayish black, plates or granular, heavy crystals, having a metallic luster and a char-acteristic odor. It is freely soluble in diethyl ether, soluble in ethanol (95), sparingly soluble in chloroform, and very slightly soluble in water. It dissolves in potassium iodide TS. It sublimes at room temperature.
(1) A solution of Iodine in ethanol (95) (1 in 50) shows a red-brown color. A solution of Iodine in chloroform (1 in 1000) shows a red-purple to purple color. Add 0.5 mL of starch TS to 10 mL of a saturated so-lution of Iodine: a dark blue color is produced. When the mixture is boiled, the color disappears, and it reappears on cooling. ‌Purity (1) Non-volatile residue—Sublime 2.0 g of Iodine on a water bath, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg. (2) Chloride or bromide—Mix 1.0 g of finely powdered Iodine with 20 mL of water, and filter the mixture. To 10 mL of the filtrate add dropwise diluted sulfurous acid solu-tion (1 in 5) until the yellow color disappears. Add 1 mL of ammonia TS, followed by 1 mL of silver nitrate TS in small portions, and add water to make 20 mL. Shake well, filter, and after discarding the first 2 mL of the filtrate, take 10 mL of the subsequent filtrate. To the filtrate add 2.0 mL of nitric acid and water to make 20 mL: the solution so ob-tained has no more turbidity than the following control solu-tion. Control solution: To 0.20 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of water, 2.5 mL of ammonia TS, 1 mL of silver nitrate TS, 2.0 mL of nitric acid and water to make 20 mL.
Place 1 g of potassium iodide and 1 mL of water in a glass-stoppered flask, weigh accurately, add about 0.3 g of Iodine to the flask, and weigh accurately again. Dissolve the iodine by gentle shaking, add 20 mL of water and 1 mL of dilute hydrochloric acid, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 12.69 mg of I
Iodoform, when dried, contains not less than 99.
Iodoform occurs as lustrous, yellow crystals or crystalline powder. It has a characteristic odor. It is freely soluble in diethyl ether, sparingly soluble in ethanol (95), and practically insoluble in water. It is slightly volatile at ordinary temperature. Melting point: about 1209C (with decomposition).
Heat 0.1 g of Iodoform: a purple gas is evolved.
(1) Water-soluble colored substances and acidity or alkalinity—Shake well 2.0 g of Iodoform, previously powdered, with 5 mL of water for 1 minute, allow to stand, and filter the supernatant liquid: the filtrate is colorless and neutral. Chloride <1.03>—Shake well 3.0 g of Iodoform, pre- viously powdered, with 75 mL of water for 1 minute, allow to stand, and filter the supernatant liquid. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Sulfate <1.14>—To 25 mL of the filtrate obtained in ‌JP XVIII Official Monographs / Iohexol 1175 (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.017z). Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Iodoform, previ-ously dried, in a 500-mL glass-stoppered flask, and dissolve it in 20 mL of ethanol (95). Add exactly 30 mL of 0.1 mol/L silver nitrate VS and 10 mL of nitric acid, stopper the flask, shake well, and allow to stand in a dark place over 16 hours. Add 150 mL of water, and titrate <2.50> the excess silver ni-trate with 0.1 mol/L ammonium thiocyanate VS (indicator: 5 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 13.12 mg of CHI3
Iohexol is a mixture of endo- and exo-products of iohexol. It contains not less than 98.
Iohexol occurs as a white powder. It is very soluble in water, freely soluble in methanol and sparingly soluble in ethanol (99.5). It dissolves in a solution of sodium hydroxide (1 in 20). A solution of Iohexol (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Iohexol (13 in 1,000,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Io-hexol, previously dried at 1059C for 6 hours, as directed in the potassium bromide disk method under Infrared Spectro- photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.1 g of Iohexol in 10 mL of methanol, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (50:25:11) to a distance of about 12 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the number of prin-cipal spots obtained from the sample solutions is two, and their Rf values are about 0.2 and about 0.3, respectively.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Iohexol in 5 mL of water is clear and colorless. Aromatic primary amine—Conduct this procedure using light-resistant vessels. Dissolve 0.20 g of Iohexol in 15 mL of water, cool in ice for 5 minutes, add 1.5 mL of 6 mol/L hydrochloric acid TS and 1 mL of a solution of so-dium nitrite (1 in 50), prepared before use, stir, and cool in ice for 4 minutes. Add 1 mL of a solution of amidosulfuric acid (standard reagent) (1 in 25), stir, and cool in ice for 1 minute. Then, add 0.5 mL of a solution, prepared by dis-solving 0.3 g of N-1-naphthylethylenediamine dihydrochlo-ride in diluted propylene glycol (7 in 10) to make 100 mL, and add water to make exactly 25 mL. Perform the test with this solution as directed under Ultraviolet-visible Spectro-photometry <2.24> within 20 minutes, using a solution pre-pared in the same manner with 15 mL of water as the blank: the absorbance at 495 nm is not more than 0.21. Chloride <1.03>—Perform the test with 2.0 g of Io- hexol. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.007z). Iodine and iodide—Dissolve 1.0 g of Iohexol in 4 mL of water, add 1 mL of dilute sulfuric acid, and allow to stand for 10 minutes while occasional shaking. Add 5 mL of chlo-roform, shake well, and allow to stand: the chloroform layer is colorless. Then, add 1 mL of sodium nitrite solution (1 in 50), prepared before use, shake, allow to stand, and deter-mine the absorbance of collected chloroform layer as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a chloroform layer prepared in the same manner with 4.0 mL of water as the blank: the absorbance at 510 nm is not larger than that of chloroform layer obtained from the following control solution. Control solution: Dissolve exactly 0.131 g of potassium iodide in water to make exactly 100 mL. Pipet 1 mL of this solution, and add water to make exactly 100 mL. Pipet 3 mL of this solution, add 1 mL of water and 1 mL of dilute sulfu-ric acid, then proceed in the same manner. Heavy metals <1.07>—Proceed with 2.0 g of Iohexol according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). 3-Chloro-1,2-propanediol—To exactly 1.0 g of Io-hexol, add exactly 2 mL of diethyl ether, and sonicate for 10 minutes under cooling. Centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve exactly 0.50 g of 3-chloro-1,2-propanediol in diethyl ether to make exactly 50 mL. Pipet 1 mL of this solution, and add diethyl ether to make exactly 100 mL. Pipet 5 mL of this solution, add diethyl ether to make exactly 25 mL, and use this solu-tion as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as di- rected under Gas Chromatography <2.02>, and determine the peak areas, AT and AS, of 3-chloro-1,2-propanediol in each solution: AT is not larger than 2.5 times AS. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.25 mm in inside diameter and 30 m in length, coated the inside surface with a layer about 0.25 mm thick of 5z diphenyl-95z dimethyl-polysiloxane for gas chromatography. Column temperature: A constant temperature of about 709C. Injection port and detector temperature: A constant tem-perature of about 2309C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of 3-chloro-1,2-propanediol is about 7 minutes. Split ratio: 1:40. System suitability— ‌System performance: To 1 mL of a solution of 3-chloro-1,2-propanediol in diethyl ether (1 in 200) and 1 mL of a so-lution of 1-hexanol in diethyl ether (1 in 800) add diethyl ether to make 200 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, 1-hex-anol and 3-chloro-1,2-propanediol are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of 3-chloro-1,2-propanediol is not more than 15z. Related substance—(i) Dissolve 1.0 g of Iohexol in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add methanol to make exactly 25 mL. Pipet 1 mL of this solution, add the methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ace-tone, 2-propanol, ammonia solution (28) and methanol (10:7:4:4) to a distance about 14 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot, other than the principal spot obtained from the sam-ple solution, having the relative Rf value of 1.4 to the spot from the standard solution, is not more intense than the spot from the standard solution. (ii) Dissolve 0.15 g of Iohexol in water to make 100 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine the peak area by the automatic integration method, and calculate the amounts by the area percentage method: the total amount of O-alkyl substances, having the relative retention time between 1.2 and 1.5 to the second principal peak (having bigger retention time) among the two principal peaks of iohexol, is not more than 0.6z, the amount of the peaks, which are eluted after the peak of iohexol and other than O-alkyl substances, is not more than 0.1z, respectively, and the total amount of the peaks, which are eluted after iohexol and other than O-alkyl substances, is not more than 0.3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Acetonitrile. Mobile phase B: Water. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 1 1 – 46 1 → 10 99 → 90 Flow rate: Adjust so that the retention time of the second principal peak (iohexol exo-product) is about 19 minutes. Time span of measurement: About 2 times as long as the retention time of iohexol exo-product. System suitability— Test for required detectability: To 1 mL of the sample so-lution add water to make 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solu-tion for system suitability test, and add water to make ex-actly 20 mL. Confirm that the peak area of iohexol exo-product obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the resolution between the adjacent two peaks, which appear at the retention time of about 18 minutes, is not less than 1.5. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of iohexol exo-product is not more than 3.0z. Water <2.48> Not more than 4.0z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Iohexol, dissolve in 25 mL of a solution of sodium hydroxide (1 in 20), add 0.5 g of zinc powder, boil under a reflux condenser for 30 minutes, and filter after cooling. Wash the flask and filter paper with 200 mL of water, combine the washings and filter, add 5 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L silver nitrate VS (indicator: 1 mL of tetrabromo-phenolphthalein ethyl ester TS) until the color of the precipi-tate changes from yellow to green. Each mL of 0.1 mol/L silver nitrate VS ＝ 27.37 mg of C19H26I3N3O9
Iopamidol, when dried, contains not less than 99.
Iopamidol occurs as a white crystalline pow-der. It is very soluble in water, sparingly soluble in methanol, and very slightly soluble in ethanol (99.5).
(1) To 50 mg of Iopamidol add 5 mL of hydrochloric acid, heat for 10 minutes in a water bath: the test solution responds to Qualitative Tests <1.09> for primary aromatic amines. Heat 0.1 g of Iopamidol over a flame: a purple gas is evolved. Determine the infrared absorption spectrum of Iopamidol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Optical rotation <2.49> [a]20 : －4.6 – －5.29(after drying, 4 g, water, warm, after cooling, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Iopamidol in 10 mL of water: the solution is clear and col-orless. Primary aromatic amines—Dissolve 0.60 g of Iopamidol in 8 mL of water, add 1 mL of a solution of so-dium nitrite (1 in 50) and 12 mL of 2 mol/L hydrochloric acid TS, shake, and allow to stand for 2 minutes. Add 1 mL of a solution of ammonium amidosulfate (1 in 10), shake well, allow to stand for 1 minute, and add 1 mL of naphthylethylenediamine TS and water to make exactly 50 mL. Determine the absorbance of this solution at 495 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution, prepared in the same manner, as the blank: the absorbance is not more than 0.12 (not more than 0.020z). Iodine—Dissolve 2.0 g of Iopamidol in 25 mL of water, add 5 mL of 1 mol/L sulfuric acid TS and 5 mL of toluene, shake well, and allow to stand: the toluene layer is colorless. Free iodine ion—Weigh accurately about 5 g of Iopamidol, dissolve in 70 mL of water, and adjust the pH to about 4.5 with dilute acetic acid. To this solution add 2 mL of 0.1 mol/L sodium chloride TS, and titrate <2.50> with 0.001 mol/L silver nitrate VS (potentiometric titration). ‌Each mL of 0.001 mol/L silver nitrate VS ＝ 0.1269 mg of I Content of iodine ion in Iopamidol is not more than 0.001z. Heavy metals <1.07>—Moisten 1.0 g of Iopamidol with a small quantity of sulfuric acid, heat gradually to almost incinerate by a possibly lower temperature. After cooling, moisten again with a small quantity of sulfuric acid, heat gradually until white fumes no longer are evolved, and incinerate by ignition between 450 to 5509C. Proceed as di-rected in Method 2, and perform the test. Prepare the con-trol solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Iopamidol in water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylamino-2, 4,6-triiodoisophthalamide in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of the both solutions by the automatic in-tegration method: each area of the peaks other than the peak of iopamidol from the sample solution is not larger than the peak area of the standard solution, and the total of these areas is not larger than 2.5 times of the peak area of the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Water. Mobile phase B: A mixture of water and methanol (3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phase A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 6 6 – 18 92 → 65 8 → 35 18 – 30 65 → 8 35 → 92 30 – 34 8 92 Flow rate: 1.5 mL per minute. Time span of measurement: About 4.3 times as long as the retention time of iopamidol. System suitability— System performance: Dissolve 1 mL of the sample solution and 10 mg of N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylamino-2,4,6-triiodoisophthalamide in water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylami-no-2,4,6-triiodoisophthalamide and iopamidol are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the peak areas of N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylamino-2,4,6-triiodoisophthalamide is not more than 1.0z. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Iopamidol, previ-ously dried, transfer to a saponification flask, dissolve in 40 mL of sodium hydroxide TS, add 1 g of zinc powder, boil for 30 minutes under a reflux condenser, cool, and filter. Wash the flask and the filter paper with 50 mL of water, and combine the washing with the filtrate. Add 5 mL of acetic acid (100) to this solution, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration). Each mL of 0.1 mol/L sliver nitrate VS ＝ 25.90 mg of C17H22I3N3O8
Iotalamic Acid, when dried, contains not less than 99.
Iotalamic Acid occurs as a white powder. It is odorless. It is sparingly soluble in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether. It dissolves in sodium hydroxide TS. It gradually colored by light.
(1) Heat 0.1 g of Iotalamic Acid over a flame: a purple gas is evolved. Determine the infrared spectrum of Iotalamic Acid, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 2.0 g of Iotalamic Acid in 10 mL of sodium hydroxide TS: the so-lution is clear and colorless. Primary aromatic amines—To 0.50 g of Iotalamic ‌1180 Acid add 15 mL of water, and dissolve it in 1 mL of sodium hydroxide TS while ice-cooling. Add 4 mL of a solution of sodium nitrite (1 in 100) to the solution, immediately add 12 mL of 1 mol/L hydrochloric acid TS, and shake gently. Then allow the mixture to stand for exactly 2 minutes, add 8 mL of ammonium amidosulfate TS, and shake occasionally for 5 minutes. Add 3 drops of a solution of 1-naphthol in ethanol (95) (1 in 10), allow to stand for 1 minute, add 3.5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), mix, and immediately add water to make 50 mL. De-termine within 20 minutes the absorbance of this solution at 485 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution, prepared in the same man-ner, as the blank: the absorbance is not more than 0.25. Soluble halides—Dissolve 0.5 g of Iotalamic Acid in 20 mL of diluted ammonia TS (1 in 40), add 6 mL of dilute nitric acid, shake, allow to stand for 5 minutes, and filter. Transfer the filtrate to a Nessler tube, wash the residue with 20 mL of water, combine the filtrate and the washings, and add water to make 50 mL. Proceed as directed for Chloride Limit Test <1.03> using this solution as the test solution. Pre-pare the control solution as follows: to 0.10 mL of 0.01 mol/L hydrochloric acid VS and add 20 mL of diluted am-monia TS (1 in 40), 6 mL of dilute nitric acid and water to make 50 mL. Iodine—Dissolve 0.20 g of Iotalamic Acid in 2.0 mL of sodium hydroxide TS, add 2.5 mL of 0.5 mol/L sulfuric acid TS, and allow to stand for 10 minutes with occasional shaking. Add 5 mL of chloroform, shake well, and allow to stand: the chloroform layer remains colorless. Heavy metals <1.07>—Proceed with 1.0 g of Iotalam- ic Acid according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.6 g of Iotalamic Acid according to Method 3, and perform the test (not more than 3.3 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Iotalamic Acid, pre-viously dried, place it in a saponification flask, dissolve in 40 mL of sodium hydroxide TS, add 1 g of zinc powder, and heat for 30 minutes under a reflux condenser. Cool, filter, wash the flask and the filter paper with 50 mL of water, and combine the washings and the filtrate. Add 5 mL of acetic acid (100) to this solution, and titrate <2.50> with 0.1 mol/L silver nitrate VS, until the color of the precipitate changes from yellow to green (indicator: 1 mL of tetrabromophenol-phthalein ethyl ester TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 20.46 mg of C11H9I3N2O4
Iotroxic Acid contains not less than 98.
Iotroxic Acid occurs as a white crystalline powder. It is soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in water and in diethyl ether. It is gradually colored by light.
(1) Heat 0.1 g of Iotroxic Acid over a flame: a purple gas evolves. (2) Dissolve a suitable amount of Iotroxic Acid in a suit-able amount of methanol, evaporate the methanol under reduced pressure, and determine the infrared absorption spectrum of the residue so obtained as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Iotroxic Acid in 10 mL of diluted sodium hydroxide TS (1 in 5): the solution is clear and colorless. Primary aromatic amines—Dissolve 0.20 g of Iotro-xic Acid in 5 mL of water and 1 mL of sodium hydroxide TS, add 4 mL of a solution of sodium nitrite (1 in 100) and 10 mL of 1 mol/L hydrochloric acid TS, mix, and allow to stand for 2 minutes. Add 5 mL of ammonium amidosulfate TS, shake well, allow to stand for 1 minute, then add 0.4 mL of a solution of a -naphthol in ethanol (95) (1 in 10), 15 mL of sodium hydroxide TS and water to make exactly 50 mL. Read the absorbance of this solution at 485 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a blank solution obtained in the same manner as above: the absorbance is not more than 0.22. Iodine—Dissolve 0.20 g of Iotroxic Acid in 2.0 mL of sodium hydrogen carbonate TS, add 5 mL of toluene, mix well, and allow to stand: the toluene layer is colorless. Free iodine ion—Weigh accurately about 5.0 g of Iotroxic Acid, dissolve in 12 mL of a solution of meglumine (3 in 20), add water to make 70 mL, and adjust the pH to about 4.5 with acetic acid (100). To this solution add 2 mL of 0.1 mol/L sodium chloride TS, and titrate <2.50> with 0.001 mol/L silver nitrate VS (potentiometric titration). Each mL of 0.001 mol/L silver nitrate ＝ 0.1269 mg of I Content of iodine ion in Iotroxic Acid, calculated on the anhydrous basis, is not more than 0.004z. Heavy metals <1.07>—Heat strongly 1.0 g of Iotroxic ‌JP XVIII Official Monographs / Ipratropium Bromide Hydrate 1181 Acid as directed under Residue on Ignition Test <2.44>, then proceed according to Method 2, and perform the test. Pre-pare the control solution with 1.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 0.15 g of Iotroxic Acid in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of toluene, acetone and formic acid (6:4:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 1.0 – 2.0z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Iotroxic Acid, dis-solve in 40 mL of sodium hydroxide TS in a saponification flask, add 1 g of zinc powder, and boil for 30 minutes under a reflux condenser. After cooling, filter, wash the flask and the filter paper with 50 mL of water, and combine the wash-ings to the filtrate. To this solution add 5 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration). Each mL of 0.1 mol/L silver nitrate VS ＝ 20.26 mg of C22H18I6N2O9
Ipratropium Bromide Hydrate occurs as a white crystalline powder. It is freely soluble in water, soluble in ethanol (99.5), slightly soluble in acetonitrile and in acetic acid (100), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Ipratropium Bromide Hy-drate in 20 mL of water is between 5.0 and 7.5. Melting point: about 2239C (with decomposition, after drying).
(1) To 5 mg of Ipratropium Bromide Hy-drate add 0.5 mL of fuming nitric acid, and evaporate on a water bath to dryness. After cooling, dissolve the residue in 5 mL of acetone, and add 2 drops of potassium hydroxide-ethanol TS: a purple color develops. Determine the absorption spectrum of a solution of Ipratropium Bromide Hydrate in 0.01 mol/L hydrochloric acid TS (3 in 2000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ipratropium Bromide Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. The solution of Ipratropium Bromide Hydrate (1 in 100) responds to Qualitative Tests <1.09> for bromide.
(1) Clarity and color of solution—Dissolve 1.0 g of Ipratropium Bromide Hydrate in 20 mL of water: the so-lution is clear and colorless. Sulfate <1.14>—Perform the test with 1.0 g of Ipratropium Bromide Hydrate. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Ipratro- pium Bromide Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Ipratropium Bromide Hydrate according to Method 3, and perform the test. Use a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) (not more than 1 ppm). Isopropylatropine bromide—Dissolve 25 mg of Ipratropium Bromide Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Perform the test with 25 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine the peak area, Aa, of ipratropium and the peak area, Ab, having the relative reten-tion time to ipratropium about 1.3 by the automatic integra-tion method: Ab/(Aa ＋ Ab) is not more than 0.01, and no peak other than the peak of ipratropium and the peak having the relative retention time to ipratropium about 1.3 appears within about 14 minutes of the retention time after the sol-vent peak. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 10 to 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: Room temperature. Mobile phase: A mixture of diluted phosphoric acid (1 in 200), acetonitrile and methanesulfonic acid (1000:120:1). Flow rate: Adjust so that the retention time of ipratropi-um is about 7 minutes. Selection of column: Heat a solution of Ipratropium Bromide Hydrate in 1 mol/L hydrochloric acid TS (1 in 100) at 1009C for 1 hour, and cool. To 2.5 mL of this solution add the mobile phase to make 100 mL. Proceed with 25 mL of this solution under the above operating conditions, and calculate the resolution. Use a column showing a resolution not less than 3 between the peak of ipratropium and the peak ‌1182 having the relative retention time to ipratropium about 0.6. Detection sensitivity: Adjust so that the peak height of ipratropium obtained with 25 mL of the sample solution composes 50 to 80z of the full scale. Apo-compounds—Dissolve 0.14 g of Ipratropium Bromide Hydrate in 0.01 mol/L hydrochloric acid TS to make 100 mL. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, A1 and A2, at 246 nm and 263 nm, respectively: A1/A2 is not more than 0.91. Loss on drying <2.41> 3.9 – 4.4z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Ipratropium Bromide Hydrate, previously dried, dissolve in 40 mL of acetic acid (100), add 40 mL of 1,4-dioxane and 2.5 mL of bismuth nitrate TS, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.24 mg of C20H30BrNO3
Ipriflavone, when dried, contains not less than 98.
Ipriflavone occurs as white to yellowish white, crystals or crystalline powder. It is soluble in acetonitrile, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. It gradually turns yellow on exposure to light.
(1) Determine the absorption spectrum of a solution of Ipriflavone in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ipriflavone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ipriflavone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ipriflavone RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ipriflavone according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ipriflavone according to Method 4, and perform the test. Prepare the test solution with 10 mL of dilute hydrochloric acid instead of using 3 mL of hydrochloric acid. Prepare the standard color with 1.0 mL of Standard Arsenic Solution (not more than 1 ppm). Related substances—Dissolve 30 mg of Ipriflavone in 50 mL of acetonitrile. To 5 mL of this solution add aceto-nitrile to make 50 mL, and use this solution as the sample so-lution. Pipet 1 mL of this solution, add acetonitrile to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of both solutions by the au-tomatic integration method: the area of each peak other than the peak of ipriflavone obtained from the sample solution is not larger than 1/2 times the peak area of ipriflavone from the standard solution, and the total area of the peaks other than the peak of ipriflavone from the sample solution is not larger than the peak area of ipriflavone from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of ipriflavone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add acetonitrile to make exactly 20 mL. Con-firm that the peak area of ipriflavone obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ipriflavone are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ipriflavone to that of the internal standard is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 30 mg each of Ipriflavone and Ipriflavone RS, previously dried, dissolve separately in acetonitrile to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution and acetonitrile to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ipriflavone to that of the internal standard. Amount (mg) of ipriflavone (C18H16O3) ＝ MS × QT/QS MS: Amount (mg) of Ipriflavone RS taken Internal standard solution—A solution of di-n-butyl phtha- ‌JP XVIII Official Monographs / Ipriflavone 1183‌ late in acetonitrile (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and water (3:2). Flow rate: Adjust so that the retention time of ipriflavone is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, ipriflavone and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ipriflavone to that of the internal standard is not more than 1.0z.
Irbesartan occurs as a white crystalline pow-der. It is freely soluble in acetic acid (100), sparingly soluble in methanol, slightly soluble in ethanol (99.5) and practically insoluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Irbesartan in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Irbe-sartan as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve the Irbesartan in methanol, evaporate the solvent, dry the residue, and perform the test using the residue.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ir-besartan according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Related substances—Dissolve 50 mg of Irbesartan in 50 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 1 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak, having a relative retention time of about 0.8 to irbesartan, obtained from the sample solution is not larger than 1.5 times the peak area of irbesartan from the standard solution, the area of the peak other than irbesartan and the peak mentioned above from the sample solution is not larger than the peak area of irbesartan from the standard solution, and the total area of the peaks other than irbesartan from the sample solu-tions is not larger than 2 times the peak area of irbesartan from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. ‌Mobile phase: To 5.5 mL of phosphoric acid add 950 mL of water, and adjust to pH 3.2 with triethylamine. To 670 mL of this solution add 330 mL of acetonitrile for liquid chromatography. Flow rate: 1.0 mL per minute. Time span of measurement: About 1.4 times as long as the retention time of irbesartan, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add methanol to make exactly 10 mL. Confirm that the peak area of irbesartan obtained with 10 mL of this solution is equivalent to 35 to 65z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of irbesartan are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irbesartan is not more than 3.0z. Azides—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, back titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Irbesartan, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 42.85 mg of C25H28N6O
Irinotecan Hydrochloride Hydrate contains not less than 99.
Irinotecan Hydrochloride Hydrate occurs as pale yellow to light yellow, crystals or crystalline powder. It is sparingly soluble in methanol, and slightly soluble in water and in ethanol (99.5). It is gradually colored to yellow-brown and decomposed by light. Melting point: about 2559C (with decomposition). It shows crystal polymorphism. Identiˆcation (1) Determine the absorption spectrum of a solution of Irinotecan Hydrochloride Hydrate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Irinotecan Hydrochloride Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. To 1 g of Irinotecan Hydrochloride Hydrate add 50 mL of water, dissolve by heating, and cool: the solution re- 20 25 2 5 6 6 3 sponds to Qualitative Tests <1.09> (2) for chloride. ＝ MS × AT/AS × V?/V × 1/25 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in (1). System suitability— System performance: To 10 mL of the irbesartan standard stock solution obtained in (1) and 2 mL of the amlodipine besilate standard stock solution add 0.02 mol/L phosphate buffer solution (pH 3.0) to make 20 mL. When the proce-dure is run with 5 mL of this solution under the above operat-ing conditions, amlodipine and irbesartan are eluted in this Optical rotation <2.49> [a]20: + 64 – + 699(0.5 g calcu-lated on the anhydrous basis, water, heat, after cooling, 50 mL, 100 mm). D pH <2.54> Dissolve 1 g of Irinotecan Hydrochloride Hy-drate in 50 mL of water by heating, and cool: the pH of this solution is between 3.5 and 4.5.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Irinotecan Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Irinotecan Hydrochloride Hydrate in a suitable amount of a mixture of diluted 0.1 mol/L potassium dihydrogen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) and 1 mL of 1 mol/L hydrochloric acid TS, and add a mixture of diluted 0.1 mol/L potassium dihydrogen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) to make 20 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add a mixture of diluted 0.1 mol/L potassium dihydrogen phosphate TS (1 in 10), methanol and aceto-nitrile (6:4:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak areas of the related sub-stances A and B, having the relative retention times of about 0.8 to irinotecan, and related substances C and D, having the relative retention times of about 1.6, obtained from the sam-ple solution are not larger than 1/5 times the peak area of irinotecan from the standard solution, and the area of the peak other than irinotecan and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of irinotecan from the standard solution. Further-more, the total area of the peaks other than irinotecan from the sample solution is not larger than 4/5 times the peak area of irinotecan from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.22 g of sodium 1-decanesul-fonate in a mixture of diluted 0.1 mol/L potassium dihydro-gen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) to make 1000 mL. Flow rate: Adjust so that the retention time of irinotecan is about 12 minutes. Time span of measurement: About 3 times as long as the retention time of irinotecan. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of diluted 0.1 mol/L potassium dihydrogen phosphate TS (1 in 10), methanol and aceto-nitrile (6:4:3) to make exactly 20 mL. Conˆrm that the peak area of irinotecan obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of irinotecan are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irinotecan is not more than 2.0z. Enantiomer—Being speciˆed separately when the drug is granted approval based on the Law. Water <2.48> 7.5 – 9.5z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.44 g of Irinotecan Hydro- chloride Hydrate, dissolve in 120 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.16 mg of C33H38N4O6.HCl
Irsogladine Maleate, when dried, contains not less than 99.
Irsogladine Maleate occurs as white, crystals or crystalline powder. It has a slightly bitter taste. It is sparingly soluble in acetic acid (100) and in ethylene-glycol, slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Dissolve 20 mg of Irsogladine Maleate in methanol to make 20 mL. Take 2 mL of this solution, and add water to make 20 mL. To 2 mL of this solution add water to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Irso-gladine Maleate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 10 mg of Irsogladine Maleate in 1 mL of dilute hydrochloric acid and 4 mL of water, and add 3 drops of potassium permanganate TS: the color of the solution is discharged immediately.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Irsogladine Maleate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Irsogladine Maleate in 10 mL of ethylene glycol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethylene glycol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of each peak other than the peaks of maleic acid and irsogladine ob-tained from the sample solution is not larger than 1/10 times the peak area of irsogladine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame- ‌JP XVIII Official Monographs / Irsogladine Maleate Fine Granules 1191 ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanesulfonic acid solution (1 in 1000) and methanol (4:1). Flow rate: Adjust so that the retention time of irsogladine is about 8 minutes. Time span of measurement: About 3 times as long as the retention time of irsogladine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add ethylene glycol to make exactly 10 mL. Confirm that the peak area of irsogladine obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of irsogladine are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of irsogladine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Irsogladine Male-ate, previously dried, dissolve in 25 mL of acetic acid (100), add 25 mL of acetic anhydride, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 18.61 mg of C9H7Cl2N5.C4H4O4
Isepamicin Sulfate is the sulfate of a derivative of gentamycin B, an aminoglycoside substance, having antibacterial activity produced by the growth of Micromonospora purpurea. It contains not less than 680 mg (potency) and not more than 780 mg (potency) per mg, calculated on the anhydrous basis.
Isepamicin Sulfate occurs as a white to pale yellow-white powder. It is very soluble in water, and practically insoluble in methanol and in ethanol (95). It is hygroscopic.
(1) Dissolve 20 mg of Isepamicun Sulfate in l mL of water, add 3 mL of anthrone TS, shake, and allow to stand: a blue-purple color develops. Dissolve 10 mg each of Isepamicin Sulfate and Isepamicin Sulfate RS in 5 mL of water, and use these solu-tions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia water (28), ethanol (99.5), 1-butanol and chlo-roform (5:5:4:2) to a distance of about 15 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at about 1009C for about 10 minutes: the principal spots from the sample so-lution and the spot from the standard solution exhibit a red-brown color and show the same Rf value. Dissolve 10 mg of Isepamicin Sulfate in 1 mL of water, and add 1 drop of barium chloride TS: a white pre-cipitate is produced. D Optical rotation <2.49> [a]20: ＋100 – ＋1209(0.25 g calcu-lated on the anhydrous bases, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.5 g of Isepamicin Sulfate in 5 mL of water: the pH of the solution is between 5.5 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g ‌1194 of Isepamicin Sulfate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Isepami- cin Sulfate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL Standard Lead So-lution (not more than 10 ppm). Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of the sample solution by the automatic integration method, and calculate the amounts of their peaks by the area percentage method: the amount of HAPA-gentamine-B equivalent to about 0.4 of the relative retention time to isepamicin is not more than 5.0z, and gen-tamicin B equivalent to about 1.3 of that is not more than 3.0z. For the peak area of gentamicin B, multiply the cor-rection factor, 1.11. Operating conditions— Apparatus, detector, column, column temperature, reac-tion coil, mobile phase, reagent, reaction temperature, flow rate of mobile phase, and flow rate of reagent: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of isepamicin. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the sample so-lution, add water to make 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solu-tion for system suitability test, and add water to make ex-actly 10 mL. Confirm that the peak area of isepamicin ob-tained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suitability test. Water <2.48> Not more than 12.0z (0.2 g, volumetric titration, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Residue on ignition <2.44> Not more than 1.0z (1 g).
Weigh accurately an amount of Isepamicin Sulfate and Isepamicin Sulfate RS, equivalent to about 20 mg (po-tency), dissolve each in water to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of isepamicin in each solution. Amount [ mg (potency)] of isepamicin (C22H43N5O12) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Isepamicin Sulfate RS taken Operating conditions— Apparatus: Consist of two pumps for the mobile phase and the reagent transport, inject port, column, reaction coil, detector and recorder. Use a reaction coil with thermostat. Detector: A fluorophotometer (excitation wavelength: 360 nm, detection wavelength: 440 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Reaction coil: A column 0.25 mm in inside diameter and 5 m in length. Mobile phase: Dissolve 28.41 g of anhydrous sodium sul-fate and 5.23 g of sodium 1-pentane sulfonate in 900 mL of water, add 1 mL of acetic acid (100), and add water to make exactly 1000 mL. Reagent: To 500 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 10.0) add 5 mL of a solution of o-phthalaldehyde in ethanol (95) (2 in 25), 1 mL of 2-mercaptoethanol and 2 mL of a solution of lauroma-crogol (1 in 4). Reaction temperature: A constant temperature of about 459C. Flow rate of mobile phase: About 0.6 mL per minute. Flow rate of reagent: About 0.5 mL per minute. System suitability— System performance: Dissolve 2 mg of gentamicin B in 10 mL of the standard solution. When the procedure is run with 5 mL of this solution under the above operating conditions, isepamicin and gentamicin B are eluted in this order with the resolution between these peaks being not less than 1.0. System repeatability: When the test is repeated 5 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas of isepamicin is not more than 3.0z.
Isoflurane occurs as a clear, colorless fluid liq-uid. tegration method: the area of the peak other than isoflurane from sample solution is not larger than the peak area of ‌1196 isoflurane from the standard solution, and the total area of the peaks other than isoflurane from the sample solution is not larger than 3 times the peak area of isoflurane from the standard solution. Operating conditions— Detector, column, column temperature, carrier gas, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of isoflurane. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add o-xylene to make exactly 2 mL. Con-firm that the peak area of isoflurane obtained with 5 mL of this solution is equivalent to 35 to 65z of that with 5 mL of the standard solution. Peroxide—Take 10 mL of Isoflurane in a Nessler tube add 1 mL of a freshly prepared solution of potassium iodide (1 in 10), shake vigorously, and allow to stand in a dark place for 1 hour: the water layer is not yellow. Residue on evaporation—Pipet 65 mL of Isoflurane, evaporate on a water bath, and dry the residue at 1059C for 1 hour: not more than 1.0 mg. Water <2.48> Not more than 0.1z (2 g, Coulometric titra-tion).
(1) The test solution obtained by Oxygen Flask Combustion Method <1.06> with 50 mL of Isoflurane, using 40 mL of water as the absorbing liquid, responds to Qualitative Tests <1.09> for chloride and fluoride. (2) Determine the infrared absorption spectrum of Isoflurane as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Isoflurane RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 20: 1.500 – 1.520
(1) Acidity or alkalinity—To 10 mL of Isoflurane add 5 mL of freshly boiled and cooled water, and shake for 1 minute: the water layer is neutral. Soluble chloride—To 60 g of Isoflurane add 40 mL of water, shake thoroughly, and separate the water layer. To 20 mL of the layer add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test with this solution as di-rected under Chloride Limit Test <1.03>. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 3 ppm). Soluble fluoride—To 6 g of Isoflurane add 12 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20), and shake for 10 minutes. Transfer 4.0 mL of the water layer into a Nessler tube, add 30 mL of a mixture of alizarin com-plexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1), add water to make 50 mL, allow to stand for 60 minutes, and use this so-lution as the sample solution. Separately, to 0.4 mL of the fluorine standard solution and 4.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20) in a Nessler tube add 30 mL of the mixture of alizarin complexone TS, acetic acid-potas-sium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1), then proceed in the same manner as for the preparation of the sample solution, and use the solution so obtained as the standard solution. Determine the absor-bances of the sample solution and standard solution at 600 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, obtained by proceeding in the same manner as above with 4.0 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20), as the blank: the absorbance of the sample solution is not more than that of the standard solu-tion (not more than 2 ppm). Fluorine standard solution: Dissolve exactly 2.21 g of so-dium fluoride in water to make exactly 1000 mL. Pipet 10 mL of this solution, and add water to make exactly 1000 mL. Each mL of this solution contains 0.01 mg of fluorine (F). Related substances—Use Isoflurane as the sample so-lution. To exactly 1 mL of the sample solution add o-xylene to make exactly 100 mL. Pipet 1 mL of this solution, add o-xylene to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under lated on the anhydrous basis. 3 2 5 Gas Chromatography <2.02> according to the following con- ditions, and determine each peak area by the automatic in- Description Isoflurane occurs as a clear, colorless fluid liq-uid. tegration method: the area of the peak other than isoflurane from sample solution is not larger than the peak area of ‌1196 isoflurane from the standard solution, and the total area of the peaks other than isoflurane from the sample solution is not larger than 3 times the peak area of isoflurane from the standard solution. Operating conditions— Detector, column, column temperature, carrier gas, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of isoflurane. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add o-xylene to make exactly 2 mL. Con-firm that the peak area of isoflurane obtained with 5 mL of this solution is equivalent to 35 to 65z of that with 5 mL of the standard solution. Peroxide—Take 10 mL of Isoflurane in a Nessler tube add 1 mL of a freshly prepared solution of potassium iodide (1 in 10), shake vigorously, and allow to stand in a dark place for 1 hour: the water layer is not yellow. Residue on evaporation—Pipet 65 mL of Isoflurane, evaporate on a water bath, and dry the residue at 1059C for 1 hour: not more than 1.0 mg. Water <2.48> Not more than 0.1z (2 g, Coulometric titra-tion).
To exactly 5 mL each of Isoflurane and Isoflurane RS (separately determined the water <2.48> in the same man-ner as Isoflurane), add exactly 3 mL of ethyl acetate as the internal standard, then add o-xylene to make 50 mL each. To 5 mL each of these solutions add o-xylene to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and calculate the ratios, QT and QS, of the peak area of isoflurane to that of the internal standard. Amount (mg) of isoflurane (C3H2ClF5O) in 5 mL of Isoflurane ＝ VS × QT/QS × 1000 × 1.506 VS: Amount (mL) of Isoflurane RS taken, calculated on the anhydrous basis 1.506: Specific gravity (d 20) of isoflurane Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A stainless steel column 3 mm in inside diameter and 3.5 m in length, packed with siliceous earth for gas chro-matography (125 – 149 mm in particle diameter), coated in 10z with nonylphenoxypoly(ethyleneoxy)ethanol for gas chromatography and in 15z with polyalkylene glycol for gas chromatography. Column temperature: A constant temperature of about 809C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of isoflurane is about 7 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, isoflurane and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of isoflurane is not more than 1.0z.
Containers—Tight containers. Storage—At a temperature not exceeding 309C.
Isomalt Hydrate is a mixture of 6-O-a-D-gluco-pyranosyl-D-sorbitol and 1-O-a-D-glucopyranosyl-D-mannitol. It contains not less than 98.
Isomalt Hydrate occurs as a white, powder or grains. It is freely soluble in water, and practically insoluble in ethanol (95). D Optical rotation [a]20: about ＋ 929(1 g calculated on the anhydrous basis, water, 100 mL, 100 mm).◆ JP XVIII Official Monographs / Isomalt Hydrate 1199 Identiˆcation ◇(1) To 1 mL of a solution of Isomalt Hy-drate (1 in 100) add 1 mL of a solution of catechol (1 in 10) prepared before use, shake thoroughly, add 2 mL of sulfuric acid rapidly, and shake: a reddish purple to red-purple color develops.◇ (2) Perform the test with 20 mL each of the sample solu-tion and standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and compare the chromatograms ob-tained from these solutions: the two principal peaks in the chromatogram obtained from the sample solution are similar in retention time to respective two peaks from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay.
◇(1) Heavy metals <1.07>—Proceed with 2.0 g of Isomalt Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◇ Nickel—Weigh exactly an amount of Isomalt Hy-drate, equivalent to 10.0 g calculated on the anhydrous basis, dissolve in 30 mL of 2 mol/L acetic acid TS, and add water to make exactly 100 mL. Add exactly 2 mL of a solu-tion of ammonium pyrrolidinedithiocarbamate (1 in100) and exactly 10 mL of water-saturated 4-methyl-2-pentanone, and shake for 30 seconds protected from light. Allow the layers to separate, and use the 4-methyl-2-pentanone layer as the sample solution. Separately, take in three vessels three exact portions of Isomalt Hydrate, each equivalent to 10.0 g calcu-lated on the anhydrous basis, dissolve in 30 mL of 2 mol/L acetic acid TS, then add exactly 0.5 mL, 1.0 mL and 1.5 mL respectively of Standard Nickel Solution for Atomic Absorp-tion Spectrophotometry, and add water to make them ex-actly 100 mL. Then, proceed in the same manner as the sam-ple solution, and use the solutions so obtained as the stand-ard solutions. Separately, prepare 4-methyl-2-pentanone layer by proceeding in the same manner as the sample solu-tion but omitting the substance to be examined, and use this solution as the blank solution. Perform the test with the sample solution and standard solution as directed in the standard addition method under Atomic Absorption Spec-trophotometry <2.23> according to the following conditions. The blank solution is used to set the zero of the instrument, and to ascertain that the readings return to zero after rinsing the sample introduction system with water between each measurement: the amount of nickel is not more than 1 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Nickel hollow-cathode lamp. Wavelength: 232.0 nm. Related substances—Weigh exactly 0.20 g of Isomalt Hydrate, dissolve in water to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh ex-actly 10.0 mg of D-sorbitol and 10.0 mg of D-mannitol, dis-solve in water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak areas of D-mannitol, having the relative retention time of about 1.6 to 1-O-a-D-glucopyranosyl-D-mannitol, and D-sorbitol, having the rela-tive retention time of about 2.0, obtained from the sample solution are not larger than the area of the corresponding peak from the standard solution (not more than 0.5z), and the area of the peak other than 1-O-a-D-glucopyranosyl-D-mannitol and 6-O-a-D-glucopyranosyl-D-sorbitol having the relative retention time of about 1.2 and the peaks mentioned above from the sample solution is not larger than the peak area of D-sorbitol from the standard solution (not more than 0.5z). In addition, the total area of the peaks other than 1-O-a-D-glucopyranosyl-D-mannitol and 6-O-a-D-glucopyranosyl-D-sorbitol from the sample solution is not larger than 4 times the peak area of D-sorbitol from the standard solution (not more than 2.0z). However, the peaks which area is not larger than 1/5 times the peak area of D-sorbitol from the standard solution are disregarded (not more than 0.1z). Operating conditions— Detector, column, column temperature, mobile phase, and ‰ow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of 1-O-a-D-glucopyranosyl-D-mannitol. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the stand- ard solution, and add water to make exactly 10 mL. Conˆrm that the peak area of D-sorbitol obtained with 20 mL of this solution is equivalent to 14 to 26z of that with 20 mL the standard solution.◇ System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of D-mannitol and D-sorbitol are not more than 2.0z, respectively.◇ Reducing sugars—Dissolve 3.3 g of Isomalt Hydrate in 10 mL of water with the aid of gentle heat, cool, and add 20 mL of copper (II) citrate TS. Add a few amount of boil-ing chips, heat so that the boiling begins after 4 minutes, and maintain boiling for 3 minutes. Cool rapidly, add 100 mL of a solution of acetic acid (100) (3 in 125) and exactly 20 mL of 0.025 mol/L iodine VS. With continuous shaking, add 25 mL of a mixture of water and hydrochloric acid (47:3). When the precipitate has dissolved, titrate <2.50> the excess of iodine with 0.05 mol/L sodium thiosulfate VS, until the blue color due to 1 mL of soluble starch TS added at near of the end point disappears: not less than 12.8 mL of 0.05 mol/L sodium thiosulfate VS is required (not more than 0.3z as glucose). Conductivity <2.51> Dissolve 20 g of Isomalt Hydrate in a suitable amount of freshly boiled and cooled water with the aid of gentle heat at 40 – 509C, cool, add the same water to make exactly 100 mL, and use this solution as the sample so-lution. Measure the conductivity (259C) of the sample solu-tion at 25 ± 0.19C while gently stirring with a magnetic stir-rer: not more than 20 mS･cm－1. Water <2.48> Not more than 7.0z (0.3 g, volumetric titra-tion, direct titration. Use a mixture of methanol for water determination and formamide for water determination (1:1) heated at 50 ± 59C instead of methanol for water determi-nation).
Weight accurately about 0.2 g of Isomalt Hydrate, dissolve in water to make exactly 10 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 0.2 g of Isomalt RS (separately determine the water <2.48> in the same manner as Isomalt Hydrate), dissolve in ‌1200 water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, ATa and ATb, and ASa and ASb, of 1-O-a-D-glucopyranosyl-D-mannitol and 6-O-a-D-glucopyranosyl-D-sorbitol in each solution. Amount (g) of 1-O-a-D-glucopyranosyl-D-mannitol (C12H24O11) ＝MS × Ka/100 × ATa/ASa Amount (g) of 6-O-a-D-glucopyranosyl-D-sorbitol Isoniazid
Isoniazid, when dried, contains not less than 98.
Isoniazid occurs as colorless crystals or a white crystalline powder. It is odorless. It is freely soluble in water and in acetic acid (100), spar-ingly soluble in ethanol (95), slightly soluble in acetic anhy-dride, and very slightly soluble in diethyl ether.
(1) Dissolve about 20 mg of Isoniazid in water to make 200 mL. To 5 mL of the solution add 1 mL of 0.1 mol/L hydrochloric acid TS and water to make 50 mL. Determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Isoniazid, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Isoniazid in 10 mL of freshly boiled and cooled water: the pH of this solution is between 6.5 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Isoniazid in 20 mL of water: the solution is clear and col-orless. Heavy metals <1.07>—Proceed with 1.0 g of Isoniazid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Isoniazid according to Method 3, and perform the test. In this case, add 10 mL of a solution of magnesium nitrate hex-ahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydro-gen peroxide (30), and ignite the ethanol to burn (not more than 5 ppm). Hydrazine—Dissolve 0.10 g of Isoniazid in 5 mL of water, add 0.1 mL of a solution of salicylaldehyde in ethanol (95) (1 in 20), shake immediately, and allow to stand for 5 minutes: no turbidity is produced. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Isoniazid, previ-ously dried, dissolve in 50 mL of acetic acid (100) and 10 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L per- ‌JP XVIII Official Monographs / Isoniazid Tablets 1201‌ chloric acid VS until the color of the solution changes from yellow to green (indicator: 0.5 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 13.71 mg of C6H7N3O
Isopropanol is a clear, colorless liquid. It has a characteristic odor. It is miscible with water, with methanol, with ethanol (95), and with diethyl ether. It is flammable and volatile.
(1) To 1 mL of Isopropanol add 2 mL of iodine TS and 2 mL of sodium hydroxide TS, and shake: a light yellow precipitate is formed. (2) To 5 mL of Isopropanol add 20 mL of potassium dichromate TS and 5 mL of sulfuric acid with caution, and warm gently on a water bath: the produced gas has the odor of acetone, and the gas turns the filter paper, previously wet-ted with a solution of salicylaldehyde in ethanol (95) (1 in 10) and with a solution of sodium hydroxide (3 in 10), to red-brown. Specific gravity <2.56> d 20: 0.785 – 0.788
(1) Clarity of solution—To 2.0 mL of Isopropanol add 8 mL of water, and shake: the solution is clear. Acidity—To 15.0 mL of Isopropanol add 50 mL of freshly boiled and cooled water and 2 drops of phenol-phthalein TS, and add 0.40 mL of 0.01 mol/L sodium hy-droxide VS: a red color develops. Residue on evaporation—Evaporate 20.0 mL of Isopropanol on a water bath to dryness, and dry at 1059C for 1 hour: the mass of the residue is not more than 1.0 mg. Water <2.48> Not more than 0.75 w/vz (2 mL, volumetric titration, direct titration). Distilling range <2.57> 81 – 839C, not less than 94 volz.
Containers—Tight containers. Storage—Remote from fire.
Isosorbide Dinitrate occurs as white, crystals or crystalline powder. It is odorless or has a faint odor like that of nitric acid. It is very soluble in N,N-dimethylformamide and in ace-tone, freely soluble in chloroform and in toluene, soluble in methanol, in ethanol (95) and in diethyl ether, and practi-cally insoluble in water. It explodes if heated quickly or subjected to percussion.
(1) Dissolve 10 mg of Isosorbide Dinitrate in 1 mL of water, and dissolve by adding 2 mL of sulfuric acid cautiously. After cooling, superimpose 3 mL of iron (II) sulfate TS, and allow to stand for 5 to 10 minutes: a brown ring is produced at the zone of contact. (2) Dissolve 0.1 g of Isosorbide Dinitrate in 6 mL of diluted sulfuric acid (1 in 2) by heating in a water bath. After cooling, add 1 mL of a solution of potassium permanganate (1 in 30), stir well, and heat in a water bath until the color of potassium permanganate disappears. Add 10 mL of 2,4-dinitro-phenylhydrazine TS, and heat in a water bath: an orange precipitate is produced. D Optical rotation <2.49> [a]20: ＋134 – ＋1399(1 g calculated on the anhydrous basis, ethanol (95), 100 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Isosorbide Dinitrate in 10 mL of acetone: the solution is clear and colorless. Sulfate <1.14>—Dissolve 1.5 g of Isosorbide Dinitrate in 15 mL of N,N-dimethylformamide, add 60 mL of water, cool, and filter. Wash the filter paper with three 20-mL por-tions of water, combine the washings with the filtrate, and add water to make 150 mL. To 40 mL of this solution add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.048z). Nitrate—Dissolve 50 mg of Isosorbide Dinitrate in 30 mL of toluene, and extract with three 20-mL portions of water. Combine the aqueous layers, and wash with two 20-mL portions of toluene. To the aqueous layer add water to make 100 mL, and use this solution as the sample solu-tion. Pipet 5.0 mL of Standard Nitric Acid Solution and 25 mL of the sample solution in each Nessler tube, and add water to make 50 mL, respectively. To each of them add 60 mg of Griss-Romijin's nitric acid reagent, stir well, allow to stand for 30 minutes, and observe from the side of the Nessler tube: the sample solution has no more color than the standard solution. sodium hydroxide (1 in 2) through the funnel, cautiously rinse the funnel with 20 mL of water, immediately close the clamp attached to the rubber tubing, then begin the distilla-tion with steam gradually, and continue the distillation until the distillate measures 100 mL. Remove the absorption flask, rinse the end of the condenser tube with a small quantity of water, and titrate <2.50> the distillate and the rinsings with 0.1 mol/L sodium hydroxide VS until the color of the solu- tion changes from red through light red-purple to light blue-green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sulfuric acid VS ＝ 11.81 mg of C6H8N2O8
Weigh accurately about 0.1 g of Isosorbide Dinitrate in a Kjeldahl flask as described under the Nitrogen Determi-nation <1.08>, dissolve in 10 mL of methanol, add 3 g of Devarda's alloy and 50 mL of water, and connect the flask with the distillation apparatus as described under the Nitro-gen Determination <1.08>. Measure exactly 25 mL of 0.05 mol/L sulfuric acid VS in an absorption flask, add 5 drops hydrous basis. 6 8 2 8 of bromocresol green-methyl red TS, and immerse the lower end of the condenser tube in it. Add 15 mL of a solution of Description Isosorbide Dinitrate occurs as white, crystals or crystalline powder. It is odorless or has a faint odor like that of nitric acid. It is very soluble in N,N-dimethylformamide and in ace-tone, freely soluble in chloroform and in toluene, soluble in methanol, in ethanol (95) and in diethyl ether, and practi-cally insoluble in water. It explodes if heated quickly or subjected to percussion. Identification (1) Dissolve 10 mg of Isosorbide Dinitrate in 1 mL of water, and dissolve by adding 2 mL of sulfuric acid cautiously. After cooling, superimpose 3 mL of iron (II) sulfate TS, and allow to stand for 5 to 10 minutes: a brown ring is produced at the zone of contact. (2) Dissolve 0.1 g of Isosorbide Dinitrate in 6 mL of diluted sulfuric acid (1 in 2) by heating in a water bath. After cooling, add 1 mL of a solution of potassium permanganate (1 in 30), stir well, and heat in a water bath until the color of potassium permanganate disappears. Add 10 mL of 2,4-dinitro-phenylhydrazine TS, and heat in a water bath: an orange precipitate is produced. D Optical rotation <2.49> [a]20: ＋134 – ＋1399(1 g calculated on the anhydrous basis, ethanol (95), 100 mL, 100 mm). Purity (1) Clarity and color of solution—Dissolve 1.0 g of Isosorbide Dinitrate in 10 mL of acetone: the solution is clear and colorless. Sulfate <1.14>—Dissolve 1.5 g of Isosorbide Dinitrate in 15 mL of N,N-dimethylformamide, add 60 mL of water, cool, and filter. Wash the filter paper with three 20-mL por-tions of water, combine the washings with the filtrate, and add water to make 150 mL. To 40 mL of this solution add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.048z). Nitrate—Dissolve 50 mg of Isosorbide Dinitrate in 30 mL of toluene, and extract with three 20-mL portions of water. Combine the aqueous layers, and wash with two 20-mL portions of toluene. To the aqueous layer add water to make 100 mL, and use this solution as the sample solu-tion. Pipet 5.0 mL of Standard Nitric Acid Solution and 25 mL of the sample solution in each Nessler tube, and add water to make 50 mL, respectively. To each of them add 60 mg of Griss-Romijin's nitric acid reagent, stir well, allow to stand for 30 minutes, and observe from the side of the Nessler tube: the sample solution has no more color than the standard solution. sodium hydroxide (1 in 2) through the funnel, cautiously rinse the funnel with 20 mL of water, immediately close the clamp attached to the rubber tubing, then begin the distilla-tion with steam gradually, and continue the distillation until the distillate measures 100 mL. Remove the absorption flask, rinse the end of the condenser tube with a small quantity of water, and titrate <2.50> the distillate and the rinsings with 0.1 mol/L sodium hydroxide VS until the color of the solu- tion changes from red through light red-purple to light blue-green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sulfuric acid VS ＝ 11.81 mg of C6H8N2O8
Isosorbide Mononitrate 70z/Lactose 30z oc-curs as a white, powder, crystalline powder, or masses. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) Shake thoroughly 1 g of Isosorbide Mononitrate 70z/Lactose 30z with 30 mL of ethyl acetate, and filter. Wash the residue with a small quantity of ethyl acetate, combine the filtrate and the washings, evaporate to dryness on a water bath, then dry in vacuum at room tem-perature for 4 hours. Determine the infrared absorption spectrum of the crystals obtained as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum of isosorbide mononitrate: both spectra exhibit ＝ MS × AT/AS × 1/10 6 8 2 8 similar intensities of absorption at the same wave numbers. (2) Dry the residue obtained in (1) at 809C for 2 hours. MS: Amount (mg) of isosorbide dinitrate for assay taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol (11:9). Flow rate: Adjust so that the retention time of isosorbide dinitrate is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of isosorbide dinitrate are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isosorbide dinitrate is not more than 1.0z.
(1) Nitrate—Dissolve an exact quantity of Isosor-bide Mononitrate 70z/Lactose 30z, equivalent to 50 mg of isosorbide mononitrate (C6H9NO6), in water to make exactly 100 mL. Pipet 25 mL of this solution, add water to make ex-actly 50 mL, and use this solution as the sample solution. Separately, to exactly 5 mL of Standard Nitric Acid Solution add water to make exactly 150 mL. Pipet 25 mL of this solu-tion, add water to make exactly 150 mL, and use this solu-tion as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak area of nitric acid of each solution by the automatic integration method: the peak area of nitric acid obtained from the sam-ple solution is not larger than the peak area of nitric acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with gel type strong basic ion-exchange resin for liquid chromatography (10 mm in particle JP XVIII Official Monographs / Isosorbide Mononitrate 70%/Lactose 30% 1207 diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 16.0 g of sodium gluconate, 18.0 g of boric acid, 25.0 g of sodium tetraborate decahydrate, and 250 mL of glycerin in water to make 1000 mL. To 20 mL of this solution add 20 mL of 1-butanol, 120 mL of acetonitrile, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of nitric acid is about 5.3 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nitric acid are not less than 800 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitric acid is not more than 2.0z. Heavy metals <1.07>—Proceed with 1.0 g of Isosor- bide Mononitrate 70z/Lactose 30z according to Method 1, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Isosorbide—To an amount of Isosorbide Mononi-trate 70z/Lactose 30z, equivalent to 1.0 g of isosorbide mononitrate (C6H9NO6), add 10 mL of acetone, shake well, centrifuge, and filter the supernatant liquid through a mem-brane filter with a pore size not exceeding 0.5 mm. To the residue add 2 mL of acetone and proceed in the same man-ner, and combine the filtrates. Evaporate the combined fil-trate to dryness on a water bath, and further dry the residue in vacuum for 30 minutes. Dissolve the residue in the mobile phase to make 10 mL, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this so-lution, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas by the automatic integration method: the peak area of isosorbide, having the relative retention time of about 0.2 to isosorbide mononitrate, obtained from the sample solution is not larger than the peak area of isosorbide mononitrate from the standard solution. Operating conditions— Detector: A differential refractometer. Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (9:1). Flow rate: Adjust so that the retention time of isosorbide mononitrate is about 16 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of isosorbide mononitrate are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times bide Mononitrate 70z/Lactose 30z, equivalent to 50 mg of isosorbide mononitrate (C6H9NO6), in 5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than isosorbide mononitrate obtained from the sample solution is not larger than 1/2 times the peak area of isosorbide mononitrate from the standard solution, and the total area of the peaks other than isosorbide mononitrate from the sample solution is not larger than the peak area of isosorbide mononitrate from the standard solution. For the area of the peak, having the relative retention time of about 4.5 to isosorbide mononitrate, multiply the correction fac-tor, 0.62. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of isosorbide mononitrate, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 1 mL of the standard solution, and add water to make exactly 10 mL. Confirm that the peak area of isosorbide mononitrate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isosorbide mononitrate is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Water <2.48> Between 1.0z and 2.0z (0.4 g, volumetric titration, direct titratioin. Use a mixture of methanol for water determination and formamide for water determination (2:1) instead of methanol for water determination). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately an amount of Isosorbide Mononi-trate 70z/Lactose 30z, previously dried, equivalent to about 0.2 g of isosorbide mononitrate (C6H9NO6), and dis-solve in water to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 20 mL of the internal standard solu-tion, then add water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of isosorbide mononitrate for assay, previously dried, and dissolve in 60 mL of water, add exactly 20 mL of the in-ternal standard solution, then, add water to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of isosorbide mononitrate to that of the internal standard. Amount (mg) of isosorbide mononitrate (C H NO ) with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ＝ MS × QT/QS × 5 6 9 6 area of isosorbide mononitrate is not more than 4.0z. Related substances—Dissolve an amount of Isosor- MS: Amount (mg) of isosorbide mononitrate for assay taken ‌1208 Internal standard solution—A solution of benzyl alcohol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about size not exceeding 0.45 mm, discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of isosorbide mononitrate for assay, previously dried in vacuum (silica gel) for 4 hours, add 30 mL of water and exactly 10 mL of the in-ternal standard solution, then add water to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of isosorbide mononitrate (C H NO ) 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in ＝ MS × QT/QS × V/100 6 9 6 1000) and methanol (4:1). Flow rate: Adjust so that the retention time of isosorbide mononitrate is about 4.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, isosorbide mononitrate and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of isosorbide mononitrate to that of the inter-nal standard is not more than 1.0z.
Isosorbide occurs as white, crystals or masses. It is odorless, or has a faint, characteristic odor, and has a bitter taste. It is very soluble in water and in methanol, freely soluble in ethanol (95), and slightly soluble in diethyl ether. It is hygroscopic.
(1) To 0.1 g of Isosorbide add 6 mL of diluted sulfuric acid (1 in 2), and dissolve by heating in a water bath. After cooling, shake well with 1 mL of a solu-tion of potassium permanganate (1 in 30), and heat in a water bath until the color of potassium permanganate disap-pears. To this solution add 10 mL of 2,4-dinitrophenyl-hydrazine TS, and heat in a water bath: an orange precipi-tate is formed. To 2 g of Isosorbide add 30 mL of pyridine and 4 mL of benzoyl chloride, boil under a reflux condenser for 50 minutes, cool, and pour gradually the solution into 100 mL of cold water. Filter the formed precipitate by suction through a glass filter (G3), wash with water, recrystallize twice from ethanol (95), and dry in a desiccator (in vacuum, silica gel) for 4 hours: it melts <2.60> between 1029C and 1039C. Determine the infrared absorption spectrum of Isosorbide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋45.0 – ＋46.09(5 g calcu-lated on the anhydrous basis, water, 50 mL, 100 mm).
(1) Clarity and color of solution—Take 25 g of Isosorbide in a Nessler tube, and dissolve in 50 mL of water: the solution is clear, and has no more color than the follow-ing control solution. Control solution: To a mixture of 1.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add water to make 10.0 mL. To 3.0 mL of this solution add water to make 50 mL. Sulfate <1.14>—Perform the test with 2.0 g of Isosor-bide. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 5.0 g of Isosor-bide according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Isosorbide according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Isosorbide in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 2 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (95) and cyclohexane (1:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly a mixture of ethanol (95) and sulfuric acid (9:1) on the plate, and heat the plate at 1509C for 30 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Water <2.48> Not more than 1.5z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 10 g of Isosorbide, calcu-lated on the anhydrous basis, and dissolve in water to make exactly 100 mL. Determine the optical rotation <2.49>, aD, of this solution at 20 ± 19C in a 100-mm cell. Amount (g) of isosorbide (C6H10O4) ＝ aD × 2.1978
Isoxsuprine Hydrochloride, when dried, contains not less than 99.
Isoxsuprine Hydrochloride occurs as a white, Test for required detectability: Pipet 1 mL of the standard ‌1210 solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of isoxsuprine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: To 1 mL of the sample solution add 2.5 mL of a solution of methyl parahydroxybenzoate (1 in 25,000) and the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, methyl parahydroxybenzoate and isox-suprine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak MS: Amount (mg) of isoxsuprine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Isoxsuprine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Isoxsuprine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of isoxsuprine hydrochlo- area of isoxsuprine is not more than 2.5z. ride (C 18H 23NO3.HCl), and use this solution as the sample Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.2z (1 g).
(1) Determine the absorption spectrum of a solution of Isoxsuprine Hydrochloride (1 in 20,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Isox-suprine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.5 g of Isoxsuprine Hydrochloride in 50 mL of water by warming, and cool: the solution responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 0.5 g of Isoxsuprine Hydrochloride in 50 mL of water by warming, and cool: the pH of the solu-tion is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve 0.1 g of Isoxsuprine Hydrochloride in 10 mL of water, warm if necessary, and cool: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Isoxsu- prine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Isoxsuprine Hydrochloride in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: each peak area other than isoxsuprine obtained from the sample solution is not larger than the peak area of isoxsuprine from the stand-ard solution, and the total area of the peaks other than the peak of isoxsuprine from the sample solution is not larger than 2 times the peak area of isoxsuprine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 269 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.3 g of diammonium hydrogen phosphate and 3.2 g of sodium 1-pentane sulfonate in water to make 1000 mL, and adjust to pH 2.5 with phosphoric acid. To 770 mL of this solution add 230 mL of acetonitrile. Flow rate: Adjust so that the retention time of isoxsuprine is about 18 minutes. Time span of measurement: About 3 times as long as the retention time of isoxsuprine, beginning after the solvent isoxsuprine hydrochloride (C 18H23 NO3.HCl). peak. System suitability— Description Isoxsuprine Hydrochloride occurs as a white, Test for required detectability: Pipet 1 mL of the standard ‌1210 solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of isoxsuprine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: To 1 mL of the sample solution add 2.5 mL of a solution of methyl parahydroxybenzoate (1 in 25,000) and the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, methyl parahydroxybenzoate and isox-suprine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak MS: Amount (mg) of isoxsuprine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Isoxsuprine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Isoxsuprine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of isoxsuprine hydrochlo- area of isoxsuprine is not more than 2.5z. ride (C 18H 23NO3.HCl), and use this solution as the sample Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Isoxsuprine Hydro-chloride, previously dried, dissolve in 5 mL of formic acid, add 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 33.78 mg of C18H23NO3.HCl
Itraconazole occurs as a white powder. It is soluble in N,N-dimethylformamide, very slightly soluble in ethanol (99.5), and practically insoluble in water and in 2-propanol. A solution of Itraconazole in N,N-dimethylformamide (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Itraconazole in 2-propanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Itraconazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Itraconazole as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Itraconazole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Itraconazole in 10 mL of a mixture of methanol and tetrahydrofuran (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of methanol and tetrahydrofuran (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add the mixture of methanol and tetrahydrofu-ran (1:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of each solu-tion by the automatic integration method: the area of each peak other than itraconazole obtained from the sample solu-tion is not larger than the peak area of itraconazole from the standard solution. Furthermore, the total area of the peaks other than itraconazole from the sample solution is not larger than 2.5 times the peak area of itraconazole from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: A solution of tetrabutylammonium hydrogensulfate (17 in 625). Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 4-yl}methoxy)phenyl]piperazin-1-yl}phenyl)-2-[(1RS )-1-methylpropyl]-2,4-dihydro-3H-1,2,4-triazol-3-one Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) [84625-61-6] Itraconazole contains not less than 98.5z and not more 0 – 20 80 → 50 20 → 50 20 – 25 50 50 ‌1212‌ Flow rate: 1.5 mL per minute. Time span of measurement: About 2 times as long as the retention time of itraconazole, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mixture of methanol and tetra-hydrofuran (1:1) to make exactly 10 mL. Confirm that the peak area of itraconazole obtained with 10 mL of this solu-tion is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 1 mg of Itraconazole and 1 mg of miconazole nitrate in 20 mL of the mixture of metha-nol and tetrahydrofuran (1:1). When the procedure is run with 10 mL of this solution under the above operating condi-tions, miconazole and itraconazole are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of itraconazole is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Itraconazole, dis-solve in 70 mL of a mixture of 2-butanone and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS dried basis. The potency of Josamycin is expressed as mass (potency) of josamycin (C42H69NO15). Description Josamycin occurs as a white to yellowish white powder. It is very soluble in methanol and in ethanol (99.5), and very slightly soluble in water. Identification (1) Determine the absorption spectrum of a solution of Josamycin in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Josamycin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve 5 mg each of Josamycin and Josamycin RS in 1 mL of methanol, add diluted methanol (1 in 2) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and compare the chromatograms obtained from these solutions: the retention time of the main peak from the sample solution is the same as that of the peak of josamycin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2). Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Josamycin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). ＝ 35.28 mg of C35H38 Cl2N8O4 (2) Related substances—Dissolve 50 mg of Josamycin in 5 mL of methanol, add diluted methanol (1 in 2) to make 50
ジョサマイシンプロピオン酸エステル
Josamycin Propionate is a derivative of josamycin. It contains not less than 843 mg (potency) and not more than 1000 mg (potency) per mg, calculated on the dried basis.
Josamycin Propionate occurs as a white to light yellow-white crystalline powder. It is very soluble in acetonitrile, freely soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Josamycin Propionate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Josamycin Propionate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve 5 mg each of Josamycin Propionate and Josamycin Propionate RS in 50 mL of diluted acetonitrile (1 in 2), and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and compare the chromatograms ob-tained from these solutions: the retention time of the peak of josamycin propionate from the sample solution is the same with that of the peak of josamycin propionate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Josamycin Propionate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 30 ppm). Related substances—Dissolve 50 mg of Josamycin Propionate in the mobile phase to make 50 mL, and use this System performance: Dissolve 5 mg of Josamycin Pro-pionate and 2 mg of josamycin in 50 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, josamycin and josamycin propionate are eluted in this order with the resolu-tion between these peaks being not less than 25. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of josamycin propionate is not more than 1.5z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium ii in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.9 to 8.1 after sterilization. Standard solutions—Weigh accurately an amount of Josamycin Propionate RS, equivalent to about 20 mg (po-tency), dissolve in 10 mL of methanol, add 1/15 mol/L phosphate buffer solution (pH 5.6) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below, and use within 3 days. Take exactly a suitable amount of the standard stock solution before use, add 1/15 mol/L phosphate buffer solu-tion (pH 5.6) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as ‌JP XVIII Official Monographs / Kainic Acid and Santonin Powder 1215‌‌ the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Josamycin Propionate, equivalent to about 20 mg (potency), dissolve in 10 mL of methanol, add 1/15 mol/L phosphate buffer solution (pH 5.6) to make exactly 50 mL. Take ex-actly a suitable amount of this solution, add 1/15 mol/L phosphate buffer solution (pH 5.6) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Josamycin is a macrolide substance having antibac-terial activity produced by the growth of Streptomyces narbonensis var. josamyceticus. It contains not less than 900 mg (potency) and not more than 1100 mg (potency) per mg, calculated on the mL, and use this solution as the sample solution.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium ii in 3) Medium for other organisms under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 7.9 to 8.1 after sterilization. Standard solutions—Weigh accurately an amount of Josamycin RS, equivalent to about 30 mg (potency), dissolve in 5 mL of methanol, add water to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below, and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add water to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Josamycin, equivalent to about 30 mg (potency), dissolve in 5 mL of methanol, and add water to make exactly 100 mL. Take exactly a suitable amount of this solution, add water to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concen-tration sample solution and the low concentration sample so-lution, respectively.
Juzentaihoto Extract contains not less than 1.
Juzentaihoto Extract is a light brown to brown powder or black-brown viscous extract. It has a slight odor and a sweet and bitter taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 15 mL of sodium hydroxide TS, centrifuge, and separate the supernatant liquid. To the liquid add 10 mL of 1-butanol, shake, centrifuge, and separate the 1-butanol layer. To the 1-butanol layer add 10 mL of water, shake, centrifuge, and separate the 1-butanol layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzalde-hyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the dark brown spot from the standard solu-tion (Ginseng). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 15 mL of sodium hydroxide TS, centrifuge, and separate the supernatant liquid. To the liquid add 10 mL of solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-brown spot from the standard solution (Astragalus Root). For preparation prescribed Atractylodes Rhizome—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 5 mL of diethyl ether, shake, and centrifuge. Use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—Shake 5.0 g of the dry extract (or 15.0 g of the vis-cous extract) with 10 mL of water, add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 40 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylaminobenzalde-hyde TS for spraying, heated at 1059C for 5 minutes, and al-lowed to cool (Atractylodes Lancea Rhizome). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 15 mL of water and 5 mL of 0.1 mol/L hydrochloric acid TS, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, use (Z)-ligustilide TS for thin-layer chromato-graphy as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromato-graphy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 Juzentaihoto Extract / Crude Drugs and Related Drugs nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solution (Cnidium Rhizome; Japanese Angelica Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Peony Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 30 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sam-ple solution. Perform the test with the sample solution as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of water, methanol and 1-butanol (1:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: a dark green spot is observed at an Rf value of about 0.6 (Rehmannia Root). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 50 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, diethyl ether and methanol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample solution. Separately, dissolve 1 mg of (E )-2-methoxycinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air- dry the plate. Examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution. Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Ginsenoside Rb1—Weigh accurately about 2 g of the dry extract (or an amount of the viscous extract, equivalent to about 2 g of the dried substance), add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 15 mL of diluted methanol (3 in 5), and repeat the same proce-dure. Combine the supernatant liquids, add diluted metha-nol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solu-tion, add 3 mL of sodium hydroxide TS, allow to stand for 30 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 20 mL. Apply exactly 5 mL of this solution to a column (about 10 mm in inside diameter and packed with 0.36 g of octadecylsilanized silica gel for pre-treatment (55 – 105 mm in particle size), washed just be-fore use with methanol and then with diluted methanol (3 in 10)), and wash the column in sequence with 2 mL of diluted methanol (3 in 10), 1 mL of sodium carbonate TS and 10 mL of diluted methanol (3 in 10). Finally, elute with methanol to collect exactly 5 mL, and use this as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of gin-senoside Rb1 in each solution. JP XVIII Crude Drugs and Related Drugs / Juzentaihoto Extract 2045 Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS × 1/5 with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak MS: Amount (mg) of Ginsenoside Rb1 lated on the anhydrous basis Operating conditions— RS taken, calcu- area of paeoniflorin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with carbamoyl groups bound silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (400:100:1). Flow rate: 1.0 mL per minute (the retention time of gin-senoside Rb1 is about 16 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ginsenoside Rb1 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z. (2) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this ‌2046‌ Kakkonto Extract / Crude Drugs and Related Drugs solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Kainic Acid Hydrate, when dried, contains not less than 99.
Kainic Acid Hydrate occurs as white, crystals or crystalline powder. It is odorless, and has an acid taste. It is sparingly soluble in water and in warm water, very slightly soluble in acetic acid (100) and in ethanol (95), and practically insoluble in diethyl ether. It dissolves in dilute hydrochloric acid and in sodium hy-droxide TS. The pH of a solution of 1.0 g of Kainic Acid Hydrate in 100 mL of water is between 2.8 and 3.5. Melting point: about 2529C (with decomposition).
(1) To 5 mL of a solution of Kainic Acid Hydrate (1 in 5000) add 1 mL of ninhydrin TS, and warm in a water bath at a temperature between 609C and 709C for 5 minutes: a yellow color is produced. Dissolve 50 mg of Kainic Acid Hydrate in 5 mL of acetic acid (100), and add 0.5 mL of bromine TS: the color of bromine disappears immediately. D Optical rotation <2.49> [a]20: －13 – －179(0.5 g, water, 50 directed above (not more than 0.021z). Sulfate <1.14>—Dissolve 0.5 g of Kainic Acid Hy-drate in 40 mL of water by warming. Cool, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Take 0.25 g of Kainic Acid Hy- drate, and perform the test. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of Kainic Acid Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of Kainic Acid Hy- drate in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 2 ppm). Amino acid and other imino acid—Dissolve 0.10 g of Kainic Acid Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test as directed under Thin-layer Chromatography <2.03> with these solu-tions. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with the supernatant liquid of a mix-ture of water, 1-butanol and acetic acid (100) (5:4:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and dry the plate at 809C for 5 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> 6.5 – 8.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.4 g of Kainic Acid Hy-drate, previously dried, and dissolve in 50 mL of warm water, cool and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (indicator: 10 drops of bromothymol blue TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 21.32 mg of C10H15NO4
Kakkonto Extract contains not less than 7 mg and not more than 21 mg (for preparation prescribed 3 g of Ephedra Herb) or not less than 10 mg and not more than 30 mg (for preparation prescribed 4 g of Ephedra Herb) of total alkaloids (ephedrine and pseudoephe-drine), not less than 14 mg and not more than 56 mg (for preparation prescribed 2 g of Peony Root) or not less than 21 mg and not more than 84 mg (for prepara-tion prescribed 3 g of Peony Root) of paeoniflorin (C23H28O11: 480.
Kakkonto Extract occurs as a light brown to brown powder or black-brown viscous extract. It has a char-acteristic odor, and a sweet first, then hot, and slightly bitter taste.
(1) To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Puerarin RS or puerarin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution (Pueraria Root). To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (4:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot is observed at an Rf value of about 0.5 (Ephedra Herb). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect an apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract), add 10 mL of water, shake, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample so-lution. Separately, dissolve 1 mg of (E )-2-methoxycinnamal-dehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu- JP XVIII Crude Drugs and Related Drugs / Kakkonto Extract 2047 tion. To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatog-raphy in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Peony Root). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed in the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Total alkaloids (ephedrine and pseudoephe-drine)—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centri-fuge, and separate the diethyl ether layer. In addition, repeat twice in the same manner for the aqueous layer using 1.0 mL of ammonia TS and 20 mL of diethyl ether. Combine all the extracts, evaporate the solvent under low pressure (in va-cuo), dissolve the residue in diluted methanol (1 in 2) to make exactly 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of ephedrine hydrochloride for assay of crude drugs, previously dried at 1059C for 3 hours, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine the peak areas, ATE and ATP, of ephe-drine and pseudoephedrine obtained with the sample solu-tion, and the peak area, AS, of ephedrine with the standard solution. Amount (mg) of total alkaloids (ephedrine and pseudoephe-drine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drugs taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, and add 650 mL of water and 1 mL of phosphoric acid to dissolve sodium lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of ephe-drine is about 27 minutes). System suitability— System performance: Dissolve 1 mg each of ephedrine hy-drochloride for assay of crude drugs and pseudoephedrine hydrochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephe-drine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent Kakkonto Extract / Crude Drugs and Related Drugs to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, flow through in a column packed with 2 g of polyamide for column chromatography, elute with 20 mL of water, add 1 mL of acetic acid (100) and water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be- Amount (mg) of paeoniflorin (C23 ＝ MS × AT/AS × 5/8 H28O 11) tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave- glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about ‌JP XVIII Crude Drugs and Related Drugs / Kakkontokasenkyushin'i Extract 2049 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Kakkontokasenkyushin'i Extract contains not less than 9.
Kakkontokasenkyushin'i Extract occurs as a light brown to brown powder or black-brown viscous ex-tract, having a characteristic order, and a sweet first, then a bitter and hot taste.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Puerarin RS or puerarin for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the stand-ard solution (Pueraria Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solution as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (4:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot is observed at an Rf value of about 0.5 (Ephedra Herb). Perform the test according to the following (i) or (ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect the apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E)-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromato-graphy <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample so-lution. Separately, dissolve 1 mg of (E)-2-methoxycinnamal-dehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu-tion. Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzoaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple to purple spot from the standard solution (Peony Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample Kakkontokasenkyushin'i Extract / Crude Drugs and Related Drugs solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 15 mL of water and 5 mL of 0.1 mol/L hydrochloric acid TS, and then shake with 25 mL of diethyl ether. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solu-tion. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Cnidium Rhizome). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, to 1 g of powdered magnolia flower add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate and hexane (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the brown spot (Rf value: about 0.4) from the standard solution (Magnolia Flower).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of dried substance) as directed in Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Total alkaloids (ephedrine and pseudoephed-rine)—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. In addition, repeat twice in the same manner for the aqueous layer using 1.0 mL of am-monia TS and 20 mL of diethyl ether. Combine the extracts, evaporate the solvent under low pressure (in vacuo), dissolve the residue in diluted methanol (1 in 2) to make exactly 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of ephe-drine hydrochloride for assay of crude drugs, previously dried at 1059C for 3 hours, dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, ATE and ATP, of ephedrine and pseudoephedrine with the sample solution, and peak area, AS, of ephedrine with standard solution. Amount (mg) of total alkaloids (ephedrine and pseudoe-phedrine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drugs taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, then add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of ephe-drine is about 27 minutes). JP XVIII Crude Drugs and Related Drugs / Kakkontokasenkyushin'i Extract 2051 System suitability— System performance: Dissolve 1 mg each of ephedrine hy-drochloride for assay of crude drugs and pseudoephedrine hydrochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephe-drine are eluted in this order with the resolution between these peaks being not less than 1.5. mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) area of ephedrine is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, flow through in a column packed with 2 g of polyamide for column chromatography, elute with 20 mL of water, add 1 mL of acetic acid (100) to the ef-fluent, then add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dis-solve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 5/8 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. ‌2052 Kamikihito Extract / Crude Drugs and Related Drugs Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Magnoflorine—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether, shake, add 3.0 mL of diluted sodium hydroxide TS (1 in 10), shake for 10 minutes, centrifuge, and remove the diethyl ether layer. Add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 3.0 mL of 0.1 mol/L hydrochloric acid TS and 20 mL of diluted methanol (1 in 2), shake for 15 minutes, centrifuge, and separate the superna-tant liquid. To the residue add 20 mL of diluted methanol (1 in 2) shake for 15 minutes, centrifuge, and separate the su-pernatant liquid. Combine the previous supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of magnoflorine iodide for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of magnoflorine in each solution. Amount (mg) of magnoflorine [as magnoflorine iodide (C20H24INO4)] ＝ MS × AT/AS × 1/20 MS: Amount (mg) of magnoflorine iodide for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 303 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, then add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of mag-noflorine is about 20 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of magnoflorine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of magnoflorine is not more than 1.5z.
Kallidinogenase is an enzyme obtained from healthy porcine pancreas, and has kinin-releasing activity based on cleavage of kininogen. It contains not less than 25 Kallidinogenase Units per mg.
Kallidinogenase occurs as a white to light brown powder. It is odorless or has a faint, characteristic odor. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. The pH of a solution of Kallidinogenase (1 in 300) is be-tween 5.5 and 7.5.
(1) Weigh accurately an appropriate amount of Kallidinogenase according to the labeled Units, and dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to prepare a solution containing 10 Kallidinogenase Units per mL. Pipet 5 mL of this solution, and add exactly 1 mL of trypsin inhibitor TS and 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Pipet 4 mL each of this solution into two separate test tubes, add exactly 1 mL each of aprotinin TS and 0.05 mol/L phosphate buffer solu-tion (pH 7.0) separately to each test tube, allow them to stand at room temperature for 20 minutes, and use these so-lutions as the sample solutions 1 and 2. Separately, pipet 1 mL of trypsin inhibitor TS, and add 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Pipet 4 mL each of this solution into two separate test tubes, add exactly 1 mL each of aprotinin TS and 0.05 mol/L phosphate buffer solution (pH 7.0) separately to each tube, allow them to stand at room temperature for 20 minutes, and use these so-lutions as the sample solutions 3 and 4. Then, pipet 2.5 mL of substrate TS for kallidinogenase assay (1), previously warmed at 30.0 ± 0.59C for 5 minutes, place in a 1-cm cell, add exactly 0.5 mL of the sample solution 1 warmed at 30.0 ± 0.59C for 5 minutes, and start simultaneously a chrono-graph. Perform the test at 30.0 ± 0.59C as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the blank, and determine the absorbances at 405 nm, A1–2 and A1–6, of this solution, after having allowed it to stand for exactly 2 and 6 minutes. Perform the same test with the sample solutions 2, 3 and 4, and determine the absorbances, A2–2, A2–6, A3–2, A3–6, A4–2 and A4–6, of these solutions. Calculate I by using the following equation: the value of I does not exceed 0.2. I ＝ (A1–6 － A1–2) － (A3–6 － A3–2) (A2–6 － A2–2) － (A4–6 － A4–2) (2) Pipet 2.9 mL of substrate TS for kallidinogenase assay (2), previously warmed at 30.0 ± 0.59C for 5 minutes, place in a 1-cm cell, add exactly 0.1 mL of the sample solu-tion obtained in the Assay, and start simultaneously a chronograph. Perform the test at 30.0 ± 0.59C as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the change of the absorbance at 253 nm for 4 to 6 minutes. Separately, pipet 1 mL of trypsin inhibitor TS, and add 0.05 mol/L phosphate buffer solution (pH 7.0) to make JP XVIII Official Monographs / Kallidinogenase 1217 exactly 10 mL. Add exactly 0.1 mL of this solution to exactly 2.9 mL of substrate TS for kallidinogenase assay (2), previ-ously warmed at 30.0 ± 0.59C for 5 minutes, and use this solution as the blank. If the rate of change in the absorbance remains constant, determine the change of absorbance per 1 minute, A, and calculate R by using the following equation: the value of R is between 0.12 and 0.16. R ＝ A/0.0383 × 1/(a × b) a: Amount (mg) of Kallidinogenase in 1 mL of the sample solution b: Amount (Unit) of kallidinogenase in 1 mg of Kalli-dinogenase obtained in the Assay Specific activity Perform the test with Kallidinogenase as directed under Nitrogen Determination <1.08> to determine the nitrogen content, convert 1mg of nitrogen (N:14.01) into 6.25 mg of protein, and calculate the specific activity using the amount (Units) of Kallidinogenase obtained in the Assay: it is not less than 100 Kallidinogenase Units per 1mg of protein.
(1) Fat—To 1.0 g of Kallidinogenase add 20 mL of diethyl ether, extract with occasional shaking for 30 minutes, and filter. Wash the residue with 10 mL of diethyl ether, combine the washing with the filtrate, evaporate the diethyl ether, and dry the residue at 1059C for 2 hours: the mass of phate buffer solution for microplate washing, and remove. Repeat this procedure more 4 times, take off the washings thoroughly, add 100 mL of substrate solution for peroxidase determination, and allow to stand at a constant temperature of about 259C for exactly 30 minutes while protecting from light. Then add 100 mL of diluted sulfuric acid (23 in 500), shake, and determine the absorbance at 490 – 492 nm. Sepa-rately, dissolve a suitable amount of bradykinin in gelatin-phosphate buffer solution (pH 7.0) to make solutions con-taining exactly 100 ng, 25 ng, 6.25 ng, 1.56 ng, 0.39 ng and 0.098 ng of bradykinin per mL, and use these solutions as the standard solution (1), the standard solution (2), the standard solution (3), the standard solution (4), the standard solution (5) and the standard solution (6), respectively. Use 1 mL of gelatin-phosphate buffer solution (pH 7.0) as the standard solution (7). To each of the well add 50 mL each of the standard solutions and 100 mL of trichloroacetic acid-gelatin-tris buffer solution, and proceed in the same manner as for the sample solution and for the control solution. Prepare the calibration curve from the amounts of bradykinin in the standard solutions and their absorbances, and calculate the amounts of bradykinin, BT (pg) and BS (pg), of the sample solution and the control solution. (vi) Judgment: The value R calculated by the following equation is not less than 0.8. R ＝ B /B the residue is not more than 1 mg. T S Kininase— Bradykinin solution: Weigh an appropriate amount of bradykinin, and dissolve in gelatin-phosphate buffer solu-tion (pH 7.4) to prepare a solution containing 0.200 mg of bradykinin per mL. Kallidinogenase solution: Weigh accurately a suitable amount of Kallidinogenase according to the labeled unit, dis-solve in gelatin-phosphate buffer solution (pH 7.4) to make a solution containing 1 unit of kallidinogenase per mL. Sample solution: Pipet 0.5 mL of bradykinin solu-tion, warm at 30 ± 0.59C for 5 minutes, then add exactly 0.5 mL of kallidinogenase solution previously warmed at 30 ± 0.59C for 5 minutes, and mix immediately. After allowing this solution to stand at 30 ± 0.59C for exactly 150 seconds, add exactly 0.2 mL of a solution of trichloroacetic acid (1 in 5), and shake. Boil for 3 minutes, then cool in ice immedi-ately, centrifuge, and allow to stand at a room temperature for 15 minutes. Pipet 0.5 mL of the supernatant liquid, add exactly 0.5 mL of gelatin-tris buffer solution (pH 8.0), and shake. Pipet 0.1 mL of this solution, add exactly 0.9 mL of trichloroacetic acid-gelatin-tris buffer solution, and mix. Pipet 0.2 mL of this solution, add exactly 0.6 mL of trichloroacetic acid-gelatin-tris buffer solution, shake, and use this solution as the sample solution. Control solution: Proceed with 0.5 mL of gelatin-phosphate buffer solution (pH 7.4) as described in (iii), and use the solution so obtained as the control solution. Procedure: Add 0.1 mL of anti-bradykinin antibody TS to goat anti-rabbit IgG antibody-coated wells of a 96-well microplate, shake, and allow to stand at a constant tempera-ture of about 259C for 1 hour. Remove the anti-bradykinin antibody TS, add 0.3 mL of phosphate buffer solution for microplate washing to the wells, then remove. Repeat this procedure 3 times, take off the washings thoroughly, then add 100 mL each of the sample solution and control solution, and 50 mL of gelatin-phosphate buffer solution (pH 7.0), shake, and allow to stand at a constant temperature of about 259C for 1 hour. Then add 50 mL of peroxidase-labeled bradykinin TS, shake, and allow to stand in a cold place for a night. Take off the reaction solution, add 0.3 mL of phos- Trypsin-like substances—Pipet 4 mL of the sample stock solution prepared for the Assay, add exactly 1 mL of trypsin inhibitor TS and 0.05 mol/L phosphate buffer solu-tion (pH 7.0) to make exactly 10 mL, and use this solution as the sample solution. Pipet 2.5 mL of substrate TS for kal-lidinogenase assay (1), previously warmed at 30 ± 0.59C for 5 minutes, place in a 1-cm cell, add exactly 0.5 mL of the sample solution, warmed at 30 ± 0.59C for 5 minutes, and start simultaneously a chronograph. Perform the test at 30 ± 0.59C as directed under Ultraviolet-visible Spectropho-tometry <2.24> using water as the blank, and determine the absorbances at 405 nm, A2 and A6, of this solution after hav-ing allowed it to stand for exactly 2 and 6 minutes. Sepa-rately, pipet 4 mL of the sample stock solution prepared for the Assay, add 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL, and use this solution as the con-trol solution. Perform the same test with the control solu-tion, and determine the absorbances, A?2 and A?6. Calculate T by using the following equation: the value of T does not exceed 0.05. T ＝ {(A?6 － A?2 ) － (A6 － A2)}/(A?6 － A?2 ) Protease—Weigh accurately an appropriate amount of Kallidinogenase according to the labeled Units, dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to prepare a solution containing 1 Kallidinogenase Unit per mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, place in a test tube, and allow to stand at 35 ± 0.59C for 5 minutes. Then, pipet 5 mL of substrate TS for kallidinogenase assay (3), previously warmed to 35 ± 0.59C, add quickly to the sample solution in the test tube, and allow to stand at 35 ± 0.59C for exactly 20 minutes. Then add ex-actly 5 mL of trichloroacetic acid TS, shake well, allow to stand at room temperature for 1 hour, and filter through a membrane filter (5 mm in pore size). Discard the first 3 mL of the filtrate, and determine the absorbance, A, of the sub-sequent filtrate at 280 nm within 2 hours as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Separately, pipet 1 mL of the sample solution, add exactly 5 mL of trichloroacetic acid TS, shake well, and ‌1218 add exactly 5 mL of the substrate TS for kallidinogenase assay (3). Proceed in the same manner as described for the sample solution, and determine the absorbance, A0, of this solution. Calculate the value of (A–A0): it is not more than 0.2. Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu- buffer solution (pH 7.0) to make exactly 10 mL. Pipet 0.5 mL of this solution, perform the test in the same manner as described for the sample solution, and determine the absor-bances, AO2 and AO6, of the solution after allowing to stand for exactly 2 and 6 minutes. Units per 1 mg of Kallidinogenase um, phosphorus (V) oxide, 4 hours). (A － A ) － (A － A ) M 1 ＝ T6 T2 O6 O2 × S × Residue on ignition <2.44> Not more than 3z (0.5 g, 650 – 7509C). Kinin-releasing activity Kallidinogenase solution: Weigh accurately a suitable amount of Kallidinogenase, according to the labeled unit, dissolve in 0.02 mol/L phosphate buffer solution (pH 8.0) to make a solution containing 0.1 unit of kallidinogenase per mL. Perform this procedure by using glassware. Sample solution: Pipet 0.5 mL of kininogen TS, warm at 30 ± 0.59C for 5 minutes, then add exactly 0.5 mL of kallidinogenase solution previously warmed at 30 ± 0.59C for 5 minutes, and mix immediately. After allow this solution to stand at 30 ± 0.59C for exactly 2 minutes, add exactly 0.2 mL of a solution of trichloroacetic acid (1 in 5), and shake. Boil for 3 minutes, then cool in ice immediately, centrifuge, and allow to stand at a room temperature for 15 minutes. Pipet 0.5 mL of the supernatant liquid, add exactly 0.5 mL of gelatin-tris buffer solution (pH 8.0), and shake. Pipet 0.1 mL of this solution, add exactly 1.9 mL of trichloroacetic acid-gelatin-tris buffer solution, shake, and use this solution as the sample solution. Procedure: Perform the test with the sample solu-tion as directed in the Purity (2), and determine the amount, B (pg), of kinin per well. The kinin-releasing activity per 1 unit of Kallidinogenase calculated by the following equation is not less than 500 ng bradykinin equivalent/minute/unit. Kinin-releasing activity (ng bradykinin equivalent/minute/ unit) per 1 unit of Kallidinogenase ＝ B × 4.8
Weigh accurately an appropriate amount of Kal-lidinogenase according to the labeled Units, dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to prepare a solu-tion containing about 10 Kallidinogenase Units per mL, and use this solution as the sample stock solution. Pipet 4 mL of the sample stock solution, add exactly 1 mL of trypsin inhi-bitor TS and 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL, and use this solution as the sample solution. Pipet 2.5 mL of substrate TS for kallidinogenase assay (1), previously warmed at 30 ± 0.59C for 5 minutes, place in a 1-cm cell, add exactly 0.5 mL of the sample solu-tion, warmed at 30 ± 0.59C for 5 minutes, and start simul-taneously a chronograph. Perform the test at 30 ± 0.59C as directed under the Ultraviolet-visible Spectrophotometry <2.24> using water as the blank, and determine the absor- bances at 405 nm, AT2 and AT6, of this solution after allowing to stand for exactly 2 and 6 minutes. Separately, dissolve Kallidinogenase RS in 0.05 mol/L phosphate buffer solution (pH 7.0) to make a solutin so that each mL contains exactly 10 Units, and use this solution as the standard stock solution. Pipet 4 mL of the stock solution, add exactly 1 mL of trypsin inhibitor TS and 0.05 mol/L phosphate buffer so-lution (pH 7.0) to make exactly 10 mL, and use this solution as the standard solution. Take exactly 0.5 mL of the stand-ard solution, perform the test in the same manner as described for the sample solution, and determine the absor-bances, AS2 and AS6, of the solution after allowing to stand for exactly 2 and 6 minutes. Separately, take exactly 1 mL of the trypsin inhibitor TS, and add 0.05 mol/L phosphate (AS6 － AS2) － (AO6 － AO2) a b MS: Amount (Units) of Kallidinogenase RS taken a: Volume (mL) of the standard stock solution b: Amount (mg) of Kallidinogenase in 1 mL of the sample stock solution Cantainers and storage Containers—Tight containers. Kanamycin Monosulfate
Kamikihito Extract contains not less than 0.
Kamikihito Extract is a light yellow-brown to brown powder or black-brown viscous extract. It has a slightly odor, and a slightly sweet, hot and bitter taste.
(1) To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL of sodium hydroxide TS, shake, centrifuge, and separate the supernatant liquid. To the liquid add 10 mL of 1-butanol, shake, centrifuge, and separate the 1-butanol layer. To this layer add 10 mL of water, shake, centrifuge, and separate the 1-butanol layer. Evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of methanol, and use the solution as the sample solution. Separately, dissolve 1 mg of ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) JP XVIII Crude Drugs and Related Drugs / Kamikihito Extract 2053 (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). For preparation prescribed Atractylodes Rhizome—To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of diethyl ether, and use the solution as the sample solution. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the red to red-purple spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), then dissolve the residue in 2 mL of hexane, and use the solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark violet spot is observed at an Rf value of about 0.5, and this spot exhibits greenish brown when the plate is sprayed evenly 4-dimethylaminoben-zaldehyde TS for spraying, heated the plate at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhi-zome). To 3.0 g of the dry extract (or 9.0 g of the viscous ex-tract) add 15 mL of sodium hydroxide TS, shake, centrifuge, and separate the supernatant liquid. To the liquid add 10 mL of 1-butanol, shake, centrifuge, and separate 1-butanol layer. To the 1-butanol layer add 10 mL of water, shake, centrifuge, separate the 1-butanol layer, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue in 2 mL of methanol, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of astragaloside IV for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 2-propanol, water and ammonia solution (28) (9:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to blue-purple spot from the standard solution (Astragalus Root). To 3.0 g of the dry extract (or 9.0 g of the viscous ex tract) add 15 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then add 2 mL of diethyl ether to the residue, and use the solution as the sam-ple solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromato-graphy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of butyl acetate and hexane (2:1) to a distance of about 7 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Japanese Angelica Root). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 30 mL of 1 mol/L hydrochloric acid TS, and heat for 10 minutes. After cooling, to 10 mL of this solution add 10 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the sample solution. Separately, to 2.0 g of a powder of polygala root add 30 mL of 1 mol/L hydrochloric acid TS, and heat for 10 minutes. After cooling, to 10 mL of this solution add 10 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol and acetic acid (100) (7:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, heat the plate at 1059C for 1 minute, and observe while hot: one of the several spots obtained from the sample solution has the same color tone and Rf value with the pur-plish red spot (at an Rf value of about 0.5) from the stand-ard solution (Polygala Root). To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL of sodium hydroxide TS, shake, centri-fuge, and separate the supernatant liquid. To the liquid add 10 mL of 1-butanol, shake, centrifuge, and separate the 1-butanol layer. To this layer add 10 mL of water, shake, cen-trifuge, and separate the 1-butanol layer. Evaporate the sol-vent under low pressure (in vacuo), dissolve the residue in 2 mL of methanol, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminoben-zaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography Kamikihito Extract / Crude Drugs and Related Drugs <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol, ammonia solution (28) (6:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzalde-hyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minute: one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solution (Garde-nia Fruit). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL of water, shake, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, to 1.0 g of a powder of saus-surea root add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 2 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (Saus-surea Root). To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL of water, shake, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with of water, shake, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, dissolve 1 mg of paeonol for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane and diethyl ether (5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the orange spot from the standard solution (Moutan Bark).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not less than 8.0z, calculated on the dried basis. However, for the dry extract prepared by adding Light Anhydrous Silicic Acid, between 9.0z and 18.0z.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. Centrifuge, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the same manner as above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centri-fuge, and separate the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, cen-trifuge, and separate the supernatant liquid. Combine all the supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of saikosaponin b2 in each solution. these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution and 5 mL of Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). For preparation prescribed Moutan Bark—To 3.0 g of the dry extract (or 9.0 g of the viscous extract) add 15 mL CS: Concentration (mg/mL) of saikosaponin b2 in sai-kosaponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. JP XVIII Crude Drugs and Related Drugs / Kamishoyosan Extract 2055 Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (retention time of sai-kosaponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. ‌factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Geniposide—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of geniposide for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. Amount (mg) of geniposide ＝ MS × AT/AS × 1/2 MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (900:100:1). Flow rate: 1.0 mL per minute (retention time of genipo-side is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of geniposide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Kamishoyosan Extract contains not less than 28 mg and not more than 84 mg of paeoniflorin (C23H28O11: 480.
Kamishoyosan Extract occurs as a yellow-brown to brown powder or black-brown viscous extract. It has slightly a characteristic odor, and a sweet, slightly hot, then bitter taste. Identiˆcation (1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultrav-iolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solution (Japanese Angelica Root). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of metha-nol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of albiflorin in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool for more than 30 minutes, and examine under ultravio-let light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange fluorescent spot from the standard solution (Peony Root). For preparation prescribed Atractylodes Rhizome—To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sam-ple solution. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the red spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying, heat the plate at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhi-zome). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of sai-kosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and examine under ul-traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 15 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane and diethyl ether (5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange spot from the stand-ard solution (Moutan Bark). To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the JP XVIII Crude Drugs and Related Drugs / Kamishoyosan Extract 2057 sample solution. Separately, dissolve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, methanol and ammonia solution (28) (6:3:2) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Gardenia Fruit). To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of diluted phosphoric acid (1 in 30), shake, then add 15 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the sample solution. Sepa-rately, shake 0.2 g of pulverized mentha herb with 10 mL of diluted phosphoric acid (1 in 30), add 15 mL of ethyl acetate, shake, centrifuge, and use the ethyl acetate layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, ethyl acetate, water and acetic acid (100) (10:10:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-brown spot (around Rf value 0.4) from the standard solution (Mentha Herb).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Geniposide—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of geniposide for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu- ‌2058 Keishibukuryogan Extract / Crude Drugs and Related Drugs tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr- Amount (mg) of geniposide ＝ MS × AT/AS × 1/2 rhizic acid is about 15 minutes). System suitability— MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (900:100:1). Flow rate: 1.0 mL per minute (the retention time of geniposide is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of geniposide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as de-scribed above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Kanamycin Monosulfate is the sulfate of an amino-glycoside substance having antibacterial activity pro-duced by the growth of Streptomyces kanamyceticus. It contains not less than 750 mg (potency) and not more than 832 mg (potency) per mg, calculated on the dried basis.
Kanamycin Monosulfate occurs as a white crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 50 mg of Kanamycin Monosulfate in 3 mL of water, and add 6 mL of anthrone TS: a blue-purple color develops. Dissolve 20 mg each of Kanamycin Monosulfate and Kanamycin Monosulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with the su-pernatant layer of a mixture of chloroform, ammonia solu-tion (28) and methanol (2:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of 0.2z nin- ‌JP XVIII Official Monographs / Kanamycin Sulfate 1219 hydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show a purple-brown color and the same Rf value. To a solution of Kanamycin Monosulfate (1 in 5) add 1 drop of barium chloride TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋112 – ＋1239(0.2 g calcu-lated on the dried basis, water, 20 mL, 100 mm). Sulfuric acid Weigh accurately about 0.25 g of Kanamycin Monosulfate, dissolve in 100 mL of water, adjust the pH to 11.0 with ammonia solution (28), add exactly 10 mL of 0.1 mol/L barium chloride VS, and titrate <2.50> with 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution, blue-purple, disappears (indicator: 0.5 mg of phthalein purple). At a near of the end-point add 50 mL of ethanol (99.5). Perform a blank determination in the same manner. The amount of sulfuric acid (SO4) is not less than 15.0z and not more than 17.0z, calculated on the dried basis. Each mL of 0.1 mol/L barium chloride VS ＝ 9.606 mg of SO4
(1) Heavy metals <1.07>—Proceed with 2.0 g of Kanamycin Monosulfate according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Kanamycin Monosulfate according to Method 4, and per-form the test (not more than 1 ppm). Related substances—Dissolve 0.30 g of Kanamycin Monosulfate in water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 45 mg of Kanamycin Monosulfate RS in water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in 1-butanol (1 in 100) on the plate, and heat the plate at 1109C for 10 minutes: the spot other than the the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Kanamycin Monosulfate, equivalent to about 20 mg (po-tency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Kanamycin Sulfate is the sulfate of an aminoglyco-side substance having antibacterial activity produced by the growth of Streptomyces kanamyceticus. It contains not less than 690 mg (potency) and not more than 740 mg (potency) per mg, calculated on the dried basis.
Kanamycin Sulfate occurs as a white to yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 20 mg each of Kanamycin Sul-fate and Kanamycin Monosulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, ammonia solution (28) and methanol (2:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show a purple-brown color and the same Rf value. (2) A solution of Kanamycin Sulfate (1 in 10) responds ‌1220 to Qualitative Test <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: ＋103 – ＋1159(0.5 g calcu-lated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Kanamycin Sulfate in 20 mL of water is between 6.0 and 7.5.
(1) Clarity and color of solution—Dissolve 1.5 g of Kanamycin Sulfate in 5 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.15. Heavy metals <1.07>—Proceed with 1.0 g of Kanamy- cin Sulfate according to Method 4, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Kanamycin Sulfate according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.30 g of Kanamycin Sulfate in water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 9.0 mg of Kanamycin Monosulfate RS in water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potas-sium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of nin-hydrin in 1-butanol (1 in 100) on the plate, and heat the plate at 1109C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Kanamycin Monosulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution between 5 and 159C and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solu-tion (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as Description Kanamycin Sulfate occurs as a white to yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5). Identification (1) Dissolve 20 mg each of Kanamycin Sul-fate and Kanamycin Monosulfate RS in 1 mL of water, and use these solutions as the sample solution and standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, ammonia solution (28) and methanol (2:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show a purple-brown color and the same Rf value. (2) A solution of Kanamycin Sulfate (1 in 10) responds ‌1220 to Qualitative Test <1.09> (1) for sulfate. D Optical rotation <2.49> [a]20: ＋103 – ＋1159(0.5 g calcu-lated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Kanamycin Sulfate in 20 mL of water is between 6.0 and 7.5. Purity (1) Clarity and color of solution—Dissolve 1.5 g of Kanamycin Sulfate in 5 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.15. Heavy metals <1.07>—Proceed with 1.0 g of Kanamy- cin Sulfate according to Method 4, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Kanamycin Sulfate according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.30 g of Kanamycin Sulfate in water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 9.0 mg of Kanamycin Monosulfate RS in water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potas-sium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of nin-hydrin in 1-butanol (1 in 100) on the plate, and heat the plate at 1109C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer having pH 7.8 to 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Kanamycin Monosulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 5 to 159C and use within 30 days. Take exactly a suitable amount of the standard stock solution be-fore use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Kanamycin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Aluminum Silicate Hydrate with Silicon Diox-ide occurs as white to light red powdered crystalline masses, which becomes easily fine powder on crushing. The powder is roughish and easily adheres to skin, and becomes slightly darken and obtains plasticity when moisten with water. It has a characteristic odor and almost tasteless. It feels like as sand of fine grains by chewing. Under a microscope <5.01>, the powder of Aluminum Sili- cate Hydrate with Silicon Dioxide, thoroughly grained be-tween a slide glass and a cover glass together with mounting medium, shows numbers of round to polygonal crystals not smaller than 10 mm in diameter.
To 0.5 g of powdered Aluminum Silicate Hydrate with Silicon Dioxide add 3 mL of diluted sulfuric acid (1 in 3), heat until white vapors evolve, then after cool-ing add 20 mL of water, and filter. The filtrate neutralized to be a weak acidity with ammonia TS responds to Qualitative Tests <1.09> (1), (2) and (4) for aluminum salt.
(1) Heavy metals <1.07>—To 1.5 g of Aluminum Silicate Hydrate with Silicon Dioxide add 50 mL of water and 5 mL of hydrochloric acid, and boil gently for 20 minutes while thorough shaking. After cooling, centrifuge, and separate the supernatant liquid. Wash the precipitate twice with 10 mL portions of water, centrifuging each time, and combine the supernatant liquids. Add ammonia solution (28) dropwise to the combined liquid until a slight precipitate form, then add, while shaking vigorously, dilute hydrochlo-ric acid dropwise to dissolve the precipitate. Add 0.45 g of hydroxylammonium chloride to this solution, heat, then after cooling add 0.45 g of sodium acetate trihydrate and 6 mL of dilute acetic acid, and add water to make 150 mL. Perform the test with 50 mL of this solution as the test solu-tion. Prepare the control solution by adding to 2.0 mL of Standard Lead Solution, 0.15 g of hydroxylammonium chlo-ride, 0.15 g of sodium acetate trihydrate and 2 mL of dilute acetic acid, and add water to make 50 mL (not more than 40 ppm). (2) Arsenic <1.11>—To 1.0 g of Aluminum Silicate Hy- drate with Silicon Dioxide add 5 mL of dilute hydrochloric acid, heat gently until boiling begins while shaking thor-oughly, then cool quickly, and centrifuge. To the precipitate add 5 mL of dilute hydrochloric acid, shake thoroughly, cen-trifuge, and take the supernatant liquid. Repeat this opera-tion with 10 mL of water, combine all the extracts, and con-centrate the extract to make 5 mL by heating on a water bath. Perform the test using this solution as the test solution (not more than 2 ppm).
Keishibukuryogan Extract contains not less than 0.
Keishibukuryogan Extract is a light brown to brown powder or black-brown viscous extract. It has a char-acteristic odor and has a taste slightly sweet first then bitter later.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample solution. Separately, dissolve 1 mg of (E )- JP XVIII Crude Drugs and Related Drugs / Keishibukuryogan Extract 2059 cinnamic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of hexane, ethyl acetate, for-mic acid and water (60:40:4:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Cinna-mon Bark). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample so-lution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and diethyl ether (5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange spot from the stand-ard solution (Moutan Bark). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of methanol, filter, and use the filtrate as the sample solution. Separately, dissolve 2 mg of amygda-lin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-propanol, ethyl acetate and water (4:4:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the green-brown spot from the standard solution (Peach Kernel). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sam-ple solution. Separately, dissolve 1 mg of albiflorin in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (6:3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool for more than 30 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the orange fluorescent spot from the standard solution (Peony Root).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis. However, for the dry extract prepared by adding Light Anhydrous Silicic Acid, between 9.0z and 18.0z.
(1) (E)-Cinnamic acid—Conduct this procedure using light-resistant vessels. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of (E)-cinnamic acid for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of (E)-cin-namic acid in each solution. Amount (mg) of (E)-cinnamic acid ＝ MS × AT/AS × 1/20 MS: Amount (mg) of (E)-cinnamic acid for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (750:250:1). Flow rate: 1.0 mL per minute (the retention time of (E)-cinnamic acid is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of (E)-cinnamic acid are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of (E)-cinnamic acid is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), ‌2060 Koi / Crude Drugs and Related Drugs dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Amygdalin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. Amount (mg) of amygdalin ＝ MS × AT/AS MS: Amount (mg) of amygdalin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amygdalin is not more than 1.5z.
Ketamine Hydrochloride occurs as white, crys-tals or crystalline powder. It is very soluble in formic acid, freely soluble in water and in methanol, sparingly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in acetic anhydride and in diethyl ether. A solution of Ketamine Hydrochloride (1 in 10) shows no optical rotation. Melting point: about 2589C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ketamine Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 3000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Keta-mine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Ketamine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> (2) for chloride. Arsenic <1.11>—Prepare the test solution with 1.0 g of Ketamine Hydrochloride, according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.5 g of Ketamine Hy-drochloride in 10 mL of methanol and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of cyclohexane and isopropylamine (49:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, dry the plate, and then spray evenly hydrogen peroxide TS: the spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more then 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Ketamine Hydro-chloride, previously dried, dissolve in 1 mL of formic acid, add 70 mL of a mixture of acetic anhydride and acetic acid (100) (6:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.42 mg of C13H16ClNO.HCl
Ketoconazole Lotion is an emulsion lotion. It contains not less than 93.
Ketoconazole Lotion occurs as a white emul-sion.
Shake well and take an amount of Ketocona-zole Lotion, equivalent to 0.1 g of Ketoconazole, add 20 mL of 2-propanol, shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 25 mg of ketoconazole in 5 mL of 2-propanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, methanol, water and am-monia solution (28) (40:40:25:2:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution.
Shake well and weigh accurately an amount of Keto-conazole Lotion, equivalent to about 25 mg of ketoconazole (C26H28Cl2N4O4), dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 4 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of ketoconazole for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add ex-actly 4 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of ketocona-zole to that of the internal standard. Amount (mg) of ketoconazole (C26H28Cl2N4O4) ＝ MS × QT/QS MS: Amount (mg) of ketoconazole for assay taken Internal standard solution—A solution of xanthone in meth-anol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To ammonium acetate solution (1 in 200) add acetic acid (100) to adjust the pH to 5.0. To 250 mL of this solution add 750 mL of methanol. Flow rate: Adjust so that the retention time of ketocona-zole is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ketoconazole are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ketoconazole to that of the internal stand-ard is not more than 1.0z.
Ketoconazole, when dried, contains not less than 99.
Ketoconazole occurs as a white to light yellow-white powder. It is soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Ketoconazole in methanol (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Ketoconazole in methanol (3 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: ‌1222 both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ketoconazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Ketoconazole as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Clarity and color of solution—Dissolve 1.0 g of Ketamine Hydrochloride in 5 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Keta- mine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Description Ketoconazole occurs as a white to light yellow-white powder. It is soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Ketoconazole in methanol (1 in 20) shows no optical rotation. Identification (1) Determine the absorption spectrum of a solution of Ketoconazole in methanol (3 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: ‌1222 both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ketoconazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Ketoconazole as directed under Flame Coloration Test <1.04> (2): a green color appears. Melting point <2.60> 148 – 1529C Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Ketoconazole according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related Substances—Dissolve 0.10 g of Ketoconazole in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 5 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of the peak other than ketoconazole obtained from the sample so-lution is not larger than 2/5 times the peak area of ketocona-zole from the standard solution, and the total area of the peaks other than ketoconazole from the sample solution is not larger than the peak area of ketoconazole from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Acetonitrile for liquid chromatography. Mobile phase B: A solution of tetrabutylammonium hydrogensulfate (17 in 5000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 5 → 50 95 → 50 10 – 15 Flow rate: 2.0 mL per minute. Time span of measurement: For 15 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add methanol to make exactly 20 mL. Confirm that the peak area of ketoconazole obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ketoconazole are not less than 40,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ketoconazole is not more than 2.5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Ketoconazole, pre-viously dried, dissolve in 70 mL of a mixture of 2-butanone and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.57 mg of C26H28Cl2N4O4
Ketoprofen, when dried, contains not less than 99.
Ketoprofen occurs as a white crystalline pow-der. It is very soluble in methanol, freely soluble in ethanol (95) and in acetone, and practically insoluble in water. A solution of Ketoprofen in ethanol (99.5) (1 in 100) shows no optical rotation. It is colored to pale yellow by light.
(1) Determine the absorption spectrum of a solution of Ketoprofen in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ketoprofen, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Ketoprofen in 10 mL of acetone: the solution is clear, and has no more color than the following control solution. Control solution: To a mixure of 0.6 mL of Cobalt (II) Chloride CS and 2.4 mL of Iron (III) Chloride CS add diluted hydrochloric acid (1 in 10) to make 10 mL. To 5.0 mL of this solution add diluted hydrochloric acid (1 in 10) to make 100 mL. Heavy metals <1.07>—Proceed with 2.0 g of Ketoprofen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure with a minimum of exposure to light, using light-resistant vessels. Dissolve 20 mg of Ketoprofen in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make ex-actly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the areas of the peaks, having the relative retention time of about 1.5 and about 0.3 to ketoprofen obtained from the sample solution, are not larger ‌JP XVIII Official Monographs / Ketotifen Fumarate 1225 than 4.5 times and not larger than 2 times the peak area of ketoprofen from the standard solution, respectively, the area of the peak other than ketoprofen and the peaks mentioned above from the sample solution is not larger than the peak area of ketoprofen from the standard solution, and the total area of these peaks is not larger than 2 times the peak area of ketoprofen from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 233 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Ketotifen Fumarate
Ketotifen Fumarate, when dried, contains not less than 99.
Ketotifen Fumarate occurs as a white to light yellow-white crystalline powder. It is sparingly soluble in methanol and in acetic acid (100), and slightly soluble in water, in ethanol (99.5) and in acetic anhydride. Melting point: about 1909C (with decomposition).
(1) Prepare the test solution with 30 mg of Ketotifen Fumarate as directed under Oxygen Flask Com-bustion Method <1.06> using 20 mL of water as the absorb-ing liquid: the test solution responds to Qualitative Tests <1.09> for sulfate. Determine the absorption spectrum of a solution of Ketotifen Fumarate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ketotifen Fumarate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.6 g of Ketotifen Fumarate in 2.5 mL of sodium carbonate TS in a crucible, heat on a water bath to dryness, and ignite at about 5009C. Dissolve the residue in 15 mL of water, filter if necessary, neutralize with diluted nitric acid (3 in 10), and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.25 mL of 0.01 mol/L hydrochloric acid VS add 2.5 mL of sodium carbonate TS, the used amount of diluted nitric acid (3 in 10) for the neutralization, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.015z). Heavy metals <1.07>—Proceed with 1.0 g of Ketotifen Fumarate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Ketotifen Fumarate in 10 mL of a mixture of methanol and ammonia TS (99:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of methanol ‌1226 and ammonia TS (99:1) to make exactly 25 mL. Pipet 1 mL of this solution, add a mixture of methanol and ammonia TS (99:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetonitrile, water and ammonia solution (28) (90:10:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying and then hydrogen peroxide TS on the plate: the number of the spot other than the principal spot obtained from the sam-ple solution is not more than four, and they are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Ketoprofen, previ-ously dried, dissolve in 25 mL of ethanol (95), add 25 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 25.43 mg of C16H14O3
Kitasamycin Acetate is a derivative of kitasamycin. It contains not less than 680 mg (potency) and not more than 790 mg (potency) per mg, calculated on the anhydrous basis.
Kitasamycin Acetate occurs as a white to light yellow-white powder. It is very soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Kitasamycin Acetate in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Kitasamycin Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Water <2.48> Not more than 5.0z (0.1 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solution—Weigh accurately an amount of Leucomycin A5 RS equivalent to about 30 mg (potency), dis-solve in 10 mL of methanol, add water to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 3 days. Take exactly a suitable amount of the stand-ard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solution—Weigh accurately an amount of Kitasamycin Acetate equivalent to about 30 mg (potency), dissolve in 25 mL of methanol, add water to make exactly 50 mL, shake well, and allow to stand at 37 ± 29C for 24 hours. Take exactly a suitable amount of the solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solu-tions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Kitasamycin Tartrate is the tartrate of kitasamycin. It contains not less than 1300 mg (potency) and not more than 1500 mg (potency) per mg, calculated on the anhydrous basis.
Kitasamycin Tartrate occurs as a white to light yellow-white powder. It is very soluble in water, in methanol and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Kitasamycin Tartrate in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Kitasamycin Tartrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 1 g of Kitasamycin Tartrate in 20 mL of water, add 3 mL of sodium hydroxide TS, add 20 mL of n-butyl acetate, shake well, and discard the n-butyl acetate layer. To the aqueous layer add 20 mL of n-butyl acetate, and shake well. The aqueous layer so obtained responds to Qualitative Tests <1.09> (1) for tartrate. pH <2.54> Dissolve 3.0 g of Kitasamycin Tartrate in 100 mL of water: the pH of the solution is between 3.0 and 5.0. Content ratio of the active principle Dissolve 20 mg of Kitasamycin Tartrate in diluted acetonitrile (1 in 2) to make 20 mL, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine the peak areas by the automatic in-tegration method, and calculate the amounts of leucomycin A5, leucomycin A4 and leucomycin A1 by the area percentage method: the amount of leucomycin A5 is 40 – 70z, leucomy-cin A4 is 5 – 25z, and leucomycin A1 is 3 – 12z. The rela-tive retention times of leucomycin A4 and leucomycin A1 to leucomycin A5 are about 1.2 and about 1.5, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To a suitable amount of a solution of am-monium acetate (77 in 5000) add diluted phosphoric acid (1 in 150) to adjust the pH to 5.5. To 370 mL of this solution add 580 mL of methanol and 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of leucomycin A5 is about 8 minutes. Time span of measurement: About 3 times as long as the retention time of leucomycin A5. System suitability— System performance: Dissolve about 20 mg each of Leuco-mycin A5 RS and Josamycin RS in 20 mL of diluted aceto-nitrile (1 in 2). When the procedure is run with 5 mL of this solution under the above operating conditions, leucomycin A5 and josamycin are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of leucomycin A5 is not more than 1.0z.
(1) Clarity and color of solution—Dissolve 1.0 g of Kitasamycin Tartrate in 10 mL of water: the solution is clear and colorless or light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Kitasamycin Tartrate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 30 ppm). Water <2.48> Not more than 3.0z (0.1 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of ‌1230‌ Leucomycin A5 RS, equivalent to about 30 mg (potency), dissolve in 10 mL of methanol, add water to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at not exceeding 59C, and use within 3 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer so-lution (pH 8.0) to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Kitasamycin Tartrate, equivalent to about 30 mg (potency), and dissolve in water to make exactly 100 mL. Take exactly a suitable amount of this solution, add phosphate buffer solu-tion (pH 8.0) to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration sample solution and the low concen-tration sample solution, respectively.
Kitasamycin is a mixture of macrolide substances having antibacterial activity produced by the growth of Streptomyces kitasatoensis. It contains not less than 1450 mg (potency) and not more than 1700 mg (potency) per mg, calculated on the anhydrous basis.
Kitasamycin occurs as a white to light yellow-white powder. It is very soluble in acetonitrile, in methanol and in ethanol (95), and practically insoluble in water.
Determine the absorption spectrum of a so-lution of Kitasamycin in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Content ratio of the active principle Dissolve 20 mg of Kitasamycin in diluted acetonitrile (1 in 2) to make 20 mL, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and measure each peak area by the automatic integra-tion method. Calculate the amounts of leucomycin A5, leu-comycin A4 and leucomycin A1 by the area percentage method: the amounts of leucomycin A5, leucomycin A4 and leucomycin A1 are 40 to 70z, 5 to 25z and 3 to 12z, re-spectively. Relative retention times of leucomycin A4 and leucomycin A1 to leucomycin A5 are about 1.2 and about 1.5, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To a volume of a solution of ammonium acetate (77 in 5000) add diluted phosphoric acid (1 in 150) to adjust to pH 5.5. To 370 mL of this solution add 580 mL of methanol and 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of leucomycin A5 is about 8 minutes. Time span of measurement: About 3 times as long as the retention time of leucomycin A5. System suitability— System performance: Dissolve about 20 mg each of Leu-comycin A5 RS and Josamycin RS in 20 mL of diluted aceto-nitrile (1 in 2). When the procedure is run with 5 mL of this solution under the above operating conditions, leucomycin A5 and josamycin are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of leucomycin A5 is not more than 1.0z. Water <2.48> Not more than 3.0z (0.1 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Leucomycin A5 RS equivalent to about 30 mg (potency), dis-solve in 10 mL of methanol, add water to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 3 days. Take exactly a suitable amount of the stand-ard stock solution before use, add phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration standard solution and the low concentra-tion standard solution, respectively. Sample solutions—Weigh accurately an amount of Kitasamycin equivalent to about 30 mg (potency), dissolve in 10 mL of methanol, and add water to make exactly 100 mL. Take exactly a suitable amount of the solution, add phos-phate buffer solution (pH 8.0) to make solutions so that each mL contains 30 mg (potency) and 7.5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Koi is a saccharized substance obtained by hydroly-sis of the starch of Zea mays Linneá(Gramineae), Manihot esculenta Crantz (Euphorbiaceae), Solanum tuberosum Linneá (Solanaceae), Ipomoea batatas Poiret (Convolvulaceae) or Oryza sativa Linneá (Gramineae), or the seed of Oryza sativa Linneáfrom which the seed coat is removed. Koi is prepared by the following processes 1 or 2, and contains mainly maltose, sometimes glucose and maltotriose also.
Koi 1: A white crystalline powder. It is odorless and has a sweet taste. Koi 2: Colorless or brown, clear or semi-translucent, masses or viscous liquid. It is odorless and has a sweet taste.
Dissolve exactly 0.50 g of Koi in a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve ex-actly 20.0 mg of maltose hydrate in a mixture of water and methanol (1:1) to make exactly 5 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography in equal size of circular spot each other. Develop the plate with a mixture of 2-butanone, water and acetic acid (100) (3:1:1) to a distance of about 7 cm, and dry at 1059C for 10 minutes the plate. Spray evenly 2,3,5-triphenyl-2H-tetrazolium chlo-ride-methanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the orange spot obtained from the standard solution, and it is larger and more intense than the spot from the standard solution.
(1) Clarity of solution—A solution obtained by dissolving 2.0 g of Koi in 20 mL of hot water is practically clear. Heavy metals <1.07> Koi 1: Proceed with 1.0 g of Koi 1 according to Method ‌JP XVIII Crude Drugs and Related Drugs / Purified Lanolin 2061‌ 1, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Koi 2: Proceed with 1.0 g of Koi 2 according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). ‌Arsenic <1.11>—Prepare the test solution with 1.0 g of Koi according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <5.01> Koi 1: Not more than 3.0z (1 g, 809C, 4 hours). Koi 2: Not more than 15.0z (1 g, 809C, 4 hours). In the case where the sample is in masses, crush the masses, weigh accurately the mass, and put in a desiccator. In the case in viscous liquid, put in a weighing bottle to spread about 1 mm thick, weigh accurately the mass, and put the bottle in a desiccator. Total ash <5.01> Not more than 0.5z.
L-Aspartic Acid, when dried, contains not less than 98.
L-Aspartic Acid occurs as white, crystals or crystalline powder. It is sparingly soluble in water, and practically insoluble in ethanol (99.5). It dissolves in dilute hydrochloric acid and in 0.2 mol/L sodium hydroxide TS.
Determine the infrared absorption spectrum of L-Aspartic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋24.0 – ＋26.09(2 g, after drying, 6 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> Dissolve 0.4 g of L-Aspartic Acid in 100 mL of water by warming, and allow to cool: between 2.5 and 3.5. Amount (mg) of calcium pantothenate (C H CaN O )
(1) Clarity and color of solution—Dissolve 1.0 g ＝ MS × QT/QS × 1/10 18 32 2 10 of L-Aspartic Acid in 20 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. MS: Amount (mg) of Calcium Pantothenate RS taken, cal-culated on the dried basis Internal standard solution—A solution of acetaminophen (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 200 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile for liquid chromatogra-phy (97:3). Flow rate: Adjust so that the retention time of pantothenic acid is about 3 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, pantothenic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of pantothenic acid to that of the internal standard is not more than 1.0z.
Weigh accurately about 0.15 g of L-Aspartic Acid, previously dried, dissolve in 50 mL of water by warming. After cooling, titrate <2.50> with 0.1 mol/L sodium hydrox-ide VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 13.31 mg of C4H7NO4
L-Carbocisteine, when dried, contains not less than 98.
L-Carbocisteine occurs as a white crystalline powder. It is odorless, and has a slightly acid taste. It is very slightly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid or in sodium hy-droxide TS. Melting point: about 1869C (with decomposition).
(1) To 0.2 g of L-Carbocisteine add 1 mL of lead acetate TS and 3 mL of water, shake, add 0.2 g of so-dium hydroxide, and heat over a flame for 1 minute: a dark brown to black precipitate is formed. (2) Determine the infrared absorption spectrum of L-Carbocisteine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －33.5 – －36.59Weigh accu-rately about 5 g of L-Carbocisteine, previously dried, dis-solve in 20 mL of water and a suitable amount of a solution of sodium hydroxide (13 in 100), and adjust the pH with 1 mol/L hydrochloric acid TS or 0.1 mol/L hydrochloric acid TS to 6.0, and add water to make exactly 50 mL. Determine the optical rotation of this solution in a 100-mm cell.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Carbocisteine in 10 mL of sodium hydroxide TS: the so-lution is clear and colorless. Chloride <1.03>—Dissolve 0.20 g of L-Carbocisteine in 10 mL of water and 20 mL of nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows. To 0.40 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of nitric acid and water to make 50 mL (not more than 0.071z). Ammonium <1.02>—Perform the test with 0.25 g of L-Carbocisteine using the distillation under reduced pres-sure. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Carbo-cisteine according to Method 2, and perform the test. Pre- pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of L-Carbocisteine according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.30 g of L-Carbo-cisteine in 10 mL of 0.2 mol/L sodium hydroxide TS, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add 0.2 mol/L sodium hydroxide TS to make exactly 100 mL. Pipet 1 mL of this solution, and add 0.2 mol/L sodium hydroxide TS to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution, in 15 mm length along the starting line on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of L-Carbocisteine, previously dried, dissolve in exactly 20 mL of 0.1 mol/L per-chloric acid VS and 50 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium ace- tate VS (potentiometric titration). Perform a blank determi-nation in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 17.92 mg of C5H9NO4S
L-Cystine, when dried, contains not less than 99.
L-Cystine occurs as white, crystals or crystal-line powder. It is practically insoluble in water and in ethanol (99.5). It dissolves in 1 mol/L hydrochloric acid TS.
Determine the infrared absorption spectrum of L-Cystine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －215 – －2259(after drying, 1 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm). ‌JP XVIII Official Monographs / Cytarabine 827
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of L-Cystine in 10 mL of 2 mol/L hydrochloric acid TS is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of L-Cystine in 10 mL of dilute nitric acid, add 10 mL of hydrogen peroxide (30), and heat in a water bath for 10 minutes. After cooling, add water to make 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 0.6 g of L-Cystine in 5 mL of dilute hydrochloric acid, add water to make 45 mL, and perform the test using this solution as the test solution. Pre-pare the control solution as follows: To 0.35 mL of 0.005 mol/L sulfuric acid VS add 5 mL of dilute hydrochloric acid and water to make 45 mL. To both the test and control solu-tions add 5 mL of barium chloride TS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Cystine, using the distillation under reduced pressure. Pre-pare the control solution with 5.0 mL of Standard Ammoni-um Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Cystine according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Cystine according to Method 3, and perform the test accord- ing to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of L-Cystine in 20 mL of 1 mol/L hydrochloric acid TS, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add water to make exactly 10 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of 1-propanol and am-monia solution (28) (67:33) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly a solution of ninhydrin in a mixture of methanol and acetic acid (100) (97:3) (1 in 100) on the plate, and heat the plate at 809C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 30 mg of L-Cystine, previ-ously dried, and perform the test as directed under Nitrogen Determination <1.08>. Each mL of 0.005 mol/L sulfuric acid VS ＝ 1.202 mg of C6H12N2O4S2
L-Glutamic Acid contains not less than 99.
L-Glutamic acid occurs as white, crystals or crystalline powder. It has a slight characteristic and acid taste. It is slightly soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 2 mol/L hydrochloric acid TS. It shows crystal polymorphism.
Determine the infrared absorption spectrum of L-Glutamic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spec-tra, dissolve L-Glutamic Acid in a small amount of water, evaporate water at 609C under reduced pressure, and per-form the test in the same manner with the dried residue. Optical rotation <2.49> [a]20: ＋31.5 – ＋32.59(2.5 g calcu-lated on the dried basis, 2 mol/L hydrochloric acid TS, 25 mL, 100 mm). D pH <2.54> The pH of a solution prepared by dissolving 0.7 g of L-Glutamic Acid in 100 mL of water by warming and then cooling is 2.9 to 3.9.
(1) Clarity and color of solution—Dissolve 1.0 g of L-Glutamic Acid in 10 mL of 2 mol/L hydrochloric acid TS: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of L-Glutamic Acid in 6 mL of dilute nitric acid and 20 mL of water, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 0.6 g of L-Glutamic Acid in 5 mL of dilute hydrochloric acid and 30 mL of water, and add water to make 45 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution from 0.35 mL of 0.005 mol/L sulfuric acid VS and 5 mL of dilute hydrochloric acid, and dilute with water to 45 mL. Prepare the test solution and the control solution with 5 mL of barium chloride TS, respectively (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Glutamic Acid. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Dissolve 1.0 g of L-Glutamic Acid in 20 mL of water and 7 mL of a solution of sodium hydroxide (1 in 25) by warming, cool, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution from 1.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Glutamic Acid according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Weigh accurately about 0.5 g of L-Glutamic Acid, and dissolve in 0.5 mL of hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount, equivalent to 2.5 mmol, of L-aspartic acid, L-threonine, L-serine, L-glutamic acid, glycine, L-alanine, L-cystine, L-valine, L-methionine, L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammonium chloride, L-histidine and L-argi-nine, dissolve them in 0.1 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, and add 0.02 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 6 mL of this solution, add 0.02 mol/L hydro-chloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the mass percentage of each amino acid, using the mass of amino acid other than glu-tamic acid in 1 mL of the sample solution obtained from the height of the peaks obtained from the sample solution and standard solution: the amount of each amino acid other than glutamic acid is not more than 0.2z, and the total amount of these amino acids is not more than 0.6z. Operating conditions— Detector: A visible absorption photometer (wavelength: ‌1078 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (Na type) com-posed with a sulfonated polystyrene (3 mm in particle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Reaction time: About 1 minute. Mobile phase: Prepare the mobile phases A, B, C, D and E according to the following table, and add 0.1 mL each of caprylic acid. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.12 g of L-Glutamic Acid, dissolve in 40 mL of water by warming, cool, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 14.71 mg of C5H9NO4
l-Isoprenaline Hydrochloride, when dried, contains not less than 98.
l-Isoprenaline Hydrochloride occurs as a white, crystalline powder. It is odorless. It is freely soluble in water, sparingly soluble in ethanol (95), and practically insoluble in acetic acid (100), in acetic anhydride, in diethyl ether and in chloroform. It gradually changes in color by air and by light.
(1) Dissolve 10 mg of l-Isoprenaline Hy-drochloride in 5 mL of water, and add 1 drop of iron (III) chloride TS: a deep green color develops, and changes through yellow-green to brown on standing. Dissolve 1 mg each of l-Isoprenaline Hydrochloride in 1 mL of water in the test tubes A and B. Add 10 mL of potassium hydrogen phthalate buffer solution (pH 3.5) to A, and add 10 mL of phosphate buffer solution (pH 6.5) to B. To each of the test tubes add 1 mL of iodine TS, allow to stand for 5 minutes, and add 2 mL each of sodium thiosul-fate TS: a red color develops in the test tube A, and a deep red color develops in the test tube B. Dissolve 10 mg of l-Isoprenaline Hydrochloride in 1 mL of water, and add 1 mL of phosphotungstic acid TS: a light brown precipitate is produced. Determine the absorption spectrum of a solution of l-Isoprenaline Hydrochloride in 0.1 mol/L hydrochloric acid TS (1 in 20,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of l-Isoprenaline Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －36 – －419(after drying, 0.25 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.10 g of l-Isoprenaline Hydrochloride in 10 mL of water: the pH of the solution is between 4.5 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of l-Isoprenaline Hydrochloride in 20 mL of 0.1 mol/L hy-drochloric acid TS: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.10 g of l- ‌JP XVIII Official Monographs / Isopropylantipyrine 1203‌ Isoprenaline Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.192z). Heavy metals <1.07>—Proceed with 1.0 g of l- Isoprenaline Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Isoproterenone—Dissolve 50 mg of l-Isoprenaline Hydrochloride in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and determine the absorbance of the solution at 310 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>: not more than 0.040. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of l-Isoprenaline Hy-drochloride, previously dried, dissolve in 100 mL of a mix-ture of acetic acid (100) and acetic anhydride (3:2) by warm-ing, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.77 mg of C11H17NO3.HCl
L-Lactic Acid is a mixture of L-lactic acid and L-lactic anhydride. It contains not less than 85.
L-Lactic Acid occurs as a clear, colorless or light yellow, viscous liquid. It is odorless or has a faint, no unpleasant odor. It is miscible with water, with ethanol (99.5) and with diethyl ether. It is hygroscopic. Specific gravity d 20: about 1.20
A solution of L-Lactic Acid (1 in 50) changes the color of blue litmus paper to red, and responds to Qualitative Tests <1.09> for lactate. D Optical rotation <2.49> [a]20: －46 – －529 Weigh accu-rately an amount of L-Lactic Acid, equivalent to about 2 g of L-lactic acid (C3H6O3), add exactly 25 mL of 1 mol/L so-dium hydroxide VS, cover with a watch glass, and heat on a water bath for 15 minutes. Cool, and adjust to pH 7.0 with 1 mol/L hydrochloric acid VS. Dissolve 5.0 g of hexaammoni-um heptamolybdate tetrahydrate in this solution, add water to make exactly 50 mL, and determine the optical rotation using a 100-mm cell.
(1) Chloride <1.03>—Perform the test with 1.0 g of L-Lactic Acid. Prepare the control solution with 1.0 mL of ‌JP XVIII Official Monographs / Anhydrous Lactose 1233 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Sulfate <1.14>—Perform the test with 2.0 g of L-Lac-tic Acid. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Heavy metals <1.07>—To 2.0 g of L-Lactic Acid add 10 mL of water and 1 drop of phenolphthalein TS, and add ammonia TS dropwise until a pale red color appears. Add 2 mL of dilute acetic acid and water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution from 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 4.0 g of L-Lactic Acid according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 5 ppm). Sugars—To 1.0 g of L-Lactic Acid add 10 mL of Anhydrous Lactose
l-Menthol occurs as colorless crystals. It has a characteristic and refreshing odor and a burning taste, fol-lowed by a cool taste. It is very soluble in ethanol (95) and in diethyl ether, and very slightly soluble in water. It sublimes gradually at room temperature.
(1) Triturate l-Menthol with an equal amount of camphor, chloral hydrate or thymol: the mixture liquefies. (2) Shake 1 g of l-Menthol with 20 mL of sulfuric acid: the mixture becomes turbid with a yellow-red color. Allow to stand for 3 hours: a clear, oily layer which possesses no aro-ma of menthol is separated. D Optical rotation <2.49> [a]20: －45.0 – －51.09 (2.5 g, ethanol (95), 25 mL, 100 mm).
(1) Non-volatile residue—Volatilize 2.0 g of l-Menthol on a water bath, and dry the residue at 1059C for 2 hours: the residue weighs not more than 1.0 mg. Thymol—Add 0.20 g of l-Menthol to a cold mixture of 2 mL of acetic acid (100), 6 drops of sulfuric acid and 2 drops of nitric acid: no green to blue-green color immedi-ately develops. Nitromethane or nitroethane—To 0.5 g of l-Menthol placed in a flask add 2 mL of a solution of sodium hydroxide (1 in 2) and 1 mL of hydrogen peroxide (30), connect a reflux condenser to the flask, and boil the mixture gently for 10 ‌minutes. After cooling, add water to make exactly 20 mL, and filter. Take 1 mL of the filtrate in a Nessler tube, add water to make 10 mL, neutralize with dilute hydrochloric acid, add 1 mL of dilute hydrochloric acid, and cool. To the mixture add 1 mL of a solution of sulfanilic acid (1 in 100), allow to stand for 2 minutes, and then add 1 mL of a solu-tion of N,N-diethyl-N?-1-naphthylethylenediamine oxalate (1 in 1000) and water to make 25 mL: no red-purple color immediately develops.
Weigh accurately about 2 g of l-Menthol, add ex-actly 20 mL of a mixture of dehydrated pyridine and acetic anhydride (8:1), connect a reflux condenser, and heat on a water bath for 2 hours. Wash the condenser with 20 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 5 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 1 mol/L sodium hydroxide VS ＝ 156.3 mg of C10H20O
Labetalol Hydrochloride occurs as a white crystalline powder. It is freely soluble in methanol, and sparingly soluble in water and in ethanol (99.5). It dissolves in 0.05 mol/L sulfuric acid TS. Melting point: about 1819C (with decomposition).
(1) Determine the absorption spectrum of a solution of Labetalol Hydrochloride in 0.05 mol/L sulfuric acid TS (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Labetalol Hydrochloride as directed in the potassium chlo-ride disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Labetalol Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution prepared by dissolving 0.5 g of Labetalol Hydrochloride in 50 mL of water is between 4.0 and 5.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Labetalol Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.8 g of Labetalol Hy-drochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 2-propanol, water, and ammonia solution (28) (25:15:8:2) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 30 minutes: the spots other than the princi-pal spot obtained from the sample solution do not exceed 2 in number and are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 5 mg of Labetalol Hydrochloride in 0.7 mL of a solution of n-butylboronic acid in dehydrated pyridine (3 in 250), allow to stand for 20 minutes, and use this solution as the sample solution. Perform the test with 2 mL of the sample solution as directed under Gas Chromatog-raphy <2.02> according to the following conditions. Deter-mine the areas of two adjacent peaks, Aa and Ab, where Aa is the peak area of the shorter retention time and Ab is the peak area of the longer retention time, using the automatic in-tegration method: the ratio Ab/(Aa ＋ Ab) is between 0.45 and 0.55. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 25 m in length, coated inside with methyl silicone poly-mer for gas chromatography in 5 mm thickness. Column temperature: A constant temperature of about 2909C. Injection port temperature: A constant temperature of about 3509C. Detector temperature: A constant temperature of about 3509C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of the peak showing earlier elution of the two peaks of labetalol is about 9 minutes. System suitability— System performance: Proceed with 2 mL of the sample so-lution under the above conditions: the resolution between the two labetalol peaks is not less than 1.5. System repeatability: Repeat the test 6 times under the above conditions with 2 mL of the sample solution: the rela-tive standard deviation of the ratio of the peak area of labetalol with the shorter retention time to that of the longer ‌JP XVIII Official Monographs / Labetalol Hydrochloride Tablets 1231 retention time is not more than 2.0z.
Weigh accurately about 0.3 g of Labetalol Hydro-chloride, previously dried, dissolve in 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.49 mg of C19H24N2O3.HCl
Lactic Acid is a mixture of lactic acid and lactic an-hydride. It contains not less than 85.
Lactic Acid occurs as a clear, colorless or light yellow, viscous liquid. It is odorless or has a faint, unpleas-ant odor. It is miscible with water, with ethanol (95) and with diethyl ether. It is hygroscopic. Specific gravity d 20: about 1.20
A solution of Lactic Acid (1 in 50) changes blue litmus paper to red and responds to Qualitative Tests <1.09> for lactate.
(1) Chloride <1.03>—Perform the test with 1.0 g of Lactic Acid. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Sulfate <1.14>—Perform the test with 2.0 g of Lactic Acid. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Heavy metals <1.07>—To 2.0 g of Lactic Acid add 10 mL of water and 1 drop of phenolphthalein TS, and add am-monia TS dropwise until a pale red color appears. Add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution from 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 4.0 g of Lactic Acid according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 5 ppm). Sugars—To 1.0 g of Lactic Acid add 10 mL of water, and neutralize with sodium hydroxide TS. Boil the mixture with 10 mL of Fehling's TS for 5 minutes: no red precipitate is produced. Citric, oxalic, phosphoric and L-tartaric acid—To 1.0 g of Lactic Acid add 1.0 mL of water, followed by 40 mL of calcium hydroxide TS. Boil the mixture for 2 minutes: no change occurs. Glycerin or mannitol—Shake 10 mL of Lactic Acid with 12 mL of diethyl ether: no turbidity is produced. Volatile fatty acids—Warm Lactic Acid: it does not produce any acetic acid-like or butyric acid-like odor. Cyanide—Transfer 1.0 g of Lactic Acid to a Nessler tube, add 10 mL of water and 1 drop of phenolphthalein TS, add dropwise a solution of sodium hydroxide (1 in 10) by shaking until a pale red color develops, add 1.5 mL of a solution of sodium hydroxide (1 in 10) and water to make 20 mL, and heat in a water bath for 10 minutes. Cool, add dropwise dilute acetic acid until a red color of the solution disappears, add 1 drop of dilute acetic acid, add 10 mL of phosphate buffer solution (pH 6.8), and 0.25 mL of sodium toluensulfonchloramide TS, stopper immediately, mix gently, and allow to stand for 5 minutes. To the solution add 15 mL of pyridine-pyrazolone TS and water to make 50 mL, and allow to stand at 259C for 30 minutes: the solution has no more color than the following control solution. Control solution: Pipet 1.0 mL of Standard Cyanide Solu-tion, and add water to make exactly 20 mL. Transfer 1.0 mL of this solution to a Nessler tube, add 10 mL of water and 1 drop of phenolphthalein TS, and then proceed as described above. Readily carbonizable substances—Superimpose slowly 5 mL of Lactic Acid, previously kept at 159C, upon 5 mL of sulfuric acid for readily carbonizable substances, pre-viously kept at 159C, and allow to stand at 159C for 15 minutes: no dark color develops at the zone of contact. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 3 g of Lactic Acid, transfer in a conical flask, add accurately measured 40 mL of 1 mol/L sodium hydroxide VS, invert a watch glass over the flask, and heat on a water bath for 10 minutes. Titrate <2.50> the excess sodium hydroxide with 0.5 mol/L sulfuric acid VS immediately (indicator: 2 drops of phenolphthalein TS). Per-form a blank determination in the same manner. Each mL of 1 mol/L sodium hydroxide VS ＝ 90.08 mg of C3H6O3
Lactose Hydrate occurs as white, crystals, powder or granulated powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).◆
Determine the infrared absorption spectrum of Lactose Hydrate, previously dried, as directed in the po-tassium bromide disk method under Infrared Spectropho- ‌JP XVIII Official Monographs / Lactulose 1235 tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Lactose for Identification RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋54.4 – ＋55.99. Weigh ac-curately about 10 g of Lactose Hydrate, calculated on the anhydrous basis, dissolve in 80 mL of water warmed to 509C, and add 0.2 mL of ammonia TS after cooling. After standing for 30 minutes, add water to make exactly 100 mL, and determine the optical rotation of this solution in a 100-mm cell.
(1) Clarity and color of solution—Dissolve 1.0 g of Lactose Hydrate in 10 mL of hot water, allow to cool, and determine the turbidity of this solution as directed under Turbidity Measurement <2.61>: it is clear, and has no more color than the following control solution. Determine the ab-sorbance at 400 nm of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, using water as the control solution: not more than 0.04. Control solution: To 2.5 mL of Cobalt (II) Chloride CS, 6.0 mL of Iron (III) Chloride CS and 1.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity or alkalinity—Dissolve 6 g of Lactose Hy-drate by heating in 25 mL of freshly boiled and cooled water, and after cooling, add 0.3 mL of phenolphthalein TS: the solution is colorless, and not more than 0.4 mL of 0.1 mol/L sodium hydroxide VS is required to produce a pale red color or red color. (3) Heavy metals <1.07>—Dissolve 4.0 g of Lactose Hy- drate in 20 mL of warm water, add 1 mL of 0.1 mol/L hy-drochloric acid TS and water to make 50 mL. Proceed with this solution according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution and 1 mL of 0.1 mol/L hydrochloric acid TS (not more than 5 ppm).◇ (4) Proteins and light absorbing substances—Dissolve 1.0 g of Lactose Hydrate in water to make 100 mL, and use this solution as the sample solution. Determine the absor-bances of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the control solution: not more than 0.25 at between 210 nm and 220 nm, and not more than 0.07 at between 270 nm and 300 nm. Loss on drying <2.41> Not more than 0.5z. For the granu-lated powder, not more than 1.0z (1 g, 809C, 2 hours). Water <2.48> 4.5 – 5.5z. ◇For the granulated powder, 4.0 – 5.5z◇ (1 g, volumetric titration, direct titration. Use a mixture of methanol for water determination and for-mamide for water determination (2:1) instead of methanol for water determination). Residue on ignition <2.44> Not more than 0.1z (1 g). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/g and 5 × 101 CFU/g, respectively. Salmonella and◇ Escherichia coli are not observed.
Lactulose is a solution of lactulose prepared by isomerizing lactose under the existing of alkaline and purified by ion-exchange resin. It contains not less than 50.
Lactulose occurs as a clear, colorless or light yellow, viscous liquid. It is odorless, and has a sweet taste. It is miscible with water and with formamide.
(1) To 0.7 g of Lactulose add 10 mL of water, 10 mL of a solution of hexaammonium heptamolyb-date tetrahydrate (1 in 25) and 0.2 mL of acetic acid (100), and heat in a water bath for 5 to 10 minutes: a blue color de-velops. (2) Mix 0.3 g of Lactulose and 30 mL of water, add 16 mL of 0.5 mol/L iodine TS, then immediately add 2.5 mL of 8 mol/L sodium hydroxide TS, allow to stand for 7 minutes, and add 2.5 mL of diluted sulfuric acid (3 in 20). To this so-lution add a saturated solution of sodium sulfite heptahy-drate until the solution turns light yellow, then add 3 drops of methyl orange TS, neutralize with a solution of sodium hydroxide (4 in 25), and add water to make 100 mL. To 10 mL of this solution add 5 mL of Fehling's TS, and boil for 5 minutes: a red precipitate is produced. pH <2.54> To 2.0 g of Lactulose add 15 mL of water: the pH of the solution is between 3.5 and 5.5. Specific gravity <2.56> d 20: 1.320 – 1.360
(1) Heavy metals <1.07>—Proceed with 5.0 g of Lactulose according to Method 4, and perform the test. Pre-pare the control solution with 2.5 mL of Standard Lead So-lution (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Lactulose according to Method 1, and perform the test (not more than 2 ppm). Galactose and lactose—Determine the heights of the peaks corresponding to galactose and lactose respectively, on the chromatogram obtained in Assay from the sample solu-tion and the standard solution, and calculate the ratios of the peak heights of galactose and lactose to that of the internal standard from the sample solution, QTa and QTb, and then from the standard solution, QSa and QSb: it contains galac-tose of not more than 11z, and lactose of not more than 6z. Amount (mg) of galactose (C6H12O6) ＝ MS × QTa/QSa ‌1236 MS: Amount (mg) of galactose taken Amount (mg) of lactose (C ＝ MS × QTb/QSb 12H22O11.H2O) Lafutidine
Lafutidine, when dried, contains not less than 99.
Lafutidine occurs as a white to pale yellow-white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble Amount (mg) of lactulose (C ＝ MS × QT/QS 12H22 O11) in water. A solution of Lafutidine in methanol (1 in 100) shows no optical rotation. MS: Amount (mg) of Lactulose RS taken Internal standard solution—A solution of D-mannitol (1 in 20). Operating conditions— Detector: A differential refractometer. Column: A stainless steel column 8 mm in inside diameter and 50 cm in length, packed with gel type strongly acidic ion-exchange resin for liquid chromatography (degree of crosslinkage: 6z) (11 mm in particle diameter). Column temperature: A constant temperature of about 759C. Mobile phase: Water. Flow rate: Adjust so that the retention time of lactulose is about 18 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lactulose and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak heights of lactulose, galactose and lactose to the height of the internal standard are not more than 2.0z, re-spectively.
(1) Determine the absorption spectrum of a solution of Lafutidine in methanol (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Lafutidine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Lafutidine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Lafutidine in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak, having the relative retention time of about 0.85 to lafutidine, obtained from the sample solution is not larger than 3/10 times the peak area of lafutidine from the standard solution, the area of the peak other than lafutidine and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of lafutidine from the standard solution, and the total area of the peaks other than lafutidine from the sample solution is not larger than 2/5 times the peak area of lafutidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌JP XVIII Official Monographs / Lafutidine Tablets 1237 Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.87 g of sodium 1-pentanesul-fonate in 1000 mL of diluted phosphoric acid (1 in 1000). To 850 mL of this solution add 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of lafutidine is about 15 minutes. Time span of measurement: About 6 times as long as the retention time of lafutidine. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of lafutidine obtained with 5 mL of this solution is equivalent to 3.5 to 6.5z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of lafutidine are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of lafutidine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Lactulose, add ex-actly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.5 g of Lactulose RS, about 80 mg of D-galactose and about 40 mg of lactose monohydrate, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of lactulose to that of the internal standard. C22H29N3O4S: 431.55 2-[(RS)-Furan-2-ylmethylsulfinyl]-N-{4-[4-(piperidin- ylmethyl)pyridin-2-yl]oxy-(2Z)-but-2-en-1-yl}acetamide [206449-93-6] Lafutidine, when dried, contains not less than 99.0z and not more than 101.0z of lafutidine (C22H29N3O4S). Description Lafutidine occurs as a white to pale yellow-white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble Amount (mg) of lactulose (C ＝ MS × QT/QS 12H22 O11) in water. A solution of Lafutidine in methanol (1 in 100) shows no optical rotation. MS: Amount (mg) of Lactulose RS taken Internal standard solution—A solution of D-mannitol (1 in 20). Operating conditions— Detector: A differential refractometer. Column: A stainless steel column 8 mm in inside diameter and 50 cm in length, packed with gel type strongly acidic ion-exchange resin for liquid chromatography (degree of crosslinkage: 6z) (11 mm in particle diameter). Column temperature: A constant temperature of about 759C. Mobile phase: Water. Flow rate: Adjust so that the retention time of lactulose is about 18 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lactulose and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak heights of lactulose, galactose and lactose to the height of the internal standard are not more than 2.0z, re-spectively.
Lanoconazole, when dried, contains not less than 98.
Lanoconazole occurs as white to pale yellow, crystals or crystalline powder. It is soluble in acetone, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. It is gradually colored to yellow by light. A solution of Lanoconazole in acetone (1 in 25) shows no optical rotation. Identiˆcation (1) To 0.1 g of Lanoconazole add 0.5 g of sodium hydroxide, heat gradually to melt, and carbonize. After cooling, add 10 mL of dilute hydrochloric acid: the gas evolved darkens moistened lead (II) acetate paper. Perform the test with Lanoconazole as directed under Flame Coloration Test <1.04> (2): a green color appears. Determine the absorption spectrum of a solution of Lanoconazole in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Lanoconazole RS prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Lanoconazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Lanoconazole RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Lanoconazole according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ‌(2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.10 g of Lanoconazole in 100 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area by the automatic integration method: the total area of the peaks other than lanoconazole obtained from the sample solution is not larger than 1/2 times the peak area of lanoconazole from the standard solu-tion. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 0.576 g of sodium 1-nonanesul-fonate in 1000 mL of a mixture of methanol, water and acetic acid (100) (55:44:1). Flow rate: Adjust so that the retention time of lanocona-zole is about 7 minutes. Time span of measurement: About 3 times as long as the retention time of lanoconazole, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, and add methanol to make exactly 50 mL. Conˆrm that the peak area of lanoconazole obtained with 5 mL of this solution is equivalent to 3.5 to 6.5z of that with 5 mL of the standard solution. System performance: Put 20 mL of the sample solution in a colorless vessel, and expose to ultraviolet light (main wave-length: 365 nm) for 30 minutes. When the procedure is run with 5 mL of this solution under the above operating condi-tions, the resolution between the peak having the relative retention time of about 0.8 to lanoconazole and the peak of lanoconazole is not less than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of lanoconazole is not more than 1.0z. Loss on drying <2.41> Not more than 0.4z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately about 50 mg each of Lanoconazole and Lanoconazole RS, both previously dried, and dissolve each in methanol to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard so-lution, add methanol to make 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lanoconazole to that of the internal standard. Amount (mg) of lanoconazole (C14H10ClN3S2) ＝MS × QT/QS MS: Amount (mg) of Lanoconazole RS taken Internal standard solution—A solution of diisopropyl 1,3- dithiolan-2-ylidenemalonate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of methanol and water (11:9). Flow rate: Adjust so that the retention time of lanocona- zole is about 9 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, lanoconazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of lanoconazole to that of the internal standard is not more than 1.0z.
Lansoprazole occurs as a white to brownish white crystalline powder. It is freely soluble in N, N-dimethylformamide, soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Lansoprazole in N, N-dimethylformamide (1 in 10) shows no optical rotation. Melting point: about 1669C (with decomposition). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Lansoprazole in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, Amount (mg) of lanoconazole (C H ClN S ) and compare the spectrum with the Reference Spectrum or ＝ MS × QT/QS 14 10 3 2 the spectrum of a solution of Lansoprazole RS prepared in the same manner as the sample solution: both spectra exhibit MS: Amount (mg) of Lanoconazole RS taken Internal standard solution—A solution of diisopropyl 1,3-dithiolan-2-ylidenemalonate in methanol (1 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Lanoconazole. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lanoconazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lanoconazole to that of the internal stand-ard is not more than 1.0z.
(1) Clarity and color of solution—Dissolve 1.0 g of Lansoprazole in 20 mL of N, N-dimethylformamide: the solution is clear and not more colored than Matching Fluid G. Heavy metals <1.07>—Proceed with 1.0 g of Lan- soprazole in a platinum crucible according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Lansoprazole in a platinum crucible according to Method 3, using a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 5), and perform the test. Prepare the stand-ard color with 1.0 mL of Standard Arsenic Solution (not more than 1 ppm). Related substances—Dissolve 50 mg of Lansoprazole in a mixture of dilute sodium hydroxide TS and methanol (3:1) to make 20 mL. To 2 mL of this solution add a mixture of dilute sodium hydroxide TS and methanol (3:1) to make 20 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of dilute sodium hydroxide TS and methanol (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having a relative retention time of about ‌to lansoprazole, obtained from the sample solution is not larger than 2/5 times the peak area of lansoprazole from the standard solution, and the area of the peak other than lan-soprazole and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of lan-soprazole from the standard solution. Furthermore, the total area of the peaks other than lansoprazole from the sample solution is not larger than 3/5 times the peak area of lan-soprazole from the standard solution. For the area of the peaks, having the relative retention time of about 0.8, about 1.1 and about 1.2, multiply their correction factors, 0.8, 1.2, and 1.3, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Water. Mobile phase B: A mixture of acetonitrile, water and triethylamine (160:40:1), adjusted to pH 7.0 with phosphoric acid. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 40 90 → 20 10 → 80 40 – 50 Flow rate: About 0.8 mL per minute (the retention time of lansoprazole is about 29 minutes). Time span of measurement: About 1.7 times as long as the retention time of lansoprazole. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of dilute sodium hydroxide TS and methanol (3:1) to make exactly 20 mL. Confirm that the peak area of lansoprazole obtained with 40 mL of this solu-tion is equivalent to 4z to 6z of that with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of lansoprazole are not less than 150,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lansoprazole is not more than 3.0z. Water <2.48> Not more than 0.10z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 50 mg each of Lansoprazole and Lansoprazole RS (separately determine the water <2.48> in the same manner as Lansoprazole), and dissolve each in exactly 10 mL of the internal standard solution. To 1 mL each of both solutions add diluting solution to make 50 mL, and use these solutions as the sample solution and the stand- ard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of lansoprazole to that of the internal stand-ard. Amount (mg) of lansoprazole (C16H14F3N3O2S) ＝ MS × QT/QS MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 4?-ethoxya-cetophenone in diluting solution (1 in 400). Diluting solution: A mixture of water, acetonitrile and triethylamine (60:40:1), adjusted to pH 11.0 with phosphoric acid. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized sili-con polymer coated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and triethylamine (60:40:1), adjusted to pH 7.0 with phosphoric acid. Flow rate: Adjust so that the retention time of lansopra-zole is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lansoprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lansoprazole to that of the internal standard is not more than 1.0z.
Lauromacrogol is a colorless or light yellow, clear liquid or a white, petrolatum-like or waxy solid. It has a characteristic odor, and a somewhat bitter and slightly irritative taste. It is very soluble in ethanol (95) and in diethyl ether. It is freely soluble or dispersed as fine oily drops in water.
(1) Shake throughly 0.5 g of Lauromacrogol with 10 mL of water and 5 mL of ammo-nium thiocyanate-cobalt nitrate TS, then shake with 5 mL of 1-butanol, and allow to stand: the 1-butanol layer becomes blue in color. (2) Warm Lauromacrogol to melt, if necessary, and de-termine the infrared absorption spectrum as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of be-tween 3500 cm－1 and 3400 cm－1, and about 2920 cm－1, 1350 cm－1, 1250 cm－1, and 1115 cm－1.
(1) Acidity—Transfer 10.0 g of Lauromacrogol into a flask, and add 50 mL of neutralized ethanol. Heat on a water bath nearly to boil, shaking once or twice while heat-ing. Cool, and add 5.3 mL of 0.1 mol/L sodium hydroxide VS and 5 drops of phenolphthalein TS: a red color develops. (2) Unsaturated compound—Shake 0.5 g of Lauro-macrogol with 10 mL of water, and add 5 drops of bromine TS: the color of the solution does not disappear. Residue on ignition <2.44> Not more than 0.2z (1 g).
Lenampicillin Hydrochloride is the hydrochloride of ampicillin methyloxodioxolenylmethyl ester. It contains not less than 653 mg (potency) and not more than 709 mg (potency) per mg, calculated on the anhydrous basis and corrected by the amount of the residual solvents.
Lenampicillin Hydrochloride occurs as a white to light yellow-white powder. It is very soluble in water, in methanol and in ethanol (95), and freely soluble in N,N-dimethylformamide.
(1) Determine the infrared absorption spec-trum of Lenampicillin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Lenampicillin Hydrochlo-ride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) To 1 mL of a solution of Lenampicillin Hydrochlo-ride (1 in 100) add 0.5 mL of dilute nitric acid and 1 drop of JP XVIII Official Monographs / Lenampicillin Hydrochloride 1247 silver nitrate TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋174 – ＋1949(0.2 g calcu-lated on the anhydrous basis and corrected on the amount of residual solvent, ethanol (95), 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Lenampicillin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Lenampicillin Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Free ampicillin—Weigh accurately about 0.1 g of Lenampicillin Hydrochloridein, dissolve in exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately an amount of Ampicillin RS, equivalent to about 25 mg (potency), and dis-solve in water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solu-tion, and use this solution as the standard solution. The sam-ple solution should be used to the following test immediately after the solution is prepared. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak height of ampicillin to that of the internal standard: the amount of ampicillin is not more than 1.0z. Amount (z) of ampicillin (C16H19N3O4S) ＝ MS/MT × QT/QS × 2 MS: Amount [mg (potency)] of Ampicillin RS taken MT: Amount (mg) of Lenampicillin Hydrochloride taken Internal standard solution—A solution of anhydrous caffeine in the mobile phase (1 in 50,000). mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction: the amount of penicilloic acid (C16H21N3O5S: 367.42) is not more than 3.0z. Each mL of 0.01 mol/L sodium thiosulfate VS ＝ 0.45 mg of C16H21N3O5S (5) Residual solvent <2.46>—Weigh accurately about 0.25 g of Lenampicillin Hydrochloride, dissolve in exactly 1 mL of the internal standard solution, add N,N-dimethylfor-mamide to make 5 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of 2-propanol and about 0.12 g of ethyl acetate, and add N,N-dimethylformamide to make exactly 100 mL. Pipet 1 mL and 3 mL of this solution, add exactly 1 mL each of the in-ternal standard solution, add N,N-dimethylformamide to make 5 mL, and use these solutions as the standard solution (1) and the standard solution (2), respectively. Perform the test with 4 mL each of the sample solution, standard solution (1) and (2) as directed under Gas Chromatography <2.02> ac- cording to the following conditions, and calculate the ratios, QTa and QTb, of the peak height of 2-propanol and ethyl ace-tate to that of the internal standard of the sample solution, the ratios, QSa1 and QSb1, of the peak height of 2-propanol and ethyl acetate to that of the internal standard of the standard solution (1) and the ratios, QSa2 and QSb2, of the peak height of 2-propanol and ethyl acetate to that of the internal standard of the standard solution (2). Calculate the amounts of 2-propanol and ethyl acetate by the following equations: not more than 0.7z and not more than 1.7z, respectively. Amount (z) of 2-propanol ＝ MSa/MT × (2QTa － 3QSa1 ＋ QSa2)/(QSa2 － QSa1) Amount (z) of ethyl acetate Operating conditions— Detector: An ultraviolet absorption photometer (wave- ＝ MSb/MT × (2QTb － 3Q Sb1 ＋ QSb2 )/(Q Sb2 － QSb1) length: 230 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.22 g of potassium dihydrogen phosphate in water to make 900 mL, and add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of ampicillin is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak height of ampicillin to that of the internal stand-ard is not more than 5z. Penicilloic acid—Weigh accurately about 0.1 g of Lenampicillin Hydrochloride, dissolve in water to make ex-actly 100 mL, and use this solution as the sample solution. Pipet 10 mL of the sample solution, add 10 mL of potassium hydrogen phthalate buffer solution (pH 4.6) and exactly 10 mL of 0.005 mol/L iodine VS, allow to stand for exactly 15 minutes while protecting from exposure to light, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS (indicator: 1 MSa: Amount (g) of 2-propanol taken MSb: Amount (g) of ethyl acetate taken MT: Amount (g) of the Lenampicillin Hydrochloride taken Internal standard solution—A solution of cyclohexane in N,N-dimethylformamide (1 in 1000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 3 m in length, packed with siliceous earth for gas chromatogra-phy (180 – 250 mm in particle diameter) coated with tetra-kishydroxypropylethylenediamine for gas chromatography at the ratio of 10 to 15z. Column temperature: A constant temperature of about 809C. Injection port temperature: A constant temperature of about 1609C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of the internal standard is about 1 minute. System suitability— System performance: When the procedure is run with 4 mL of the standard solution (2) under the above operating condi-tions, the internal standard, ethyl acetate and 2-propanol are eluted in this order, and the resolution between the peaks of the internal standard and ethyl acetate is not less than 2.0. System repeatability: When the test is repeated 3 times with 4 mL of the standard solution (2) under the above oper-ating conditions, the relative standard deviation of the ratios of the peak height of ethyl acetate to that of the internal standard is not more than 5.0z. ‌1248 Water <2.48> Not more than 1.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately an amount of Lenampicillin Hy-drochloride and Lenampicillin Hydrochloride RS, equivalent to about 0.1 g (potency), dissolve each in the internal stand-ard solution to make exactly 10 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lenampicillin to that of the internal standard. Amount [ mg (potency)] of ampicillin (C16H19N3O4S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Lenampicillin Hydrochlo-ride RS taken Internal standard solution—A solution of ethyl aminobenzo-ate in the mobile phase (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 9.53 g of potassium dihydrogen phosphate in water to make exactly 700 mL, and add aceto-nitrile to make exactly 1000 mL. Flow rate: Adjust so that the retention time of lenampicil-lin is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, lenampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of lenampicillin to that of the internal standard is not more than 1.0z.
Lenograstim (Genetical Recombination) is an aque-ous solution in which a desired product is a recom-binant human granulocyte colony-stimulating factor produced in Chinese hamster ovary cells. It is a glycoprotein (molecular mass: ca. 20,000) consisting of 174 amino acid residues. It contains not less than 0.
Lenograstim (Genetical Recombination) oc-curs as a clear and colorless liquid.
(1) Use Lenograstim (Genetical Recombi-nation) and Lenograstim RS as the sample solution and the standard solution, respectively. Perform the test with a volume each of the sample solution and standard solution, equivalent to 20 mg of protein, as directed under Liquid Chromatography <2.01> according to the following condi-tions: the retention times of the two peaks of lenograstim in the chromatogram obtained from the sample solution and of those in the chromatogram obtained from the standard solu-tion are the same. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 7.5 cm in length, packed with diethylaminoethyl group binding synthetic polymer for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: 0.02 mol/L tris buffer solution (pH 7.4). Mobile phase B: 0.02 mol/L tris buffer solution (pH 7.4) containing 0.5 mol/L sodium chloride. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. JP XVIII Official Monographs / Lenograstim (Genetical Recombination) 1249 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) System suitability— System performance: When the procedure is run with the standard solution under the above operating conditions, the 0 – 35 100 → 80 0 → 20 35 – 40 80 20 Flow rate: Adjust so that the retention time of the first ap-peared peak of lenograstim is about 27 minutes. System suitability— System performance: When the procedure is run with a volume of the standard solution, equivalent to 20 mg of pro-tein, under the above operating conditions, the resolution between the two peaks of lenograstim is not less than 4. (2) Desalt 2 mL each of Lenograstim (Genetical Recom-bination) and Lenograstim RS by a suitable method, and as-sign them as the desalted sample and the desalted reference standard, respectively. Add the desalted sample and the desalted reference standard in 100 mL each of a mixture of water and 1-propanol (3:2), add 4 mL of urea-EDTA TS, and allow them to stand at 379C for 18 hours. Then, add 10 mL of 2-mercaptoethanol to them, and allow to stand at 379C for 4 hours. To these solutions add a solution of 27 mg of iodoacetic acid in 150 mL of sodium hydroxide TS, and react at 379C for 15 minutes, avoiding exposure to light. Re-move the reagents from these reaction solution by a suitable method, and assign obtained these substances as the reduced carboxymethylated sample and the reduced carboxymethy-lated reference standard. To these substances add 100 mL each of a mixture of water and 1-propanol (3:2), and add 1 mL of 0.05 mol/L ammonium hydrogen carbonate solution. Add 20 mL each of a solution of V8 protease in 0.05 mol/L ammonium hydrogen carbonate solution (1 in 1000), and react at 379C for 18 hours. To each reaction solution add 50 mL of diluted trifluoroacetic acid (1 in 10) to stop the reaction, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 100 mL – 150 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and compare the chromatograms from these solutions: both chromatograms show the similar peaks at the corresponding retention time. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile for liq-uid chromatography and trifluoroacetic acid (950:50:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, water, and trifluoroacetic acid (800:200:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. resolution between the first appeared peak and the second appeared peak is not less than 15. Monosaccharide composition Put exactly 2 mL of Leno-grastim (Genetical Recombination) into a precolumn, packed with 0.36 g of octadecylsilanized silica gel for pretreatment, wash the column with 5 mL of a mixture of water, acetonitrile and trifluoroacetic acid (600:400:1), then elute with a mixture of acetonitrile, water and trifluoroacetic acid (800:200:1), and collect exactly 5 mL of the first eluate. Pipet 1.5 mL of the eluate in a test tube, add exactly 20 mL of the internal standard solution, and lyophilize. Dissolve the lyophilized substance in 250 mL of a mixture of methanol and acetyl chloride (9:1), seal the tube, and heat at 909C for 2 hours. After cooling, open the tube, and dry the content under reduced pressure. To the residue add 200 mL of metha-nol, and evaporate to dryness under reduced pressure. Dis-solve the residue in 200 mL of a solution of pyridine in meth-anol (1 in 10) and 50 mL of acetic anhydride, stopper the tube tightly, and allow to stand for 10 minutes. Evaporate the so-lution to dryness at about 509C under reduced pressure, add 200 mL of methanol to the residue, and evaporate to dryness at 509C under reduced pressure. To the residue add 50 mL of a mixture of pyridine, 1,1,1,3,3,3-hexamethyldisilazane and chlorotrimethylsilane (10:2:1), stopper tightly, shake vigor-ously for 30 seconds, and warm at 509C for 10 minutes. After cooling, add 300 mL of pentane, stir gently, then add 300 mL of water, and stir gently. Separate the upper layer, evaporate to concentrate to about 10 mL under a stream of nitrogen, and use this as the sample solution. Separately, weigh accurately about 54 mg of D-galactose and about 33 mg of N-acetylgalactosamine, dissolve them separately in water to make exactly 20 mL each, and use these solutions as D-galactose solution and N-acetylgalactosamine solution, respectively. Weigh accurately about 9.3 mg of N-acetyl-neuraminic acid, add exactly 1 mL of the D-galactose solu-tion and exactly 2 mL of the N-acetylgalactosamine solution to dissolve, and add water to make exactly 20 mL. Pipet 1 mL of this solution, add exactly 1 mL of the internal stand-ard solution, and freeze-dry 40 mL of this solution. Dissolve the freeze-dried substance in 250 mL of a mixture of metha-nol and acetyl chloride (9:1), then proceed in the same man-ner as the sample solution, and use the solution obtained as the monosaccharide standard solution. Perform the test with 2 mL each of the sample solution and the monosaccharide standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios of each peak area of D-galactose, N-acetylgalac-tosamine and N-acetylneuraminic acid to that of the internal standard, QT and QS. Calculate the amount (mol/mol of lenograstim) of each monosaccharide by the following for-mula: the amounts of D-galactose, N-acetylgalactosamine and N-acetylneuraminic acid are between 0.7 and 1.2, be-tween 0.7 and 1.2, and between 1.0 and 2.0, respectively. Time after injection Mobile phase A Mobile phase B Amount (mol/mol of lenograstim) of each monosaccharide of sample (min) (volz) (volz) ＝ M/(Mm × DS) × QT/QS × 18,667/C × 5/3 0 – 120 100 → 20 0 → 80 M: Amount (mg) of each monosaccharide taken 120 – 140 20 → 0 80 → 100 Mm: Molecular mass of each monosaccharide 140 – 150 D-galactose: 180.16 N-acetylgalactosamine: 221.21 Flow rate: Adjust so that the retention time of the first ap-peared peak is about 33 minutes. N-acetylneuraminic acid: 309.27 DS: Dilution rate of each monosaccharide D-galactose: 20,000 N-acetylgalactosamine and: 10,000 N-acetylneuraminic acid: 1000 C: Protein concentration (mg/mL) of Lenograstim (Genetical Recombination) 18,667: Molecular mass of protein moiety of lenograstim Internal standard solution—Dissolve 48 mg of myoinositol in water to make 50 mL. To 1 mL of this solution add water to make 20 mL. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.25 mm in inside diameter and 30 m in length, coated the inside surface with 7z cyanopropyl-7z phenyl-methyl silicon polymer for gas chro-matography 0.25 mm in thickness. Column temperature: Rise the temperature at a rate of 109C per minute from 1109C to 1859C, then at a rate of 29C per minute to 2109C, and to 2609C at a rate of 89C per minute, and maintain 2609C for 15 minutes. Carrier gas: Helium. Flow rate: Adjust so that the retention time of the internal standard is about 24 minutes. System suitability— System performance: When the procedure is run with 2 mL of the monosaccharide standard solution under the above operating conditions, D-galactose, the internal standard, N-acetylgalactosamine and N-acetylneuraminic acid are eluted in this order, and the resolution between the peaks of the in-ternal standard and N-acetylgalactosamine is not less than 10. pH <2.54> 7.7 – 8.3
(1) Related substances—Perform the test with a volume of Lenograstim (Genetical Recombination), equiva-lent to 30 mg of protein, as directed under Liquid Chroma- number of theoretical plates of the peak of lenograstim is not less than 2700. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law.
(1) Protein—Use Lenograstim (Genetical Recom-bination) and Lenograstim RS as the sample solution and the standard solution, respectively. Perform the test with exactly 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of lenograstim in each solution. Amount (mg) of protein in 1 mL of Lenograstim (Genetical Recombination) ＝ CS × AT/AS CS: Concentration (mg/mL) of protein in Lenograstim RS Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile for liq-uid chromatography and trifluoroacetic acid (600:400:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, water and trifluoroacetic acid (800:200:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method, Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) and calculate the amount of these peaks by the area percen-tage method excluding the area of the solvent peak: the total amount of the peaks other than lenograstim is not more than 1.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with porous silica gel for liq-uid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.4 g of anhydrous disodium hydrogen phosphate and 5.8 g of sodium chloride in water to make 1000 mL (Solution A). Separately, dissolve 1.6 g of so-dium dihydrogen phosphate dihydrate and 5.8 g of sodium chloride in water to make 1000 mL (Solution B). Adjust the pH of Solution A to 7.4 with Solution B. Flow rate: Adjust so that the retention time of lenograstim is about 21 minutes. Time span of measurement: About 2 times as long as the retention time of lenograstim. System suitability— Test for required detectability: When the procedure is run with 60 mL of diluted Lenograstim RS with the solvent of Lenograstim (Genetical Recombination) containing 0.1 volz polysorbate 20 (1 in 500) under the above operating conditions, the peak of lenograstim is detectable. System performance: When the procedure is run using Lenograstim RS under the above operating conditions, the 0 – 40 80 → 30 20 → 70 Flow rate: Adjust so that the retention time of lenograstim is about 35 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the number of theoretical plates of the peak of lenograstim is not less than 2900. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lenograstim is not more than 4.0z. (2) Specific activity—Dilute Lenograstim (Genetical Recombination) with FBS-IMDM so that each mL contains an estimate amount of 7.69 units, 10.0 units and 13.0 units, and name them as the sample solution (1), the sample solu-tion (2) and the sample solution (3), respectively. Separately, dilute Lenograstim RS with FBS-IMDM so that each mL contains 7.69 units, 10.0 units and 13.0 units, and name them as the standard solution (1), the standard solution (2) and the standard solution (3), respectively. Put exactly 100 mL each of the sample solutions and standard solutions in wells of a sterile disposable multiple well plate, add 50 mL each of NFS-60 cell suspension (prepared by adding FBS-IMDM so the each mL contains about 5 ×105 cells) to each well and mix to make homogenize, and place the plate in a CO2 incubator at 379C. After incubation for 22 hours, add 15 mL of resazurin solution to each well, and determine the JP XVIII Official Monographs / L-Leucine 1251 ‌absorbances at 570 nm, AT1 and AS1, and at 600 nm, AT2 and AS2. From the reaction values at each concentration of the standard solution and sample solution [deference of absor-bance (AS1 － AS2 and AT1 － AT2)], determine the rate of po-tency (Pr) of the sample solution to the standard solution by the parallel assay, and calculate the potency (unit) per 1 mg of protein of Lenograstim (Genetical Recombination). Pr ＝ anti ln (M ) M ＝ (PT － PS)/db PT ＝ T1 ＋ T2 ＋ T3 PS ＝ S1 ＋ S2 ＋ S3 b ＝ HL(LS ＋ LT)/Inh HL ＝ 12n/(d3 － d) LS ＝ 1S1 ＋ 2S2 ＋ 3S3 － 1/2(d ＋ 1)PS LT ＝ 1T1 ＋ 2T2 ＋ 3T3 － 1/2(d ＋ 1)PT d ＝ 3 I ＝ ln 1.3 n ＝ 3 h ＝ 2 T1: Mean of reaction values of the sample solution (1) T2: Mean of reaction values of the sample solution (2) T3: Mean of reaction values of the sample solution (3) S1: Mean of reaction values of the standard solution (1) S2: Mean of reaction values of the standard solution (2) S3: Mean of reaction values of the standard solution (3) Specific activity (unit/mg of protein) of lenograstim ＝ S × Pr × D /D /C the pH of this solution is between 5.5 and 6.5. Purity (1) Clarity and color of solution—Dissolve 0.5 g of L-Leucine in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of L-Leucine in 40 mL of water and 6 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 0.6 g of L-Leucine in 40 mL of water and 1 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Leucine. Prepare the control solution with 5.0 mL of Standard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 1.0 g of L-Leu-cine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of L-Leucine according to Method 2, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of L-Leucine in water by warming, after cooling, add water to make 25 mL, and use this solution as the sample solution. Pipet 1 mL of T S the sample solution, and add water to make exactly 50 mL. S: Potency (unit/mL) of Lenograstim RS DT: Dilution rate of the sample solution (3) DS: Dilution rate of the standard solution (3) C: Concentration (mg/mL) of protein of sample
Containers—Tight containers. Storage—At a temperature not exceeding －209C.
Leuprorelin Acetate contains not less than 96.
Leuprorelin Acetate occurs as a white to yel-lowish white powder. It is very soluble in water and in acetic acid (100), freely soluble in methanol, and sparingly soluble in ethanol (99.5). It is hygroscopic.
Determine the infrared absorption spectrum of Leuprorelin Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Leuprorelin Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －38 – －419(0.25 g calcu-lated on the anhydrous and residual acetic acid-free basis, diluted acetic acid (100) (1 in 100), 25 mL, 100 mm). pH <2.54> The pH of a solution of 0.10 g of Leuprorelin Acetate in 10 mL of water is 5.5 to 7.5. Constituent amino acids When hydrolyzed by Method 1 described in ``1. Hydrolysis of Protein and Peptide'' and performed the test by Method 1 described in ``2. Methodol-ogies of Amino Acid Analysis'' under Amino Acid Analysis of Proteins <2.04>, histidine, glutamic acid, proline, tyrosine and arginine is 1 and leucine is 2, respectively. Procedure Hydrolysis Weigh accurately about 50 mg of Leuprorelin Acetate, and dissolve in 1 mL of water. Put 0.1 mL of this solution in a test tube for hydrolysis, freeze-dry the content, and add 2 mL of a solution of phenol in 6 mol/L hydrochloric acid (1 in 200). Freeze the solution, seal the tube in vacuum, and heat the tube at 1109C for 24 hours. After cooling, open the tube, take out 0.1 mL of the hydrolyzate, add 1 mL of water, and freeze-dry. Dissolve the residue in 7.8 mL of diluting solution, and use this solution Column: A stainless steel column 4.6 mm in inside diame-ter and 6 cm in length, packed with strongly acidic ion-ex-change resin (Na type) for liquid chromatography (3 mm in particle diameter). Column temperature: Maintain a constant temperature of about 589C for 18 minutes after injection, then maintain a constant temperature of about 709C for a further 20 minutes. Reaction vessel temperature: A constant temperature of about 1359C. Mobile phase A Mobile phase B Mobile phase C Mobile phase D Mobile phase E Citric acid monohy- 19.80 g drate 22.00 g 12.80 g 6.10 g — Trisodium citrate 6.19 g dihydrate 7.74 g 13.31 g 26.67 g — Sodium 5.66 g chloride 7.07 g 3.74 g 54.35 g — Sodium — hydroxide — — — 8.00 g Ethanol 130 mL (99.5) 20.0 mL 4.0 mL — 100 mL Thiodiglycol 5.0 mL 5.0 mL 5.0 mL — — Benzyl — alcohol — — 5.0 mL — Lauro- macrogol 4.0 mL 4.0 mL 4.0 mL 4.0 mL 4.0 mL Mobile phase: Prepare the mobile phases A, B, C, D and E according to the following table, then add 0.1 mL of caprylic acid to each mobile phase. solution (1 in 4) a sufficient a sufficient a sufficient a sufficient a sufficient as the sample solution. Separately, weigh exactly 0.45 mg of L-alanine, 0.66 mg of L-aspartic acid, 1.05 mg of L-arginine Water amount amount amount amount amount hydrochloride, 0.74 mg of L-glutamic acid, 0.38 mg of gly-cine, 1.05 mg of L-histidine hydrochloride monohydrate, 0.66 mg of L-isoleucine, 0.66 mg of L-leucine, 0.58 mg of Total amount 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL L-proline, 0.53 mg of L-serine, 0.60 mg of L-threonine and 0.91 mg of L-tyrosine, dissolve in diluting solution to make exactly 100 mL, and use this solution as the standard solu-tion (1). Separately, dissolve 1 mg of L-tryptophan and 0.4 mg of ethylamine hydrochloride in diluting solution to make 100 mL, and use this solution as the standard solution (2). Amino acid analysis Perform the test with exactly 100 mL each of the sample solution and the standard solu-tions (1) and (2) as directed under Liquid Chromatography Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B, C, D and E as directed in the follow-ing table. Time after Mobile Mobile Mobile Mobile Mobile injection of phase A phase B phase C phase D phase E sample (min) (volz) (volz) (volz) (volz) (volz) 0 – 1.6 1.6 – 4.5 4.5 – 13.5 13.5 – 27.0 27.0 – 33.0 Official Monographs / Leuprorelin Acetate 1253 Reaction reagent: Dissolve an appropriate amount of lithi-um acetate dihydrate, acetic acid (100) and 1-methoxy-2-propanol in water to make 1000 mL, and use this solution as solution A. Separately, dissolve an appropriate amount of ninhydrin and sodium borohydride in 1-methoxy-2-propanol to make 1000 mL, and use this solution as solution B. Mix equal parts of solutions A and B before use. Flow rate of mobile phase: About 0.40 mL per minute. Flow rate of reaction reagent: About 0.35 mL per minute. System suitability— System performance: When the procedure is run with 100 mL of the standard solution (1) under the above operating conditions, the resolutions between the peaks of threonine and serine, glycine and alanine, and isoleucine and leucine are not less than 1.2, respectively. System repeatability: When the test is repeated 5 times with 100 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviation of the peak area of arginine, aspartic acid, proline and serine is not more than 4.0z.
Related substances—Dissolve 0.10 g of Leuprorelin Acetate in the mobile phase to make 100 mL, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks, having the relative retention time of about 0.65, about 0.77, about 0.78 and about 0.90 to leuprorelin, ob-tained from the sample solution is not larger than 1/2 times the peak area of leuprorelin from the standard solution, and the total area of the peaks other than leuprorelin from the sample solution is not larger than 2 times the peak area of leuprorelin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of leuprorelin, beginning after the solvent peak. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of leuprorelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. Water <2.48> Not more than 5.0z (0.1 g, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (0.5 g). Acetic acid Weigh accurately about 0.1 g of Leuprorelin Acetate, dissolve in the mobile phase to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of acetic acid (100), add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of acetic acid in each solution, and calculate the amount of acetic acid by the following equation: 4.7 – 8.0z. Amount (z) of acetic acid ＝ MS/MT × AT/AS × 10 MS: Amount (g) of acetic acid (100) taken MT: Amount (g) of Leuprorelin Acetate taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 0.7 mL of phosphoric acid add water to make 1000 mL, and adjust to pH 3.0 with a solution of so-dium hydroxide (21 in 50). To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of acetic acid is 3 to 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the symmetry factor of the peak of acetic acid is not more than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of acetic acid is not more than 2.0z.
Weigh accurately about 0.1 g each of Leuprorelin Acetate and Leuprorelin Acetate RS (separately determine the water <2.48> and acetic acid in the same manner as Leuprorelin Acetate), dissolve separately in the mobile phase to make exactly 100 mL. To exactly 5 mL each of these solu-tions add the mobile phase to make them exactly 100 mL, and use so obtained solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of leuprorelin in each solution. Amount (mg) of leuprorelin (C59H84N16O12) ＝ MS × AT/AS MS: Amount (mg) of Leuprorelin Acetate RS taken, calcu-lated on the anhydrous and de-acetic acid basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 15.2 g of triethylamine in 800 mL of water, adjust to pH 3.0 with phosphoric acid, and add ‌1254 water to make 1000 mL. To 850 mL of this solution add 150 mL of a mixture of acetonitrile and 1-propanol (3:2). Flow rate: Adjust so that the retention time of leuprorelin is 41 to 49 minutes (1.0 – 1.5 mL per minute). System suitability— ‌System performance: Dissolve about 0.1 g of Leuprorelin Acetate RS in 100 mL of the mobile phase. To 5 mL of this solution add water to make 50 mL. To 5 mL of this solution add 0.1 mL of sodium hydroxide TS, stopper the vessel, shake vigorously, then heat at 1009C for 60 minutes. After cooling, add 50 mL of 1 mol/L phosphoric acid solution, and shake vigorously. When the procedure is run with 20 mL of this solution under the above operating conditions, a peak having the relative retention time of about 0.90 to leuprore-lin and leuprorelin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of leuprorelin is not more than 1.5z.
Levallorphan Tartrate, when dried, contains not less than 98.
Levallorphan Tartrate occurs as a white to pale yellow crystalline powder. It is odorless. It is soluble in water and in acetic acid (100), sparingly soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Levallorphan Tartrate in 0.01 mol/L hydrochlo-ric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Levallorphan Tartrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Levallorphan Tartrate (1 in 30) re-sponds to Qualitative Tests <1.09> (1) and (2) for tartrate. D Optical rotation <2.49> [a]20: －37.0 – －39.29(after dry-ing, 0.2 g, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.2 g of Levallorphan Tartrate in 20 Each mL of 0.1 mol/L perchloric acid VS ＝ 43.35 mg of C19H25NO.C4H6O6
(1) Clarity and color of solution—Dissolve 0.2 g of Levallorphan Tartrate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Levallor- phan Tartrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Levallorphan Tartrate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and ammonia TS (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.10z (1 g).
Weigh accurately about 0.5 g of Levallorphan Tar-trate, previously dried, dissolve in 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indica-tor: 2 drops of crystal violet TS). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. C19H25NO.C4H6O6: 433.49 17-Allylmorphinan-3-ol monotartrate [71-82-9] Levallorphan Tartrate, when dried, contains not less than 98.5z of levallorphan tartrate (C19H25NO. C4H6O6). Description Levallorphan Tartrate occurs as a white to pale yellow crystalline powder. It is odorless. It is soluble in water and in acetic acid (100), sparingly soluble in ethanol (95), and practically insoluble in diethyl ether. Identification (1) Determine the absorption spectrum of a solution of Levallorphan Tartrate in 0.01 mol/L hydrochlo-ric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Levallorphan Tartrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Levallorphan Tartrate (1 in 30) re-sponds to Qualitative Tests <1.09> (1) and (2) for tartrate. D Optical rotation <2.49> [a]20: －37.0 – －39.29(after dry-ing, 0.2 g, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.2 g of Levallorphan Tartrate in 20 Each mL of 0.1 mol/L perchloric acid VS ＝ 43.35 mg of C19H25NO.C4H6O6
Levodopa, when dried, contains not less than 98.
Levodopa occurs as white or slightly grayish white, crystals or crystalline powder. It is odorless. It is freely soluble in formic acid, slightly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid. The pH of a saturated solution of Levodopa is between 5.0 and 6.5. Melting point: about 2759C (with decomposition).
(1) To 5 mL of a solution of Levodopa (1 in 1000) add 1 mL of ninhydrin TS, and heat for 3 minutes in a water bath: a purple color develops. To 2 mL of a solution of Levodopa (1 in 5000) add 10 mL of 4-aminoantipyrine TS, and shake: a red color de-velops. Dissolve 3 mg of Levodopa in 0.001 mol/L hydro-chloric acid TS to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. 1 cm Absorbance <2.24> E 1z (280 nm): 136 – 146 (after drying, 30 mg, 0.001 mol/L hydrochloric acid TS, 1000 mL). D Optical rotation <2.49> [a]20: －11.5 – －13.09(after dry-ing, 2.5 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Levodopa in 20 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of Levodopa in 6 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 0.40 g of Levodopa in 1 mL of dilute hydrochloric acid and 30 mL of water, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.005 mol/L sulfuric acid VS (not more than 0.030z). Heavy metals <1.07>—Proceed with 1.0 g of Levo- dopa according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of Levodopa in 5 mL of dilute hydrochloric acid, and perform the test with this so-lution as the test solution (not more than 2 ppm). Related substances—Dissolve 0.10 g of Levodopa in 10 mL of sodium disulfite TS, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add so-dium disulfite TS to make exactly 25 mL. Pipet 1 mL of this solution, add sodium disulfite TS to make exactly 20 mL, ‌1256 and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water, acetic acid (100) and methanol (10:5:5:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate and heat the plate at 909C for 10 minutes: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.30z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Levodopa, previ-ously dried, dissolve in 3 mL of formic acid, add 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-green to green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.72 mg of C9H11NO4
Levofloxacin Hydrate occurs as light yellowish white to yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in water and in methanol, and slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. It gradually turns dark light yellow-white on exposure to light. Melting point: about 2269C (with decomposition).
(1) Determine the absorption spectrum of a solution of Levofloxacin Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 150,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Levofloxacin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －92 – －999(0.1 g calculated on the anhydrous basis, methanol, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Levofloxacin Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Levofloxacin Hy-drate in 10 mL of a mixture of water and methanol (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and metha-nol (1:1) to make exactly 10 mL. Pipet 1 mL of this solution, add a mixture of water and methanol (1:1) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area of both solutions by the automatic integration method: the area of the peak of the enantiomer having the relative retention time of about 1.2 to levofloxa-cin obtained from the sample solution is not larger than 2/5 times the peak area of levofloxacin from the standard solu-tion, and the area of each peak other than the peaks of levofloxacin and the enantiomer from the sample solution is not larger than 1/5 times the peak area of levofloxacin from the standard solution. Furthermore, the total area of the peaks other than levofloxacin and the peak of the enantiom-er from the sample solution is not larger than 3/10 times the peak area of levofloxacin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 1.76 g of L-valine, 7.71 g of am-monium acetate and 1.25 g of Copper (II) sulfate pentahy-drate in water to make 1000 mL. To this solution add 250 mL of methanol. Flow rate: Adjust so that the retention time of levofloxa-cin is about 22 minutes. Time span of measurement: About 2 times as long as the retention time of levofloxacin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and methanol (1:1) to make exactly 20 mL. Confirm that the peak area of levoflox-acin obtained with 10 mL of this solution is equivalent to 4 to 6z of that with 10 mL of the standard solution. System performance: Dissolve 10 mg of ofloxacin in 20 mL of a mixture of water and methanol (1:1). To 1 mL of this solution add a mixture of water and methanol (1:1) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolu-tion between the peaks of levofloxacin and the enantiomer is not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌JP XVIII Official Monographs / Levofloxacin Fine Granules 1257 ing conditions, the relative standard deviation of the peak area of levofloxacin is not more than 3.0z. Water <2.48> 2.1 – 2.7z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.3 g of Levofloxacin Hy-drate, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.14 mg of C18H20FN3O4
Levomepromazine Maleate, when dried, contains not less than 98.
Levomepromazine Maleate occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in acetic acid (100), soluble in chlo-roform, sparingly soluble in methanol, slightly soluble in ethanol (95) and in acetone, very slightly soluble in water, and practically insoluble in diethyl ether. Melting point: 184 – 1909C (with decomposition).
(1) Dissolve 5 mg of Levomepromazine Maleate in 5 mL of sulfuric acid: a red-purple color de-velops, which slowly becomes deep red-purple. To this solu-tion add 1 drop of potassium dichromate TS: a brownish yel-low-red color is produced. To 0.2 g of Levomepromazine Maleate add 5 mL of sodium hydroxide TS and 20 mL of diethyl ether, and shake
Weigh accurately about 1 g of Levomepromazine Maleate, previously dried, and dissolve in a mixture of 40 mL of acetic acid (100) and 20 mL of acetone for nona-queous titration. Titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from red-purple through blue-purple to blue (indicator: 5 drops of bromocresol green-methylrosaniline chloride TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.45 mg of C H N OS.C H O 19 24 2 4 4 4 Levomepromazine Maleate, when dried, contains not less than 98.0z of levomepromazine maleate (C19H24N2OS.C4H4O4). Description Levomepromazine Maleate occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in acetic acid (100), soluble in chlo-roform, sparingly soluble in methanol, slightly soluble in ethanol (95) and in acetone, very slightly soluble in water, and practically insoluble in diethyl ether. Melting point: 184 – 1909C (with decomposition). Identification (1) Dissolve 5 mg of Levomepromazine Maleate in 5 mL of sulfuric acid: a red-purple color de-velops, which slowly becomes deep red-purple. To this solu-tion add 1 drop of potassium dichromate TS: a brownish yel-low-red color is produced. To 0.2 g of Levomepromazine Maleate add 5 mL of sodium hydroxide TS and 20 mL of diethyl ether, and shake
Lidocaine, when dried, contains not less than 99.
Lidocaine occurs as white to pale yellow, crys-tals or crystalline powder. It is very soluble in methanol and in ethanol (95), soluble in acetic acid (100) and in diethyl ether, and practically in-soluble in water. It dissolves in dilute hydrochloric acid.
(1) Dissolve 40 mg of Lidocaine in 10 mL of 1 mol/L hydrochloric acid TS, and add water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Lidocaine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Lidocaine in 2 mL of dilute hydrochloric acid, and add water to make 10 mL: the solution is clear and colorless to light yellow. Chloride <1.03>—Dissolve 0.6 g of Lidocaine in 6 mL of dilute nitric acid, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.70 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.041z). Sulfate <1.14>—Dissolve 0.5 g of Lidocaine in 5 mL of dilute hydrochloric acid, add water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 1.0 mL of 0.005 mol/L sulfu-ric acid VS, 5 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.096z). Heavy metals <1.07>—Carbonize 2.0 g of Lidocaine by gentle ignition. After cooling, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and fire the ethanol to burn. After cooling, add 1 mL of sulfuric acid, proceed according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Lidocaine in 2 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog- ‌1264 raphy. Develop the plate with a mixture of ethyl acetate, 2-butanone, water and formic acid (5:3:1:1) to a distance of about 10 cm, air-dry the plate, and dry more at 809C for 30 minutes. After cooling, examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. ‌Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 0.5 g of Lidocaine, previously dried and accurately weighed, in 20 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 1 drop of crystal violet TS) until the color of the solution changes from purple to blue-green through blue. Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 23.43 mg of C14H22N2O
Light Anhydrous Silicic Acid contains not less than 98.
Light Anhydrous Silicic Acid occurs as a white to bluish white, light, fine power. It is odorless and tasteless, and smooth to the touch. It is practically insoluble in water, in ethanol (95), and in diethyl ether. It dissolves in hydrofluoric acid, in hot potassium hydrox-ide TS and in hot sodium hydroxide TS, and does not dis-solve in dilute hydrochloric acid.
(1) Dissolve 0.1 g of Light Anhydrous Silicic Acid in 20 mL of sodium hydroxide TS by boiling, and add 12 mL of ammonium chloride TS: a white, gelatinous precipitate is produced. The precipitate does not dissolve in dilute hydrochloric acid. To the precipitate obtained in (1) add 10 mL of a so-lution of methylene blue trihydrate (1 in 10,000), and wash with water: the precipitate has a blue color. Prepare a bead by fusing ammonium sodium hydrogenphosphate tetrahydrate on a platinum loop. Bring the hot, transparent bead into contact with Light Anhydrous Silicic Acid, and fuse again: an insoluble matter is percepti-ble in the bead. The resulting bead, upon cooling, becomes opaque and acquires a reticulated appearance.
(1) Chloride <1.03>—Dissolve 0.5 g of Light An-hydrous Silicic Acid in 20 mL of sodium hydroxide TS by boiling, cool, filter if necessary, and wash with 10 mL of water. Combine the filtrate and washings, add 18 mL of di-lute nitric acid, shake, and add water to make 50 mL. Per-form the test using this solution as the test solution. To 0.15 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of sodi-um hydroxide TS, 18 mL of dilute nitric acid and water to make 50 mL, and use this solution as the control solution (not more than 0.011z). Heavy metals <1.07>—Dissolve 0.5 g of Light Anhy- drous Silicic Acid in 20 mL of sodium hydroxide TS by boil- ing, cool, add 15 mL of acetic acid (31), shake, filter if nec-essary, wash with 10 mL of water, combine the filtrate and washings, and add water to make 50 mL. Perform the test using this solution as the test solution. Add acetic acid (31) to 20 mL of sodium hydroxide TS and 1 drop of phenol-phthalein TS until the color of this solution disappears, add 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL, and use this solution as the control solution (not more than 40 ppm). Iron <1.10>—To 0.040 g of Light Anhydrous Silicic Acid add 10 mL of dilute hydrochloric acid, and heat for 10 minutes in a water bath while shaking. After cooling, add 0.5 g of L-tartaric acid to dissolve by shaking. Prepare the test solution with this solution according to Method 2, and perform the test according to Method B. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 500 ppm). Aluminum—Dissolve 0.5 g of Light Anhydrous Silicic Acid in 40 mL of sodium hydroxide TS by boiling, cool, add sodium hydroxide TS to make 50 mL, and filter. Measure 10 mL of the filtrate, add 17 mL of acetic acid (31), shake, add 2 mL of aluminon TS and water to make 50 mL, and allow to stand for 30 minutes: the color of this solution is not deeper than that of the following control solution. Control solution: Dissolve 0.176 g of aluminum potassium sulfate dodecahydrate in water, and add water to make 1000 mL. To 15.5 mL of this solution add 10 mL of sodium hy-droxide TS, 17 mL of acetic acid (31), 2 mL of aluminon TS and water to make 50 mL. Calcium—Dissolve 1.0 g of Light Anhydrous Silicic Acid in 30 mL of sodium hydroxide TS by boiling, cool, add 20 mL of water, 1 drop of phenolphthalein TS and dilute nitric acid until the color of this solution disappears, imme-diately add 5 mL of dilute acetic acid, shake, add water to make 100 mL, and obtain a clear liquid by centrifugation or filtration. To 25 mL of this liquid add 1 mL of oxalic acid TS and ethanol (95) to make 50 mL, immediately shake, and allow to stand for 10 minutes: the turbidity of this solution is not deeper than that of the following control solution. Control solution: Dissolve 0.250 g of calcium carbonate, previously dried at 1809C for 4 hours, in 3 mL of dilute hy-drochloric acid, and add water to make 100 mL. To 4 mL of this solution add 5 mL of dilute acetic acid and water to make 100 mL. To 25 mL of this solution add 1 mL of oxalic acid TS and ethanol (95) to make 50 mL, and shake. Arsenic <1.11>—Dissolve 0.40 g of Light Anhydrous Silicic Acid in 10 mL of sodium hydroxide TS by boiling in a porcelain crucible, cool, add 5 mL of water and 5 mL of di-lute hydrochloric acid, shake, and perform the test with this solution as the test solution (not more than 5 ppm). Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 4 hours). Loss on ignition <2.43> Not more than 12.0z (1 g, 850 – 9009C, constant mass).
Weigh accurately about 1 g of Light Anhydrous Silicic Acid, add 20 mL of hydrochloric acid, and evaporate to dryness on a sand bath. Moisten the residue with hydro-chloric acid, evaporate to dryness, and heat between 1109C and 1209C for 2 hours. Cool, add 5 mL of dilute hydrochlo-ric acid, and heat. Allow to cool to room temperature, add 20 to 25 mL of hot water, filter rapidly, and wash the residue with warm water until the last washing becomes negative to Qualitative Tests <1.09> (2) for chloride. Transfer the residue together with the filter paper to a platinum crucible, ignite to ash, and continue the ignition for 30 minutes. Cool, weigh ‌the crucible, and designate the mass as a (g). Moisten the residue in the crucible with water, add 6 mL of hydrofluoric acid and 3 drops of sulfuric acid, and evaporate to dryness. Heat strongly for 5 minutes, cool, weigh the crucible, and designate the mass as b (g). Amount (g) of silicon dioxide (SiO2) ＝ a － b
Light Liquid Paraffin is a clear, colorless oily liquid, nearly free from fluorescence. It is odorless and taste-less. It is freely soluble in diethyl ether, and practically insolu-ble in water and in ethanol (95). Boiling point: above 3009C.
(1) Heat Light Liquid Paraffin strongly in a porcelain dish, and fire: it burns with a bright flame and the odor of paraffin vapor is perceptible. (2) Heat 0.5 of Light Liquid Paraffin with 0.5 g of sulfur with shaking carefully: the odor of hydrogen sulfide is per-ceptible. Specific gravity <2.56> d 20: 0.830 – 0.870 Viscosity <2.53> Less than 37 mm2/s (Method 1, 37.89C).
(1) Odor—Transfer a suitable amount of Light Liquid Paraffin to a small beaker, and heat on a water bath: no foreign odor is perceptible. Acidity or alkalinity—Shake vigorously 10 mL of Light Liquid Paraffin with 10 mL of hot water and 1 drop of phenolphthalein TS: no red color develops. Shake this solu-tion with 0.20 mL of 0.02 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Ignite 2.0 g of Light Liquid Paraffin in a crucible, first moderately until charred, then between 4509C and 5509C to ash. Cool, add 2 mL of hydro-chloric acid, and evaporate on a water bath to dryness. To the residue add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and lead (II) oxide in a solution of sodium hydroxide (1 in 5), and mix 2 drops of this clear solution with 4.0 mL of Light Liquid Paraffin and 2 mL of ethanol (99.5). Heat at 709C for 10 minutes with frequent shaking, and cool: no dark brown color develops. Polycyclic aromatic hydrocarbons—Take 25 mL of Light Liquid Paraffin by a 25-mL measuring cylinder, trans-fer to a 100-mL separator, and wash out the cylinder with 25 mL of hexane for ultraviolet-visible spectrophotometry. Combine the washings with the liquid in the separator, and shake vigorously. Shake this solution vigorously for 2 minutes with 5.0 mL of dimethylsulfoxide for ultraviolet-visible spectrophotometry, and allow to stand for 15 minutes. Transfer the lower layer to a 50-mL separator, add 2 mL of hexane for ultraviolet-visible spectrophotometry, shake vigorously for 2 minutes, and allow to stand for 2 minutes. Transfer the lower layer to a glass-stoppered 10-mL centrifuge tube, and centrifuge between 2500 revolutions per minute and 3000 revolutions per minute for about 10 minutes, and use the clear solution so obtained as the sample solution. Separately, transfer 25 mL of hexane for ultravio-let-visible spectrophotometry to a 50-mL separator, add 5.0 mL of dimethylsulfoxide for ultraviolet-visible spectropho-tometry, shake vigorously for 2 minutes, and allow to stand for 2 minutes. Transfer the lower layer to a glass-stoppered 10-mL centrifuge tube, centrifuge between 2500 revolutions per minute and 3000 revolutions per minute for about 10 minutes, and use the clear solution so obtained as a control solution. Immediately determine the absorbance of the sam-ple solution using the control solution as the blank as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.10 at the wavelength region between 260 nm and 350 nm. Readily carbonizable substances—Transfer 5 mL of Light Liquid Paraffin to a Nessler tube, and add 5 mL of sulfuric acid for readily carbonizable substances. After heat-ing in a water bath for 2 minutes, remove the tube from the water bath, and immediately shake vigorously and vertically for 5 seconds. Repeat this procedure four times: the liquid paraffin layer remains unchanged in color, and sulfuric acid layer has no more color than the following control solution. Control solution: Mix 3.0 mL of Iron (III) Chloride CS with 1.5 mL of Cobalt (II) Chloride CS and 0.50 mL of Copper (II) Sulfate CS.
Limaprost Alfadex is a a-cyclodextrin clathrate compound of limaprost. It contains not less than 2.
Limaprost Alfadex occurs as a white powder. It is freely soluble in water, slightly soluble in methanol, very slightly soluble in ethanol (99.5), and practically insolu- ble in ethyl acetate. It is hygroscopic.
(1) Dissolve 20 mg of Limaprost Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, and use the upper layer as the sample solution (1). Sepa-rately, to 20 mg of Limaprost Alfadex add 5 mL of ethyl acetate, shake, centrifuge, and use the supernatant liquid as the sample solution (2). Evaporate the solvent of the sample solutions (1) and (2) under reduced pressure, add 2 mL of sulfuric acid to each of the residue, and shake them for 5 minutes: the solution obtained from the sample solution (1) develops an orange-yellow color while the solution obtained from the sample solution (2) does not develop any color. Dissolve 20 mg of Limaprost Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, and evaporate the solvent of the upper layer under reduced pres-sure. Dissolve the residue in 2 mL of ethanol (95), 5 mL of 1,3-dinitrobenzene TS, add 5 mL of a solution of potassium hydroxide in ethanol (95) (17 in 100) while ice-cooling, and allow to stand in a dark place while ice-cooling for 20 minutes: a purple color develops. To 50 mg of Limaprost Alfadex add 1 mL of iodine TS, dissolve by heating in a water bath, and allow to stand: a dark blue precipitate is formed. Determine the absorption spectrum of a solution of Limaprost Alfadex in dilute ethanol (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it does not exhibit a maximum between 200 nm and 400 nm. To 10 mL of this solution add 1 mL of potassium hydroxide-ethanol TS, and allow to stand for 15 minutes. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. D Optical rotation <2.49> [a]20: ＋125 – 1359(0.1 g calculated on the anhydrous basis, dilute ethanol, 20 mL, 100 mm).
Related substances—Perform the test immediately after preparation of the sample solution. Dissolve 0.10 g of Limaprost Alfadex in 2 mL of water, add 1 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dilute ethanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 3 mL of the standard solution (1), add dilute ethanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 3 mL each of the sample solution and standard solutions (1) and (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak of 17-epi-isomer, having the relative retention time of about 1.1 to limaprost, and the area of the peak of 11-deoxy sub-stance, having the relative retention time of about 2.1, ob-tained from the sample solution are not larger than the peak area of limaprost from the standard solution (2), and the area of the peak other than the principal peak and the peaks mentioned above from the sample solution is not larger than 1/3 times the peak area of limaprost from the standard solu-tion (2). The total area of the peaks other than limaprost from the samples solution is not larger than the peak area of limaprost from the standard solution (1). Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of limaprost beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution (1) add dilute ethanol to make exactly 10 mL. Confirm that the peak area of limaprost obtained with 3 mL of this solution is equivalent to 8 to 12z of that with 3 mL of the standard solution (1). System repeatability: When the test is repeated 6 times with 3 mL of the standard solution (1) under the above condi-tions, the relative standard deviation of the peak area of limaprost is not more than 2.0z. Water <2.48> Not more than 6.0z (0.2 g, volumetric titra-tion, direct titration).
To an exactly measured volume of Lidocaine Injec-tion, equivalent to about 0.1 g of lidocaine hydrochloride (C14H22N2O.HCl), add exactly 10 mL of the internal stand-ard solution and 0.001 mol/L hydrochloric acid TS to make (2E )-7-{(1R,2R,3R)-3-Hydroxy-2-[(1E,3S,5S )-3- hydroxy-5-methylnon-1-en-1-yl]- 5-oxocyclopentyl}hept-2-enoic acid-a-cyclodextrin [100459-01-6, limaprost:alfadex ＝ 1:1; clathrate compound] Limaprost Alfadex is a a-cyclodextrin clathrate compound of limaprost. It contains not less than 2.8z and not more than 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 85 mg of lidocaine for assay, 3.2z of limaprost (C the anhydrous basis. 22H36 O5: 380.52), calculated on JP XVIII Official Monographs / Limaprost Alfadex 1265 Description Limaprost Alfadex occurs as a white powder. It is freely soluble in water, slightly soluble in methanol, very slightly soluble in ethanol (99.5), and practically insolu- ble in ethyl acetate. It is hygroscopic. Identification (1) Dissolve 20 mg of Limaprost Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, and use the upper layer as the sample solution (1). Sepa-rately, to 20 mg of Limaprost Alfadex add 5 mL of ethyl acetate, shake, centrifuge, and use the supernatant liquid as the sample solution (2). Evaporate the solvent of the sample solutions (1) and (2) under reduced pressure, add 2 mL of sulfuric acid to each of the residue, and shake them for 5 minutes: the solution obtained from the sample solution (1) develops an orange-yellow color while the solution obtained from the sample solution (2) does not develop any color. Dissolve 20 mg of Limaprost Alfadex in 5 mL of water, add 5 mL of ethyl acetate, shake, centrifuge, and evaporate the solvent of the upper layer under reduced pres-sure. Dissolve the residue in 2 mL of ethanol (95), 5 mL of 1,3-dinitrobenzene TS, add 5 mL of a solution of potassium hydroxide in ethanol (95) (17 in 100) while ice-cooling, and allow to stand in a dark place while ice-cooling for 20 minutes: a purple color develops. To 50 mg of Limaprost Alfadex add 1 mL of iodine TS, dissolve by heating in a water bath, and allow to stand: a dark blue precipitate is formed. Determine the absorption spectrum of a solution of Limaprost Alfadex in dilute ethanol (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it does not exhibit a maximum between 200 nm and 400 nm. To 10 mL of this solution add 1 mL of potassium hydroxide-ethanol TS, and allow to stand for 15 minutes. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. D Optical rotation <2.49> [a]20: ＋125 – 1359(0.1 g calculated on the anhydrous basis, dilute ethanol, 20 mL, 100 mm). Purity Related substances—Perform the test immediately after preparation of the sample solution. Dissolve 0.10 g of Limaprost Alfadex in 2 mL of water, add 1 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dilute ethanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 3 mL of the standard solution (1), add dilute ethanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 3 mL each of the sample solution and standard solutions (1) and (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak of 17-epi-isomer, having the relative retention time of about 1.1 to limaprost, and the area of the peak of 11-deoxy sub-stance, having the relative retention time of about 2.1, ob-tained from the sample solution are not larger than the peak area of limaprost from the standard solution (2), and the area of the peak other than the principal peak and the peaks mentioned above from the sample solution is not larger than 1/3 times the peak area of limaprost from the standard solu-tion (2). The total area of the peaks other than limaprost from the samples solution is not larger than the peak area of limaprost from the standard solution (1). Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of limaprost beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution (1) add dilute ethanol to make exactly 10 mL. Confirm that the peak area of limaprost obtained with 3 mL of this solution is equivalent to 8 to 12z of that with 3 mL of the standard solution (1). System repeatability: When the test is repeated 6 times with 3 mL of the standard solution (1) under the above condi-tions, the relative standard deviation of the peak area of limaprost is not more than 2.0z. Water <2.48> Not more than 6.0z (0.2 g, volumetric titra-tion, direct titration). Assay Weigh accurately about 0.1 g of Limaprost Afladex, dissolve in 5 mL of water, add exactly 5 mL of the internal standard solution, and use this solution as the sample solu-tion. Separately, weigh accurately about 3 mg of Limaprost RS, dissolve in exactly 5 mL of the internal standard solu-tion, add 5 mL of water, and use this solution as the stand-ard solution. Perform the test with 3 mL each of the sample solution and the standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of limaprost to that of the internal standard. Amount (mg) of limaprost (C22H36O5) ＝ MS × QT/QS MS: Amount (mg) of Limaprost RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in ethanol (95) (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogen phosphate TS, acetonitrile for liquid chromatogra-phy and 2-propanol for liquid chromatography (9:5:2). Flow rate: Adjust so that the retention time of limaprost is about 12 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, the internal standard and limaprost are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of limaprost to that of the internal standard is not more than 1.0z.
Lincomycin Hydrochloride Hydrate is the hydro-chloride of a substance having antibacterial activity produced by the growth of Streptomyces lincolnensis var. lincolnensis. It contains not less than 850 mg (potency) and not more than 930 mg (potency) per mg, calculated on the anhydrous basis.
Lincomycin Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is freely soluble in water and in methanol, sparingly soluble in ethanol (95).
(1) Determine the infrared absorption spec-trum of Lincomycin Hydrochloride Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Lincomycin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of Lincomycin Hydrochloride Hydrate (1 in 100) responds to Qualitative Tests <1.09> (2) for chlo-ride. D Optical rotation <2.49> [a]20: ＋135 – ＋1509(0.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Lincomycin Hydrochloride Hydrate in 1 mL of water: 3.0 – 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Lincomycin Hydrochloride Hydrate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Linco- mycin Hydrochloride Hydrate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 5 ppm). Lincomycin B—Use the sample solution obtained in the Assay as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas of lincomycin and lincomycin B, having the relative retention time of about 0.5 to lincomycin obtained from the sample solution, by the automatic integra-tion method: the peak area of lincomycin B is not more than 2.0z of the sum of the peak areas of lincomycin and linco-mycin B. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 1 mL of the sample solution, and add the mobile phase to make ex-actly 50 mL. Confirm that the peak area of lincomycin ob-tained from 20 mL of this solution is equivalent to 1.4 to 2.6z of that obtained from 20 mL of the sample solution. Water <2.48> 3.0 – 6.0z (0.5 g, volumetric titration, direct titration).
Weigh accurately an amount of Lincomycin Hydro-chloride Hydrate and Lincomycin Hydrochloride RS, equivalent to about 10 mg (potency), dissolve each in the mobile phase to make exactly 10 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of lincomycin in each solution. Amount [ mg (potency)] of lincomycin (C18H34N2O6S) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Lincomycin Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 469C. Mobile phase: To 13.5 mL phosphoric acid add 1000 mL of water, and adjust the pH to 6.0 with ammonia TS. To 780 mL of this solution add 150 mL of acetonitrile and 150 mL of methanol. Flow rate: Adjust so that the retention time of lincomycin is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of lincomycin are not less than 4000 and not more than 1.3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lincomycin is not more than 2.0z.
Liquefied Phenol is Phenol maintained in a liquid condition by the presence of 10z of Water, Purified Water or Purified Water in Containers. It contains not less than 88.
Liquefied Phenol is a colorless or slightly red-dish liquid. It has a characteristic odor. It is miscible with ethanol (95), with diethyl ether and with glycerin. A mixture of equal volumes of Liquefied Phenol and glyc-erin is miscible with water. The color changes gradually to dark red on exposure to light or air. It cauterizes the skin, turning it white. Specific gravity d 20: about 1.065
(1) Add 1 drop of iron (III) chloride TS to 10 mL of a solution of Liquefied Phenol (1 in 100): a blue-purple color develops. (2) Add bromine TS dropwise to 5 mL of a solution of Liquefied Phenol (1 in 10,000): a white precipitate is pro-duced, which at first dissolves with shaking, but becomes permanent as excess of the reagent is added. Boiling point <2.57> Not more than 1829C.
(1) Clarity and color of solution and acidity or alkalinity—Dissolve 1.0 g of Liquefied Phenol in 15 mL of water: the solution is clear, and neutral or only faintly acid. Add 2 drops of methyl orange TS: no red color develops. (2) Residue on evaporation—Weigh accurately about 5 g of Liquefied Phenol, evaporate on a water bath, and dry the residue at 1059C for 1 hour: the mass is not more than 0.05z of the mass of the sample.
Dissolve about 1.7 g of Liquefied Phenol, accurately weighed, in a water to make exactly 1000 mL. Transfer ex-actly 25 mL of this solution to an iodine flask, add exactly 30 mL of 0.05 mol/L bromine VS, then 5 mL of hydrochloric acid, and immediately stopper the flask. Shake the flask repeatedly for 30 minutes, allow to stand for 15 minutes, then add 7 mL of potassium iodide TS, at one stopper the flask tightly, and shake well. Add 1 mL of chloroform, stop-per the flask, and shake thoroughly. Titrate <2.50> the liber-ated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 1.569 mg of C H O mediately stopper the flask. Shake the flask repeatedly for 6 6 30 minutes, allow to stand for 15 minutes, then add 7 mL of potassium iodide TS, at once stopper the flask, and shake well. Add 1 mL of chloroform, stopper the flask, and shake thoroughly. Titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 1.569 mg of C6H6O
Liquid Paraffin is a colorless, transparent, oily liquid, nearly free from fluorescence. It is odorless and taste-less. It is freely soluble in diethyl ether, very slightly soluble in ethanol (99.5), and practically insoluble in water and in ethanol (95). Boiling point: above 3009C.
(1) Heat Liquid Paraffin strongly in a por-celain dish, and fire: it burns with a bright flame and the odor of paraffin vapor is perceptible. (2) Heat 0.5 of Liquid Paraffin with 0.5 g of sulfur with shaking carefully: the odor of hydrogen sulfide is percepti-ble. Specific gravity <2.56> d 20: 0.860 – 0.890 Viscosity <2.53> Not less than 37 mm2/s (Method 1, 37.89C).
(1) Odor—Transfer a suitable amount of Liquid Paraffin to a small beaker, and heat on a water bath: a for-eign odor is not perceptible. Acidity or alkalinity—Shake vigorously 10 mL of Liquid Paraffin with 10 mL of hot water and 1 drop of phe-nolphthalein TS: no red color develops. Shake this solution with 0.20 mL of 0.02 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Ignite 2.0 g of Liquid Paraffin in a crucible, first moderately until charred, then between 4509C and 5509C to ash. Cool, add 2 mL of hydrochloric acid, and evaporate on a water bath to dryness. To the residue add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Liquid Paraffin, according to Method 3 except that after addition of 10 mL of a solution of magnesium nitrate hexa-hydrate in ethanol (95) (1 in 50), add 1.5 mL of hydrogen peroxide (30), fire to burn, and perform the test (not more than 2 ppm). Solid paraffin—Transfer 50 mL of Liquid Paraffin, previously dried at 1059C for 2 hours, to a Nessler tube, and cool in ice water for 4 hours: the turbidity produced, if any, is not deeper than that of the following control solution. Control solution: To 1.5 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL, add 1 mL of silver nitrate TS, and allow to stand for 5 minutes. Sulfur compounds—Prepare a saturated solution of lead (II) oxide in a solution of sodium hydroxide (1 in 5), and mix 2 drops of this clear solution with 4.0 mL of Liquid Paraffin and 2 mL of ethanol (99.5). Heat at 709C for 10 minutes with frequent shaking, and cool: no dark brown color develops. Polycyclic aromatic hydrocarbons—Take 25 mL of Liquid Paraffin by a 25-mL measuring cylinder, transfer to a 100-mL separator, and wash out the cylinder with 25 mL of hexane for ultraviolet-visible spectrophotometry. Combine the washings with the liquid in the separator, and shake vig-orously. Shake this solution vigorously for 2 minutes with 5.0 mL of dimethylsulfoxide for ultraviolet-visible spectro-photometry, and allow to stand for 15 minutes. Transfer the lower layer to a 50-mL separator, add 2 mL of hexane for ultraviolet-visible spectrophotometry, shake vigorously for 2 minutes, and allow to stand for 2 minutes. Transfer the lower layer to a 10-mL glass-stoppered centrifuge tube, and centrifuge between 2500 revolutions per minute and 3000 revolutions per minute for about 10 minutes, and use the clear solution obtained as the sample solution. Transfer 25 mL of hexane for ultraviolet-visible spectrophotometry to another 50-mL separator, shake vigorously for 2 minutes with 5.0 mL of dimethylsulfoxide for ultraviolet-visible spec-trophotometry, and allow to stand for 2 minutes. Transfer the lower layer to a 10-mL glass-stoppered centrifuge tube, centrifuge between 2500 revolutions per minute and 3000 revolutions per minute for about 10 minutes, and use the clear solution thus obtained as a control solution. Immedi-ately determine the absorbance of the sample solution using the control solution as the blank as directed under Ultravio-let-visible Spectrophotometry <2.24>: not more than 0.10 at the wavelength region between 260 nm and 350 nm. Readily carbonizable substances—Transfer 5 mL of Liquid Paraffin to a Nessler tube, and add 5 mL of sulfuric acid for readily carbonizable substances. After heating in a ‌water bath for 2 minutes, remove the tube from the water bath, and immediately shake vigorously and vertically for 5 seconds. Repeat this procedure 4 times: the Liquid Paraffin layer remains unchanged in color, and the sulfuric acid layer has no more color than the following control solution. Control solution: Mix 3.0 mL of Iron (III) Chloride CS with 1.5 mL of Cobalt (II) Chloride CS and 0.50 mL of Copper (II) Sulfate CS.
Lisinopril Hydrate contains not less than 98.
Lisinopril Hydrate occurs as a white crystalline powder, having a slight characteristic odor. It is soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (99.5). Melting point: about 1609C (with decomposition).
(1) Determine the absorption spectrum of a solution of Lisinopril Hydrate in methanol (1 in 1000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Lisinopril Hydrate as directed in the paste method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: －43.0 – －47.09(0.25 g cal-culated on the anhydrous basis, 0.25 mol/L zinc acetate buffer solution (pH 6.4), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Lisinopril Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve about 0.10 g of Lisinopril Hydrate in 50 mL of water, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having the relative retention time of about 1.2 to lisinopril obtained from the sample solution, is not larger than 1/5 times the peak area of lisinopril from the standard solution, the area of the peak other than lisinopril and the peak mentioned above from the sample solution, is not larger than 2/15 times the peak area of lisinopril from the standard solution, and the total area of the peaks other than lisinopril from the sample solution, is not larger than the peak area of lisinopril from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). ‌Column: A stainless steel column 4.0 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase A: Diluted 0.05 mol/L sodium dihydrogen phosphate TS (1 in 2). Mobile phase B: A mixture of diluted 0.05 mol/L sodium dihydrogen phosphate TS (1 in 2) and acetonitrile for liquid chromatography (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 90 → 50 10 → 50 10 – 25 Flow rate: About 1.5 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of lisinopril, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 2.5 mL of the standard solution, and add water to make exactly 50 mL. Confirm that the peak area of lisinopril obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System performance: To 10 mg of lisinopril hydrate and 2 mL of a solution of anhydrous caffeine (1 in 1000) add water to make 200 mL. When the procedure is run with 15 mL of this solution under the above operating conditions, lisinopril and caffeine are eluted in this order with the resolution be-tween these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lisinopril is not more than 2.0z. Water <2.48> Not less than 8.0z and not more than 9.5z (0.3 g, volumetric titration, back titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.66 g of Lisinopril Hy-drate, dissolve in 80 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 40.55 mg of C21H31N3O5
Lithium Carbonate, when dried, contains not less than 99.
Lithium Carbonate occurs as a white crystal-line powder. It is odorless. It is sparingly soluble in water, slightly soluble in hot water, and practically insoluble in ethanol (95) and in diethyl ether. It dissolves in dilute acetic acid. The pH of a solution dissolved 1.0 g of Lithium Car-bonate in 100 mL of water is between 10.9 and 11.5.
(1) Perform the test as directed under Flame Coloration Test <1.04> (1) with Lithium Carbonate: a persistent red color appears. Dissolve 0.2 g of Lithium Carbonate in 3 mL of dilute hydrochloric acid, and add 4 mL of sodium hydroxide TS and 2 mL of disodium hydrogen phosphate TS: a white precipitate is produced. To the precipitate add 2 mL of dilute hydrochloric acid: it dissolves. A solution of Lithium Carbonate (1 in 100) responds to Qualitative Tests <1.09> for carbonate.
(1) Clarity and color of solution—Dissolve 0.10 g of Lithium Carbonate in 10 mL of water by warming: the so-lution is clear and colorless. Acetic acid-insoluble substances—Take 1.0 g of Lithium Carbonate, dissolve in 40 mL of dilute acetic acid, filter the insoluble substances using filter paper for quantita-tive analysis, wash with five 10-mL portions of water, and ignite the insoluble substances together with the filter paper to incinerate: the mass of the residue is not more than 1.5 mg. Chloride <1.03>—To 0.40 g of Lithium Carbonate add 10 mL of water and 7 mL of dilute nitric acid, and dis-solve by heating to boil. After cooling, add 6 mL of dilute nitric acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.022z). Sulfate <1.14>—To 0.40 g of Lithium Carbonate add 10 mL of water and 4 mL of dilute hydrochloric acid, and dissolve by heating to boil. After cooling, add 1 mL of dilute hydrochloric acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.048z). Heavy metals <1.07>—To 4.0 g of Lithium Carbonate add 5 mL of water, gradually add 10 mL of hydrochloric acid while mixing, and dissolve. Evaporate the solution on a water bath to dryness. To the residue add 10 mL of water, and dissolve. Place the solution in a Nessler tube, add 1 drop of phenolphthalein TS, add ammonia TS until the solution shows a slight red color, then add 2 mL of dilute acetic acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: Evaporate 10 mL of hydrochloric acid on a water bath to dryness. To the residue add 10 mL of water, and dis-solve. Place the solution in a Nessler tube, add 1 drop of phenolphthalein TS, add ammonia TS until the solution shows a pale red color, then add 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to make 50 mL (not more than 5 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Lithium Carbonate according to Method 2 using 11 mL of dilute hydrochloric acid, and perform the test according to Method B. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Aluminum—To 5.0 g of Lithium Carbonate add 20 mL of water, add gradually 15 mL of hydrochloric acid while stirring, and evaporate to dryness on a water bath. To the residue add 50 mL of water to dissolve, filter if neces-sary, and assign this solution as solution A. Separately, evaporate 15 mL of hydrochloric acid to dryness on a water bath, then proceed in the same manner, and assign the solu-tion so obtained as solution B. To 10 mL of solution A add 10 mL of water and 5 mL of acetic acid-sodium acetate buffer solution (pH 4.5), and shake. Add 1 mL of a solution of L-ascorbic acid (1 in 100), 2 mL of aluminon TS and water to make 50 mL, shake well, and allow to stand for 10 minutes: the solution has no more color than the following control solution. Control solution: Dissolve 0.1758 g of aluminum potas-sium sulfate dodecahydrate in water to make 1000 mL. To 1.0 mL of this solution add 10 mL of solution B and water to make 20 mL, add 5 mL of acetic acid-sodium acetate buffer solution (pH 4.5), and proceed in the same manner. Barium—To 20 mL of solution A obtained in (7) add 6 mL of water, 0.5 mL of dilute hydrochloric acid, 3 mL of ethanol (95) and 2 mL of potassium sulfate TS, and allow to stand for 1 hour: the solution has no more turbidity than the following control solution. Control solution: Dissolve 17.8 mg of barium chloride di-hydrate in water to make 1000 mL. To 6 mL of this solution add 20 mL of solution B obtained in (7), 0.5 mL of dilute hydrochloric acid and 3 mL of ethanol (95), and proceed in the same manner. Calcium—Weigh accurately about 5 g of Lithium Carbonate, add 50 mL of water and 15 mL of hydrochloric acid, and dissolve. Remove carbon dioxide from the solution by boiling, add 5 mL of ammonium oxalate TS, then make alkaline with ammonia TS, and allow to stand for 4 hours. Filter the produced precipitate through a glass filter (G4), wash with warm water until the turbidity of the washing is not produced with calcium chloride TS within 1 minute. Transfer the precipitate and the glass filter into a beaker, add water until the glass filter is covered with water, then add 3 mL of sulfuric acid, heat between 709C and 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS until a pale red color persists for 30 seconds: the amount of calcium (Ca: 40.08) is not more than 0.05z. Each mL of 0.02 mol/L potassium permanganate VS ＝ 2.004 mg of Ca Magnesium—To 3.0 mL of solution A obtained in (7) add 0.2 mL of a solution of titan yellow (1 in 1000) and water to make 20 mL, then add 5 mL of sodium hydroxide (3 in 20), and allow to stand for 10 minutes: the solution has no more color than the following control solution. Control solution: Dissolve 49.5 mg of magnesium sulfate heptahydrate, previously dried at 1059C for 2 hours and heated at 4509C for 3 hours, in water to make 1000 mL. To 6 mL of this solution add 3 mL of solution B obtained in (7), 0.2 mL of a solution of titanium yellow (1 in 1000) and water to make 20 mL, and proceed in the same manner. Potassium—Dissolve 1.0 g of Lithium Carbonate in water to make 100 mL, and use this solution as the sample solution. To 5 mL of the sample solution add 1.0 mL of dilute acetic acid, shake, add 5 mL of a solution of sodium tetraphenylborate (1 in 30), shake immediately, and allow to stand for 10 minutes: the solution has no more turbidity than the following control solution. Control solution: Dissolve 9.5 mg of potassium chloride in water to make 1000 mL. To 5 mL of this solution add 1.0 mL of dilute acetic acid, shake, and proceed in the same manner. Sodium—Weigh accurately about 0.8 g of Lithium Carbonate, dissolve in water to make exactly 100 mL, and use this solution as the sample stock solution. Measure ex-actly 25 mL of the sample stock solution, add water to make exactly 100 mL, and use this solution as the sample solution (1). Separately, weigh accurately 25.4 mg of sodium chlo-ride, dissolve in water to make exactly 1000 mL, and use this solution as the standard solution. Measure exactly 25 mL of the sample stock solution, add exactly 20 mL of the standard solution, then add water to make exactly 100 mL, and use this solution as the sample solution (2). Determine emission intensities of sodium using a flame photometer with the sample solution (1) and the sample solution (2) under the fol-lowing conditions. Adjust the wavelength dial to 589 nm, atomize the sample solution (2) into the flame, then adjust the sensitivity so that the emission intensity LS shows 100 adjustment, and determine emission intensity LT of the sam-ple solution (1). Then, make the other conditions identical, change the wavelength dial to 580 nm, determine emission intensity LB of the sample solution (1): the amount of so-dium, calculated from the following equation, is not more than 0.05z. Amount (z) of sodium (Na) ＝ (LT － LB)/(LS － LT) × M?/M × 100 M: Amount (mg) of the sample in 25 mL of the sample stock solution M?: Amount (mg) of sodium in 20 mL of the standard so- lution ‌JP XVIII Official Monographs / Lorazepam 1273‌ Arsenic <1.11>—Prepare the test solution with 1.0 g of Lithium Carbonate, add 2 mL of water and 3 mL of hy-drochloric acid, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately the mass of not less than 20 Lisinopril Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 5 mg of lisinopril (C21H31N3O5), add exactly 25 mL of the internal standard solution, shake for 20 minutes, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of lisinopril for assay (separately de-termined the water <2.48> in the same manner as Lisinopril Hydrate), add exactly 50 mL of the internal standard solu-tion to dissolve, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of lisinopril to that of the internal standard. Amount (mg) of lisinopril (C21H31N3O5) ＝ MS × QT/QS × 1/2 MS: Amount (mg) of lisinopril for assay taken, calculated on the anhydrous basis Description Lithium Carbonate occurs as a white crystal-line powder. It is odorless. It is sparingly soluble in water, slightly soluble in hot water, and practically insoluble in ethanol (95) and in diethyl ether. It dissolves in dilute acetic acid. The pH of a solution dissolved 1.0 g of Lithium Car-bonate in 100 mL of water is between 10.9 and 11.5. Identification (1) Perform the test as directed under Flame Coloration Test <1.04> (1) with Lithium Carbonate: a persistent red color appears. Dissolve 0.2 g of Lithium Carbonate in 3 mL of dilute hydrochloric acid, and add 4 mL of sodium hydroxide TS and 2 mL of disodium hydrogen phosphate TS: a white precipitate is produced. To the precipitate add 2 mL of dilute hydrochloric acid: it dissolves. A solution of Lithium Carbonate (1 in 100) responds to Qualitative Tests <1.09> for carbonate. Purity (1) Clarity and color of solution—Dissolve 0.10 g of Lithium Carbonate in 10 mL of water by warming: the so-lution is clear and colorless. Acetic acid-insoluble substances—Take 1.0 g of Lithium Carbonate, dissolve in 40 mL of dilute acetic acid, filter the insoluble substances using filter paper for quantita-tive analysis, wash with five 10-mL portions of water, and ignite the insoluble substances together with the filter paper to incinerate: the mass of the residue is not more than 1.5 mg. Chloride <1.03>—To 0.40 g of Lithium Carbonate add 10 mL of water and 7 mL of dilute nitric acid, and dis-solve by heating to boil. After cooling, add 6 mL of dilute nitric acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.022z). Sulfate <1.14>—To 0.40 g of Lithium Carbonate add 10 mL of water and 4 mL of dilute hydrochloric acid, and dissolve by heating to boil. After cooling, add 1 mL of dilute hydrochloric acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.048z). Heavy metals <1.07>—To 4.0 g of Lithium Carbonate add 5 mL of water, gradually add 10 mL of hydrochloric acid while mixing, and dissolve. Evaporate the solution on a water bath to dryness. To the residue add 10 mL of water, and dissolve. Place the solution in a Nessler tube, add 1 drop of phenolphthalein TS, add ammonia TS until the solution shows a slight red color, then add 2 mL of dilute acetic acid, and dilute with water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: Evaporate 10 mL of hydrochloric acid on a water bath to dryness. To the residue add 10 mL of water, and dis-solve. Place the solution in a Nessler tube, add 1 drop of phenolphthalein TS, add ammonia TS until the solution shows a pale red color, then add 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to make 50 mL (not more than 5 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Lithium Carbonate according to Method 2 using 11 mL of dilute hydrochloric acid, and perform the test according to Method B. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Aluminum—To 5.0 g of Lithium Carbonate add 20 mL of water, add gradually 15 mL of hydrochloric acid while stirring, and evaporate to dryness on a water bath. To the residue add 50 mL of water to dissolve, filter if neces-sary, and assign this solution as solution A. Separately, evaporate 15 mL of hydrochloric acid to dryness on a water bath, then proceed in the same manner, and assign the solu-tion so obtained as solution B. To 10 mL of solution A add 10 mL of water and 5 mL of acetic acid-sodium acetate buffer solution (pH 4.5), and shake. Add 1 mL of a solution of L-ascorbic acid (1 in 100), 2 mL of aluminon TS and water to make 50 mL, shake well, and allow to stand for 10 minutes: the solution has no more color than the following control solution. Control solution: Dissolve 0.1758 g of aluminum potas-sium sulfate dodecahydrate in water to make 1000 mL. To 1.0 mL of this solution add 10 mL of solution B and water to make 20 mL, add 5 mL of acetic acid-sodium acetate buffer solution (pH 4.5), and proceed in the same manner. Barium—To 20 mL of solution A obtained in (7) add 6 mL of water, 0.5 mL of dilute hydrochloric acid, 3 mL of ethanol (95) and 2 mL of potassium sulfate TS, and allow to stand for 1 hour: the solution has no more turbidity than the following control solution. Control solution: Dissolve 17.8 mg of barium chloride di-hydrate in water to make 1000 mL. To 6 mL of this solution add 20 mL of solution B obtained in (7), 0.5 mL of dilute hydrochloric acid and 3 mL of ethanol (95), and proceed in the same manner. Calcium—Weigh accurately about 5 g of Lithium Carbonate, add 50 mL of water and 15 mL of hydrochloric acid, and dissolve. Remove carbon dioxide from the solution by boiling, add 5 mL of ammonium oxalate TS, then make alkaline with ammonia TS, and allow to stand for 4 hours. Filter the produced precipitate through a glass filter (G4), wash with warm water until the turbidity of the washing is not produced with calcium chloride TS within 1 minute. Transfer the precipitate and the glass filter into a beaker, add water until the glass filter is covered with water, then add 3 mL of sulfuric acid, heat between 709C and 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS until a pale red color persists for 30 seconds: the amount of calcium (Ca: 40.08) is not more than 0.05z. Each mL of 0.02 mol/L potassium permanganate VS ＝ 2.004 mg of Ca Magnesium—To 3.0 mL of solution A obtained in (7) add 0.2 mL of a solution of titan yellow (1 in 1000) and water to make 20 mL, then add 5 mL of sodium hydroxide (3 in 20), and allow to stand for 10 minutes: the solution has no more color than the following control solution. Control solution: Dissolve 49.5 mg of magnesium sulfate heptahydrate, previously dried at 1059C for 2 hours and heated at 4509C for 3 hours, in water to make 1000 mL. To 6 mL of this solution add 3 mL of solution B obtained in (7), 0.2 mL of a solution of titanium yellow (1 in 1000) and water to make 20 mL, and proceed in the same manner. Potassium—Dissolve 1.0 g of Lithium Carbonate in water to make 100 mL, and use this solution as the sample solution. To 5 mL of the sample solution add 1.0 mL of dilute acetic acid, shake, add 5 mL of a solution of sodium tetraphenylborate (1 in 30), shake immediately, and allow to stand for 10 minutes: the solution has no more turbidity than the following control solution. Control solution: Dissolve 9.5 mg of potassium chloride in water to make 1000 mL. To 5 mL of this solution add 1.0 mL of dilute acetic acid, shake, and proceed in the same manner. Sodium—Weigh accurately about 0.8 g of Lithium Carbonate, dissolve in water to make exactly 100 mL, and use this solution as the sample stock solution. Measure ex-actly 25 mL of the sample stock solution, add water to make exactly 100 mL, and use this solution as the sample solution (1). Separately, weigh accurately 25.4 mg of sodium chlo-ride, dissolve in water to make exactly 1000 mL, and use this solution as the standard solution. Measure exactly 25 mL of the sample stock solution, add exactly 20 mL of the standard solution, then add water to make exactly 100 mL, and use this solution as the sample solution (2). Determine emission intensities of sodium using a flame photometer with the sample solution (1) and the sample solution (2) under the fol-lowing conditions. Adjust the wavelength dial to 589 nm, atomize the sample solution (2) into the flame, then adjust the sensitivity so that the emission intensity LS shows 100 adjustment, and determine emission intensity LT of the sam-ple solution (1). Then, make the other conditions identical, change the wavelength dial to 580 nm, determine emission intensity LB of the sample solution (1): the amount of so-dium, calculated from the following equation, is not more than 0.05z. Amount (z) of sodium (Na) ＝ (LT － LB)/(LS － LT) × M?/M × 100 M: Amount (mg) of the sample in 25 mL of the sample stock solution M?: Amount (mg) of sodium in 20 mL of the standard so- lution ‌JP XVIII Official Monographs / Lorazepam 1273‌ Arsenic <1.11>—Prepare the test solution with 1.0 g of Lithium Carbonate, add 2 mL of water and 3 mL of hy-drochloric acid, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Assay Weigh accurately about 1 g of Lithium Carbonate, previously dried, add exactly 100 mL of water and 50 mL of 0.5 mol/L sulfuric acid VS, remove carbon dioxide by boil-ing gently, cool, and titrate <2.50> the excess sulfuric acid with 1 mol/L sodium hydroxide VS until the color of the so-lution changes from red to yellow (indicator: 3 drops of methyl red TS). Perform a blank determination in the same manner. Each mL of 0.5 mol/L sulfuric acid VS ＝ 36.95 mg of Li2CO3
Depressed ellipsoidal aril, 1 – 2 cm in length, about 1 cm in width; yellow-red-brown to black-brown; soft in texture and mucous; when immersed in water, bell-shaped, with the tip split in several parts. Odor, characteristic; taste, sweet. Under a microscope <5.01>, a transverse section reveals the outmost layer composed of an epidermis, beneath this ob-served parenchyma consisting of depressed parenchyma cells; the innermost layer composed of slightly thick-walled epidermis; parenchyma contains red-brown to brown con-tents as well as solitary crystals, amorphous crystals and sand crystals of calcium oxalate.
To 1 g of coarse cuttings of Longan Aril, add 10 mL of water, shake thoroughly, and filter. To 3 mL of the filtrate, add 3 mL of Fehling solution, and heat on a water bath: a red precipitate is produced. Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: Not less than 75.0z.
Lorazepam, when dried, contains not less than 98.
Lorazepam occurs as a white crystalline pow-der. It is odorless. It is sparingly soluble in ethanol (95) and in acetone, slightly soluble in diethyl ether, and practically insoluble in water. It is gradually colored by light.
(1) To 0.02 g of Lorazepam add 15 mL of dilute hydrochloric acid, boil for 5 minutes, and cool: the so-lution responds to Qualitative Tests <1.09> for primary aro-matic amines. Determine the absorption spectrum of a solution of Lorazepam in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of ‌1274 Lorazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Lorazepam as directed under Flame Coloration Test <1.04> (2): a green color appears. Losartan Potassium
低置換度ヒドロキシプロピルセルロース
Low Substituted Hydroxypropylcellulose is a low substituted hydroxypropyl ether of cellulose. It contains not less than 5.
Low Substituted Hydroxypropylcellulose occurs as a white to yellowish white, powder or granules. It is practically insoluble in ethanol (99.5). It dissolves in a solution of sodium hydroxide (1 in 10), and produces a viscous solution. It swells in water, in sodium carbonate TS and in 2 mol/L hydrochloric acid TS.◆
Shake thoroughly 0.1 g of Low Substituted Hydrox-ypropylcellulose with 10 mL of water: it does not dissolve. To the dispersed solution obtained in (1) add 1 g of sodium hydroxide, and shake until the solution becomes uniform. Transfer 5 mL of this solution to a suitable vessel, add 10 mL of a mixture of acetone and methanol (4:1), and shake: a white, flocculent precipitate is formed. Determine the infrared absorption spectrum of Low Substituted Hydroxypropylcellulose as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. pH <2.54> To 1.0 g of Low Substituted Hydroxypropylcel-lulose add 100 mL of freshly boiled and cooled water, and shake: the pH of the solution is between 5.0 and 7.5.
◇Heavy metals <1.07>—Proceed with 2.0 g of Low Substituted Hydroxypropylcellulose according to Method 2, ‌JP XVIII Official Monographs / Hydroxyzine Hydrochloride 1125 and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◇ Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.8z (1 g).
(i) Apparatus—Reaction vial: A 5-mL pressure-tight serum vial, 20 mm in outside diameter and 50 mm in height, 20 mm in outside diameter and 13 mm in inside di-ameter at the neck, equipped with a septum made of butyl-rubber whose surface is processed with fluoroplastics, which can be fixed to the vial and stoppered tightly with an aluminum cap or another system providing an equivalent air-tightness. Heater: A square-shaped aluminum block, having holes 20 mm in diameter and 32 mm in depth, adopted to the reaction vial. Capable of stirring the content of the reaction vial by means of a magnetic stirrer or of a reciprocal shaker about 100 times per minute. (ii) Procedure—Weigh accurately about 65 mg of Low Substituted Hydroxypropylcellulose, transfer to a reaction vial, add 0.06 to 0.10 g of adipic acid, 2.0 mL of the internal standard solution and 2.0 mL of hydroiodic acid, immedi-ately stopper the vial tightly, and weigh accurately. Using a magnetic stirrer equipped in the heating module, or using a shaker, mix for 60 minutes while heating so that the temper-ature of the vial content is 130 ± 29C. In the case when a magnetic stirrer or shaker is not available, heat for 30 minutes with repeated shaking at 5-minutes intervals by hand, and continue heating for an additional 30 minutes. Allow the vial to cool, and again weigh accurately. If the mass loss is less than 26 mg and there is no evidence of a leak of the content, use the upper layer of the mixture as the sam-ple solution. Separately, put 0.06 to 0.10 g of adipic acid, 2.0 mL of the internal standard solution and 2.0 mL of hydroiodic acid in a reaction vial, immediately stopper the vial tightly, and weigh accurately. Add 15 to 22 mL of isopropyl iodide for assay through the septum using a micro-syringe, and weigh accurately. Shake the reaction vial thor-oughly, and use the upper layer of the content as the stand-ard solution. Perform the test with 1 to 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of isopropyl iodide to that of the internal standard. Amount (z) of hydroxypropoxy group (C H O ) Carrier gas: Helium. Flow rate: 4.3 mL per minute (the retention time of the in-ternal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 to 2 mL of the standard solution under the above operating con-ditions, isopropyl iodide and the interal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 1 to 2 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the ratio of the peak area of isopropyl iodide to that of the internal standard is not more than 2.0z.
Lysozyme Hydrochloride is the hydrochloride of lysozyme obtained from albumen of hen's egg, and is a protein consisting of 129 amino acid residues. It contains not less than 0.
Lysozyme Hydrochloride occurs as white, crystals, or crystalline or amorphous powder. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic. The pH of a solution of 3 g of Lysozyme Hydrochloride in 200 mL of water is between 3.0 and 5.0.
(1) To 5 mL of a solution of Lysozyme Hy-drochloride in acetate buffer solution (pH 5.4) (1 in 500) add 1 mL of ninhydrin TS, and heat for 10 minutes: a blue-purple color develops. (2) Determine the absorption spectrum of a solution of Lysozyme Hydrochloride in acetate buffer solution (pH 5.4) (1 in 10,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity of solution—To 5 mL of a solution of Lysozyme Hydrochloride (3 in 200) add, if necessary, dilute hydrochloric acid to adjust the pH to 3: the solution is clear. (2) Heavy metals <1.07>—Proceed with 1.0 g of Lyso- zyme Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 8.0z (0.1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 2.0z (0.5 g). Nitrogen Perform the test as directed under Nitrogen De-termination <1.08>: the amount of nitrogen (N: 14.01) is be-tween 16.8z and 18.6z, calculated on the dried basis.
Weigh accurately an amount of Lysozyme Hydro-chloride, equivalent to about 25 mg (potency), dissolve in phosphate buffer solution (pH 6.2) to make exactly 100 mL. Pipet 2 mL of this solution, add phosphate buffer solution (pH 6.2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Lysozyme RS (separately determine its loss on drying <2.41> under the same condition as Lysozyme Hydrochloride), equivalent to about 25 mg (potency), and dissolve in phos-phate buffer solution (pH 6.2) to make exactly 100 mL. Pipet 1 mL and 2 mL of this solution, add phosphate buffer solution (pH 6.2) to them to make exactly 50 mL, and use these solutions as the standard solution (1) and the solution (2), respectively. Keep the sample solution and the standard solutions in an ice-bath. Pipet 4 mL of substrate solution for lysozyme hydrochloride, previously warmed in a water bath of 359C for about 5 minutes, add exactly 100 mL of the sam-ple solution, previously warmed in a water bath of 359C for about 3 minutes, and allow to stand at 359C for exactly 10 minutes, then add exactly 0.5 mL of 1 mol/L hydrochloric acid TS, and immediately shake. Determine the absorbance under Ultraviolet-visible Spectrophotometry <2.24>, AT, of this solution at 640 nm, using water as the blank. Determine the absorbances, AS1 and AS2, of the solutions obtained with the standard solution (1) and the standard solution (2) in the same manner as the sample solution. Amount [mg (potency)] of lysozyme per mg, calculated on the dried basis ＝ MS/2MT ×{(AS1 － AT)/(AS1 － AS2) ＋ 1} MS: Amount (mg) of Lysozyme RS taken, calculated on the dried basis. MT: Amount (mg) of the sample taken, calculated on the dried basis.
Maltose Hydrate, when dried, contains not less than 98.
Maltose Hydrate occurs as white, crystals or crystalline powder. It has a sweet taste. It is freely soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Dissolve 0.5 g of Maltose Hydrate in 5 mL of water, add 5 mL of ammonia TS, and heat for 5 minutes on a water bath: an orange color develops. (2) Add 2 to 3 drops of a solution of Maltose Hydrate (1 in 50) to 5 mL of boiling Fehling TS: a red precipitate is formed. D Optical rotation <2.49> [a]20: ＋126 – ＋1319Weigh accu-rately about 10 g of Maltose Hydrate, previously dried, dis-solve in 0.2 mL of ammonia TS and water to make exactly 100 mL, and determine the optical rotation of this solution in a 100-mm cell. pH <2.54> The pH of a solution of 1.0 g of Maltose Hy-drate in 10 mL of water is between 4.5 and 6.5.
(1) Clarity and color of solution—Put 10 g of Mal-tose Hydrate in 30 mL of water in a Nessler tube, warm at 609C in a water bath to dissolve, and after cooling, add water to make 50 mL: the solution is clear, and has no more color than the following control solution. Control solution: Add water to a mixture of 1.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS to make 10.0 mL. To 1.0 mL of this solution add water to make 50 mL. Chloride <1.03>—Perform the test with 2.0 g of Mal-tose Hydrate. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Sulfate <1.14>—Perform the test with 2.0 g of Mal-tose Hydrate. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 5.0 g of Maltose Hydrate according to Method 1, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 4 ppm). Arsenic <1.11>—Dissolve 1.5 g of Maltose Hydrate in 5 mL of water, add 5 mL of dilute sulfuric acid and 1 mL of bromine TS, heat on a water bath for 5 minutes, then heat to concentrate to 5 mL, and use this solution as the test solu-tion after cooling. Perform the test (not more than 1.3 ppm). Dextrin, soluble starch and sulfite—Dissolve 1.0 g of Maltose Hydrate in 10 mL of water, and add 1 drop of iodine TS: a yellow color appears, and the color changes to a blue by adding 1 drop of starch TS. Nitrogen—Weigh accurately about 2 g of Maltose Hydrate, and perform the test as directed under Nitrogen Determination <1.08> using 10 mL of sulfuric acid for the decomposition and 45 mL of a solution of sodium hydroxide (2 in 5) for the addition: the amount of nitrogen (N: 14.01) is not more than 0.01z. Related substances—Dissolve 0.5 g of Maltose Hy-drate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following oper-ating conditions. Determine the peak areas from both solu-tions by the automatic integration method: the total area of the peaks which appear before the peak of maltose obtained from the sample solution is not larger than 1.5 times the peak area of maltose from the standard solution, and the total area of the peaks which appear after the peak of mal-tose from the sample solution is not larger than 1/2 times the peak area of maltose from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in the operating conditions in the Assay. Detection sensitivity: Adjust the sensitivity so that the peak height of maltose obtained with 20 mL of the standard solution is about 30 mm. Time span of measurement: About 2 times as long as the ‌retention time of maltose. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 4 hours). ‌Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g each of Maltose Hy-drate and Maltose RS, previously dried, dissolve in exactly 10 mL each of the internal standard solution, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following operat-ing conditions, and calculate the ratios, QT and QS, of the peak area of maltose to that of the internal standard. Amount (mg) of maltose hydrate (C12H22O11.H2O) ＝ MS × QT/QS MS: Amount (mg) of Maltose RS taken Internal standard solution—A solution of ethylene glycol (1 in 50). Operating conditions— Detector: A differential refractometer. Column: A stainless steel column about 8 mm in inside diameter and about 55 cm in length, packed with gel-type strong acid cation-exchange resin for liquid chromatography (degree of cross-linking: 8 z) (10 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Water. Flow rate: Adjust so that the retention time of maltose is about 18 minutes. Selection of column: Dissolve 0.25 g of maltose, 0.25 g of glucose and 0.4 g of ethylene glycol in water to make 100 mL. Proceed with 20 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of maltose, glucose and ethylene glycol in this order with the resolution of between the peaks of maltose and glucose being not less than 4.
Manidipine Hydrochloride, when dried, contains not less than 98.
Manidipine Hydrochloride occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in dimethylsulfoxide, sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practi-cally insoluble in water. A solution of Manidipine Hydrochloride in dimethylsul-foxide (1 in 100) shows no optical rotation. Manidipine Hydrochloride turns slightly brown-yellowish white on exposure to light. Melting point: about 2079C (with decomposition).
(1) Determine the absorption spectrum of a solution of Manidipine Hydrochloride in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Manidipine Hydrochloride RS prepared in the same manner as the sam-ple solution: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Manidipine Hydrochloride as directed in the potassium chlo-ride disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Manidipine Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Add 10 mL of water to 0.1 g of Manidipine Hydro-chloride, shake vigorously, and filter. Add 1 drop of ammo-nia TS to 3 mL of the filtrate, allow to stand 5 minutes, and filter. The filtrate responds to Qualitative Tests <1.09> (2) for chlorides.
(1) Heavy metals <1.07>— Proceed with 1.0 g of Manidipine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Manidipine Hydrochloride according to Method 4, and perform the test (not more than 1 ppm). Related substances—Dissolve 20 mg of Manidipine Hydrochloride in a mixture of water and acetonitrile (1:1) to make 200 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and acetonitrile (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex- actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area from both solutions by the automatic integration method: the area of the peaks other than manidipine ob-tained from the sample solution is not larger than 1/5 times the manidipine peak area from the standard solution. Fur-thermore, the total of the areas of all peaks other than manidipine from the sample solution is not larger than 7/10 times the peak area of manidipine from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. ‌Time span of measurement: About 3.5 times as long as the retention time of manidipine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, add a mixture of water and acetonitrile (1:1) to make exactly 100 mL. Confirm that the peak area of mani-dipine obtained with 20 mL of this solution is equivalent to 8 to 12z of that with 20 mL of the standard solution. System performance: Dissolve 50 mg of Manidipine Hy-drochloride in a mixture of water and acetonitrile (1:1) to make 50 mL. To 10 mL of this solution add 5 mL of a solu-tion of butyl benzoate in acetonitrile (7 in 5000) and the mix-ture of water and acetonitrile (1:1) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, manidipine and butyl benzoate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of manidipine is not more than 2.0z. Loss on drying <2.41> Not more than 1.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.1 g of Manidipine Hydro-chloride, previously dried, and dissolve in a mixture of water and acetonitrile (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solu-tion, add the mixture of water and acetonitrile (1:1) to make 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 25 mg of Manidipine Hydro-chloride RS, previously dried, and dissolve in the mixture of water and acetonitrile (1:1) to make exactly 50 mL. Pipet 20 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add the mixture of water and acetonitrile (1:1) to make 100 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of manidipine to that of the internal standard. Amount (mg) of manidipine hydrochloride (C35H38N4O6.2HCl) ＝ MS × QT/QS × 4 MS: Amount (mg) of Manidipine Hydrochloride RS taken Internal standard solution—A solution of butyl benzoate in acetonitrile (7 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 13.6 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 4.6 with diluted potassium hydroxide TS (1 in 10). To 490 mL of this solution add 510 mL of acetonitrile. Flow rate: Adjust so that the retention time of manidipine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, manidipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of manidipine to that of the internal standard is not more than 1.0z.
Maoto Extract contains not less than 15 mg and not more than 45 mg of total alkaloids (ephedrine and pseudoephedrine), not less than 48 mg and not more than 192 mg of amygdalin, and not less than 11 mg and not more than 33 mg of glycyrrhizic acid (C42H62O 16: 822.
Maoto Extract occurs as a light brown powder or black-brown viscous extract, having a slightly order, and a sweet and bitter, then a slightly astringent taste.
(1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (4:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot is observed at an Rf value of about 0.5 (Ephedra Herb). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 2 mg of amygdalin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of 1-propanol, ethyl acetate and water (4:4:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the green-brown spot from the standard solution (Apricot Ker-nel). Perform the test according to the following (i) or (ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect the apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E)-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromato-graphy <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the yellow-orange spot from the standard solution. Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample so-lution. Separately, dissolve 1 mg of (E)-2-methoxycinnamal-dehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu-tion. Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). JP XVIII Crude Drugs and Related Drugs / Maoto Extract 2071 Total ash <5.01> Not more than 13.0z, calculated on the dried basis. However, for the dry extract prepared by adding Light Anhydrous Silicic Acid, between 10.0z and 22.0z.
(1) Total alkaloids (ephedrine and pseudoephed-rine)—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. In addition, repeat twice in the same manner for the aqueous layer using 1.0 mL of ammonia TS and 20 mL of diethyl ether. Combine all the extracts, evaporate the solvent under low pressure (in va-cuo), dissolve the residue in diluted methanol (1 in 2) to make exactly 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of ephedrine hydrochloride for assay of crude drugs, previously dried at 1059C for 3 hours, dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, ATE and ATP, of ephe-drine and pseudoephedrine obtained from the sample solu-tion, and peak area, AS, of ephedrine from the standard solution. Amount (mg) of total alkaloids (ephedrine and pseudoe-phedrine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drugs taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, then add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of ephe-drine is about 27 minutes). System suitability— System performance: Dissolve 1 mg each of ephedrine hy-drochloride for assay of crude drugs and pseudoephedrine hydrochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephe-drine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 1.5z. Amygdalin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, flow through in a column packed with 2 g of polyamide for column chromatography, then elute with water to make exactly 20 mL, and use this effluent as the sample solution. Separately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desicca-tor (silica gel) for 24 hours or more, and dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. Amount (mg) of amygdalin ＝ MS × AT/AS × 4 MS: Amount (mg) of amygdalin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amygdalin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ‌2072 Mentha Herb / Crude Drugs and Related Drugs ‌409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Mentha Herb Menthae Herba
Maprotiline Hydrochloride, when dried, contains not less than 99.
Maprotiline Hydrochloride occurs as a white crystalline powder. It is soluble in methanol and in acetic acid (100), sparingly soluble in ethanol (99.5), and slightly soluble in water. Melting point: about 2449C (with decomposition). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Maprotiline Hydrochloride in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Maprotiline Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of ‌absorption at the same wave numbers. If any difference ap-pears between the spectra, recrystallize Maprotiline Hydro-chloride with ethanol (99.5), filter, dry the crystals so ob-tained, and perform the test with the crystals. To 5 mL of a solution of Maprotiline Hydrochloride (1 in 200) add 2 mL of ammonia TS, heat on a water bath for 5 minutes, cool, and filter. Acidify the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> for chloride. ‌Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Maprotiline Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Maprotiline Hydrochloride in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop with a mixture of 2-butanol, diluted ammonia solution (28) (1 in 3) and ethyl acetate (14:5:4) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the number of the spot other than the principal spot ob-tained from the sample solution is not more than 2 and they are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Maprotiline Hy-drochloride, previously dried, dissolve in 80 mL of acetic acid (100), add 8 mL of a solution of bismuth nitrate penta-hydrate in acetic acid (100) (1 in 50), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.39 mg of C20H23N.HCl
Meclofenoxate Hydrochloride contains not less than 98.
Meclofenoxate Hydrochloride occurs as white, crystals or crystalline powder. It has a faint, characteristic odor and a bitter taste. It is freely soluble in water and in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Meclofenoxate Hydro-chloride in 20 mL of water is between 3.5 and 4.5.
(1) To 10 mg of Meclofenoxate Hydrochlo-ride add 2 mL of ethanol (95), dissolve by warming if neces-sary, cool, add 2 drops of a saturated solution of hydroxyl-ammonium chloride in ethanol (95) and 2 drops of a saturat-ed solution of potassium hydroxide in ethanol (95), and heat in a water bath for 2 minutes. After cooling, render the solu-tion slightly acidic with dilute hydrochloric acid, and add 3 drops of iron (III) chloride TS: a red-purple to dark purple color develops. Dissolve 50 mg of Meclofenoxate Hydrochloride in 5 mL of water, and add 2 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Meclofenoxate Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. A solution of Meclofenoxate Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 0.5 g of Meclofenoxate Hydrochloride in 10 mL of water: the so-lution is clear and colorless. Sulfate <1.14>—Perform the test with 1.0 g of Meclofenoxate Hydrochloride. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Meclo- fenoxate Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Meclofenoxate Hydrochloride according to method 3, and perform the test (not more than 2 ppm). Organic acids—To 2.0 g of Meclofenoxate Hydro-chloride add 50 mL of diethyl ether, shake for 10 minutes, filter through a glass filter (G3), wash the residue with two 5-mL portions of diethyl ether, and combine the washings ‌with the filtrate. To this solution add 50 mL of neutralized ethanol and 5 drops of phenolphthalein TS, and neutralize with 0.1 mol/L sodium hydroxide VS: the volume of 0.1 mol/L sodium hydroxide VS consumed is not more than 0.54 mL. Water <2.48> Not more than 0.50z (1 g, volumetric titra-tion, dirct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Meclofenoxate Hy-drochloride, dissolve in 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from blue-green through yel-low-green to pale greenish yellow [indicator: 3 drops of a so-lution of malachite green oxalate in acetic acid (100) (1 in 100)]. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.42 mg of C12H16ClNO3.HCl
Mecobalamin occurs as dark red, crystals or crystalline powder. It is sparingly soluble in water, slightly soluble in ethanol (99.5), and practically insoluble in acetonitrile. It decomposes on exposure to light.
(1) Conduct this procedure without ex-posure to light, using light-resistant vessels. Determine the absorption spectrum of a solution of Mecobalamin in hydro-chloric acid-potassium chloride buffer solution (pH 2.0) (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- Water <2.48> Not more than 12z (0.1 g, volumetric titra-tion, direct titration).
(1) Clarity and color of solution—Dissolve 20 mg of Mecobalamin in 10 mL of water: the solution is clear and red color. (2) Related substances—Perform the test with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: each area of the peaks other than mecobalamin is not more than 0.5z of the peak area of mecobalamin, and the total area of the peaks other than mecobalamin is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of mecobalamin. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the sample solution add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suita-bility test. Pipet 1 mL of the solution for system suitability C H CoN O P: 1344.38 test, add the mobile phase to make exactly 10 mL. Confirm 63 91 13 14 that the peak area of mecobalamin obtained with 10 mL of Coa-[a-(5,6-Dimethyl-1H-benzimidazol-1-yl)]-Cob- methylcobamide [13422-55-4] Mecobalamin contains not less than 98.0z and not more than 101.0z of mecobalamin this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia- (C63H 91CoN13 O14 P), calculated on the anhydrous tion of the peak areas of mecobalamin is not more than 3.0z. basis. Description Mecobalamin occurs as dark red, crystals or crystalline powder. It is sparingly soluble in water, slightly soluble in ethanol (99.5), and practically insoluble in acetonitrile. It decomposes on exposure to light. Identification (1) Conduct this procedure without ex-posure to light, using light-resistant vessels. Determine the absorption spectrum of a solution of Mecobalamin in hydro-chloric acid-potassium chloride buffer solution (pH 2.0) (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- Water <2.48> Not more than 12z (0.1 g, volumetric titra-tion, direct titration).
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 50 mg of Mecobalamin and Mecobalamin RS (separately, deter-mine the water <2.48> in the same manner as Mecobalamin), dissolve each in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with exactly 10 mL of each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mecobalamin in each solution. add 10 mL of phosphate buffer solution (pH 7.0), sonicate, and add phosphate buffer solution (pH 7.0) to make 20 mL. Amount (mg) of mecobalamin (C ＝ MS × AT/AS 63H 91CoN 13O 14P) Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.8 mm. Determine the absorption spectrum of the filtrate as di- MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis Operating conditions— ‌Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 200 mL of acetonitrile add 800 mL of 0.02 mol/L phosphate buffer solution (pH 3.5), then add 3.76 g of sodium 1-hexane sulfonate to dissolve. Flow rate: Adjust so that the retention time of mecobala-min is about 12 minutes. System suitability— System performance: Dissolve 5 mg each of cyanocobala-min and hydroxocobalamin acetate in the mobile phase to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, cyanocobala-min and hydroxocobalamin are eluted in this order with the resolution between these peaks being not less than 3. And when the procedure is run with 10 mL of the standard solu-tion under the above operating conditions, the number of theoretical plates of the peak of mecobalamin is not less than rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 264 nm and 268 nm, between 339 nm and 343 nm, and between 520 nm and 524 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. Take 1 tablet of Mecobalamin Tablets, and disintegrate the tablet by adding V/5 mL of water. Add methanol to make exactly V mL so that each mL contains about 25 mg of mecobalamin (C63H91CoN13O14P). After shaking for 5 minutes, allow to stand for not less than 10 minutes. Filter thus obtained supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 25 mg of Mecobalamin RS (separately determine the water <2.48> in the same manner as Mecobalamin), and dis-solve in water to make exactly 100 mL. Pipet 5 mL of this solution, add 5 mL of water and methanol to make exactly 50 mL. Use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mecobalamin in each solution. 6000. Amount (mg) of mecobalamin (C H CoN O P) System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ＝ MS × AT/AS × V/1000 63 91 13 14 ing conditions, the relative standard deviation of the peak areas of mecobalamin is not more than 1.0z.
Medazepam, when dried, contains not less than 98.
Medazepam occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in methanol, in ethanol (99.5), in acetic acid (100) and in diethyl ether, and practically insoluble in water. It gradually turns yellow on exposure to light.
(1) Dissolve 10 mg of Medazepam in 3 mL of citric acid-acetic acid TS: a deep orange color develops. Heat in a water bath for 3 minutes: the color changes to dark red. Determine the absorption spectrum of a solution of Medazepam in methanol (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Medazepam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Medazepam as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Clarity and color of solution—Dissolve 1.0 g of Medazepam in 10 mL of methanol: the solution is clear and light yellow to yellow in color. Chloride <1.03>—Dissolve 1.5 g of Medazepam in 50 mL of diethyl ether, add 46 mL of water and 4 mL of so-dium carbonate TS, shake, and collect the water layer. Wash the water layer with two 20-mL portions of diethyl ether, and filter. To 20 mL of the filtrate add dilute nitric acid to neutralize, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Heavy metals <1.07>—Proceed with 1.0 g of Medazepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g ‌of Medazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Medazepam in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of cyclo-hexane, acetone and ammonia solution (28) (60:40:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure without exposure to light, using light-resistant vessels. Disintegrate 20 tablets of Mecobalamin Tablets with V/5 mL of water. Add methanol to make exactly V mL so that each mL contains about 50 mg of mecobalamin (C63H91CoN13O14P). After shaking for 5 minutes, allow to stand for not less than 10 minutes. Filter thus obtained supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Separately, weigh accurately about 25 mg of Mecobalamin RS (separately determine the water <2.48> in the same manner as Mecobalamin), and dissolve in water to make exactly 100 mL. To exactly 10 mL of this solution add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of mecobalamin in each solution. Amount (mg) of mecobalamin (C63H91CoN13O14P) in 1 tablet ＝ MS × AT/AS × V/10000 MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis Description Medazepam occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in methanol, in ethanol (99.5), in acetic acid (100) and in diethyl ether, and practically insoluble in water. It gradually turns yellow on exposure to light. Identification (1) Dissolve 10 mg of Medazepam in 3 mL of citric acid-acetic acid TS: a deep orange color develops. Heat in a water bath for 3 minutes: the color changes to dark red. Determine the absorption spectrum of a solution of Medazepam in methanol (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Medazepam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Medazepam as directed under Flame Coloration Test <1.04> (2): a green color appears. Melting point <2.60> 101 – 1049C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Medazepam in 10 mL of methanol: the solution is clear and light yellow to yellow in color. Chloride <1.03>—Dissolve 1.5 g of Medazepam in 50 mL of diethyl ether, add 46 mL of water and 4 mL of so-dium carbonate TS, shake, and collect the water layer. Wash the water layer with two 20-mL portions of diethyl ether, and filter. To 20 mL of the filtrate add dilute nitric acid to neutralize, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Heavy metals <1.07>—Proceed with 1.0 g of Medazepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g ‌of Medazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Medazepam in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of cyclo-hexane, acetone and ammonia solution (28) (60:40:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.4 g of Medazepam, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.08 mg of C16H15ClN2
Medicinal Carbon occurs as a black, odorless and tasteless powder.
Place 0.5 g of Medicinal Carbon in a test tube, and heat by direct application of flame with the aid of a current of air: it burns without any flame. Pass the evolved gas through calcium hydroxide TS: a white turbidity is pro-duced.
(1) Acidity or alkalinity—Boil 3.0 g of Medicinal Carbon with 60 mL of water for 5 minutes, allow to cool, dilute to 60 mL with water, and filter: the filtrate is colorless and neutral. Chloride <1.03>—Take 4.0 mL of the filtrate ob- tained in (1) in a Nessler tube, add 6 mL of dilute nitric acid and sufficient water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.80 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.142z). Sulfate <1.14>—Take 5 mL of the filtrate obtained in in a Nessler tube, add 1 mL of dilute hydrochloric acid and sufficient water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.192z). Sulfide—Boil 0.5 g of Medicinal Carbon with a mix-ture of 15 mL of dilute hydrochloric acid and 10 mL of water: lead (II) acetate paper does not become brown when held in the evolved gas within 5 minutes. Cyanogen compounds—Place a mixture of 5 g of Medicinal Carbon, 2 g of L-tartaric acid and 50 mL of water in a distilling flask connected to a condenser provided with a tightly fitting adapter, the end of which dips below the sur-face of a mixture of 2 mL of sodium hydroxide TS and 10 mL of water, contained in a small flask surrounded by ice. Heat the mixture in the distilling flask to boiling, and distil to 25 mL. Dilute the distillate with water to 50 mL. To 25 mL of the diluted distillate add 1 mL of a solution of iron (II) sulfate heptahydrate (1 in 20), heat the mixture almost to boiling, cool, and filter. To the filtrate add 1 mL of hydro-chloric acid and 0.5 mL of dilute iron (III) chloride TS: no blue color is produced. Acid soluble substances—To about 1 g of Medicinal Carbon, accurately weighed, add 20 mL of water and 5 mL of hydrochloric acid, boil for 5 minutes, filter, wash the residue with 10 mL of hot water, and add the washings to the filtrate. Add 5 drops of sulfuric acid to the filtrate, evapo-rate to dryness, and ignite the residue strongly: the mass of the residue is not more than 3.0z. Heavy metals <1.07>—Proceed with 0.5 g of Medici- nal Carbon according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 50 ppm). Zinc—Ignite 0.5 g of Medicinal Carbon to ash, add 5 mL of dilute nitric acid to the residue, boil gently for 5 minutes, filter, wash with 10 mL of water, and combine the washings and the filtrate. Add 3 mL of ammonia TS to the solution, filter again, wash with water, combine the wash-ings and the filtrate, add another washing to make 25 mL, add 1 drop of sodium sulfide TS, and allow to stand for 3 minutes: the liquid produces no turbidity. Arsenic <1.11>—Prepare the test solution with 1.0 g of Medicinal Carbon according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 15.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 4z (1 g). Adsorptive power (1) Add 1.0 g of Medicinal Carbon, previously dried, to 100 mL of water containing 120 mg of quinine sulfate hydrate, shake the mixture vigorously for 5 minutes, filter immediately, and reject the first 20 mL of the filtrate. Add 5 drops of iodine TS to 10 mL of the subse-quent filtrate: no turbidity is produced. Dissolve 250 mg of methylene blue trihydrate, exactly weighed, in water to make exactly 250 mL. Measure two 50-mL portions of this solution into each of two glass-stoppered flasks. To one flask add exactly 250 mg of Medici-nal Carbon, previously dried, and shake vigorously for 5 minutes. Filter the contents of each flask, rejecting the first 20 mL of each filtrate. Pipet 25-mL portions of the remain-ing filtrate into two 250-mL volumetric flasks. To each volu- ‌metric flask add 50 mL of a solution of sodium acetate trihy-drate (1 in 10), then add exactly 35 mL of 0.05 mol/L iodine VS with swirling. Allow them to stand for 50 minutes, shak-ing vigorously from time to time. Dilute each mixture to ex-actly 250 mL with water, allow to stand for 10 minutes, and filter each solution at a temperature not exceeding 209C, rejecting the first 30 mL of each filtrate. Titrate <2.50> the excess iodine in a 100-mL aliquot of each filtrate with 0.1 mol/L sodium thiosulfate VS. The difference between the two titrations is not less than 1.2 mL.‌
Medicinal Soap occurs as a white to light yel-low, powder or granules. It has a characteristic odor free from rancidity. Medicinal Soap is sparingly soluble in water, and slightly soluble in ethanol (95). A solution of Medicinal Soap (1 in 100) is alkaline. Fatty acid Dissolve 25 g of Medicinal Soap in 300 mL of hot water, add 60 mL of dilute sulfuric acid slowly, and warm in a water bath for 20 minutes. After cooling, filter off the precipitate, and wash with warm water until the washing no longer shows acidity to methyl orange TS. Transfer the precipitate to a small beaker, and heat on a water bath to complete separation of water and transparent fatty acids. Filter the fatty acid into a small beaker while warm, dry at 1009C for 20 minutes, and perform the test with this mate-rial as directed under Fats and Fatty Oils <1.13>. The con-gealing point of the fatty acid is between 189C and 289C. The acid value is 185 – 205. The iodine value is 82 – 92.
(1) Acidity or alkalinity—Dissolve 5.0 g of Medici-nal Soap in 85 mL of neutralized ethanol by warming on a water bath, filter while hot through absorbent cotton, and wash the filter and the residue with three 5-mL portions of hot neutralized ethanol. Combine the filtrate and the wash-ings, add hot neutralized ethanol to make exactly 100 mL, and perform the following tests quickly using this as the sample solution at 709C. Add 3 drops of phenolphthalein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS to 40 mL of the sample so-lution: a red color develops. Add 3 drops of phenolphthalein TS and 0.20 mL of 0.05 mol/L sulfuric acid VS to 40 mL of the sample solu-tion: no red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Medici- nal Soap according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Ethanol-insoluble substances—Weigh accurately about 2 g of Medicinal Soap, dissolve by warming in 100 mL of neutralized ethanol, filter the solution through a glass filter (G4), wash the residue with 100 mL of hot neutralized ethanol, and dry at 1059C for 4 hours: the mass of the residue is not more than 1.0z. Water-insoluble substances—Wash thoroughly the dried substances obtained in (3) with 200 mL of water, and dry at 1059C for 4 hours: the mass of the residue is not more than 0.15z. Alkali carbonates—To the washings obtained in (4) add 3 drops of methyl orange TS and 2 mL of 0.05 mol/L sulfuric acid VS: a red color develops. Loss on drying Not more than 5.0z in the case of the pow-der, and not more than 10.0z in the case of the granules. Weigh accurately about 0.5 g of Medicinal Soap in a tared beaker, add 10 g of sea sand (No. 1), previously dried at 1059C for 1 hour, and again weigh the beaker. Add 10 mL of ethanol (95), evaporate on a water bath to dryness with thorough stirring, and dry at 1059C for 3 hours.
メドロキシプロゲステロン酢酸エステル
Medroxyprogesterone Acetate, when dried, contains not less than 97.
Medroxyprogesterone Acetate occurs as a white crystalline powder. It is soluble in acetone, sparingly soluble in acetonitrile, slightly soluble in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Medroxyprogesterone Acetate in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of a solution of Medrox-yprogesterone Acetate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Medroxyprogesterone Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Medroxypro-gesterone Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋ 47 – ＋ 539(after drying, 0.25 g, acetone, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Medroxyprogesterone Acetate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 100 mL, ‌and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than medroxyprogesterone acetate obtained from the sample solution is not larger than the peak area of medroxyprogesterone acetate from the standard solution, and the total area of the peaks other than medroxyprogester-one acetate from the sample solution is not larger than 2 times the peak area of medroxyprogesterone acetate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.2 times as long as the retention time of medroxyprogesterone acetate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add acetonitrile to make exactly 10 mL. Con-firm that the peak area of medroxyprogesterone acetate ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of medroxyprogesterone acetate are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution, the relative standard de-viation of the peak area of medroxyprogesterone acetate is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Weigh accurately about 25 mg each of Medrox-yprogesterone Acetate and Medroxyprogesterone Acetate RS, both previously dried, dissolve in acetonitrile to make them exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of medroxyprogesterone acetate in each solution. Amount (mg) of medroxyprogesterone acetate (C24H34O4) ＝ MS × AT/AS MS: Amount (mg) of Medroxyprogesterone Acetate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (3:2). Flow rate: Adjust so that the retention time of medrox- yprogesterone acetate is about 31 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of medroxyprogesterone acetate are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution, the relative standard de-viation of the peak area of medroxyprogesterone acetate is not more than 1.0z.
Mefenamic Acid, when dried, contains not less than 99.
Mefenamic Acid occurs as a white to light yel-low powder. It is odorless and tasteless at first, but leaves a slightly bitter aftertaste. It is sparingly soluble in diethyl ether, slightly soluble in methanol, in ethanol (95) and in chloroform, and practically insoluble in water. It dissolves in sodium hydroxide TS. Melting point: about 2259C (with decomposition).
(1) Dissolve 10 mg of Mefenamic Acid in 1 mL of methanol by warming, cool, add 1 mL of a solution of 4-nitrobenzene diazonium fluoroborate (1 in 1000) and 1 mL of sodium hydroxide TS, and mix thoroughly: an orange-red color is produced. Dissolve 10 mg of Mefenamic Acid in 2 mL of sulfu-ric acid, and heat: the solution shows a yellow color and a green fluorescence. Dissolve 7 mg of Mefenamic Acid in a solution of hy-drochloric acid in methanol (1 in 1000) to make 500 mL. De-termine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Chloride <1.03>—To 1.0 g of Mefenamic Acid add 20 mL of sodium hydroxide TS, and dissolve by warm-ing. Cool, add 2 mL of acetic acid (100) and water to make 100 mL, and mix well. Remove the produced precipitate by filtration, discard the first 10 mL of the filtrate, and to sub-sequent 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.50 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of sodium hydroxide TS, 0.5 mL of acetic acid (100), 6 mL of nitric acid and water to make 50 mL (not more than 0.071z). Heavy metals <1.07>—Proceed with 2.0 g of Mefenamic Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Mefenamic Acid according to Method 3, and perform the test (not more than 2 ppm). ‌Related substances—Dissolve 0.10 g of Mefenamic Acid, in 5 mL of a mixture of chloroform and methanol (3:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (3:1) to make exactly 200 mL, pipet 10 mL of this solution, add a mixture of chloroform and methanol (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 2-methyl-1-propanol and ammonia solution (28) (3:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Mefenamic Acid, previously dried, and dissolve in 100 mL of ethanol (95), pre-viously neutralized to phenol red TS with 0.1 mol/L sodium hydroxide VS, by warming gently. Cool, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color of the solution changes from yellow through yellow-red to red-pur-ple (indicator: 2 to 3 drops of phenol red TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 24.13 mg of C15H15NO2
Mefloquine Hydrochloride contains not less than 99.
Mefloquine Hydrochloride occurs as white eter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and diluted phos-phoric acid (1 in 14) (24:1). Flow rate: Adjust so that the retention time of mefloquine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of mefloquine. System suitability— Test for required detectability: To exactly 10 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of mefloquine obtained with 10 mL of this solution is equivalent to 40 to 60z of that with 10 mL of the standard solution. ‌System performance: Dissolve 10 mg of mefloquine hy-drochloride and 5 mg of diprophylline in 50 mL of the mo-bile phase. To 2 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, diprophylline and mefloquine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mefloquine is not more than 2.0z. Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
(1) Dissolve 2 mg of Mefloquine Hydro-chloride in 1 mL of sulfuric acid: the solution shows a blue fluorescence under ultraviolet light (main wavelength: 365 nm). Determine the absorption spectrum of a solution of Mefloquine Hydrochloride in methanol (1 in 25,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mefloquine Hydrochloride, previously dried at 1059C for 2 hours, as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Mefloquine Hydrochloride (1 in 1000) add 1 mL of dilute nitric acid and 1 mL of silver nitrate TS: a white precipitate is formed, and the separated precipitate dissolves on the addition of an excess amount of ammonia TS.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Mefloquine Hydrochloride according to Method 2 using a quartz crucible, and perform the test. Prepare the control so-lution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Mefloquine Hydrochlo- ride add 10 mL of a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 10), burn the ethanol, gradually heat, and incinerate by ignition at 8009C. If a carbonized residue still retains, moisten the residue with a little amount of nitric acid, and ignite again to incinerate. After cooling, to the residue add 3 mL of hydrochloric acid, warm on a water bath to dissolve, and perform the test using this solu-tion as the test solution (not more than 2 ppm). Related substances—Dissolve 50 mg of Mefloquine Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than mefloquine and the peak eluted first obtained from the sample solution is not larger than the peak area of mefloquine from the standard solution, and the total area of the peaks other than the peak of mefloquine and the peak eluted first from the sample so-lution is not larger than 2.5 times the peak area of meflo-quine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with aminopropylsilanized silica gel for liquid chromatography (10 mm in particle diam- Description Mefloquine Hydrochloride occurs as white eter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and diluted phos-phoric acid (1 in 14) (24:1). Flow rate: Adjust so that the retention time of mefloquine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of mefloquine. System suitability— Test for required detectability: To exactly 10 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of mefloquine obtained with 10 mL of this solution is equivalent to 40 to 60z of that with 10 mL of the standard solution. ‌System performance: Dissolve 10 mg of mefloquine hy-drochloride and 5 mg of diprophylline in 50 mL of the mo-bile phase. To 2 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, diprophylline and mefloquine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mefloquine is not more than 2.0z. Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.5 g of Mefloquine Hydro-chloride, dissolve in 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.48 mg of C17H16F6N2O.HCl
Mefruside, when dried, contains not less than 98.
Mefruside occurs as a white crystalline pow-der. It is very soluble in N,N-dimethylformamide, freely solu-ble in acetone, soluble in methanol, sparingly soluble in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Mefruside in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mefruside, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Mefruside as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Dissolve 1.0 g of Mefru-side in 30 mL of acetone, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 30 mL of acetone, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Mefruside according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Mefruside in 10 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of chloroform and acetone (5:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Mefruside, previ-ously dried, dissolve in 80 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hy-droxide VS (potentiometric titration). Separately, perform a blank determination in the same manner with a solution pre-pared by adding 13 mL of water to 80 mL of N,N-dimethyl-formamide, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ylmethyl]-3-sulfamoylbenzenesulfonamide [7195-27-9] ＝ 38.29 mg of C13H19 ClN2O5S2 Mefruside, when dried, contains not less than 98.5z of mefruside (C13H19ClN2O5S2). Description Mefruside occurs as a white crystalline pow-der. It is very soluble in N,N-dimethylformamide, freely solu-ble in acetone, soluble in methanol, sparingly soluble in ethanol (95), and practically insoluble in water.
Meglumine, when dried, contains not less than 99.
Meglumine occurs as a white crystalline pow-der. It is odorless, and has a slightly bitter taste. It is freely soluble in water, and slightly soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Meglumine in 10 mL of water is between 11.0 and 12.0.
(1) To 1 mL of a solution of Meglumine (1 in 10) add 1 mL of potassium 1,2-naphthoquinone-4-sulfonate TS: a deep red color develops. To 2 mL of a solution of Meglumine (1 in 10) add 1 drop of methyl red TS, and add 0.5 mL of dilute sodium hy-droxide TS and 0.5 g of boric acid after neutralizing with 0.5 mol/L sulfuric acid TS: a deep red color develops. Dissolve 0.5 g of Meglumine in 1 mL of diluted hy-drochloric acid (1 in 3), and add 10 mL of ethanol (99.5): a white precipitate is produced. Then, rubbing the inside wall of the container with a glass rod, cool with ice and produce more precipitate. Filter the precipitate by suction through a glass filter (G3), wash the precipitate with a small volume of ethanol (99.5), and dry at 1059C for 1 hour: the residue thus obtained melts <2.60> between 1499C and 1529C. D Optical rotation <2.49> [a]20: －16.0 – －17.09(after dry-ing, 1 g, water, 10 mL, 100 mm).
Weigh accurately about 0.4 g of Meglumine, previ-ously dried, dissolve in 25 mL of water, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (indicator: 2 drops of methyl red TS). Each mL of 0.1 mol/L hydrochloric acid VS ＝ 19.52 mg of C7H17NO5
Honey is a light yellow to light yellow-brown, syrupy liquid. Usually it is transparent, but often opaque with separated crystals. It has a characteristic odor and a sweet taste. Specific gravity <2.56> Mix 50.0 g of Honey with 100 mL of water: the specific gravity of the solution is not less than d 20: 1.111.
(1) Acidity—Mix 10 g of Honey with 50 mL of water, and titrate <2.50> with 1 mol/L potassium hydroxide VS (indicator: 2 drops of phenolphthalein TS): not more than 0.5 mL is required. Sulfate—Mix 1.0 g of Honey with 2.0 mL of water, and filter. To the filtrate add 2 drops of barium chloride TS: the solution does not show any change immediately. Ammonia-coloring substances—Mix 1.0 g of Honey with 2.0 mL of water, and filter. To the filtrate add 2 mL of ammonia TS: the solution does not show any change imme- Starch or dextrin—(i) Shake 7.5 g of Honey with 15 mL of water, warm the mixture on a water bath, and add 0.5 mL of tannic acid TS. After cooling, filter, and to 1.0 mL of the filtrate add 1.0 mL of ethanol (99.5) containing 2 drops of hydrochloric acid: no turbidity is produced. (ii) To 2.0 g of Honey add 10 mL of water, warm in a water bath, mix, and allow to cool. Shake 1.0 mL of the mixture with 1 drop of iodine TS: no blue, green or red-brown color develops. Foreign matter—Mix 1.0 g of Honey with 2.0 mL of water, centrifuge the mixture, and examine the precipitate microscopically <5.01>: no foreign substance except pollen grains is observable. Total ash <5.01> Not more than 0.4z.
Melphalan occurs as a white to light yellowish white crystalline powder. It is slightly soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether. It dissolves in dilute hydrochloric acid and in dilute so-dium hydroxide TS. It is gradually colored by light. D Optical rotation [a]20: about －329(0.5 g calculated on the dried basis, methanol, 100 mL, 100 mm).
(1) To 20 mg of Melphalan add 50 mL of methanol, dissolve by warming, add 1 mL of a solution of 4-(4-nitrobenzyl)pyridine in acetone (1 in 20), and evaporate on a water bath to dryness. Dissolve the residue in 1 mL of warmed methanol and add 2 drops of ammonia solution (28): a purple color develops. Dissolve 0.1 g of Melphalan in 10 mL of dilute so-dium hydroxide TS, and heat on a water bath for 10 minutes. After cooling, add dilute nitric acid to acidify, and filter: the filtrate responds to Qualitative Tests <1.09> for chloride. Determine the absorption spectrum of a solution of Melphalan in methanol (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Ionisable chloride—Weigh accurately about 0.5 g of Melphalan, dissolve in 80 mL of diluted nitric acid (1 in 40), stir for 2 minutes, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration): the consumed volume is not more than 1.0 mL to 0.50 g of Melphalan. Heavy metals <1.07>—Proceed with 1.0 g of Melpha- lan according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Melphalan according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 7.0z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.25 g of Melphalan, add 20 mL of a solution of potassium hydroxide (1 in 5), and heat under a reflux condenser on a water bath for 2 hours. After cooling, add 75 mL of water and 5 mL of nitric acid, cool, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentio-metric titration). Make any necessary correction by using the results obtained in the Purity (1). ‌Each mL of 0.1 mol/L silver nitrate VS ＝ 15.26 mg of C13H18Cl2N2O2
Menatetrenone contains not less than 98.
Menatetrenone occurs as yellow, crystals, crys-talline powder, waxy mass or oily material. It is very soluble in hexane, soluble in ethanol (99.5), spar-ingly soluble in 2-propanol, slightly soluble in methanol, and practically insoluble in water. It decomposes and the color becomes more intense by light. Melting point: about 379C.
(1) Dissolve 0.1 g of Menatetrenone in 5 mL of ethanol (99.5) by warming, cool, and add 1 mL of a solution of potassium hydroxide in ethanol (95) (1 in 10): a blue color develops, and upon standing it changes from blue-purple to red-brown through red-purple. Determine the infrared absorption spectrum of Menatetrenone, after melting by warming if necessary, as di-rected in the liquid film method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Menatetrenone RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Menatetrenone according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Menadione—To 0.20 g of Menatetrenone add 5 mL of diluted ethanol (1 in 2), shake well, and filter. To 0.5 mL of the filtrate add 1 drop of a solution of 3-methyl-1-phenyl-5-pyrazorone in ethanol (99.5) (1 in 20) and 1 drop of ammo-nia water (28), and allow to stand for 2 hours: no blue-pur-ple color develops. 254 nm): the spot corresponding to relative Rf value 1.1 regarding to the principal spot obtained from the sample so-lution is not more intense than the spot from the standard solution. (4) Related substances—Conduct this procedure without exposure to light, using a light-resistant vessel. Dissolve 0.10 g of Menatetrenone in 100 mL of ethanol (99.5), and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of these solutions by the automatic integration method: the total area of peaks other than the peak of menatetrenone obtained from the sample solution is not larger than the peak area of menatetrenone from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 6 times as long as the retention time of menatetrenone, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 5 mL of the standard solution add ethanol (99.5) to make exactly 50 mL. Confirm that the peak area of menatetrenone obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of menatetrenone is not more than 1.0z. Water <2.48> Not more than 0.5z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure without exposure to light, using a light-resistant vessel. Weigh accurately about 0.1 g each of Menatetrenone and Menatetrenone RS (separately, determine the water <2.48> in the same manner as Menatetre-none), dissolve each in 50 mL of 2-propanol, and add ethanol (99.5) to make exactly 100 mL. Pipet 10 mL of these solutions, and add ethanol (99.5) to make exactly 100 mL. Pipet 2 mL each of these solutions, add exactly 4 mL each of the internal standard solution, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of menatetrenone to that of the internal standard. (3) cis Isomer—Dissolve 0.10 g of Menatetrenone in 10 mL of hexane, and use this solution as the sample solution. Amount (mg) of menatetrenone (C 31H40O2) ＝ M S × QT/QS Pipet 1 mL of the sample solution, add hexane to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the chromatogram with a mixture of hexane and di-n-butyl ether (17:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: MS: Amount (mg) of Menatetrenone RS taken, calculated on the dehydrated basis Internal standard solution—A solution of phytonadione in 2-propanol (1 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ‌gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Methanol. ‌Flow rate: Adjust so that the retention time of menatetre-none is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, menatetrenone and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of menatetrenone to that of the internal standard is not more than 1.0z.
Mentha Water is a clear, colorless liquid, hav-ing the odor of mentha oil.
Mepenzolate Bromide is white to pale yellow, crystals or crystalline powder. It is odorless, and has a bitter taste. It is very soluble in formic acid, freely soluble in metha-nol, soluble in hot water, slightly soluble in water and in ethanol (95), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: about 2309C (with decomposition).
(1) To 30 mg of Mepenzolate Bromide add 10 drops of sulfuric acid: a red color develops. Dissolve 10 mg of Mepenzolate Bromide in 20 mL of water and 5 mL of dilute hydrochloric acid, and to 5 mL of this solution add 1 mL of Dragendorff's TS: an orange pre-cipitate is produced. Determine the absorption spectrum of a solution of Mepenzolate Bromide in 0.01 mol/L hydrochloric acid TS (1 in 2000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Dissolve 0.5 g of Mepenzolate Bromide in 50 mL of water and 3 mL of nitric acid by heating. This solution re-sponds to Qualitative Tests <1.09> for Bromide.
(1) Heavy Metals <1.07>—Proceed with 1.0 g of Mepenzolate Bromide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not less than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Mepenzolate Bromide according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.40 g of Mepenzolate Bromide in exactly measured 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add methanol to make exactly 200 mL, and use this solution as the standard solution (1). Separately, dis-solve 40 mg of benzophenone in methanol to make exactly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution, standard solutions (1) and (2) on a plate of silica gel with fluorecent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of methanol, 1-butanol, water and acetic acid (100) (3:3:2:1) to a distance of about 10 cm, and air-dry the plate and then at 809C for 30 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than either the principal spot or the spot corresponding to benzophenone obtained from the sam-ple solution are not more intense than the spot from stand-ard solution (1), and the spot corresponding to benzophe-none from the sample solution is not more intense than the spot from standard solution (2). Spray evenly Dragendorff's TS on the plate: the spots other than the principal spot from the sample solution are not more intense than the spot from standard solution (1). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Mepenzolate Bromide, previously dried, dissolve in 2 mL of formic acid, add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 42.03 mg of C21H26BrNO3
Mepitiostane occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in triethylamine, in chloroform, in diethyl ether and in cyclohexane, soluble in diethylene glycol dimethyl ether and in petroleum ether, sparingly soluble in acetone, slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water. It is hydrolyzed in moist air.
(1) Dissolve 1 mg of Mepitiostane in 1 mL of methanol, and add 0.5 mL of palladium (II) chloride TS: an orange precipitate is formed. To this suspension add 1 mL of water and 2 mL of chloroform, shake well, and allow to stand: an orange color develops in the chloroform layer. Dissolve 0.1 g of Mepitiostane in 2 mL of diethylene glycol dimethyl ether, shake with 1 mL of 1 mol/L hydro-chloric acid TS, and filter. To the filtrate add 1.5 mL of 2,4-dinitrophenylhydrazine-diethylene glycol dimethyl ether TS and 1.5 mL of diluted ethanol (95) (2 in 3): an orange-yellow precipitate is formed. Filter the precipitate, recrystallize from ethanol (99.5), and dry in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours: the crystals melt <2.60> between 1449C and 1499C. ‌Determine the infrared absorption spectrum of Mepitiostane as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋20 – ＋239 (0.1 g, chlo-roform, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.10 g of Mepitiostane in 4 mL of petroleum ether: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 1.0 g of Mepitiostane according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Mepitiostane in exactly 5 mL of a mixture of acetone and triethylamine (1000:1), and use this solution as the sample solution. Sepa-rately, dissolve 10 mg of Epitiostanol RS in a mixture of acetone and triethylamine (1000:1) to make exactly 10 mL. Pipet 1 mL and 3 mL of this solution, to each add a mixture of acetone and triethylamine (1000:1) to make exactly 25 mL, and use these solutions as the standard solution (1) and the standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solutions (1) and (2) on a plate of silica gel with fluores-cent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (3:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 5) on the plate, heat the plate be-tween 1209C and 1309C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): the spots other than the principal spot obtained from the sample solution showing the same Rf value as the standard solutions are not more intense than the spot from the standard solution (2), and the remaining spots other than the principal spot are not more intense than the spot from the standard solution (1). Water <2.48> Not more than 0.7z (0.3 g, volumetric titra-tion, back titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.3 g of Mepitiostane, and dissolve in cyclohexane to make exactly 10 mL. Pipet 2 mL of this solution, add 10 mL of ethanol (99.5), mix with ex-actly 2 mL each of 0.01 mol/L hydrochloric acid TS and the internal standard solution, add ethanol (99.5) to make 20 mL, allow to stand at ordinary temperature for 30 minutes, and use this solution as the sample solution. Separately, weigh accurately about 45 mg of Epitiostanol RS, dissolve in exactly 2 mL of the internal standard solution, add ethanol (99.5) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of epitiostanol to that of the internal standard, respectively. Amount (mg) of mepitiostane (C25H40O2S) ＝ MS × QT/QS × 5 × 1.320 MS: Amount (mg) of Epitiostanol RS taken, calculated on the anhydrous basis Internal standard solution—A solution of n-octylbenzene in ethanol (99.5) (1 in 300). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (20:3). Flow rate: Adjust so that the retention time of epitiostanol is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, epitiostanol and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of epitiostanol to that of the internal stand-ard is not more than 1.0z.
Mepivacaine Hydrochloride, when dried, contains not less than 98.
(1) Determine the absorption spectrum of a solution of Mepivacaine Hydrochloride (1 in 2500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mepivacaine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Mepivacaine Hydrochloride (1 in 50) responds to Qualitative Tests <1š.09> for chloride. pH <2.54> Dissolve 0.2 g of Mepivacaine Hydrochloride in 10 mL of water: the pH of this solution is between 4.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Mepivacaine Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Perform the test with 0.5 g of Mepivacaine Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Mepivacaine Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Mepivacaine Hydrochloride in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this so-lution, add methanol to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of diethyl ether, meth-anol and ammonia solution (28) (100:5:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly bismuth ni-trate-potassium iodide TS on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Mepivacaine Hy-drochloride, previously dried, dissolve in 10 mL of acetic acid (100) and add 70 mL of acetic anhydride. Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.28 mg of C15H22N2O.HCl
Mequitazine occurs as white, crystals or crys-talline powder. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95), and practically insoluble in water. It is gradually colored by light. A solution of Mequitazine in methanol (1 in 50) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Mequitazine in ethanol (95) (1 in 250,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Me-quitazine, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Mequitazine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 50 mg of Mequitazine in 5 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 50 mL, then pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop with a mix-ture of ethyl acetate, methanol and diethylamine (7:2:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the num-ber of the spot other than the principal spot obtained from the sample solution is not more than 3 and they are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Mequitazine, dis-solve in 50 mL of acetic acid (100), titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.25 mg of C20H22N2S
Mercaptopurine Hydrate contains not less than 98.
Mercaptopurine Hydrate occurs as light yellow to yellow, crystals or crystalline powder. It is odorless. It is practically insoluble in water, in acetone and in diethyl ether. It dissolves in sodium hydroxide TS and in ammonia TS.
(1) Dissolve 0.6 g of Mercaptopurine Hy-drate in 6 mL of sodium hydroxide solution (3 in 100), and add slowly 0.5 mL of iodomethane with vigorous stirring. Stir well for 10 minutes, cool in an ice bath, and adjust the pH with acetic acid (31) to about 5. Collect the separated crystals by filtration, recrystallize from water, and dry at 1209C for 30 minutes: the crystals melt <2.60> between 2189C and 2229C (with decomposition). (2) Determine the absorption spectrum of a solution of Mercaptopurine Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity of solution—Dissolve 0.20 g of Mer-captopurine Hydrate in 10 mL of ammonia TS: the solution is clear. Sulfate <1.14>—Dissolve 50 mg of Mercaptopurine Hydrate in 10 mL of dilute hydrochloric acid, add 5 drops of barium chloride TS, and allow to stand for 5 minutes: no turbidity is produced. Heavy metals <1.07>—Proceed with 1.0 g of Mercap- topurine Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Hypoxanthine—Dissolve 50 mg of Mercaptopurine Hydrate in exactly 10 mL of a solution of ammonia solution (28) in methanol (1 in 10), and use this solution as the sample solution. Separately, dissolve 5.0 mg of hypoxanthine in a solution of ammonia solution (28) in methanol (1 in 10) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of methanol, chlo- Amount (mg) of mequitazine (C ＝ MS × AT/AS × 1/8 20H22N2S) roform, n-butyl formate and ammonia solution (28) (8:6:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): MS: Amount (mg) of mequitazine for assay taken
Weigh accurately the mass of not less than 20 Me-quitazine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3 mg of mequitazine (C20H22N2S), add 50 mL of a mixture of methanol and water (4:3), shake thoroughly, and add methanol to make exactly 100 mL. Centrifuge, if necessary, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard 10 mL of the first filtrate, pipet 4 mL of the subsequent filtrate, add methanol to make exactly 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 24 mg of mequitazine for assay, previously dried in vacuum at 609C using phos-phorous (V) oxide as the desiccant for 3 hours, and dissolve in methanol to make exactly 50 mL. Pipet 1 mL of this solu-tion, add methanol to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 254 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. C5H4N4S.H2O: 170.19 1,7-Dihydro-6H-purine-6-thione monohydrate [6112-76-1] Mercaptopurine Hydrate contains not less than 98.0z of mercaptopurine (C5H4N4S: 152.18), calcu-lated on the anhydrous basis. Description Mercaptopurine Hydrate occurs as light yellow to yellow, crystals or crystalline powder. It is odorless. It is practically insoluble in water, in acetone and in diethyl ether. It dissolves in sodium hydroxide TS and in ammonia TS. Identification (1) Dissolve 0.6 g of Mercaptopurine Hy-drate in 6 mL of sodium hydroxide solution (3 in 100), and add slowly 0.5 mL of iodomethane with vigorous stirring. Stir well for 10 minutes, cool in an ice bath, and adjust the pH with acetic acid (31) to about 5. Collect the separated crystals by filtration, recrystallize from water, and dry at 1209C for 30 minutes: the crystals melt <2.60> between 2189C and 2229C (with decomposition). (2) Determine the absorption spectrum of a solution of Mercaptopurine Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Purity (1) Clarity of solution—Dissolve 0.20 g of Mer-captopurine Hydrate in 10 mL of ammonia TS: the solution is clear. Sulfate <1.14>—Dissolve 50 mg of Mercaptopurine Hydrate in 10 mL of dilute hydrochloric acid, add 5 drops of barium chloride TS, and allow to stand for 5 minutes: no turbidity is produced. Heavy metals <1.07>—Proceed with 1.0 g of Mercap- topurine Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Hypoxanthine—Dissolve 50 mg of Mercaptopurine Hydrate in exactly 10 mL of a solution of ammonia solution (28) in methanol (1 in 10), and use this solution as the sample solution. Separately, dissolve 5.0 mg of hypoxanthine in a solution of ammonia solution (28) in methanol (1 in 10) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of methanol, chlo- Amount (mg) of mequitazine (C ＝ MS × AT/AS × 1/8 20H22N2S) roform, n-butyl formate and ammonia solution (28) (8:6:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): MS: Amount (mg) of mequitazine for assay taken
Meropenem Hydrate contains not less than 980 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the anhydrous basis.
Meropenem Hydrate occurs as a white to light yellow crystalline powder. It is sparingly soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It dissolves in sodium hydrogen carbonate TS.
(1) Dissolve 10 mg of Meropenem Hydrate in 2 mL of water, add 3 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 5 minutes, add 1 mL of acidic ammonium iron (III) sulfate TS, and shake: a red-brown color develops. Determine the absorption spectra of solutions of Meropenem Hydrate and Meropenem RS (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectra: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectra of Meropenem Hydrate and Meropenem RS as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectra: both spectra ex-hibit similar intensities of absorption at the same wave num-bers. D Optical rotation <2.49> [a]20: －17 – －219(0.22 g calcu-lated on the anhydrous basis, water, 50 mL, 100 mm). pH <2.54> Dissolve 0.2 g of Meropenem Hydrate in 20 mL of water: the pH of the solution is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Meropenem Hydrate in 10 mL of sodium hydrogen car-bonate TS: the solution is clear and has no more color than the following control solution. Control solution: To a mixture of 0.3 mL of Cobalt (II) Chloride CS and 1.2 mL of Iron (III) Chloride CS add 18.5 mL of diluted hydrochloric acid (1 in 40). Heavy metals <1.07>—Proceed with 2.0 g of Meropenem Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Meropenem Hydrate in 10 mL of triethylamine-phosphate buffer solu-tion (pH 5.0), and use this solution as the sample solution. Prepare the sample solution before use. Pipet 1 mL of the sample solution, and add triethylamine-phosphate buffer so-lution (pH 5.0) to make exactly 100 mL. Pipet 3 mL of this solution, add triethylamine-phosphate buffer solution (pH 5.0) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of ring-opened meropenem, having the relative retention time about 0.5 to meropenem, and the peak area of the dimmer, having the relative retention time about 2.2 to meropenem, obtained from the sample solution are not larger than the peak area of meropenem from the standard solution, the area of the peak other than meropenem and the peaks mentioned above from the sample solution is not larger than 1/3 times the peak area of meropenem from the standard solution, and the total area of the peaks other than meropenem from the sample solu-tion is not larger than 3 times the peak area of meropenem from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of triethylamine-phosphate buffer solution (pH 5.0) and acetonitrile (100:7). Flow rate: Adjust so that the retention time of meropenem is about 6 minutes. Time span of measurement: About 7 times as long as the retention time of meropenem. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add triethylamine-phosphate buffer solution (pH 5.0) to make exactly 25 mL. Confirm that the peak area of meropenem obtained with 10 mL of this solution is equiva-lent to 16 to 24z of that with 10 mL of the standard solu-tion. System performance: Warm the sample solution at 609C for 30 minutes. When the procedure is run with 10 mL of this solution under the above operating conditions, the ring-o-pened meropenem, meropenem and the dimer are eluted in this order, and the resolution between the peaks of the ring-opened meropenem and meropenem is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of meropenem is not more than 1.5z. Water <2.48> Not less than 11.4z and not more than 13.4z (0.35 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Meropenem Hy-drate and Meropenem RS, equivalent to about 50 mg (po-tency), dissolve each in exactly 10 mL of the internal stand-ard solution, add triethylamine-phosphate buffer solution (pH 5.0) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 5 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of meropenem to that of the internal standard. conditions, the relative standard deviation of the ratios of the peak area of meropenem to that of the internal standard is not more than 1.0z.
Mesalazine, when dried, contains not less than 98.
Mesalazine occurs as white, light gray or red-dish-white, crystals or crystalline powder. It is very slightly soluble in water, and practically insoluble in ethanol (99.5). It dissolves in dilute hydrochloric acid.
(1) Determine the absorption spectrum of a solution of Mesalazine in 0.1 mol/L hydrochloric acid TS (1 in 80,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Mesalazine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Perform this procedure while keeping the solution at 409C. A solution ob-tained by dissolving 0.5 g of Mesalazine in 20 mL of 1 mol/L hydrochloric acid TS is clear, and its absorbance at 440 nm and 650 nm, determined immediately as directed under Ul-traviolet-visible Spectrophotometry <2.24>, is not more than 0.15 and not more than 0.10, respectively. Chloride <1.03>—Dissolve 0.30 g of Mesalazine in 6 mL of dilute nitric acid and 40 mL of water, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.80 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.095z). Sulfate—To 1.0 g of Mesalazine add 20 mL of water, shake for 1 minute, and filter. To 15 mL of the filtrate add Amount [mg (potency)] of meropenem (C ＝ MS × QT/QS 17H25N3O5S) 0.5 mL of acetic acid (31), then add 2.5 mL of the following solution A, and use this solution as the test solution. Solu-tion A: To 3 mL of barium chloride TS add 4.5 mL of a so- MS: Amount [mg (potency)] of Meropenem RS taken Internal standard solution—A solution of benzyl alcohol in triethylamine-phosphate buffer solution (pH 5.0) (1 in 300).
Weigh accurately about 50 mg of Mesalazine, previ-ously dried, dissolve in 100 mL of hot water, cool to room temperature quickly, and titrate <2.50> with 0.1 mol/L so- dium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 15.31 mg of C7H7NO3
Mestranol, when dried, contains not less than 97.
Mestranol occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in chloroform, sparingly soluble in ethanol (99.5), and practically insoluble in water.
(1) Dissolve 2 mg of Mestranol in 1 mL of a mixture of sulfuric acid and ethanol (99.5) (2:1): a red-purple color develops with a yellow-green fluorescence. Determine the absorption spectrum of a solution of Mestranol in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mestranol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi- Amount (mg) of mesalazine (C7H7NO3) ＝ M S × QT/QS ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mes- MS: Amount (mg) of mesalazine for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 300 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 400 mL of methanol, 1 mL of phosphoric acid, 0.865 g of sodium lauryl sulfate and 0.679 g of tetrabutylammonium hydrogensulfate in water to make 1000 mL. Flow rate: Adjust so that the retention time of mesalazine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, mesalazine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. tranol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Mestranol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋1 – ＋69 (after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Mestranol according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Mestranol according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Mestranol in 20 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add chloroform to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each ‌of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (99.5) (29:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 5) on the plate, and heat the plate at 1059C for 15 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately the mass of not less than 20 Mesalazine Extended-release Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 40 mg of mesalazine (C7H7NO3), add 100 mL of diluted phos-phoric acid (1 in 1000), shake vigorously, and sonicate for 5 minutes. Add exactly 10 mL of the internal standard solu-tion, then add 90 mL of methanol and diluted phosphoric acid (1 in 1000) to make 250 mL. Filter this solution through a membrane filter with a pore size 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of mesalazine for assay, previously dried at 1059C for 2 hours, add 100 mL of diluted phosphoric acid (1 in 1000), shake vigorously, and sonicate for 5 minutes to dissolve. Add exactly 10 mL of the internal standard solution, then add 90 mL of methanol and diluted phosphoric acid (1 in 1000) to make 250 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of mesalazine to that of the internal standard. C21H26O2: 310.43 Methoxy-19-nor-17a-pregna-1,3,5(10)-trien-20-yn-17-ol [72-33-3] Mestranol, when dried, contains not less than 97.0z and not more than 102.0z of mestranol (C21H26O2). Description Mestranol occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in chloroform, sparingly soluble in ethanol (99.5), and practically insoluble in water. Identification (1) Dissolve 2 mg of Mestranol in 1 mL of a mixture of sulfuric acid and ethanol (99.5) (2:1): a red-purple color develops with a yellow-green fluorescence. Determine the absorption spectrum of a solution of Mestranol in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mestranol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi- Amount (mg) of mesalazine (C7H7NO3) ＝ M S × QT/QS ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mes- MS: Amount (mg) of mesalazine for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 300 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 400 mL of methanol, 1 mL of phosphoric acid, 0.865 g of sodium lauryl sulfate and 0.679 g of tetrabutylammonium hydrogensulfate in water to make 1000 mL. Flow rate: Adjust so that the retention time of mesalazine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, mesalazine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. tranol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Mestranol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋1 – ＋69 (after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm). Melting point <2.60> 148 – 1549C Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Mestranol according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Mestranol according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Mestranol in 20 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add chloroform to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each ‌of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (99.5) (29:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (1 in 5) on the plate, and heat the plate at 1059C for 15 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Weigh accurately about 10 mg each of Mestranol and Mestranol RS, previously dried, dissolve in ethanol (99.5) to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. De-termine the absorbances, AT and AS, of the sample solution and the standard solution at 279 nm as directed under Ultra-violet-visible Spectrophotometry <2.24>. and stir for 15 minutes. Filter the precipitate so obtained by suction through a glass filter (G4), wash with 10 mL of water, and dry at 1059C for 1 hour: it melts <2.60> between 1579C and 1619C. Determine the infrared absorption spectrum of Metenolone Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋39 – ＋429(after drying, 0.2 g, chloroform, 10 mL, 100 mm). Melting point <2.60> 141 – 1449C Purity (1) Clarity and color of solution—Dissolve 0.50 g of Metenolone Acetate in 10 mL of 1,4-dioxane: the solution is clear and colorless to pale yellow. (2) Heavy metals <1.07>—Proceed with 2.0 g of Meteno- lone Acetate according to Method 2, and perform the test. Amount (mg) of mestranol (C 21H26O2) ＝ M S × AT/AS Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). MS: Amount (mg) of Mestranol RS taken
Metenolone Acetate, when dried, contains not less than 97.
Metenolone Acetate occurs as a white to pale yellow-white crystalline powder. It is odorless. It is freely soluble in acetone, in 1,4-dioxane and in chlo-roform, soluble in methanol and in ethanol (95), sparingly soluble in diethyl ether and in sesame oil, slightly soluble in hexane and in petroleum ether, and practically insoluble in water.
(1) Dissolve 1 mg of Metenolone Acetate in 5 mL of a mixture of sulfuric acid and ethanol (95) (1:1), and heat for 30 minutes in a water bath: a red-brown color de-velops. To 10 mg of Metenolone Acetate add 0.5 mL of dilute sodium hydroxide-ethanol TS, and heat for 1 minute on a water bath. After cooling, add 0.5 mL of diluted sulfu-ric acid (1 in 2), and boil gently for 1 minute: the odor of ethyl acetate is perceptible. Dissolve 50 mg of Metenolone Acetate in 3 mL of methanol, add 0.3 mL of a solution of potassium carbonate (1 in 6), and boil for 2 hours under a reflux condenser. After cooling, add this solution gradually to 50 mL of cold water, (3) Related substances—Dissolve 35 mg of Metenolone Acetate in 20 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, dilute with chloroform to exactly 250 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and cyclohexane (1:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 10 mg of Metenolone Ace-tate, previously dried, and dissolve in methanol to make ex-actly 100 mL. Pipet 5 mL of this solution, and dilute with methanol to exactly 50 mL. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 242 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>. Amount (mg) of metenolone acetate (C22H32O3) ＝ A/391 × 10,000
Metenolone Enanthate occurs as white, crys-tals or crystalline powder. It is odorless. It is very soluble in ethanol (95), in acetone, in 1,4-dioxane and in chloroform, freely soluble in methanol, in ethyl ace-tate, in diethyl ether, in cyclohexane, in petroleum ether and in toluene, soluble in sesame oil, and practically insoluble in water.
(1) Heat 1 mg of Metenolone Enanthate with 5 mL of a mixture of sulfuric acid and ethanol (95) (1:1) on a water bath for 30 minutes: a red-brown color develops. (2) Dissolve 50 mg of Metenolone Enanthate in 3 mL of methanol, add 0.3 mL of a solution of potassium carbonate (1 in 6), boil under a reflux condenser for 2 hours, cool, add slowly this solution to 50 mL of cold water, and stir for 15 minutes. Filter the produced precipitate by suction through a glass filter (G4), wash with water until the washings become neutral, and dry at 1059C for 1 hour: it melts <2.60> between 1569C and 1629C. D Optical rotation <2.49> [a]20: ＋39 – ＋439(after drying, 0.2 g, chloroform, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Metenolone Enanthate in 10 mL of 1,4-dioxane: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Meteno- lone Enanthate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 20 mg of Metenolone Enanthate in exactly 10 mL of chloroform, and use this solu-tion as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate and cyclohexane (1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): any spot other than the principal spot does not appear. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.1 g of Metenolone Enan- thate, previously dried, and dissolve in methanol to make ex-actly 100 mL. Pipet 10 mL of this solution, and dilute with methanol to make exactly 100 mL. Pipet 10 mL of this solu-tion, and dilute again with methanol to make exactly 100 mL. Determine the absorbance, A, of this solution at the wavelength of maximum absorption at about 242 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of metenolone enanthate (C27H42O3) ＝ A/325 × 100,000
Metformin Hydrochloride, when dried, contains not less than 98.
Metformin Hydrochloride occurs as white, crystals or crystalline powder. It is freely soluble in water, sparingly soluble in acetic acid (100), and slightly soluble in ethanol (99.5). Melting point: about 2219C (with decomposition).
(1) Determine the absorption spectrum of a solution of Metformin Hydrochloride (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Met-formin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Metformin Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Metformin Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 2.5 g of Metformin not more intense than the spot from the standard solution (1), the number of them showing more intense than the spot from the standard solution (2) is not more than two, and the spot from the sample solution appeared at the position cor-responding to the spot from the standard solution (3) is not more intense than the spot from the standard solution (3). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Metformin Hydro-chloride, previously dried, dissolve in 40 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 4.141 mg of C4H11N5.HCl
Methamphetamine Hydrochloride occurs as colorless crystals or a white crystalline powder. It is odorless. It is freely soluble in water, in ethanol (95) and in chlo- roform, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Methamphetamine Hy-drochloride in 10 mL of water is between 5.0 and 6.0.
(1) To 5 mL of a solution of Metham- Amount (mg) of metformin hydrochloride (C H N .HCl) phetamine Hydrochloride (1 in 100) add 0.5 mL of hydrogen ＝ MS × QT/QS 4 11 5 hexachloroplatinate (IV) TS: an orange-yellow, crystalline precipitate is produced. MS: Amount (mg) of metformin hydrochloride for assay taken Internal standard solution—Dissolve 0.3 g of isobutyl par-ahydroxybenzoate in 100 mL of the mixture of water and acetonitrile (3:2). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.8 g of sodium lauryl sulfate in 620 mL of diluted phosphoric acid (1 in 2500), and add 380 mL of acetonitrile. Flow rate: Adjust so that the retention time of metformin is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, metformin and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of metformin to that of the internal standard is not more than 1.0z.
(1) Acidity or alkalinity—Dissolve 2.0 g of Methamphetamine Hydrochloride in 40 mL of freshly boiled and cooled water, add 2 drops of methyl red TS, and use this solution as the sample solution. To 20 mL of the sample solution add 0.20 mL of 0.01 mol/L sulfuric acid VS: a red color develops. To 20 mL of the sample solution add 0.20 mL of 0.02 mol/L sodium hydroxide VS: a yellow color develops. (2) Sulfate <1.14>—Dissolve 0.05 g of Methamphetamine Hydrochloride in 40 mL of water, add 1 mL of dilute hydro-chloric acid and 1 mL of barium chloride TS, and allow to stand for 10 minutes: the solution remains unchanged. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Methamphetamine Hydrochloride, previously dried, and dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3). Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.57 mg of C10H15N.HCl
Methotrexate is a mixture of 4-amino-10-methylfolic acid and closely related compounds. It contains not less than 94.
Methotrexate occurs as a yellow-brown crys-talline powder. It is slightly soluble in pyridine, and practically insoluble in water, in acetonitrile, in ethanol (95) and in diethyl ether. It dissolves in dilute sodium hydroxide TS and in dilute so- dium carbonate TS. It is gradually affected by light.
(1) Dissolve 1 mg of Methotrexate in 100 mL of 0.1 mol/L hydrochloric acid TS. Determine the ab-sorption spectrum of this solution as directed under Ultra- ‌violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Methotrexate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Methotrexate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Methotrexate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Water <2.48> Take 5 mL of pyridine for water determina-tion and 20 mL of methanol for water determination in a dried titration flask, and titrate with Karl Fischer TS until the end point. Weigh accurately about 0.2 g of Methotrexate, immediately place in the titration flask, and add a known excess volume of Karl Fischer TS for water determination. Mix well for 30 minutes, and perform the test: the water content is not more than 12.0z. Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 25 mg each of Methotrexate and Methotrexate RS, dissolve in the mobile phase to make exactly 250 mL, and use these solutions as the sample solu-tion and standard solution. Perform the test with exactly 10 mL each of these solutions as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of methotrexate in each solution. Amount (mg) of methotrexate (C20H22N8O5) ＝ MS × AT/AS MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 302 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of disodium hydrogen phos-phate-citric acid buffer solution (pH 6.0) and acetonitrile Methotrexate Capsules
Methoxsalen occurs as white to pale yellow, crystals or crystalline powder. It is odorless and tasteless. It is freely soluble in chloroform, slightly soluble in meth- Amount (mg) of methotrexate (C H N O ) anol, in ethanol (95) and in diethyl ether, and practically in- ＝ MS × AT/AS × 2/5 20 22 8 5 soluble in water. MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 302 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of disodium hydrogen phos-phate-citric acid buffer solution (pH 6.0) and acetonitrile (89:11). Flow rate: Adjust so that the retention time of metho-trexate is about 8 minutes. System suitability— System performance: Dissolve 10 mg each of methotrexate and folic acid in 100 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, folic acid and methotrexate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methotrexate is not more than 1.0z.
(1) To 10 mg of Methoxsalen add 5 mL of dilute nitric acid, and heat: a yellow color develops. Make this solution alkaline with a solution of sodium hydroxide (2 in 5): the color changes to red-brown. To 10 mg of Methoxsalen add 5 mL of sulfuric acid, and shake: a yellow color develops. Determine the absorption spectrum of a solution of Methoxsalen in ethanol (95) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Methoxsalen RS prepared in the same man-ner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Methoxsalen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Methoxsalen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Methoxsalen in 10 mL of chloroform, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add chloroform to make exactly 50 mL. Pipet 1 mL of this solution, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, hexane and ethyl acetate (40:10:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Methoxsalen and Methoxsalen RS, and dissolve each in ethanol (95) to ‌make exactly 100 mL. Pipet 2 mL each of these solutions, and dilute each with ethanol (95) to make exactly 25 mL. Pipet 10 mL each of these solutions, and dilute each again with ethanol (95) to make exactly 50 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Determine the absorbances, AT and AS, of the sample solution and the standard solution at 300 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. ‌Amount (mg) of methoxsalen (C12H8O4) ＝ MS × AT/AS MS: Amount (mg) of Methoxsalen RS taken, calculated on the anhydrous basis
Methyl Parahydroxybenzoate contains not less than 98.
Methyl Parahydroxybenzoate, occurs as col-orless crystals or a white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tone, and slightly soluble in water.◆
Determine the infrared absorption spectrum of Methyl Parahydroxybenzoate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Methyl Parahydroxybenzoate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Methyl Parahydroxybenzoate in ethanol (95) to make 10 mL: the solution is clear and not more intensely colored than the following control solution. Control solution: To 5.0 mL of Cobalt (II) Chloride CS, 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity—To 2 mL of the solution of Methyl Parahy-droxybenzoate obtained in (1) add 3 mL of ethanol (95), add 5 mL of freshly boiled and cooled water and 0.1 mL of bromocresol green-sodium hydroxide-ethanol TS, then add 0.1 mol/L sodium hydroxide VS until the solution shows a blue color: the volume of 0.1 mol/L sodium hydroxide VS used does not exceed 0.1 mL. (3) Heavy metals <1.07>—Dissolve 1.0 g of Methyl Par- ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute dissolve separately in 2.5 mL each of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of methyl parahydroxybenzoate in each solution. Amount (mg) of methyl parahydroxybenzoate (C H O ) acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control ＝ MS × AT/AS 8 8 3 solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm).◆ ‌Related substances—Dissolve 50.0 mg of Methyl Par-ahydroxybenzoate in 2.5 mL of methanol, and add the mo-bile phase to make exactly 50 mL. Pipet 10 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak area of parahydroxybenzoic acid having a relative retention time of about 0.6 to methyl par-ahydroxybenzoate obtained from the sample solution is not larger than the peak area of methyl parahydroxybenzoate from the standard solution (0.5z). For the peak area of parahydroxybenzoic acid, multiply the correction factor, 1.4. Furthermore, the area of the peak other than methyl parahydroxybenzoate and parahydroxybenzoic acid from the sample solution is not larger than the peak area of methyl parahydroxybenzoate from the standard solution (0.5z), and the total area of the peaks other than methyl parahydroxybenzoate from the sample solution is not larger than 2 times the peak area of methyl parahydroxybenzoate from the standard solution (1.0z). For this calculation the peak area not larger than 1/5 times the peak area of methyl parahydroxybenzoate from the standard solution is excluded (0.1z). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of methyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of methyl parahydroxyben-zoate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately and powder not less than 20 Methylergometrine Maleate Tablets. Weigh accurately a portion of the powder, equivalent to about 0.3 mg of methylergometrine maleate (C20H25N3O2.C4H4O4), transfer to a brown separator, add 15 mL of sodium hydrogen car-bonate solution (1 in 20), and extract with four 20-mL por-tions of chloroform. Filter each portion of the chloroform extracts through a pledget of absorbent cotton, previously moistened with chloroform, into another dried, brown sepa-rator, combine all the extracts, and use this extract as the sample solution. Separately, weigh accurately about 10 mg of Methylergometrine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, dissolve in water, and add water to make exactly 100 mL. Pipet 3 mL of this solution, and transfer to a brown separator, proceed in the same man-ner as the preparation of the sample solution, and use this extract as the standard solution. To each total volume of the Description Methyl Parahydroxybenzoate, occurs as col-orless crystals or a white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tone, and slightly soluble in water.◆ Identification Determine the infrared absorption spectrum of Methyl Parahydroxybenzoate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Methyl Parahydroxybenzoate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 125 – 1289C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Methyl Parahydroxybenzoate in ethanol (95) to make 10 mL: the solution is clear and not more intensely colored than the following control solution. Control solution: To 5.0 mL of Cobalt (II) Chloride CS, 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity—To 2 mL of the solution of Methyl Parahy-droxybenzoate obtained in (1) add 3 mL of ethanol (95), add 5 mL of freshly boiled and cooled water and 0.1 mL of bromocresol green-sodium hydroxide-ethanol TS, then add 0.1 mol/L sodium hydroxide VS until the solution shows a blue color: the volume of 0.1 mol/L sodium hydroxide VS used does not exceed 0.1 mL. (3) Heavy metals <1.07>—Dissolve 1.0 g of Methyl Par- ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute dissolve separately in 2.5 mL each of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of methyl parahydroxybenzoate in each solution. Amount (mg) of methyl parahydroxybenzoate (C H O ) acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control ＝ MS × AT/AS 8 8 3 solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm).◆ ‌Related substances—Dissolve 50.0 mg of Methyl Par-ahydroxybenzoate in 2.5 mL of methanol, and add the mo-bile phase to make exactly 50 mL. Pipet 10 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak area of parahydroxybenzoic acid having a relative retention time of about 0.6 to methyl par-ahydroxybenzoate obtained from the sample solution is not larger than the peak area of methyl parahydroxybenzoate from the standard solution (0.5z). For the peak area of parahydroxybenzoic acid, multiply the correction factor, 1.4. Furthermore, the area of the peak other than methyl parahydroxybenzoate and parahydroxybenzoic acid from the sample solution is not larger than the peak area of methyl parahydroxybenzoate from the standard solution (0.5z), and the total area of the peaks other than methyl parahydroxybenzoate from the sample solution is not larger than 2 times the peak area of methyl parahydroxybenzoate from the standard solution (1.0z). For this calculation the peak area not larger than 1/5 times the peak area of methyl parahydroxybenzoate from the standard solution is excluded (0.1z). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of methyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of methyl parahydroxyben-zoate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 50.0 mg each of Methyl Par-ahydroxybenzoate and Methyl Parahydroxybenzoate RS, MS: Amount (mg) of Methyl Parahydroxybenzoate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and potassium dihy-drogen phosphate solution (17 in 2500) (13:7). Flow rate: 1.3 mL per minute. System suitability— System performance: Dissolve 5 mg each of Methyl Par-ahydroxybenzoate and parahydroxybenzoic acid in the mo-bile phase to make exactly 100 mL. Pipet 1 mL of this solu-tion, and add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, parahydroxybenzoic acid and methyl parahydroxybenzoate are eluted in this order, the relative retention time of parahydroxybenzoic acid to methyl parahydroxybenzoate is about 0.6, and the resolution be-tween these peaks is not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methyl parahydroxybenzoate is not more than 0.85z.
Methyl Salicylate contains not less than 98.
Methyl Salicylate is a colorless to pale yellow liquid. It has a strong, characteristic odor. It is miscible with ethanol (95) and with diethyl ether. It is very slightly soluble in water. Specific gravity d 20: 1.182 – 1.192 Boiling point: 219 – 2249C
Shake 1 drop of Methyl Salicylate thor-oughly with 5 mL of water for 1 minute, and add 1 drop of iron (III) chloride TS: a purple color develops.
(1) Acidity—Shake 5.0 mL of Methyl Salicylate thoroughly with 25 mL of freshly boiled and cooled water and 1.0 mL of 0.1 mol/L sodium hydroxide VS for 1 minute, add 2 drops of phenol red TS, and titrate <2.50> with Amount (mg) of methylprednisolone succinate (C ＝ MS × QT/QS 26H 34O8) 0.1 mol/L hydrochloric acid VS until the red color disap-pears: not more than 0.45 mL of 0.1 mol/L sodium hydrox-ide VS is consumed. MS: Amount (mg) of Methylprednisolone Succinate RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (1:1) (3 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of 0.05 mol/L potassium dihy-drogen phosphate TS add a suitable amount of 0.05 mol/L disodium hydrogen phosphate TS to make a solution having pH 5.5. To 640 mL of this solution add 360 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of methyl- (2) Heavy metals—Shake 10.0 mL of Methyl Salicylate thoroughly with 10 mL of water, add 1 drop of hydrochloric acid, and saturate with hydrogen sulfide by passing it through the mixture: neither the oily layer nor the aqueous layer shows a dark color.
Weigh accurately about 15 mg each of Methylpred-nisolone Succinate and Methylprednisolone Succinate RS, previously dried, dissolve separately in 5 mL of methanol, and add the mixture of 0.05 mol/L phosphate buffer solu-tion (pH 3.5) and acetonitrile (1:1) to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of methylpredniso-lone succinate to that of the internal standard. Description Methyl Salicylate is a colorless to pale yellow liquid. It has a strong, characteristic odor. It is miscible with ethanol (95) and with diethyl ether. It is very slightly soluble in water. Specific gravity d 20: 1.182 – 1.192 Boiling point: 219 – 2249C Identification Shake 1 drop of Methyl Salicylate thor-oughly with 5 mL of water for 1 minute, and add 1 drop of iron (III) chloride TS: a purple color develops. Purity (1) Acidity—Shake 5.0 mL of Methyl Salicylate thoroughly with 25 mL of freshly boiled and cooled water and 1.0 mL of 0.1 mol/L sodium hydroxide VS for 1 minute, add 2 drops of phenol red TS, and titrate <2.50> with Amount (mg) of methylprednisolone succinate (C ＝ MS × QT/QS 26H 34O8) 0.1 mol/L hydrochloric acid VS until the red color disap-pears: not more than 0.45 mL of 0.1 mol/L sodium hydrox-ide VS is consumed. MS: Amount (mg) of Methylprednisolone Succinate RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (1:1) (3 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of 0.05 mol/L potassium dihy-drogen phosphate TS add a suitable amount of 0.05 mol/L disodium hydrogen phosphate TS to make a solution having pH 5.5. To 640 mL of this solution add 360 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of methyl- (2) Heavy metals—Shake 10.0 mL of Methyl Salicylate thoroughly with 10 mL of water, add 1 drop of hydrochloric acid, and saturate with hydrogen sulfide by passing it through the mixture: neither the oily layer nor the aqueous layer shows a dark color. Assay Weigh accurately about 2 g of Methyl Salicylate, add an exactly measured 50 mL of 0.5 mol/L potassium hy-droxide-ethanol VS, and heat on a water bath for 2 hours under a reflux condenser. Cool, and titrate <2.50> the excess potassium hydroxide with 0.5 mol/L hydrochloric acid VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 0.5 mol/L potassium hydroxide-ethanol VS ＝ 76.08 mg of C8H8O3
Methylbenactyzium Bromide, when dried, contains not less than 99.
Methylbenactyzium Bromide occurs as white, crystals or crystalline powder. It is odorless, and has an ex-tremely bitter taste. It is freely soluble in water and in acetic acid (100), soluble in ethanol (95), slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Methylbenactyzium Bromide in 50 mL of water is between 5.0 and 6.0.
(1) Shake 0.5 mL of a solution of Methyl-benactyzium Bromide (1 in 100) with 5 mL of phosphate buffer solution (pH 7.0), 2 to 3 drops of bromothymol blue TS and 5 mL of chloroform: a yellow color develops in the chloroform layer. To about 1 g of Methylbenactyzium Bromide add 5 mL of water and 10 mL of sodium hydroxide TS, allow to stand for 5 minutes, add 5 mL of dilute hydrochloric acid, collect the precipitate, wash well with water, recrystallize from a mixture of water and ethanol (95) (10:3), and dry at 1059C for 1 hour: the crystals melt <2.60> between 1459C and 1509C. Continue the heating up to about 2009C: a red color develops. Add 2 mL of dilute nitric acid to 5 mL of a solution of Methylbenactyzium Bromide (1 in 10): the solution re-sponds to Qualitative Tests <1.09> (1) for bromide.
(1) Clarity and color of solution—Dissolve 1.0 g of Methylbenactyzium Bromide in 10 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Perform the test with 0.5 g of Methylbenactyzium Bromide. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Methyl- benactyzium Bromide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Methylbenactyzium Bromide, previously dried, and dissolve in 80 mL of a mix-ture of acetic anhydride and acetic acid (100) (4:1). Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 42.24 mg of C21H28BrNO3
Methylcellulose is a methyl ether of cellulose. It contains not less than 26.
Methylcellulose occurs as a white to yellow-ish white, powder or grains. It is practically insoluble in ethanol (99.5). It swells, when water is added, and forms a clear or slightly turbid, viscous liquid.◆ Identiˆcation (1) Disperse evenly 1.0 g of Methylcellulose over the surface of 100 mL of water in a beaker, while gently tapping the top of the beaker, if necessary, and allow the beaker to stand: it aggregates on the surface of water. Add 1.0 g of Methylcellulose to 100 mL of hot water, and stir: it becomes a suspension. Cool the suspension to 59C, and stir: the resulting liquid is a clear or a slightly cloudy, viscous ‰uid. To 0.1 mL of the viscous ‰uid obtained in (2) add 9 mL of diluted sulfuric acid (9 in 10), shake, heat in a water bath for exactly 3 minutes, and immediately cool in ice water. Add carefully 0.6 mL of ninhydrin TS, shake, and allow to stand at 259C: the solution shows a red color, and it does not change to purple color within 100 minutes. Pour and spread out 2 to 3 mL of the viscous ‰uid obtained in (2) onto a glass plate, and allow the water to evaporate: a transparent ˆlm results. Pipet 50 mL of water, add exactly 50 mL of the vis-cous ‰uid obtained in (2), and warm to raise the temperature at a rate of 2 to 59C per minute while stirring: the tempera-ture, when a white turbidity of the solution starts to increase, is not less than 509C. Viscosity <2.53> (i) Method I: Apply to Methylcellulose having a labeled viscosity of less than 600 mPa･s. Put an exact amount of Methylcellulose, equivalent to 4.000 g, calculated on the dried basis, in a tared, wide-mouth bottle, add water (between 909C and 999C) to make 200 g, stopper the bottle, stir by mechanical means at 350 to 450 revolutions per minute for 10 to 20 minutes to get a homogeneous dis-persion. If necessary, take oŠ the sample attached on the walls of the bottle, put them in the dispersed solution, and dissolve by continuing the stirring in a water bath at not exceeding 59C for 20 to 40 minutes. Add cold water, if nec-essary, to make 200 g, and use this solution as the sample so-lution. Centrifuge the solution if necessary to expel any en-trapped air bubbles. Perform the test with the sample solu-tion at 20 ± 0.19C as directed in Method I under Viscosity Determination: not less than 80z and not more than 120z of the labeled viscosity. (ii) Method II: Apply to Methylcellulose having a labeled viscosity of not less than 600 mPa･s. Put an exact amount of Methylcellulose, equivalent to 10.00 g, calculated on the dried basis, in a tared, wide-mouth bottle, add water (be-tween 909C and 999C) to make 500 g, and prepare the sam-ple solution in the same manner as directed in Method I. Perform the test with the sample solution at 20 ± 0.19C as directed in Method II under Viscosity Determination, using a single cylinder-type rotational viscometer, according to the following operating conditions: not less than 75z and not more than 140z of the labeled viscosity. Operating conditions— Apparatus: Brookˆeld type viscometer LV model or an equivalent apparatus. Rotor No., rotation frequency, and calculation multiplier: According to the following table, depending on the labeled viscosity. Rotation of the mixture of nitric acid and sulfuric acid (5:4). Then boil gently until the solution changes to black. After cooling, add 2 mL of nitric acid, and heat until the solution changes to black. Repeat this procedure until the solution no longer changes to black, and heat strongly until dense white fumes are evolved. After cooling, add 5 mL of water, boil gently until dense white fumes are evolved, then heat until the volume of the solution becomes to 2 to 3 mL. After cooling, if the solution reveals yellow color by addition of 5 mL of water, add 1 mL of hydrogen peroxide (30), and heat until the volume of the solution becomes to 2 to 3 mL. After cool-ing, dilute the solution with 2 to 3 mL of water, transfer to a Nessler tube, add water to make 25 mL, and use this solution as the sample solution. Separately, put 2.0 mL of Standard Lead Solution in a 100-mL Kjeldahl ‰ask, add 18 mL of the mixture of nitric acid and sulfuric acid (5:4) and an amount of nitric acid equal to that used for preparation of the sam-ple solution, and heat until dense white fumes are evolved. After cooling, add 10 mL of water. In the case where hydro-gen peroxide (30) is added for the preparation of the sample solution, add the same amount of hydrogen peroxide (30), then proceed in the same manner for preparation of the sam-ple solution, and use so obtained solution as the control so-lution. Adjust the sample solution and the control solution to pH 3.0 to 4.0 with ammonia solution (28), and add water to make 40 mL, respectively. To these solutions add 1.2 mL of thioacetamide-alkaline glycerin TS, 2 mL of acetate buŠer solution (pH 3.5) and water to make 50 mL, separately. After allowing to stand for 5 minutes, observe vertically both tubes on a white background: the color obtained with the sample solution is not more intense than that with the control solution (not more than 20 ppm).◇ Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 1.5z (1 g).
Heavy metals—Put 1.0 g of Methylcellulose in a 100-mL Kjeldahl ‰ask, add a su‹cient amount of a mixture of nitric acid and sulfuric acid (5:4) to wet the sample, and heat gently. Repeat this procedure until to use totally 18 mL content is 130 ± 29C. In the case when a magnetic stirrer or shaker is not available, heat for 30 minutes with repeated shaking at 5-minute intervals by hand, and continue heating for an additional 30 minutes. Allow the vial to cool, and again weigh accurately. If the mass loss is less than 26 mg and there is no evidence of a leak of the content, use the up-per layer of the mixture as the sample solution. Separately, put 60 to 100 mg of adipic acid, 2.0 mL of the internal stand-ard solution and 2.0 mL of hydroiodic acid in a reaction vial, immediately stopper the vial tightly, and weigh accu-rately. Add 45 mL of iodomethane for assay through the sep-tum using a micro-syringe, weigh accurately, shake, and use the upper layer of the mixture as the standard solution. Per-form the test with 1 to 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iodomethane to that of the internal standard. Content (z) of methoxy group (CH3O) ＝ MS/M × QT/QS × 21.86 MS: Amount (mg) of iodomethane for assay taken M: Amount (mg) of Methylcellulose taken, calculated on the dried basis ‌Internal standard solution—A solution of n-octane in o-xylene (3 in 100). Operating conditions— Detector: A thermal conductivity detector or hydrogen ‰ame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Use a guard column, if necessary. Column temperature: Maintain the temperature at 509C for 3 minutes, raise to 1009C at a rate of 109C per minute, then raise to 2509C at a rate of 359C per minute, and main-tain at 2509C for 8 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2809C. Carrier gas: Helium. Flow rate: 4.3 mL per minute (the retention time of the in-ternal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 – 2 mL of the standard solution under the above operating con-ditions, iodomethane and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 1 – 2 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the ratio of the peak area of iodomethane to that of the internal standard is not more than 2.0z.
(i) Apparatus—Reaction vial: A 5-mL pressure-tight serum vial, having 20 mm in outside diameter and 50 mm in height, the neck 20 mm in outside diameter and 13 mm in inside diameter, equipped with a septum of butyl-rubber processed the surface with ‰uoroplastics, which can be ˆxed tightly to vial with aluminum cap, or equivalent. Heater: A square-shaped aluminum block, having holes 20 mm in diameter and 32 mm in depth, adopted to the reaction vial. Capable of stirring the content of the reaction vial by means of magnetic stirrer or of reciprocal shaker about 100 Labeled viscosity Rotor frequency Calculation times per minute. (mPa･s) No. /min multiplier (ii) Procedure—Weigh accurately about 65 mg of Methylcellulose, transfer to a reaction vial, add 60 to 100 mg of adipic acid, 2.0 mL of the internal standard solution and Not less than 600 and less than 1400 〃 〃 〃 〃 〃 〃 99,500 〃 99,500 2.0 mL of hydroiodic acid, immediately stopper the vial tightly, and weigh accurately. Using a magnetic stirrer equipped in the heating module, or using a shaker, stir for 60 minutes while heating so that the temperature of the vial Procedure of apparatus: Read the value after 2 minutes of rotation, and stop the rotation for at least 2 minutes. Repeat this procedure more two times, and average the three ob-served values. pH <2.54> The pH of the sample solution obtained in the Viscosity, measured after 5 minutes immersing the electrode in the sample solution, is between 5.0 and 8.0. Purity Heavy metals—Put 1.0 g of Methylcellulose in a 100-mL Kjeldahl ‰ask, add a su‹cient amount of a mixture of nitric acid and sulfuric acid (5:4) to wet the sample, and heat gently. Repeat this procedure until to use totally 18 mL content is 130 ± 29C. In the case when a magnetic stirrer or shaker is not available, heat for 30 minutes with repeated shaking at 5-minute intervals by hand, and continue heating for an additional 30 minutes. Allow the vial to cool, and again weigh accurately. If the mass loss is less than 26 mg and there is no evidence of a leak of the content, use the up-per layer of the mixture as the sample solution. Separately, put 60 to 100 mg of adipic acid, 2.0 mL of the internal stand-ard solution and 2.0 mL of hydroiodic acid in a reaction vial, immediately stopper the vial tightly, and weigh accu-rately. Add 45 mL of iodomethane for assay through the sep-tum using a micro-syringe, weigh accurately, shake, and use the upper layer of the mixture as the standard solution. Per-form the test with 1 to 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iodomethane to that of the internal standard. Content (z) of methoxy group (CH3O) ＝ MS/M × QT/QS × 21.86 MS: Amount (mg) of iodomethane for assay taken M: Amount (mg) of Methylcellulose taken, calculated on the dried basis ‌Internal standard solution—A solution of n-octane in o-xylene (3 in 100). Operating conditions— Detector: A thermal conductivity detector or hydrogen ‰ame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 30 m in length, coated with dimethylpolysiloxane for gas chromatography in 3 mm thickness. Use a guard column, if necessary. Column temperature: Maintain the temperature at 509C for 3 minutes, raise to 1009C at a rate of 109C per minute, then raise to 2509C at a rate of 359C per minute, and main-tain at 2509C for 8 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2809C. Carrier gas: Helium. Flow rate: 4.3 mL per minute (the retention time of the in-ternal standard is about 10 minutes). Split ratio: 1:40. System suitability— System performance: When the procedure is run with 1 – 2 mL of the standard solution under the above operating con-ditions, iodomethane and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 1 – 2 mL of the standard solution under the above oper-ating conditions, the relative standard deviation of the ratio of the peak area of iodomethane to that of the internal standard is not more than 2.0z.
Methyldopa Hydrate contains not less than 98.
Methyldopa Hydrate occurs as a white to pale grayish white crystalline powder. It is slightly soluble in water, in methanol and in acetic acid (100), very slightly soluble in ethanol (95), and practi-cally insoluble in diethyl ether. It dissolves in dilute hydrochloric acid.
(1) To 10 mg of Methyldopa Hydrate add 3 drops of ninhydrin TS, and heat in a water bath for 3 minutes: a purple color develops. Determine the absorption spectrum of a solution of Methyldopa Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 25,000) as directed under Ultraviolet-visible Spectropho-tometry <2.44>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Methyldopa RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Methyldopa Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Methyldopa RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －25 – －289(1 g calculated on the anhydrous basis, aluminum (III) chloride TS, 20 mL, 100 mm).
(1) Acidity—Shake 1.0 g of Methyldopa Hydrate with 100 mL of freshly boiled and cooled water, and add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 2 drops of methyl red TS: a yellow color develops. Chloride <1.03>—Perform the test with 0.5 g of Methyldopa Hydrate. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.028z). Heavy metals <1.07>—Proceed with 2.0 g of Methyl- dopa Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Methyldopa Hydrate in 5 mL of dilute hydrochloric acid, and perform the test (not more than 2 ppm). 3-O-Methylmethyldopa—Dissolve 0.10 g of Methyl-dopa Hydrate in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 5 mg of 3-O-methylmethyldopa for thin-layer chromatography in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (13:5:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-nitroaniline-sodium nitrite TS on the plate, and air-dry the plate, then spray evenly a solution of sodium carbonate decahydrate (1 in 4) on the plate: the spot obtained from the sample solution corresponding to that from the standard solution is not more intense than the spot from the standard solution. Water <2.48> 10.0 – 13.0z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Methyldopa Hy-drate, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green ‌(indicator: 2 to 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.12 mg of C10H13NO4
メチルエルゴメトリンマレイン酸塩
Methylergometrine Maleate, when dried, contains not less than 95.
Methylergometrine Maleate occurs as a white to pale yellow crystalline powder. It is odorless. It is slightly soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether. It gradually changes to yellow by light. Melting point: about 1909C (with decomposition).
(1) A solution of Methylergometrine Male-ate (1 in 200) shows a blue fluorescence. The colored solution obtained in the Assay develops a deep blue in color. Determine the absorption spectrum of the colored solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Methylergometrine Maleate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. To 5 mL of a solution of Methylergometrine Maleate (1 in 500) add 1 drop of potassium permanganate TS: the red color of the test solution fades immediately. D Optical rotation <2.49> [a]20: ＋44 – ＋509(after drying, 0.1 g, water, 20 mL, 100 mm).
Related substances—Conduct this procedure with-out exposure to light, using light-resistant vessels. Dissolve 8 mg of Methylergometrine Maleate in 2 mL of a mixture of ethanol (95) and ammonia solution (28) (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add a mixture of ethanol (95) and ammonia solution (28) (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test immediately with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography, and immediately de-velop the plate with a mixture of chloroform, methanol and water (75:25:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z (0.2 g, in vacu-um, phosphorus (V) oxide, 4 hours).
Weigh accurately about 10 mg each of Methyler- gometrine Maleate and Methylergometrine Maleate RS, pre-viously dried, add water to make exactly 250 mL, and use these solutions as the sample solution and the standard solu-tion. Pipet 2 mL each of the sample solution and the stand-ard solution separately into brown glassstoppered test tubes, add exactly 4 mL each of 4-dimethylaminobenzaldehyde-iron (III) chloride TS while ice cooling, warm for 10 minutes at 459C, and allow to stand for 20 minutes at room tempera-ture. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solu-tion, prepared with 2.0 mL of water in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 545 nm, respectively. Amount (mg) of methylergometrine maleate (C20H25N3O2.C4H4O4) ＝ MS × AT/AS MS: Amount (mg) of Methylergometrine Maleate RS taken
メチルプレドニゾロンコハク酸エステル
Methylprednisolone Succinate, when dried, contains not less than 97.
Methylprednisolone Succinate occurs as a white, crystals or crystalline powder. It is soluble in methanol, sparingly soluble in ethanol (95), and practically insoluble in water. Melting point: about 2359C (with decomposition). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Methylprednisolone Succinate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Methylpred-nisolone Succinate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Methylprednisolone Succinate, previously dried, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Methylprednisolone Succinate RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. In case when some differences are found between the spectra, repeat the test with residues obtained by dissolving these sub-stances in ethanol (95), evaporating to dryness, and drying. D Optical rotation <2.49> [a]25: ＋99 – ＋1039(after drying, g, ethanol (95), 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Methylprednisolone Succinate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Methylprednisolone Succinate according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 15 mg of Methylpred-nisolone Succinate in 5 mL of methanol, add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and aceto-nitrile (1:1) to make 50 mL, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add the mix-ture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (1:1) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by ‌the automatic integration method: the area of the peaks other than the peak of methylprednisolone succinate ob-tained from sample solution is not larger than 1/2 times the peak area of methylprednisolone succinate from the stand-ard solution, and the total area of the peaks other than the peak of methylprednisolone succinate is not larger than the peak area of methylprednisolone succinate from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of methylprednisolone succinate. System suitability— System performance: Proceed as directed in the System suitability in the Assay. Test for required detectability: Pipet 1 mL of the standard solution, and add the mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (1:1) to make ex-actly 10 mL. Confirm that the peak area of methylpredniso-lone succinate obtained with 5 mL of this solution is equiva-lent to 7 to 13z of that with 5 mL of the standard solution. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of methylprednisolone succinate is not more than 2.5z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). prednisolone succinate is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, methylprednisolone succinate and the internal stand-ard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of methylprednisolone succinate to that of the in-ternal standard is not more than 1.0z.
Methylprednisolone, when dried, contains not less than 96.
Methylprednisolone occurs as a white crystal-line powder. It is odorless. ‌It is sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. Melting point: 232 – 2409C (with decomposition).
(1) Add 2 mL of sulfuric acid to 2 mg of Methylprednisolone: a deep red color develops with no fluorescence. Then add 10 mL of water to this solution: the color fades, and a gray, flocculent precipitate is produced. Dissolve 10 mg of Methylprednisolone in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: a red precipi-tate is produced. Determine the absorption spectrum of a solution of Methylprednisolone in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]25: ＋93 – ＋1039(after drying, g, ethanol (99.5), 10 mL, 100 mm).
Related substances—Dissolve 50 mg of Methylpred-nisolone in 5 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of chloroform and methanol (9:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, diethyl ether, methanol and water (385:75:40:6) to a distance of about 12 cm, and air-dry the plate. Then heat the plate at 1059C for 10 minutes, cool, and spray evenly alkaline blue tetrazolium TS on the plate: the spots other than the princi-pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.2 g).
Weigh accurately about 10 mg of Methylpredniso-lone, previously dried, and dissolve in methanol to make ex-actly 100 mL. To exactly 5 mL of this solution add methanol to make exactly 50 mL, and determine the absorbance A at the wavelength of maximum absorption at about 243 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of methylprednisolone (C22H30O5) ＝ A/400 × 10,000
Methyltestosterone, when dried, contains not less than 98.
Methyltestosterone occurs as white to pale yel-low, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Methyltestosterone in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec-trum or the spectrum of a solution of Methyltestosterone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Methyltestosterone, previously dried, as directed in the po-tassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of dried Methyltestosterone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋79 – ＋859(after drying, 0.1 g, ethanol (95), 10 mL, 100 mm).
Related substances—Dissolve 40 mg of Methyltesto-sterone in 2 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 10 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 20 mg each of Methyltesto-sterone and Methyltestosterone RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 10 hours, dissolve each in methanol to make exactly 200 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, add methanol to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of methyltestosterone to that of the internal standard. Amount (mg) of methyltestosterone (C20H30O2) ＝ MS × QT/QS MS: Amount (mg) of Methyltestosterone RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in methanol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 241 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile and water (11:9). Flow rate: Adjust so that the retention time of methyltestosterone is about 10 minutes. ‌System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and methyltestosterone are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of methyltestosterone to that of the internal standard is not more than 1.0z.
Meticrane, when dried, contains not less than 98.
Meticrane occurs as white, crystals or crystal-line powder. It is freely soluble in N,N-dimethylformamide, slightly soluble in acetonitrile and in methanol, very slightly soluble in ethanol (95), and practically insoluble in water. Melting point: about 2349C (with decomposition).
(1) Determine the absorption spectrum of a solution of Meticrane in methanol (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Meti-crane, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Ammonium <1.02>—Perform the test with 0.10 g of Meticrane. Prepare the control solution with 3.0 mL of Standard Ammonium Solution (not more than 0.03z). Heavy metals <1.07>—Proceed with 1.0 g of Meti- crane according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Meticrane according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Meticrane in 50 mL of acetonitrile. To 5 mL of this solution add the mo-bile phase to make 25 mL, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo- bile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area of both so-lutions by the automatic integration method: the total area of the peaks other than meticrane obtained from the sample solution is not larger than the peak area of meticrane from the standard solution. Operating conditions 1— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (17:3). Flow rate: Adjust so that the retention time of meticrane is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of meticrane, beginning after the solvent peak. System suitability 1— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of meticrane obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 10 mg each of Meticrane and caffeine in 100 mL of acetonitrile. To exactly 2 mL of this solution add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions 1, caffeine and meticrane are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions 1, the relative standard deviation of the peak area of meticrane is not more than 2.0z. Operating conditions 2— Detector, column, and column temperature: Proceed as directed in the operating conditions 1. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of meticrane is about 2 minutes. Time span of measurement: About 10 times as long as the retention time of meticrane, beginning after the solvent peak. System suitability 2— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of meticrane obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 20 mg each of Meticrane and methyl parahydroxybenzoate in 100 mL of acetonitrile. To exactly 2 mL of this solution add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions 2, meticrane and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions 2, the relative standard deviation of the peak area of meticrane is not more than 2.0z. ‌Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Meticrane, previ-ously dried, dissolve in 50 mL of N,N-dimethylformamide, add 5 mL of water, and titrate <2.50> with 0.1 mol/L potas-sium hydroxide-ethanol VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 27.54 mg of C10H13NO4S2
Metildigoxin occurs as a white to light yellow-white crystalline powder. It is freely soluble in N,N-dimethylformamide, in pyridine and in acetic acid (100), soluble in chloroform, sparingly soluble in methanol, slightly soluble in ethanol (95) and in acetone, and very slightly soluble in water. It shows crystal polymorphism.
(1) Dissolve 2 mg of Metildigoxin in 2 mL of acetic acid (100), shake well with 1 drop of iron (III) chlo-ride TS, and add gently 2 mL of sulfuric acid to divide into two layers: a brown color develops at the interface, and a deep blue color gradually develops in the acetic acid layer. Dissolve 2 mg of Metildigoxin in 2 mL of 1,3-dinitro-benzene TS, add 2 mL of a solution of tetramethylammo-nium hydroxide in ethanol (95) (1 in 200), and shake: a pur-ple color gradually develops, and changes to blue-purple. Determine the absorption spectrum of a solution of Metildigoxin in methanol (1 in 50,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Metildigoxin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Metil-digoxin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Metildigoxin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Metildigoxin and Metildigoxin RS in acetone, respectively, then evaporate the acetone to dryness, and repeat the test on the residues. 546.1 Optical rotation <2.49> [a]20 : ＋22.0 – ＋25.59(1 g calcu-lated on the anhydrous basis, pyridine, 10 mL, 100 mm).
(1) Arsenic <1.11>—Prepare the test solution with 0.5 g of Metildigoxin according to Method 3, and perform the test (not more than 4 ppm). (2) Related substances—Dissolve 10 mg of Metildigoxin in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-butanone and chloroform (3:1) to a dis-tance of about 15 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1109C for 10 minutes: the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Acetone Weigh accurately about 0.1 g of Metildigoxin, dis-solve in exactly 2 mL of the internal standard solution, add N,N-dimethylformamide to make 10 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 0.4 g of acetone in a 50-mL volumetric flask contain-ing about 10 mL of N,N-dimethylformamide, and add N,N-dimethylformamide to make 50 mL. Pipet 5 mL of this solu-tion, add exactly 20 mL of the internal standard solution, then add N,N-dimethylformamide to make 100 mL, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as di-rected under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of acetone to that of the internal standard: the amount of acetone is between 2.0z and 5.0z. Amount (z) of acetone ＝ MS/MT × QT/QS MS: Amount (g) of acetone taken MT: amount (g) of Metildigoxin taken Internal standard solution—A solution of t-butyl alcohol in N,N-dimethylformamide (1 in 2000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column about 2 mm in inside diameter and 1 to 2 m in length, packed with porous ethylvinylben-zene-divinylbenzene copolymer for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature between 1709C and 2309C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of acetone is about 2 minutes. Selection of column: Proceed with 1 mL of the standard solution under the above operating conditions, and calculate the resolution. Use a column giving elution of acetone and t-butyl alcohol in this order with the resolution between these peaks being not less than 2.0. ‌Water <2.48> Not more than 3.0z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g). ‌Assay Weigh accurately 0.1 g each of Metildigoxin and Metildigoxin RS (separately, determine the water <2.48> in the same manner as Metildigoxin), and dissolve each in methanol to make exactly 50 mL. Pipet 5 mL each of the so-lutions, add methanol to each to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Pipet 5 mL each of the sample solu-tion and standard solution, add 15 mL of 2,4,6-trinitro-phenol-ethanol TS and 2 mL of sodium hydroxide TS to each, shake well, add methanol to make exactly 25 mL, and allow to stand at 20 ± 0.59C for 20 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution prepared by mix-ing 15 mL of 2,4,6-trinitrophenol-ethanol TS and 2 mL of sodium hydroxide TS and adding methanol to make exactly 25 mL as the blank. Determine the maximum absorbances, AT and AS, of the subsequent solutions obtained from the sample solution and the standard solution, respectively, by measuring every 5 minutes, at 495 nm. Amount (mg) of metildigoxin (C42H66O14.1/2C3H6O) ＝ MS × AT/AS MS: Amount (mg) of Metildigoxin RS taken, calculated on the anhydrous basis
Metoclopramide, when dried, contains not less than 99.
Metoclopramide occurs as white, crystals or a crystalline powder, and is odorless. It is freely soluble in acetic acid (100), soluble in methanol and in chloroform, sparingly soluble in ethanol (95), in ace-tic anhydride and in acetone, very slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid.
(1) Dissolve 10 mg of Metoclopramide in 1 mL of dilute hydrochloric acid and 4 mL of water: the solu-tion responds to Qualitative Tests <1.09> for Primary Aro-matic Amines. Dissolve 10 mg of Metoclopramide in 5 mL of dilute hydrochloric acid and 20 mL of water, and to 5 mL of this solution add 1 mL of Dragendorff's TS: a reddish orange precipitate is produced. Dissolve 0.1 g of Metoclopramide in 1 mL of 1 mol/L hydrochloric acid TS, and dilute with water to make 100 mL. To 1 mL of the solution add water to make 100 mL, determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 1.0 g of Metoclopramide in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Metoclo- pramide as directed under Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of Metoclopramide in 5 mL of 1 mol/L hydrochloric acid TS, and use this solution as the sample solution. Perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Metoclo-pramide in 10 mL of methanol, and use this solution as the sample solution. Dilute 1 mL of the sample solution, exactly measured, with methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol and ammonia solution (28) (19:1) to a distance of about 10 cm. Dry the plate, first in air and then at 809C for 30 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 0.4 g of Metoclopramide, previously dried and accurately weighed, in 50 mL of acetic acid (100), add 5 mL of acetic anhydride, and warm for 5 minutes. Allow to cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform the blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.98 mg of C14H22ClN3O2
Metoprolol Tartrate, when dried, contains not less than 99.
Metoprolol Tartrate occurs as a white crystal-line powder. It is very soluble in water, and freely soluble in methanol, in ethanol (95) and in acetic acid (100). D Optical rotation [a]20: ＋7.0 – ＋10.09(after drying, 1 g, water, 50 mL, 100 mm). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Metoprolol Tartrate in ethanol (95) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Metoprolol Tartrate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spec-tra, recrystallize Metoprolol Tartrate from a solution in ace-tone (23 in 1000), filter and dry the crystals, and perform the test with the crystals. A solution of Metoprolol Tartrate (1 in 5) responds to Qualitative Tests <1.09> (1) for tartrate. pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Metoprolol Tartrate in 10 mL of water is between 6.0 and 7.0.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Metoprolol Tartrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Metoprolol Tartrate in 5 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. After saturating the plate with the atmosphere by allowing to stand in a developing vessel, which contains the developing solvent and a glass vessel containing ammonia water (28), develop with the developing solvent, a mixture of ethyl ace-tate and methanol (4:1), to a distance of about 12 cm, and air-dry the plate. Allow to stand the plate in an iodine vapors until the spot with the standard solution appears obviously: ‌the spot other than the principal spot and other than the spot on the original point obtained from the sample solution is not more than three spots, and they are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately not less than 20 Metoclopramide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 75 mg of metoclopramide (C14H22ClN3O2), add 300 mL of 0.1 mol/L hydrochloric acid TS, shake for 1 hour, and add 0.1 mol/L hydrochloric acid TS to make exactly 500 mL. Centrifuge for 10 minutes, pipet 4 mL of the supernatant liquid, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of metoclopramide for assay, previously dried at 1059C for 3 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 500 mL. Pipet 4 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 308 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of metoclopramide (C14H22ClN3O2) ＝ MS × AT/AS MS: Amount (mg) of metoclopramide for assay taken
Metronidazole, when dried, contains not less than 99.
Metronidazole occurs as white to pale yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in ethanol (99.5) and in acetone, and slightly soluble in water. It dissolves in dilute hydrochloric acid. It is colored to yellow-brown by light.
(1) Determine the absorption spectrum of a solution of Metronidazole in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Metronidazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Metronidazole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) 2-Methyl-5-nitroimidazol—Dissolve 0.10 g of Metro-nidazole in acetone to make exactly 10 mL, and use this solu-tion as the sample solution. Separately, dissolve 20 mg of 2-methyl-5-nitroimidazole for thin-layer chromatography in acetone to make exactly 20 mL, then pipet 5 mL of this solu-tion, add acetone to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Immediately develop the plate with a mixture of acetone, water and ethyl acetate (8:1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution corresponding to the spot from the standard solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Metronidazole, pre-viously dried, and dissolve in 30 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 0.5 mL of p-naphtholbenzein TS) until the color of the solu-tion changes from orange-yellow to green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 17.12 mg of C6H9N3O3
Metyrapone, when dried, contains not less than ＝ MS × AT/AS × V?/V × 1/C × 45 98.
Metyrapone occurs as a white to pale yellow crystalline powder. It has a characteristic odor and a bitter taste. It is very soluble in methanol, in ethanol (95), in acetic an-hydride, in chloroform, in diethyl ether and in nitrobenzene, and sparingly soluble in water. It dissolves in 0.5 mol/L sulfuric acid TS.
(1) Mix 5 mg of Metyrapone with 10 mg of 1-chloro-2,4-dinitrobenzene, melt by gently heating for 5 to 6 seconds, cool, and add 4 mL of potassium hydroxide-ethanol TS: a dark red color develops. (2) Determine the absorption spectrum of a solution of Metyrapone in 0.5 mol/L sulfuric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity and color of solution—Dissolve 0.5 g of Metyrapone in 5 mL of methanol: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 2.0 g of Metyra- pone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ‌Arsenic <1.11>—Prepare the test solution with 1.0 g of Metyrapone, according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Metyrapone in 5 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and methanol (15:1) to a distance of about 10 cm, and air-dry the plate for about 15 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately the mass of not less than 20 Metronidazole Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 0.25 g of metronida-zole (C6H9N3O3), add 25 mL of a mixture of water and meth-anol (1:1), shake vigorously for 10 minutes, and add the mix-ture of water and methanol (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add a mixture of water and methanol (4:1) to make exactly 100 mL. Filter this solution through a membrane filter with pore size of 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of metronidazole for assay, previously dried in vacu- Description Metyrapone occurs as a white to pale yellow crystalline powder. It has a characteristic odor and a bitter taste. It is very soluble in methanol, in ethanol (95), in acetic an-hydride, in chloroform, in diethyl ether and in nitrobenzene, and sparingly soluble in water. It dissolves in 0.5 mol/L sulfuric acid TS. Identification (1) Mix 5 mg of Metyrapone with 10 mg of 1-chloro-2,4-dinitrobenzene, melt by gently heating for 5 to 6 seconds, cool, and add 4 mL of potassium hydroxide-ethanol TS: a dark red color develops. (2) Determine the absorption spectrum of a solution of Metyrapone in 0.5 mol/L sulfuric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Melting point <2.60> 50 – 549C Purity (1) Clarity and color of solution—Dissolve 0.5 g of Metyrapone in 5 mL of methanol: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 2.0 g of Metyra- pone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ‌Arsenic <1.11>—Prepare the test solution with 1.0 g of Metyrapone, according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Metyrapone in 5 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and methanol (15:1) to a distance of about 10 cm, and air-dry the plate for about 15 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.2 g of Metyrapone, previ-ously dried, dissolve in 10 mL of nitrobenzene and 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.31 mg of C14H14N2O
Mexiletine Hydrochloride, when dried, contains not less than 98.
Mexiletine Hydrochloride occurs as a white powder. It is freely soluble in water and in ethanol (95), and slightly soluble in acetonitrile. It dissolves in 0.01 mol/L hydrochloric acid TS. A solution of Mexiletine Hydrochloride (1 in 20) shows no optical rotation. Mexiletine Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Mexiletine Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mexiletine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mexi-letine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Mexiletine Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize Mexiletine Hydro-chloride from ethanol (95), filter, dry the crystals, and per-form the test with the crystals. A solution of Mexiletine Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Mexiletine Hydrochloride in 10 mL of water: the pH of this solution is between 3.8 and 5.8.
(1) Clarity and color of solution—Dissolve 1.0 g of Mexiletine Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy Metals <1.07>—Proceed with 2.0 g of Mexile- tine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 20 mg of Mexiletine Hydrochloride in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: each peak area other than mexiletine obtained from the sam-ple solution is not larger than the peak area of mexiletine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in the operating conditions in the Assay. Detection sensitivity: Adjust so that the peak height of mexiletine obtained from 20 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About 3 times as long as the retention time of mexiletine, beginning after the solvent peak. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Mexiletine Hydrochloride and Mexiletine Hydrochloride RS, each pre-viously dried, and dissolve each in the mobile phase to make ‌exactly 20 mL. Pipet 5 mL each of these solutions, add ex-actly 5 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of mexiletine to that of the internal standard, respectively. Identification (1) Determine the absorption spectrum of a solution of Miconazole in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Miconazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry Amount (mg) of mexiletine hydrochloride (C ＝ MS × QT/QS 11H 17NO.HCl) <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. MS: Amount (mg) of Mexiletine Hydrochloride RS taken Internal standard solution—A solution of phenetylamine hy-drochloride in the mobile phase (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (about 7 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 2.5 g of sodium lauryl sulfate and 3 g of sodium dihydrogen phosphate dihydrate in 600 mL of water, and add 420 mL of acetonitrile. Flow rate: Adjust so that the retention time of mexiletine is about 6 minutes. Selection of column: Proceed with 20 mL of the standard solution under the above conditions, and calculate the reso-lution. Use a column giving elution of the internal standard and mexiletine in this order with the resolution between these peaks being not less than 9.
Miconazole Nitrate occurs as a white crystal-line powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in methanol, slightly soluble in ethanol (95), in ace-tone and in acetic acid (100), and very slightly soluble in water and in diethyl ether. Melting point: about 1809C (with decomposition).
(1) To 2 mL of a solution of Miconazole Nitrate in methanol (1 in 100) add 2 mL of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Miconazole Nitrate in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Perform the test with a solution of Miconazole Nitrate in methanol (1 in 100) as directed under Flame Coloration Test <1.04> (2): a green color appears. A solution of Miconazole Nitrate in methanol (1 in 100) responds to Qualitative Tests <1.09> for nitrate.
(1) Clarity and color of solution—Dissolve 1.0 g of Miconazole Nitrate in 100 mL of methanol: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.10 g of Miconazole Nitrate in 6 mL of dilute nitric acid and N,N-dimethylfor-mamide to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL (not more than 0.09z). Heavy metals <1.07>—Proceed with 1.0 g of Micona- zole Nitrate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Miconazole Nitrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Miconazole Nitrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 20 mL, pipet 1 mL of this solu-tion, add methanol to make exactly 20 mL, and use this solu-tion as the standard solution. Perform the test with these so-
Weigh accurately about 0.35 g of Miconazole Nitrate, previously dried, dissolve in 50 mL of acetic acid (100) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 47.91 mg of C H Cl N O.HNO 18 14 4 2 3 Description Miconazole Nitrate occurs as a white crystal-line powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in methanol, slightly soluble in ethanol (95), in ace-tone and in acetic acid (100), and very slightly soluble in water and in diethyl ether. Melting point: about 1809C (with decomposition). Identification (1) To 2 mL of a solution of Miconazole Nitrate in methanol (1 in 100) add 2 mL of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Miconazole Nitrate in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Perform the test with a solution of Miconazole Nitrate in methanol (1 in 100) as directed under Flame Coloration Test <1.04> (2): a green color appears. A solution of Miconazole Nitrate in methanol (1 in 100) responds to Qualitative Tests <1.09> for nitrate. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Miconazole Nitrate in 100 mL of methanol: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.10 g of Miconazole Nitrate in 6 mL of dilute nitric acid and N,N-dimethylfor-mamide to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL (not more than 0.09z). Heavy metals <1.07>—Proceed with 1.0 g of Micona- zole Nitrate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Miconazole Nitrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Miconazole Nitrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 20 mL, pipet 1 mL of this solu-tion, add methanol to make exactly 20 mL, and use this solu-tion as the standard solution. Perform the test with these so-
Miconazole, when dried, contains not less than 98.
Miconazole occurs as a white to pale yellow-white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tic acid (100), soluble in diethyl ether, and practically insolu-ble in water. A solution of Miconazole in methanol (1 in 20) shows no optical rotation.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Miconazole according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Miconazole according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Miconazole in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 20 mL. Pipet 1 mL of this solution, add metha-nol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, chloroform, methanol and ammonia solution (28) (60:30:10:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 20 minutes: the spots other than the prin-cipal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 60z, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Miconazole, previ-ously dried, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of p-naphtholbenzein TS) until the color of the solution changes from light yellow-brown to light yellow-green. Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.61 mg of C18H14Cl4N2O
Microcrystalline Cellulose occurs as a white crystalline powder having ‰uidity. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It swells with sodium hydroxide TS on heating.◆ Identiˆcation (1) Dissolve 20 g of zinc chloride and 6.5 g of potassium iodide in 10.5 mL of water, add 0.5 g of iodine, and shake for 15 minutes. Place about 10 mg of Microcrystalline Cellulose on a watch glass, and disperse in 2 mL of this solution: the substance develops a blue-violet color. Determine the infrared absorption spectrum of Microcrystalline Cellulose as directed in the ATR method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the spectrum of Microcrystalline Cellulose for Identiˆcation RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If there are absorp-tions between 800 cm－1 and 825 cm－1, and between 950 cm－1 and 1000 cm－1, disregard the absorptions. Transfer about 1.3 g of Microcrystalline Cellulose, accurately weighed, to a 125-mL conical ‰ask, and add exactly 25 mL each of water and 1 mol/L cupriethylenedia-mine TS. Immediately purge the solution with nitrogen, insert the stopper, and shake on a suitable mechanical shaker to dissolve. Perform the test with a suitable amount of this solution, taken exactly, according to Method 1 under Vis-cosity Determination <2.53> using a capillary viscometer hav-ing the viscosity constant (K) of approximately 0.03, at 25 ± 0.19C, and determine the kinematic viscosity, n. Separately, perform the test with a mixture of exactly 25 mL each of water and 1 mol/L cupriethylenediamine TS in the same manner as above, using a capillary viscometer having K of approximately 0.01, and determine the kinematic viscosity, no. Calculate the relative viscosity, hrel, of Microcrystalline Cellulose by the following formula: hrel ＝ n/n0 Obtain the product, [h]C, of intrinsic viscosity [h](mL/g) and concentration C (g/100 mL) from the value hrel of the table. When calculate the degree of polymerization, P, by the following formula, P is not more than 350 ◇and within the labeled range.◇ P ＝ 95〔h〕C/MT MT: Amount (g) of the Microcrystalline Cellulose taken, calculated on the dried basis pH <2.54> Shake 5.0 g of Microcrystalline Cellulose with 40 mL of water for 20 minutes, and centrifuge: the pH of the supernatant liquid is between 5.0 and 7.5.
Micronomicin Sulfate is the sulfate of an amino-glycoside substance having antibacterial activity pro-duced by the growth of Micromonospora sagamiensis. It contains not less than 590 mg (potency) and not more than 660 mg (potency) per mg, calculated on the anhydrous basis.
Micronomicin Sulfate occurs as a white to light yellow-white powder. It is very soluble in water, sparingly soluble in ethylene glycol, and practically insoluble in methanol and in ethanol (99.5). It is hygroscopic. ‌Identification (1) Dissolve 50 mg each of Micronomicin Sulfate and Micronomicin Sulfate RS in 10 mL of water, and use these solutions as the sample solution and the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5), 1-butanol and ammonia solution (28) (10:8:7) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in a mixture of acetone and pyridine (25:1) (1 in 500), and heat the plate at 1009C for 10 minutes: the spots obtained from the sample solution and the standard solution are red-purple to red-brown and their Rf values are the same. (2) To 5 mL of a solution of Micronomicin Sulfate (1 in 100) add 1 mL of barium chloride TS: a white precipitate is formed, and it does not dissolve by addition of dilute nitric acid. D Optical rotation <2.49> [a]20: ＋110 – ＋1309(0.25 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Micronomicin Sulfate in 10 mL of water is between 3.5 and 5.5.
(1) Clarity and color of solution—Dissolve 1.5 g of Micronomicin Sulfate in 10 mL of water: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 1.0 g of Micronomicin Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.40 g of Micronomi-cin Sulfate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (99.5), 1-butanol and ammo-nia solution (28) (10:8:7) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in a mixture of acetone and pyridine (25:1) (1 in 500), and heat the plate at 1009C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, back titration). Use a mixture of methanol for water determination and ethylene glycol for water determi-nation (1:1) instead of methanol for water determination.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Micronomicin Sulfate RS, equivalent to about 20 mg (po-tency), dissolve in 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make exactly 20 mL, and use this so-lution as the standard stock solution. Keep the standard stock solution at 5 – 159C, and use within 30 days. Take ex-actly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 2 mg (potency) and 0.5 mg (potency), and use these solutions as the high concentration standard solution and the low concentra-tion standard solution, respectively. Sample solutions—Weigh accurately an amount of Micronomicin Sulfate, equivalent to about 20 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution for an-tibiotics (pH 8.0) to make exactly 20 mL. Take exactly a suit-able amount of this solution, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 2 mg (potency) and 0.5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respec-tively.
Midecamycin Acetate is a derivative of midecamy-cin. It contains not less than 950 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the dried basis.
Midecamycin Acetate occurs as white, crystals or crystalline powder. It is sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Midecamycin Acetate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Midecamycin Acetate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Midecamycin Acetate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or spectrum of dried Midecamycin Acetate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
Heavy metals <1.07>—Proceed with 1.0 g of Midecamycin Acetate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 2.0z (1 g, in vacuum not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics ‌<4.02> according to the following conditions. Test organism—Kocuria rhizophila ATCC 9341 Culture medium—Use the medium i in 3) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Midecamycin Acetate RS, previously dried, equivalent to about 25 mg (potency), and dissolve in methanol to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 5 – 159C and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phos-phate buffer solution (pH 4.5) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Midecamycin Acetate, previously dried, equivalent to about 25 mg (potency), and dissolve in methanol to make exactly 50 mL. Take exactly a suitable amount of the solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make solu-tions so that each mL contains 20 mg (potency) and 5 mg (po-tency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Midecamycin is a macrolide substance having anti-bacterial activity produced by the growth of Strep-tomyces mycarofaciens. It contains not less than 950 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the dried basis.
Midecamycin occurs as a white crystalline powder. It is very soluble in methanol, freely soluble in ethanol (95), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Midecamycin in methanol (1 in 50,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Midecamycin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Midecamycin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Midecamycin RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers. ‌
Heavy metals <1.07>—Proceed with 1.0 g of Midecamycin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Loss on drying <2.41> Not more than 2.0z (1.0 g, in vacu-um not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Midecamycin RS, previously dried, equivalent to about 20 mg (potency), dissolve in 10 mL of methanol, add water to make exactly 50 mL, and use this solution as the standard Midecamycin Acetate
Miglitol is a white to pale yellowish white pow-der. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) Determine the infrared absorption spec-trum of Miglitol, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Miglitol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Dissolve 10 mg each of Miglitol and Miglitol RS in 1 mL of water, and use these solutions as the sample solution and the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, ethyl acetate and diluted ammonia solution (28) (9 in 10) (2:2:1) to a dis-tance of about 17 cm, and dry the plate at 1059C. Allow the plate to stand in iodine vapor: the principal spot obtained from the sample solution and the spot from the standard so-lution show a brown color and the same Rf value. D Optical rotation <2.49> [a]20: － 7.3 – － 8.39(1.2 g calcu-lated on the dried basis, water, 50 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 2.5 g of Miglitol in 50 mL of water, and use this solution as the test solution. Determine the turbidity of the test solution as directed under Turbidity Measurement <2.61>: it exhibits no more turbidity than Reference suspension II, and has no more color than the following control solution. Control solution: To a mix of 0.3 mL of Cobalt (II) Chlo-ride CS and 1.2 mL of Iron (III) Chloride CS add 38.5 mL of diluted hydrochloric acid (1 in 100). Heavy metals—Dissolve 2.5 g of Miglitol in 25 mL of water, and use this solution as the sample solution. Sepa-rately, to 10 mL of a solution obtained by diluting Standard Lead Stock Solution to 50-fold with water before use, add 2 mL of the sample solution, and use this solution as the con-trol solution. To 12 mL of the sample solution and the con-trol solution add 2 mL of hydrochloric acid-ammonium ace-tate buffer solution (pH 3.5) and 1.2 mL of thioacetamide TS, mix, allow to stand for 2 minutes, and observe vertically or horizontally both Nessler tubes against a white back-ground: the color obtained with the sample solution is not more intense than that with the control solution (not more than 20 ppm). Related substances—Dissoble 0.19 g of Miglitol in 50 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of the peaks having the relative retention time of about 0.9 and about 1.5 to miglitol is not more than 0.2z, and the amount of the peaks other than miglitol and the peaks mentioned above is not more than 0.1z. The total amount of the peaks other than miglitol is not more than 0.5z. For the area of the peak, having the relative retention time of about 1.5, multiply the correction factor 4.1. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of miglitol, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of miglitol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of miglitol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia- tion of the peak area of miglitol is not more than 5.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, 609C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Miglitol and Miglitol RS (separately determine the loss on drying <2.41> under the same conditions as Miglitol), dissolve each in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 20 mL of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of miglitol in each solution. under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, ethyl acetate and diluted ammo-nia solution (28) (9 in 10) (2:2:1) to a distance of about 8 cm, and air-dry the plate. Allow the plate to stand in iodine vapors: the principal spot obtained from the sample solution and the spot from the standard solution show a brown color and the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Miglitol Tablets add 20 mL of a mixture of acetonitrile for liquid chromatography and water (4:1), soni-cate, and add a mixture of acetonitrile for liquid chromatog- ‌Amount (mg) of miglitol (C8H17NO5) ＝ M S × AT/AS raphy and water (4:1) to make exactly V mL so that each mL contains about 1 mg of miglitol (C8H17NO5). Centrifuge this MS: Amount (mg) of Miglitol RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with pentaethylenehexaami-nated polyvinyl alcohol polymer beads for liquid chromatog-raphy (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 0.6 g of potassium dihydrogen phosphate and 0.28 g of anhydrous disodium hydrogen phosphate in water to make 1000 mL. To 300 mL of this solution add 900 mL of acetonitrile for liquid chromatogra-phy. Flow rate: Adjust so that the retention time of miglitol is about 11 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of miglitol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviation of the peak area of miglitol is not more than 1.0z.
Migrenin is composed of 90 parts of antipyrine, 9 parts of caffeine, and 1 part of citric acid in mass. tive Tests <1.09> for citrate. Melting point <2.60> 104 – 1109C Purity (1) Clarity and color of solution—Dissolve 1.0 g of Migrenin in 40 mL of water: the solution is clear and col-orless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Migrenin according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay (1) Antipyrine—Weigh accurately about 0.25 g of Migrenin, previously dried in an iodine flask, dissolve in 25 mL of sodium acetate TS, add exactly 30 mL of 0.05 mol/L iodine VS, and allow to stand for 20 minutes with occasional shaking. Add 15 mL of chloroform to dissolve the precipi-tate so obtained, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 9.411 mg of C11H12N2O (2) Caffeine—To about 1 g of Migrenin, previously dried and accurately weighed, add exactly 5 mL of the inter-nal standard solution, dissolve in chloroform to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 90 mg of Caffeine RS, previously dried at 809C for 4 hours, add exactly 5 mL of the internal standard solution, dissolve in chloroform to make 10 mL, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions, and calculate the ratios, QT and QS, of the peak area of caffeine to that of the internal standard. Migrenin, when dried, contains not less than 87.
Migrenin occurs as a white, powder or crystal-line powder. It is odorless and has a bitter taste. It is very soluble in water, freely soluble in ethanol (95) MS: Amount (mg) of Caffeine RS taken Internal standard solution—A solution of ethenzamide in chloroform (1 in 50). Operating conditions— Detector: A hydrogen flame-ionization detector. ‌Column: A glass column 2.6 mm in inside diameter and 210 cm in length, packed with siliceous earth for gas chroma-tography (180 to 250 mm in particle diameter) coated with 50z phenyl-methyl silicon polymer for gas chromatography at the ratio of 15z. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethen-zamide is about 4 minutes. System suitability— System performance: Dissolve 0.9 g of antipyrine and 0.09 g of caffeine in 10 mL of chloroform. When the proce-dure is run with 1 mL of this solution under the above operat-ing conditions, caffeine and antipyrine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of caffeine to that of the internal standard is not more than 1.0z.
(1) Clarity and color of solution—Dissolve 1.0 g of Migrenin in 40 mL of water: the solution is clear and col-orless to pale yellow. (2) Heavy metals <1.07>—Proceed with 1.0 g of Migrenin according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Antipyrine—Weigh accurately about 0.25 g of Migrenin, previously dried in an iodine flask, dissolve in 25 mL of sodium acetate TS, add exactly 30 mL of 0.05 mol/L iodine VS, and allow to stand for 20 minutes with occasional shaking. Add 15 mL of chloroform to dissolve the precipi-tate so obtained, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 9.411 mg of C11H12N2O (2) Caffeine—To about 1 g of Migrenin, previously dried and accurately weighed, add exactly 5 mL of the inter-nal standard solution, dissolve in chloroform to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 90 mg of Caffeine RS, previously dried at 809C for 4 hours, add exactly 5 mL of the internal standard solution, dissolve in chloroform to make 10 mL, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions, and calculate the ratios, QT and QS, of the peak area of caffeine to that of the internal standard. Migrenin, when dried, contains not less than 87.0z and not more than 93.0z of antipyrine (C11H12N2O: Amount (mg) of caffeine (C8H10N4O2) ＝ M S × QT/QS 188.23) and not less than 8.6z and not more than 9.5z of caffeine (C8H10N4O2: 194.19). Description Migrenin occurs as a white, powder or crystal-line powder. It is odorless and has a bitter taste. It is very soluble in water, freely soluble in ethanol (95) MS: Amount (mg) of Caffeine RS taken Internal standard solution—A solution of ethenzamide in chloroform (1 in 50). Operating conditions— Detector: A hydrogen flame-ionization detector. ‌Column: A glass column 2.6 mm in inside diameter and 210 cm in length, packed with siliceous earth for gas chroma-tography (180 to 250 mm in particle diameter) coated with 50z phenyl-methyl silicon polymer for gas chromatography at the ratio of 15z. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethen-zamide is about 4 minutes. System suitability— System performance: Dissolve 0.9 g of antipyrine and 0.09 g of caffeine in 10 mL of chloroform. When the proce-dure is run with 1 mL of this solution under the above operat-ing conditions, caffeine and antipyrine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of caffeine to that of the internal standard is not more than 1.0z.
Minocycline Hydrochloride is the hydrochloride of a derivative of tetracycline. It contains not less than 890 mg (potency) and not more than 950 mg (potency) per mg, calculated on the anhydrous basis.
Minocycline Hydrochloride occurs as a yellow crystalline powder. It is freely soluble in N,N-dimethylformamide, soluble in methanol, sparingly soluble in water, and slightly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Minocycline Hydrochloride in a solution of hy-drochloric acid in methanol (19 in 20,000) (1 in 62,500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Minocycline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Minocycline Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Minocycline Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Minocycline Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Minocycline Hydrochloride in 100 mL of water: the pH of the solution is between 3.5 and 4.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Minocycline Hydrochloride in 100 mL of water: the solu-tion is clear, and when the test is performed within 1 hour after preparation of this solution, the absorbance of the so-lution at 560 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.06. Heavy metals <1.07>—Proceed with 0.5 g of Mino- cycline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Stand-ard Lead Solution (not more than 50 ppm). Related substances—Dissolve 50 mg of Minocycline Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Perform the test, immedi-ately after the preparation of the sample solution, with 20 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method. Calculate the amount of each peak area by the area percen-tage method: the amount of epiminocycline is not more than 1.2z, the amount of each peak other than minocycline and epiminocycline is not more than 1.0z, and the total area of the peaks other than minocycline and epiminocycline is not more than 2.0z. Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay. Flow rate: Adjust so that the retention time of minocy-cline is about 12 minutes. The retention time of epiminocy- cline is about 10 minutes under this condition. Time span of measurement: About 2.5 times as long as the retention time of minocycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the sample solution add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suita-bility test. Pipet 5 mL of the solution for system suitability test, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of minocycline obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Not less than 4.3z and not more than 8.0z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.5z (1 g).
Weigh accurately an amount of Minocycline Hydro-chloride and Minocycline Hydrochloride RS, equivalent to about 50 mg (potency), dissolve each in the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of minocycline in each solution. to 50 mg (potency) of Minocycline Hydrochloride, add 60 mL of the mobile phase, shake vigorously, add the mobile phase to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi- Amount [ mg (potency)] of minocycline (C ＝ MS × AT/AS × 1000 23H27N3O7) tions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak of epiminocycline, having MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken Operating conditions— ‌Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust to pH 6.5 of a mixture of a solution of ammonium oxalate monohydrate (7 in 250), N,N-dimethylformamide and 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate TS (11:5:4) with tetrabutylam-monium hydroxide TS. Flow rate: Adjust so that the retention time of minocy-cline is about 12 minutes. System suitability— System performance: Dissolve 50 mg of Minocycline Hy-drochloride in 25 mL of water. Heat 5 mL of this solution on a water bath for 60 minutes, then add water to make 25 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, epiminocycline and minocy-cline are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of peak areas of minocycline is not more than 1.0z.
Mitomycin C is a substance having antitumor activity produced by the growth of Streptomyces caespitosus. It contains not less than 970 mg (potency) and not more than 1030 mg (potency) per mg, calculated on the dried basis.
Mitomycin C occurs as blue-purple, crystals or crystalline powder. It is freely soluble in N,N-dimethylacetamide, slightly soluble in water and in methanol, and very slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Mitomycin C (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mitomycin C RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Mitomycin C as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Mitomycin C RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
Related substances—Conduct this procedure rapidly after the sample and the standard solutions are prepared. Dissolve 50 mg of Mitomycin C in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: each area of the peak other than mitomycin C obtained from the sample solution is not larger than the peak area of mitomycin C from the standard solution, and the total area of the peaks other than mitomycin C from the sample solution is not larger than 3 times the peak area of mitomycin C from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: To 20 mL of 0.5 mol/L ammonium ace-tate TS add water to make 1000 mL. To 800 mL of this solu-tion add 200 mL of methanol. Mobile phase B: To 20 mL of 0.5 mol/L ammonium ace-tate TS add water to make 1000 mL. To this solution add 1000 mL of methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 10 – 30 100 → 0 0 → 100 30 – 45 Flow rate: About 1.0 mL per minute. Time span of measurement: About 2 times as long as the retention time of mitomycin C, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add methanol to make exactly 100 mL. Confirm that the peak area of mitomycin C obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 25 mg of Mitomycin C and 40 mg of 3-ethoxy-4-hydroxybenzaldehyde in 50 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, mitomycin C and 3-ethoxy-4-hydroxybenzaldehyde are eluted in this order with the resolution between these peaks being not less than 15. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitomycin C is not more than 3.0z. Loss on drying <2.41> Not more than 1.0z (0.1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours).
Weigh accurately an amount of Mitomycin C and Mitomycin C RS, equivalent to about 25 mg (potency), dissolve each in N,N-dimethylacetamide to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of mitomycin C in each solution. Amount [ mg (potency)] of mitomycin C (C15H18N4O5) ＝ MS × AT/AS × 1000 ‌MS: Amount [mg (potency)] of Mitomycin C RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 365 nm). ‌Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with phenylated silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 40 mL of 0.5 mol/L ammonium acetate TS add 5 mL of diluted acetic acid (100) (1 in 20) and water to make 1000 mL. To 600 mL of this solution add 200 mL of methanol. Flow rate: Adjust so that the retention time of mitomycin C is about 7 minutes. System suitability— System performance: Dissolve about 25 mg of Mitomycin C RS and about 0.375 g of 3-ethoxy-4-hydroxybenzaldehyde in 50 mL of N,N-dimethylacetamide. When the procedure is run with 10 mL of this solution under the above operating conditions, mitomycin C and 3-ethoxy-4-hydroxybenzalde-hyde are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitomycin C is not more than 1.0z.
Mizoribine occurs as a white to yellowish white crystalline powder. It is freely soluble in water, and practically insoluble in methanol and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Mizoribine (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mizoribine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mizoribine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Mizoribine RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －25 – －279(0.5 g calculated on the anhydrous basis, water, 25 mL, 100 mm). ‌Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Mizoribine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Mizoribine in the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the areas of the peaks other than mizoribine obtained from the sample solution are not larger than the peak area of mizoribine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of mizoribine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 5 mL. Confirm that the peak area of mizoribine obtained with 5 mL of this solution is equivalent to 14 to 26z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of mizoribine are not less than 10,000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mizoribine is not more than 2.0z. Water <2.48> Not more than 0.5z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Mizoribine, and dis-solve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Mizoribine RS (separately determine the water <2.48> using the same manner as Mizori-bine), dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and stand-ard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mizoribine in each solution. Amount (mg) of mizoribine (C9H13N3O6) ＝ MS × AT/AS × 10 MS: Amount (mg) of Mizoribine RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 279 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Diluted phosphoric acid (1 in 1500). Flow rate: Adjust so that the retention time of mizoribine is about 9 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of mizoribine are not less than 10,000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mizoribine is not more than 1.0z.
Containers—Tight containers. Storage—At a temperature between 2 and 89C.
Morphine Hydrochloride Hydrate contains not less than 98.
Morphine Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is freely soluble in formic acid, soluble in water, spar-ingly soluble in methanol, and slightly soluble in ethanol (95). It gradually becomes yellow-brown by light.
(1) Determine the absorption spectrum of a solution of Morphine Hydrochloride Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, determine the absorp-tion spectrum of a solution of Morphine Hydrochloride Hy-drate in dilute sodium hydroxide TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mor-phine Hydrochloride Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Morphine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －111 – －1169(0.5 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Morphine Hydrochloride Hydrate in 10 mL of water is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 0.40 g of Morphine Hydrochloride Hydrate in 10 mL of water: the solution is clear. When perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, the absorbance at 420 nm is not more than 0.12. Sulfate <1.14>—Dissolve 0.20 g of Morphine Hydro-chloride Hydrate in 5 mL of water, and add 2 to 3 drops of barium chloride TS: no turbidity is produced. Meconic acid—Dissolve 0.20 g of Morphine Hydro-chloride Hydrate in 5 mL of water, and add 5 mL of dilute hydrochloric acid and 2 drops of iron (III) chloride TS: no red color develops. ‌Related substances—Dissolve 0.20 g of Morphine Hydrochloride Hydrate in 10 mL of diluted methanol (4 in 5), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted methanol (4 in 5) to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 5 mL of the standard solution (1), add diluted methanol (4 in 5) to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of acetone, ethanol (99.5) and ammonia solution (28) (21:14:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot having an Rf value of about 0.17 obtained from the sample solution is not more in-tense than the spot from the standard solution (1), and the spots other than the principal spot, the spot mentioned above and the spot of the starting point from the sample so-lution are not more intense than the spot from the standard solution (2). Water <2.48> 13 – 15z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.5 g of Morphine Hydro-chloride Hydrate, dissolve in 3.0 mL of formic acid, add 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), mix, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.18 mg of C17H19NO3.HCl
Morphine Sulfate Hydrate contains not less than 98.
Morphine Sulfate Hydrate occurs as a white, crystals or crystalline powder. It is very soluble in formic acid, soluble in water, slightly soluble in methanol, and very slightly soluble in ethanol (99.5). It dissolves in dilute sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Morphine Sulfate Hydrate (1 in 10,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the absorption spectrum of a sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mor-phine Sulfate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. A solution of Morphine Sulfate Hydrate (1 in 25) re-sponds to Qualitative Tests <1.09> (1) and (3) for sulfate. D Optical rotation <2.49> [a]20: －107 – －1129(0.2 g calcu-lated on the anhydrous basis, water, 20 mL, 100 mm).
(1) Acidity—Dissolve 0.5 g of Morphine Sulfate Hydrate in 15 mL of water, add 2 drops of methyl red TS, and neutralize with 0.02 mol/L sodium hydroxide VS: the necessary volume of 0.02 mol/L sodium hydroxide VS is not more than 0.50 mL. Ammonium—Being specified separately when the drug is granted approval based on the Law. Chloride—Dissolve 0.10 g of Morphine Sulfate Hy-drate in 10 mL of water, add 1 mL of dilute nitric acid, then add 1 mL of silver nitrate TS: no turbidity is produced. Meconic acid—Dissolve 0.20 g of Morphine Sulfate Hydrate in 5 mL of water, add 5 mL of dilute hydrochloric acid and 2 drops of iron (III) chloride TS: no red color de-velops. Related substances—Dissolve 0.20 g of Morphine Sulfate Hydrate in 10 mL of diluted methanol (4 in 5), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted methanol (4 in 5) to make ex-actly 100 mL, and use this solution as the standard solution (1). Pipet 5 mL of the standard solution (1), add diluted methanol (4 in 5) to make exactly 10 mL, and use this solu-tion as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and the standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of acetone, ethanol (99.5) and ammonia solution (28) (21:14:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot at an Rf value of about 0.17 obtained from the sample solution is not more intense than the spot from the standard solution (1), and the spot other than the principle spot, the spot at an Rf value of about 0.17 and the spot at original point from the sample so-lution is not more intense than the spot from the standard solution (2). Water <2.48> 11.0 – 13.0z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.5 g of Morphine Sulfate Hydrate, dissolve in 3 mL of formic acid, add 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 33.44 mg of (C17H19NO3)2.H2SO4
Mosapride Citrate Hydrate occurs as a white to yellowish white crystalline powder. It is freely soluble in N,N-dimethylformamide and in ace-tic acid (100), sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. A solution of Mosapride Citrate Hydrate in N,N-dimethylformamide (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Mosapride Citrate Hydrate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Mosapride Citrate Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Mosapride Citrate Hydrate in N,N-dimethylformamide (1 in 10) responds to the Qualitative Tests <1.09> (1) for citrate.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Mosapride Citrate Hydrate in a platinum crucible according to Method 4, and perform the test. Prepare the control solu-tion with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Mosapride Citrate Hydrate in 50 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 50 mL. Pipet 1 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak having the relative retention time of about 0.47 to mosapride ob-tained from the sample solution is not larger than 3 times the peak area of mosapride from the standard solution, and the area of each peak other than mosapride and the peak men-tioned above from the sample solution is not larger than the peak area of mosapride from the standard solution. Further-more, the total area of the peaks other than mosapride from the sample solution is not larger than 5 times the peak area 0 – 35 80 → 45 20 → 55 Flow rate: 1.0 mL per minute. Time span of measurement: For 35 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 4 mL of the standard solution, and add methanol to make exactly 20 mL. Confirm that the peak area of mosapride obtained with 5 mL of this solution is equivalent to 15 to 25z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of mosapride are not less than 40,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mosapride is not more than 5.0z. Water <2.48> 5.0 – 6.5z (0.5 g, volumetric titration, back titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately 0.5 g of Mosapride Citrate Hy-drate, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 61.40 mg of C21H25ClFN3O3.C6H8O7
Mukoi-Daikenchuto Extract contains not less than 1.
Mukoi-Daikenchuto Extract is a light brown powder. It has a slight odor, and has a pungent taste.
(1) Shake 2.0 g of Mukoi-Daikenchuto Ex-tract with 10 mL of water, add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sam-ple solution. Separately, shake 2.0 g of pulverized japanese zanthoxylum peel with 10 mL of water, add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, methanol and acetic acid (100) (20:20:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark purple spot (Rf value: about 0.3) from the standard solution (Japanese Zanthoxy-lum Peel). Shake 2.0 g of Mukoi-Daikenchuto Extract with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Sepa-rately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). Shake 2.0 g of Mukoi-Daikenchuto Extract with 10 mL of water, add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Sepa-rately, dissolve 1 mg of [6]-shogaol for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Processed ginger).
(1) Heavy metals <1.07>—Prepare the test solution with 2.0 g of Mukoi-Daikenchuto Extract as directed under Extracts (4), and perform the test (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 2.0 g of Mukoi-Daikenchuto Extract according to Method 3, and perform the test (not more than 1 ppm). Loss on drying <2.41> Not more than 5.9z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z.
(1) Ginsenoside Rb1—Weigh accurately about 2 g of Mukoi-Daikenchuto Extract, add 30 mL of diluted meth-anol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 15 mL of diluted methanol (3 in 5), and repeat the same procedure. Combine the supernatant liquids, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of sodium hydroxide TS, allow to stand for 30 minutes, add 3 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 20 mL. Apply exactly 5 mL of this solution to a column [10 mm in inside diameter, packed with 0.36 g of octadecylsilanized silica gel for pre-treatment (55 – 105 mm in particle size), and washed just before using with methanol and then diluted methanol (3 in 10)], and wash the column in sequence with 2 mL of diluted methanol (3 in 10), 1 mL of sodium carbonate TS and 10 mL of diluted methanol (3 in 10). Finally, elute with methanol to collect exactly 5 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and ‌2076 Mulberry Bark / Crude Drugs and Related Drugs standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of Ginsenoside Rb1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with carbamoyl group bound silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (400:100:1). Flow rate: 1.0 mL per minute (the retention time of gin-senoside Rb1 is about 16 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ginsenoside Rb1 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z. (2) [6]-Shogaol—Weigh accurately about 0.5 g of Mukoi-Daikenchuto Extract, add exactly 50 mL of diluted methanol (3 in 4), shake for 15 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of [6]-shogaol for assay, dis-solve in diluted methanol (3 in 4) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (3 in 4) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of [6]-shogaol in each solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-shogaol is not more than 1.5z.
Nabumetone occurs as white to yellowish white, crystals or a crystalline powder. It is soluble in acetonitrile, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Nabumetone in methanol (1 in 30,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Nabumetone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Nabumetone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Nabumetone RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Nabumetone according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Nabumetone in 20 mL of acetonitrile, and use this solution as the sample solution. Pipet 5 mL of the sample solution, add acetonitrile nitrile to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak area, AT and AS, of nabumetone in each so-lution. to make exactly 50 mL. Pipet 1 mL of this solution, add acetonitrile to make exactly 20 mL, and use this solution as Amount (mg) of nabumetone (C 15H 16O2) ＝ MS × AT/AS ‌the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the peak area of the related substance G obtained from the sample solution is not larger than 3/5 times the peak area of nabumetone from the standard solution, and each peak area other than nabu-metone and the related substance G is not larger than 1/5 times the peak area of nabumetone from the standard solu-tion. Furthermore, the total area of the peaks other than nabumetone from the sample solution is not larger than 1.6 times the peak area of nabumetone from the standard solu-tion. For each peak area of the related substances A, B, C, D, E, F and G, which are having the relative retention time of about 0.73, 0.85, 0.93, 1.2, 1.9, 2.6 and 2.7 to nabume- tone, multiply their correction factors, 0.12, 0.94, 0.25, 0.42, 1.02, 0.91 and 0.1, respectively. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase A: A mixture of water and acetic acid (100) (999:1). Mobile phase B: A mixture of acetonitrile and tetra-hydrofuran (7:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: Amount (mg) of Nabumetone RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 600 mL of a mixture of water and acetic acid (100) (999:1) add 400 mL of a mixture of acetonitrile and tetrahydrofuran (7:3). Flow rate: Adjust so that the retention time of nabume-tone is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nabumetone are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nabumetone is not more than 1.0z.
Nadolol, when dried, contains not less than 98.
Nadolol occurs as a white to yellowish-brown-white crystalline powder. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95), and slightly soluble in water and in chloroform. A solution of Nadolol in methanol (1 in 100) shows no op-tical rotation. Melting point: about 1379C.
(1) Determine the absorption spectrum of a solution of Nadolol in methanol (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. (2) Determine the infrared absorption spectrum of Nadolol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 1585 cm－1, 1460 cm－1, 1092 cm－1, 935 cm－1 and 770 cm－1.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Nadolol according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Dissolve 0.5 g of Nadolol in 10 mL of a mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Perform the test with ‌the sample solution as directed under Thin-layer Chromatog-raphy <2.03>. Spot 100 mL each of the sample solution and a mixture of methanol and chloroform (1:1) as a control solu-tion with 25 mm each of width at an interval of about 10 mm on the starting line of a plate 0.25 mm in thickness of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of acetone, chlo-roform and diluted ammonia TS (1 in 3) (8:1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm), and confirm the po-sitions of the principal spot and the spots other than the principal spot obtained from the sample solution. Scratch and collect the silica gel of the positions of the plate cor-responding to the principal spot and the spots other than the principal spot. To the silica gel collected from the principal spot add exactly 30 mL of ethanol (95), and to the silica gel from the spots other than the principal spot add exactly 10 mL of ethanol (95). After shaking them for 60 minutes, cen-trifuge, and determine the absorbances of these supernatant liquids at 278 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Separately, proceed in the same man-ner with each position of the silica gel from the control solu-tion corresponding to the principal spot and the spots other than the principal spot of the sample solution, and perform a blank determination to make correction. Amount of the related substances calculated by the following equation is not more than 2.0z. Amount (z) of related substances ＝ Ab/(Ab ＋ 3Aa) × 100 Aa: Corrected absorbance of the principle spot Ab: Corrected absorbance of the spots other than the prin-ciple spot Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Prepare a paste with 0.01 g of Nadolol as di-rected in the paste method under Infrared Spectrophotome-try <2.25> so that its transmittance at an absorption band at a wave number of about 1585 cm－1 is 25 to 30z, and deter- mine the infrared absorption spectrum between 1600 cm－1 Nafamostat Mesilate
Nafamostat Mesilate, when dried, contains not less than 99.
Nafamostat Mesilate occurs as a white crystal-line powder. It is freely soluble in formic acid, soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. Melting point: about 2629C (with decomposition).
(1) Determine the absorption spectrum of a solution of Nafamostat Mesilate in 0.01 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Nafamostat Mesilate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A 0.1-g portion of Nafamostat Mesilate responds to Qualitative Tests <1.09> (1) for mesilate. and 1100 cm－1. Determine the absorbances, A and A 1250, pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Nafamostat Mesilate in 50 mL of water is between from the transmittances, T1265 and T1250, at wave numbers of about 1265 cm－1 (racemic substance A) and 1250 cm－1 (racemic substance B), respectively: the ratio A1265/A1250 is between 0.72 and 1.08.
(1) Clarity and color of solution—A solution pre-pared by dissolving 1.0 g of Nafamostat Mesilate in 50 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Nafamostat Mesilate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.10 g of Nafamostat Mesi-late in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 10 mL of the sample solution, add the mobile phase to make exactly 100 mL. Then pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the area of each peak other than nafamostat ob-tained from the sample solution is not larger than 1/5 times the peak area of nafamostat from the standard solution. Furthermore, the total area of the peaks other than ‌nafamostat from the sample solution is not larger than the peak area of nafamostat from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.07 g of sodium 1-heptane sul-fonate in 1000 mL of diluted acetic acid (100) (3 in 500). To 700 mL of this solution add 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of nafamostat is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of nafamostat, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Pipet 15 mL of this solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of nafamostat obtained with 10 mL of this solution is equivalent to 1.1 to 1.9z of that with 10 mL of the standard solution. System performance: Dissolve 0.1 g of nafamostat mesi-late in the mobile phase to make 100 mL. To 10 mL of this solution add the mobile phase to make 100 mL. To 5 mL of this solution add 5 mL of a solution of 6-amidino-2-naphthol methanesulfonate in the mobile phase (1 in 20,000). When the procedure is run with 10 mL of this solu-tion under the above operating conditions, 6-amidino-2-naphthol and nafamostat are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nafamostat is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.28 g of Nadolol, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to green-blue (indicator: 3 drops of crystal violet TS). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.94 mg of C17H27NO4
Naftopidil, when dried, contains not less than 99.
Naftopidil occurs as a white crystalline pow-der. It is very soluble in acetic anhydride, freely soluble in N,N-dimethylformamide and in acetic acid (100), slightly soluble in methanol and in ethanol (99.5), and practically in-soluble in water. It is gradually colored to light brown by light. A solution of Naftopidil in N,N-dimethylformamide (1 in 10) shows no optical rotation.
(1) Dissolve 50 mg of Naftopidil in 5 mL of acetic acid (100), and add 0.1 mL of Dragendorff's TS: orange colored precipitates are produced. Determine the absorption spectrum of a solution of Naftopidil in methanol (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Naftopidil, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Naftopidil according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Naftopidil in 60 mL of methanol, add diluted 0.1 mol/L potassium dihy-drogen phosphate TS (pH 2.0) (1 in 2) to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:2) to make exactly 100 mL. Pipet 4 mL of this solution, add a mixture of methanol and water (3:2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by automatic integration method: each peak area other than naftopidil obtained from the sample solution is not larger than 3/4 times the peak area of naftopidil from the standard solution, and the total area of the peaks other than naftopidil from the sample solution is not larger than 2.5 times the peak area of naftopidil from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.80 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 4.0 with diluted phosphoric acid (1 in 10), and add water to make 1000 mL. To 450 mL of this solution add 550 mL of methanol. ‌Flow rate: Adjust so that the retention time of naftopidil is about 10 minutes. Time span of measurement: About 2 times as long as the retention time of naftopidil, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, add a mixture of methanol and water (3:2) to make exactly 10 mL. Confirm that the peak area of naftopidil obtained with 10 mL of this solution is equivalent to 17.5 to 32.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of naftopidil are not less than 2500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of naftopidil is not more than 3.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Naftopidil, previ-ously dried, dissolve in 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.25 mg of C24H28N2O3
Nalidixic Acid, when dried, contains not less than 99.
Nalidixic Acid occurs as white to light yellow, crystals or crystalline powder. It is sparingly soluble in N,N-dimethylformamide, very slightly soluble in ethanol (99.5), and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Nalidixic Acid in 0.01 mol/L sodium hydroxide dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution with 0.35 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.012z). Heavy metals <1.07>—Proceed with 1.0 g of Nalidixic Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Nalidixic Acid in 20 mL of 0.01 mol/L sodium hydroxide TS. To 5 mL of this solution, add water to make 10 mL, and use this solution as the sample solution. Pipet 2 mL of the sample so-lution, add water to make exactly 1000 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than nalidixic acid obtained from the sample solution is not larger than the peak area of nalidixic acid from the standard solution, and the total area of the peaks other than nalidixic acid from the sample solution is not larger than 2.5 times the peak area of nalidixic acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.24 g of sodium dihydrogen phos-phate dihydrate in 950 mL of water, adjust the pH to 2.8 with phosphoric acid, and add water to make 1000 mL. To 300 mL of this solution add 200 mL of methanol. Flow rate: Adjust so that the retention time of nalidixic acid is about 19 minutes. Time span of measurement: About 3 times as long as the retention time of nalidixic acid, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add water to make exactly 10 mL. Confirm that the peak area of nalidixic acid obtained with 10 mL of this solution is equivalent to 40 to 60z of that with 10 mL of the standard solution. System performance: Dissolve 25 mg of methyl parahy-droxybenzoate in 100 mL of a mixture of water and metha-nol (1:1). To 1 mL of this solution add water to make 10 mL. To 5 mL of this solution add 5 mL of the standard solu-tion. When the procedure is run with 10 mL of this solution under the above operating conditions, methyl parahydroxy- ‌benzoate and nalidixic acid are eluted in this order with the resolution between these peaks being not less than 13. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nalidixic acid is not more than 2.0z. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). ‌Assay Weigh accurately about 0.3 g of Nalidixic Acid, pre-viously dried, dissolve in 50 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethyl ammonium hy-droxide VS (potentiometric titration). Separately, to 50 mL of N,N-dimethylformamide add 13 mL of a mixture of water and methanol (89:11), perform a blank determination with the solution in the same manner, and make any neces-sary correction. Each mL of 0.1 mol/L tetramethyl ammonium hydroxide VS ＝ 23.22 mg of C12H12N2O3
Naloxone Hydrochloride contains not less than 98.
Naloxone Hydrochloride occurs as white to yellowish white, crystals or crystalline powder. It is freely soluble in water, soluble in methanol, slightly soluble in ethanol (99.5) and in acetic acid (100), and very slightly soluble in acetic anhydride. It is hygroscopic. It is gradually colored by light.
(1) Determine the absorption spectrum of a solution of Naloxone Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Naloxone Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Naloxone Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. Optical rotation <2.49> [a]25: －170 – －1819(0.25 g calcu-lated on the dried basis, water, 10 mL, 100 mm). D pH <2.54> Dissolve 0.10 g of Naloxone Hydrochloride in 10 mL of freshly boiled and cooled water: the pH of the so-lution is between 4.5 and 5.5.
Related substances—Conduct this procedure as rapidly as possible without exposure to light, using light-resistant containers. Dissolve 0.08 g of Naloxone Hydrochlo-ride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop with a mixture of ammonia-saturated 1-butanol TS and methanol (20:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly iron (III) chloride-potassium hexacyano-ferrate (III) TS on the plate: the number of the spot other than the principal spot from the sample solution is not more than 1 and it is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z [0.1 g, 1059C, 5 hours. Use a desiccator (phosphorus (V) oxide) for cool-ing]. Residue on ignition <2.44> Not more than 0.2z (0.1 g).
Weigh accurately about 0.3 g of Naloxone Hydro-chloride, dissolve in 80 mL of acetic acid (100) by warming. After cooling, add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.38 mg of C19H21NO4.HCl
Naphazoline Hydrochloride, when dried, contains not less than 98.
Naphazoline Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in water, soluble in ethanol (95) and in acetic acid (100), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: 255 – 2609C (with decomposition).
(1) To 10 mL of a solution of Naphazoline Hydrochloride (1 in 100) add 5 mL of bromine TS, and boil: a deep purple color develops. ‌To 30 mL of a solution of Naphazoline Hydrochlo-ride (1 in 100) add 2 mL of sodium hydroxide TS, and ex-tract with two 25-mL portions of diethyl ether. Evaporate the combined diethyl ether extracts to dryness with the aid of a current of air. Dry the residue at 809C for 1 hour: the residue melts <2.60> between 1179C and 1209C.‌ Dissolve 0.02 g of the residue obtained in (2) in 2 to 3 drops of dilute hydrochloric acid and 5 mL of water, and add 2 mL of Reinecke salt TS: a red-purple, crystalline pre-cipitate is formed. A solution of Naphazoline Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 0.10 g of Naphazoline Hydrochloride in 10 mL of freshly boiled and cooled water: the pH of the solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Naphazoline Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Napha- zoline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Naphazoline Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.67 mg of C14H14N2.HCl
Naphazoline Nitrate, when dried, contains not less than 98.
Naphazoline Nitrate occurs as a white crystal-line powder. It is odorless, and has a bitter taste. It is freely soluble in acetic acid (100), soluble in ethanol (95), sparingly soluble in water, slightly soluble in acetic an-hydride, and practically insoluble in diethyl ether.
(1) To 10 mL of a solution of Naphazoline Nitrate (1 in 100) add 5 mL of bromine TS, and boil: a deep purple color develops. To 20 mL of a solution of Naphazoline Nitrate (1 in 100) add 5 mL of sodium hydroxide TS, and extract with two 25-mL portions of diethyl ether. Combine the diethyl ether extracts, evaporate to dryness with the aid of a current of air, and dry the residue at 809C for 1 hour: the residue so obtained melts <2.60> between 1179C and 1209C. A solution of Naphazoline Nitrate (1 in 20) responds to Qualitative Tests <1.09> for nitrate. pH <2.54> Dissolve 0.1 g of Naphazoline Nitrate in 10 mL of freshly boiled and cooled water: the pH of the solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Naphazoline Nitrate in 50 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Napha- zoline Nitrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Naphazoline Nitrate, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (4:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.33 mg of C14H14N2.HNO3
Naproxen, when dried, contains not less than 98.
Naproxen occurs as white, crystals or crystal-line powder. It is odorless. It is freely soluble in acetone, soluble in methanol, in ethanol (99.5) and in chloroform, sparingly soluble in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Dissolve 0.01 g of Naproxen in 5 mL of methanol, add 5 mL of water, then add 2 mL of potassium iodide TS and 5 mL of a solution of potassium iodate (1 in 100), and shake: a yellow to yellow-brown color develops. To this solution add 5 mL of chloroform, and shake: a light red-purple color develops in the chloroform layer. To 1 mL of a solution of Naproxen in ethanol (99.5) (1 in 300) add 4 mL of hydroxylamine perchlorate-ethanol TS and 1 mL of N,N?-dicyclohexylcarbodiimide-ethanol TS, shake well, and allow to stand in lukewarm water for 20 minutes. After cooling, add 1 mL of iron (III) perchlorate-ethanol TS, and shake: a red-purple color develops. Determine the absorption spectrum of a solution of Naproxen in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare (C16H19 ClN2.C4H4O4) the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave- ＝ MSb × QTb/QSb × 1/25 MSa: Amount (mg) of naphazoline nitrate for assay taken MSb: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—A solution of ethenzamide in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column, about 4 mm in inside diameter and 25 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). lengths. Determine the infrared absorption spectrum of Naproxen, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]25: ＋63.0 – ＋68.59(after dry-ing, 0.1 g, chloroform, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 2.0 g of Naproxen in 20 mL of acetone: the solution is clear. Per- form the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 400 nm is not more than 0.070. ‌Heavy metals <1.07>—Proceed with 2.0 g of Napro-xen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Naproxen according to Method 3, and perform the test (not more than 1 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Naproxen in 10 mL of a mixture of ethanol (99.5) and chloroform (1:1), and use this solution as the sample so-lution. Pipet 2 mL of the sample solution, and add a mixture of ethanol (99.5) and chloroform (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of ethanol (99.5) and chloroform (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of hexane, dichloromethane, tetrahydrofuran and acetic acid (100) (50:30:17:3) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5g of Naproxen, previ-ously dried, add 100 mL of diluted methanol (4 in 5), dis-solve by gentle warming if necessary, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 3 drops of phe-nolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 23.03 mg of C14H14O3
Nartograstim (Genetical Recombination) oc-curs as a clear and colorless, liquid.
(1) To a suitable amount of Nartograstim (Genetical Recombination) add tris-sodium chloride buffer solution (pH 8.0) so that each mL contains 1 mg of protein, and use this solution as the sample solution. Put 0.1 mL of the sample solution in the well of a 96-well microplate, allow to stand at 59C for not less than 10 hours, then remove the liquid, and wash the well. Then to the well add 0.25 mL of blocking TS for nartograstim test, and allow to stand at room temperature for 1 hour. Remove the blocking TS, add 0.1 mL of rabbit anti-nartograstim antibody TS to the well, and shake gently at room temperature for 3 hours. Remove the rabbit anti-nartograstim antibody TS, and wash the well. Then, add 0.1 mL of peroxidase labeled anti-rabbit antibody TS, shake gently at room temperature for 2 hours, remove the TS, and wash the well. Then, add 0.1 mL of 2,2?-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammoni-um salt TS, allow to stand at room temperature for 10 minutes, add 0.1 mL of a solution of oxalic acid dihydrate (1 in 50), and name this well as the sample well. Separately, proceed with 0.1 mL of tris-sodium chloride buffer solution (pH 8.0) in the same manner as for the sample solution, and name the well so obtained as the control well. When com-pare the sample well and the control well, the sample well re-veals a green color, while the control well reveals no color. Washing procedure of well: To the well add 0.25 mL of washing fluid for nartograstim test, allow to stand for 3 minutes, and remove the washing fluid. Repeat this proce-dure 2 times more. (2) To a suitable amount of Nartograstim (Genetical Recombination) add water so that each mL contains 1 mg of protein. Replace the solvent of 2 mL of this solution with tris-calcium chloride buffer solution (pH 6.5). To 0.5 mL of the solution so obtained add 0.5 mL of tris-calcium chloride buffer solution (pH 6.5) and 5 mL of thermolysin solution (1 in 1000), allow to stand at 379C for 21 hours, and use this solution as the sample solution. Separately, proceed with 2 mL of Nartograstim RS in the same manner as for the sam-ple solution, and use the solution so obtained as the standard solution. Perform the test with 20 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and compare these chromatograms: the similar peaks appear at the same retention times. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: A mixture of water and trifluoroacetic acid (1000:1). Mobile phase B: A mixture of acetonitrile, water and trifluoroacetic acid (900:100:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 5 5 – 90 100 → 40 0 → 60 Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above conditions, the number of the peak which shows not less than 1.6 of the resolution between the adjacent peaks is not less than 15. Molecular mass To a suitable amount of Nartograstim (Genetical Recombination) add reduction buffer solution for nartograstim sample so that each mL contains about 0.5 mg of protein, and use this solution as the sample solution. Separately, to 50 mL of molecular mass marker for nar-tograstim test add reduction buffer solution for nartograstim sample to make 1.0 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution, both previously warmed at 409C for 15 minutes, by SDS polyacrylamide gel electropho-resis, using buffer solution for SDS polyacrylamide gel elec-trophoresis and polyacrylamide gel for nartograstim. After electrophoresis, immerse the gel in a solution of coomassie brilliant blue R-250 in a mixture of water, ethanol (95) and acetic acid (100) (5:4:1) (1 in 1000), and stir gently at room temperature for not less than 12 hours. Then, destain the gel with a mixture of water, ethanol (95) and acetic acid (100) (13:5:2), and dry the gel under reduced pressure. Prepare a calibration curve from the migration distance of the molecu-lar mass markers of the standard solution by plotting the migration distance on the horizontal axis and logarithm of the molecular mass on the vertical axis. Calculate the molecular mass of the sample solution from the calibration curve: the molecular mass of the main band is between 17,000 and 19,000. Compositions ratio of related substance Being specified separately when the drug is granted approval based on the Law. pH <2.54> 7.0 – 7.5
(1) Related substances—To a suitable amount of Nartograstim (Genetical Recombination) add the buffer so-lution for nartograstim sample so that each mL contains about 0.5 mg of protein, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the buffer solution for nartograstim sample to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution by SDS polyacrylamide gel electrophore-sis, using buffer solution for SDS polyacrylamide gel elec-trophoresis and polyacrylamide gel for nartograstim. After electrophoresis, immerse the gel in a solution of coomassie brilliant blue R-250 in a mixture of water, ethanol (95) and acetic acid (100) (5:4:1) (1 in 1000), and stir gently at room temperature for not less than 12 hours. Then, destain the gel with a mixture of water, ethanol (95) and acetic acid (100) (13:5:2), and dry the gel under reduced pressure. Determine the areas of the colored bands obtained from the sample so-lution and standard solution by a densitometer at the meas-ure wavelength 560 nm and the control wavelength 400 nm: the total area of the band other than the principal band ob-tained from the sample solution is not larger than the band area from the standard solution. Host cell protein—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 0.62 EU/mg.
(1) Protein content—To exactly V1 mL of Nar-tograstim (Genetical Recombination) add exactly V2 mL of water so that each mL contains about 0.5 mg of protein, and centrifuge. Determine the absorbance, A, of the supernatant liquid at the absorption maximum at about 280 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of protein in 1 mL of Nartograstim (Genetical Recombination) ＝ A/8.71 × (V1 ＋ V2)/V1 × 10 8.71: Specific absorbance (2) Specific activity—To a suitable exact amount of Nar-tograstim (Genetical Recombination) add potency measuring medium for nartograstim test so that the potency is equivalent to 50z to 150z of the relative potency of the standard solution according to the expected potency, and use this solution as the sample solution. Separately, to a suitable exact amount of Nartograstim RS add an exact amount of the potency measuring medium for nartograstim test so that each mL contains exactly 1.2 × 104 units of nartograstim, and use this solution as the standard solution. Culture NFS-60 cells with subculture medium for nartograstim test, centrifuge the medium, remove the supernatant liquid by suction, and wash the cells with the potency measuring me-dium for nartograstim test. Repeat the washing procedure twice more, prepare two cell suspensions, containing 8 ×105 cells per mL and 4 × 105 cells per mL in the potency measur- ing medium for nartograstim test, and use these solutions as the cell suspension (1) and (2), respectively. In 8 wells of the 12th column of a 8×12 well-microplate put 50 mL each of the cell suspension (1), and in all wells of the 1st to 11th columns put 50 mL each of the cell suspension (2). Where, the wells of the 1st and 8th lines are not used for the test. To the wells of the 2nd to 4th lines of the 12th column add 50 mL each of the standard solution, and to the wells of 5th to 7th lines of the 12th column add 50 mL each of the sample solution. From the wells of the 12th column take 50 mL each of the content liquid and transfer to the corresponding wells of the 1st column. Then, from the wells of the 1st column take 50 mL each of the content liquid and transfer to the cor-responding wells of the 2nd column. Proceed in the same way sequentially to the 10th column to prepare two-fold seri-al dilution wells. The wells of the 11th column are not per-formed any process. Incubate the plate under the at-mosphere of 5 volz carbon dioxide at 379C for about 40 hours. After incubation, add to the all wells 10 mL each of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bro-mide TS, and allow to stand under the atmosphere of 5 volz carbon dioxide at 379C for 4 – 6 hours. Add 0.125 mL of dimethylsulfoxide, shake for 5 to 10 minutes, then determine the absorbances of all wells at 550 nm and 660 nm, A1 and A2, using a spectrophotometer for microplate, and calculate the difference, (A1 – A2). Divide by 6 the total of the differ-ences (A1 – A2) of six wells of the 11th and the 1st column, which were added the standard solution, and use the value so obtained as the 50z absorbance, AM. Determine the dilution index numbers (column number) of the two serial wells of the sample solution and standard solution, they are corre-sponding to just the before and after of the 50z absorbance (AM), nT1, nT2 and nS1, nS2, respectively, where nT1 ＜ nT2 and ‌nS1 ＜ nS2. Differences of the absorbance of the serial wells are named as AT1, AT2 and AS1, AS2, respectively. Calculate the relative potencies of each sample solution by the follow-ing equation using the mean value of the three standard solu-tions, and average them. Perform the same procedure by reversing the place of the sample solution and the standard solution. Then, calculate the mean relative potency by aver-aging both values. Relative potency of the sample solution ＝ 2a S2b × 1 a: nT1 ＋ (AT1 － AM)/(AT1 － AT2) b: nS1 ＋ (AS1 － AM)/(AS1 － AS2) Obtain the potency per mL by the following equation, and calculate the potency per mg of protein using the protein content obtained in (1). Amount (unit) of nartograstim per mL of Nartograstim (Ge-netical Recombination) ＝ S × mean relative potency of the sample solution × d S: Concentration (unit/mL) of the standard solution d: Dilution factor for the sample solution System suitability— The absorbance difference of the individual wells of the standard solution of the 3rd column should be not less than AM, and that of the individual wells of the 8th column should be not more than AM. If they do not meet the require-ments, prepare the standard solution of the range of 1.0 × 103 to 1.6 × 104 units, and perform the test.
Pimaricin occurs as white to yellow-white crys-talline powder. It is slightly soluble in methanol and in acetic acid (100), and practically insoluble in water and in ethanol (99.5).
(1) To 3 mg of Pimaricin add 1 mL of hy-drochloric acid, and mix: a blue-purple color appears. (2) Dissolve 5 mg of Pimaricin in a solution of acetic acid (100) in methanol (1 in 100) to make 1000 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pimaricin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]20: ＋243 – ＋2599(0.1 g, acetic acid (100), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Pimaricin according to Method 4, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm). (2) Related substances—Dissolve 20 mg of Pimaricin in methanol to make 100 mL, and use this solution as the sam-ple solution. Perform the test with 10 mL of the sample solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the total area of the peaks other than pimaricin by the automatic inte-gration method. Calculate the amount of the peaks by the area percentage method: not more than 4.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 303 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of ammonium acetate in 1000 mL of a mixture of water, methanol and tetrahydrofu-ran (47:44:2). Flow rate: Adjust so that the retention time of pimaricin is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of pimaricin. System suitability— Test for required detectability: Measure exactly 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add methanol to make exactly 10 mL. Confirm that the peak area of pimaricin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pimaricin are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of pimaricin is not more than 2.0z. Water <2.48> Between 6.0z and 9.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately an amount of Pimaricin and Pimaricin RS, equivalent to about 25 mg (potency), and dis-solve each in methanol to make exactly 100 mL. Pipet 2 mL each of these solutions, add a solution of acetic acid (100) in methanol (1 in 100) to make exactly 100 mL, and use these solutions as the sample solution and standard solution. De-termine the absorbances at 295.5 nm, AT1 and AS1, at 303 nm, AT2 and AS2, and at 311 nm, AT3 and AS3, of the sample ‌solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. uid Chromatography <2.01> according to the following con-ditions. Determine each peak area of both solutions by the Amount [ mg (potency)] of pimaricin (C H NO ) automatic integration method: the area of the peak other ＝ MS × AT2 － AT1 ＋ AT3 A ＋ A × 1000 33 47 13 than the peak of pimozide obtained from the sample solu-tion is not larger than the peak area of pimozide from the standard solution, and the total area of the peaks other than the peak of pimozide from the sample solution is not larger A － S1 S3 S2 2 MS: Amount [mg (potency)] of Pimaricin RS taken
Containers—Tight containers. Storage—Light resistant.
Nateglinide, when dried, contains not less than 98.
Nateglinide occurs as a white crystalline pow-der. It is freely soluble in methanol and in ethanol (99.5), spar-ingly soluble in acetonitrile, and practically insoluble in water. It dissolves in dilute sodium hydroxide TS. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Nateglinide in methanol (1 in 1000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Nateglinide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nateglinide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Nateglinide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the reference standard according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals. D Optical rotation <2.49> [a]20: －36.5 – －40.09(after drying, 0.2 g, dilute sodium hydroxide TS, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Nateglinide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.25 g of Nateglinide in 20 mL of acetonitrile. To 4 mL of this solution add the mo-bile phase to make 25 mL, and use this solution as the sam-ple solution. Pipet 2.5 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than nateglinide obtained from the sample so-lution is not larger than the peak area of nateglinide from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and ‌flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of nateglinide, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nateglinide are not less than 6000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nateglinide is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Nateglinide, previ-ously dried, and dissolve in acetonitrile to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Nateglinide RS, previously dried, and dissolve in acetonitrile to make exactly 20 mL. Pipet 10 mL of this solution, add exactly 5 mL of the inter-nal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nateglinide to that of the internal standard. Amount (mg) of nateglinide (C19H27NO3) ＝ MS × QT/QS × 2 MS: Amount (mg) of Nateglinide RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter).
Fradiomycin Sulfate occurs as a white to light yellow powder. It is freely soluble in water, and practically insoluble in ethanol (95). It is hygroscopic.
(1) Dissolve 50 mg each of Fradiomycin Sulfate and Fradiomycin Sulfate RS in 1 mL of water, and use these solutions as the sample solution and the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, ammonia solution (28) and dichloro-methane (3:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 1109C for 15 minutes: the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. sterilization to 7.8 – 8.0 with sodium hydroxide TS. Standard solutions—Weigh accurately an amount of Fradiomycin Sulfate RS, previously dried, equivalent to about 50 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 14 days. Take exactly a suitable amount of the stand-ard stock solution before use, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, re-spectively. Sample solutions—Weigh accurately an amount of Fradiomycin Sulfate, previously dried, equivalent to about 50 mg (potency), dissolve in 0.1 mol/L phosphate buffer so-lution for antibiotics (pH 8.0) to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L ‌1046‌ phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Fradiomycin Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Fradiomycin Sulfate according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.63 g of Fradiomycin Sulfate in 5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, ammonia solution (28) and dichloro-methane (3:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 1109C for 15 minutes: the spot at around Rf value 0.4 obtained from the sample so-lution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 8.0z (0.2 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Glucose 1.0 g Peptone 6.0 g Meat extract 1.5 g Yeast extract 3.0 g Sodium chloride 2.5 g Agar 15.0 g Water 1000 mL Agar medium for seed and base layer mass (potency) of fradiomycin (C 23H46 N6O 13: 614.64). Mix all the ingredients and sterilize. Adjust the pH after Description Fradiomycin Sulfate occurs as a white to light yellow powder. It is freely soluble in water, and practically insoluble in ethanol (95). It is hygroscopic. Identification (1) Dissolve 50 mg each of Fradiomycin Sulfate and Fradiomycin Sulfate RS in 1 mL of water, and use these solutions as the sample solution and the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, ammonia solution (28) and dichloro-methane (3:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 1109C for 15 minutes: the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. sterilization to 7.8 – 8.0 with sodium hydroxide TS. Standard solutions—Weigh accurately an amount of Fradiomycin Sulfate RS, previously dried, equivalent to about 50 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 14 days. Take exactly a suitable amount of the stand-ard stock solution before use, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, re-spectively. Sample solutions—Weigh accurately an amount of Fradiomycin Sulfate, previously dried, equivalent to about 50 mg (potency), dissolve in 0.1 mol/L phosphate buffer so-lution for antibiotics (pH 8.0) to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L ‌1046‌ phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Neostigmine Methylsulfate, when dried, contains not less than 98.
Neostigmine Methylsulfate occurs as a white crystalline powder. It is very soluble in water, and freely soluble in acetonitrile and in ethanol (95).
(1) Determine the absorption spectrum of a solution of Neostigmine Methylsulfate (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Neostigmine Methylsulfate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of ‌Neostigmine Methylsulfate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Neostigmine Methylsulfate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Neostigmine Methylsulfate in 10 mL of freshly boiled and cooled water: the pH of the so-lution is between 3.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Neostigmine Methylsulfate in 10 mL of water: the solu-tion is clear and colorless. Sulfate—Dissolve 0.20 g of Neostigmine Methylsul-fate in 10 mL of water, add 1 mL of dilute hydrochloric acid and 1 mL of barium chloride TS: no turbidity is produced immediately. Dimethylaminophenol—Dissolve 0.10 g of Neostig-mine Methylsulfate in 5 mL of water, add 1 mL of sodium hydroxide TS, and while cooling with ice, add 1 mL of diazobenzenesulfonic acid TS: no color develops. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 25 mg each of Neostigmine Methylsulfate and Neostigmine Methylsulfate RS, previously dried, dissolve each in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT areas of neostigmine methylsulfate is not more than 1.0z.
Nicardipine Hydrochloride occurs as a slightly greenish yellow crystalline powder. It is freely soluble in methanol and in acetic acid (100), sparingly soluble in ethanol (99.5), and slightly soluble in water, in acetonitrile and in acetic anhydride. A solution of Nicardipine Hydrochloride in methanol (1 in 20) shows no optical rotation. It is gradually affected by light.
(1) Determine the absorption spectrum of a solution of Nicardipine Hydrochloride in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Nicardipine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.02 g of Nicardipine Hydrochloride in 10 mL of water and 3 mL of nitric acid: the solution responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Nicardipine Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Nicardipine Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL, then take exactly 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than nicardipine obtained from the sample solution is not larger than the peak area of nicardipine from the standard solution, and the total area of each peak other than nicardipine is not larger than 2 times the peak area of nicardipine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of a solution of perchloric acid (43 in 50,000) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of nicardipine is about 6 minutes. Time span of measurement: About 4 times as long as the retention time of nicardipine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of nicardipine obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 mL of the standard solution. System performance: Dissolve 2 mg each of Nicardipine Hydrochloride and nifedipine in 50 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, nicardipine and nifedipine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of nicardipine is not more than 3z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 0.9 g of Nicardipine Hydrochloride, previously dried, dissolve in 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 51.60 mg of C26H29N3O6.HCl
Nicergoline, when dried, contains not less than 98.
Nicergoline occurs as white to light yellow, crystals or crystalline powder. It is soluble in acetonitrile, in ethanol (99.5) and in acetic anhydride, and practically insoluble in water. It is gradually colored to light brown by light. Melting point: about 1369C (with decomposition). ‌Identification (1) Determine the absorption spectrum of a solution of Nicergoline in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nicer-goline as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋5.2 – ＋6.29(after drying, 0.5 g, ethanol (95), 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Nicergoline according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Nicergoline in 25 mL of acetonitrile, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add aceto-nitrile to make exactly 100 mL. Pipet 10 mL of this solution, add acetonitrile to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having the relative retention time of about 0.5 to nicergoline obtained from the sample solution, is not larger than 4 times the peak area of nicergoline from the standard solution, and the area of the peak other than nicergoline and the peak mentioned above from the sample solution is not larger than 2.5 times the peak area of nicergoline from the standard solution. The peak which area is larger than the peak area of nicergoline from the standard solution is not more than two peaks, and the total area of the peaks other than nicergoline from the sample solution is not larger than 7.5 times the peak area of nicergoline from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 288 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Adjust the pH of 0.05 mol/L potassium di-hydrogen phosphate TS to 7.0 with triethylamine. To 350 mL of this solution add 350 mL of methanol and 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of nicergoline is about 25 minutes. Time span of measurement: About 2 times as long as the retention time of nicergoline, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add acetonitrile to make exactly 50 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add aceto-nitrile to make exactly 100 mL. Confirm that the peak area of nicergoline obtained with 20 mL of this solution is equiva-lent to 3 to 7z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of nicergoline are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nicergoline is not more than 4.0z. Loss on drying <2.41> Not more than 0.5z (2 g, in vacu-um, 609C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Nicergoline, previ-ously dried, add 10 mL of acetic anhydride, and warm to dissolve. After cooling, add 40 mL of nitrobenzene, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes to blue-green from red through a blue-purple (indicator: 10 drops of neutral red TS). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.22 mg of C24H26BrN3O3
Niceritrol, when dried, contains not less than 99.
Niceritrol occurs as a white to pale yellow-white powder. It is odorless, and has a slightly bitter taste. It is freely soluble in chloroform, soluble in N,N-dimethylformamide, very slightly soluble in ethanol (95), and practically insoluble in water and in diethyl ether.
(1) Determine the absorption spectrum of a solution of Niceritrol in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Niceritrol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—To 2.0 g of Niceritrol add 50 mL of water, and warm at 709C for 20 minutes, while shak-ing occasionally. After cooling, filter, and to 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 1.0 g of Niceritrol according to Method 2, and perform the test. Pre- ‌pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Niceritrol according to Method 3, and perform the test. Use 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) (not more than 2 ppm). Pyridine—Dissolve 0.5 g of Niceritrol in N,N-dimethylformamide to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of pyridine, and add N,N-dimethylformamide to make exactly 100 mL. Pipet 1 mL of this solution, add N,N-dimethylformamide to make exactly 100 mL, then pipet 0.5 mL of this solution, add N,N-dimethylformamide to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solu-tion and standard solution as directed under Gas Chroma-tography <2.02> according to the following conditions. Determine each peak area of pyridine in both solutions: the peak area of pyridine obtained from the sample solution is not larger than the peak area of pyridine from the standard solution. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with polyethylene glycol 20M for gas chroma-tography coated at the ratio of 10z on acid-treated and silanized siliceous earth for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature of about 1609C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of pyridine is about 2 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the number of theoretical plates of the peak of pyri-dine is not less than 1500. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas of pyridine is not more than 3.0z. Free acids—Transfer about 1 g of Niceritrol, weighed accurately, to a separator, dissolve in 20 mL of chloroform, and extract with 20 mL and then 10 mL of water while shak-ing well. Combine the whole extracts, and titrate <2.50> with 0.01 mol/L sodium hydroxide VS (indicator: 3 drops of phe-nolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction, and calcu-late the amount of free acid by the following equation: it is not more than 0.1z. Each mL of 0.01 mol/L sodium hydroxide VS ＝ 1.231 mg of C6H5NO2 Related substances—Dissolve 0.10 g of Niceritrol in 10 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add chlo-roform to make exactly 20 mL. Pipet 2 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi- pal spot obtained from the sample solution are not more in-tense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Niceritrol, previously dried, add exactly 25 mL of 0.5 mol/L sodium hydroxide VS, boil gently for 20 minutes under a reflux condenser with a carbon dioxide absorber (soda lime). After cooling, titrate <2.50> immediately the excess sodium hydroxide with 0.5 mol/L hydrochloric acid VS (indicator: 3 drops of phenol-phthalein TS). Perform a blank determination in the same manner. Each mL of 0.5 mol/L sodium hydroxide VS ＝ 69.57 mg of C29H24N4O8
Nicomol, when dried, contains not less than 98.
Nicomol occurs as a white crystalline powder. It is odorless and tasteless. It is soluble in chloroform, and practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid and in dilute nitric acid.
(1) Mix 0.01 g of Nicomol with 0.02 g of 1-chloro-2,4-dinitrobenzene, add 2 mL of dilute ethanol, heat in a water bath for 5 minutes, cool, and add 4 mL of potas-sium hydroxide-ethanol TS: a dark red color develops. Dissolve 0.1 g of Nicomol in 5 mL of dilute hydro-chloric acid, and add 5 drops of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Nicomol in 1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Nicomol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Nicomol in 10 mL of 1 mol/L hydrochloric acid TS: the solution is clear and colorless. Acidity—To 1.0 g of Nicomol add 50 mL of freshly boiled and cooled water, shake for 5 minutes, filter, and to 25 mL of the filtrate add 0.60 mL of 0.01 mol/L sodium hy-droxide VS and 2 drops of phenolphthalein TS: a red color develops. Chloride <1.03>—Dissolve 0.6 g of Nicomol in 15 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.01 mol/L hydro-chloric acid VS add 15 mL of dilute nitric acid and water to make 50 mL (not more than 0.024z). ‌Heavy metals <1.07>—Proceed with 1.0 g of Nicomol according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not Identification To a portion of powdered Nicomol Tablets, equivalent to 0.5 g of Nicomol, add 20 mL of chloroform, shake, and filter. Evaporate the filtrate on a water bath to dryness. Proceed with the residue as directed in the Identifi-cation (1) and (2) under Nicomol. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Nicomol Tablets is not less than 75z. Start the test with 1 tablet of Nicomol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g so that each mL contains about 18 mg of nicomol (C 34H 32N4 of Nicomol according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Nicomol in 20 mL of chloroform, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add chloroform to make exactly 20 mL. Pipet 2 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dichloromethane, ethanol (95), acetonitrile and ethyl acetate (5:3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1.5 g of Nicomol, previ-ously dried, add exactly 40 mL of 0.5 mol/L sodium hydrox-ide VS, and boil gently under a reflux condenser connected to a carbon dioxide absorption tube (soda lime) for 10 minutes. After cooling, titrate <2.50> immediately the excess sodium hydroxide with 0.25 mol/L sulfuric acid VS (indica-tor: 3 drops of phenolphthalein TS). Perform a blank deter-mination in the same manner. Each mL of 0.5 mol/L sodium hydroxide VS ＝ 80.08 mg of C34H32N4O9
Nicorandil occurs as white crystals. It is freely soluble in methanol, in ethanol (99.5) and in acetic acid (100), soluble in acetic anhydride, and sparingly soluble in water. Melting point: about 929C (with decomposition).
(1) Determine the absorption spectrum of a solution of Nicorandil (1 in 50,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nicorandil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—Dissolve 2.0 g of Nicorandil in 20 mL of dilute ethanol, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS, 20 mL of dilute ethanol and 1 mL of dilute hydrochloric acid, and dilute with water to make 50 mL (not more than 0.010z). Heavy metals <1.07>—Proceed with 2.0 g of Nicoran- dil according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 20 mg of Nicorandil in 10 mL of the mobile phase, and use this solution as the sam-ple solution. Perform the test with 10 mL of the sample solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak area of N-(2-hydroxyethyl)isonicotinamide nitric ester, having the relative retention time of about 0.86 to nicoran-dil, is not more than 0.5z of the peak area of nicorandil, the area of all other peaks is less than 0.1z, and the sum area of the peaks other than nicorandil and N-(2-hydrox-yethyl)isonicotinamide nitric ester is not more than 0.25z of the total peak area. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, tetrahydrofuran, triethylamine and trifluoroacetic acid (982:10:5:3). sample solution. When the procedure is run with 10 mL of this solution under the above operating conditions, N-(2-hydroxyethyl)isonicotinamide nitric ester and nicorandil are eluted in this order with the resolution between these peaks being not less than 3.0. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of nicorandil is not more than 1.5z. Water <2.48> Not more than 0.1z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Nicorandil, dissolve in 30 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 21.12 mg of C8H9N3O4
Nicotinamide, when dried, contains not less than 98.
Nicotinamide occurs as white, crystals or crys-talline powder. It is odorless, and has a bitter taste. It is freely soluble in water and in ethanol (95), and slightly soluble in diethyl ether.
(1) Mix 5 mg of Nicotinamide with 0.01 g of 1-chloro-2,4-dinitrobenzene, heat gently for 5 to 6 seconds, and fuse the mixture. Cool, and add 4 mL of potas-sium hydroxide-ethanol TS: a red color is produced. ‌To 0.02 g of Nicotinamide add 5 mL of sodium hydroxide TS, and boil carefully: the gas evolved turns moistened red litmus paper blue. Dissolve 0.02 g of Nicotinamide in water to make 1000 mL. Determine the absorption spectrum of the solul-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Nicotinamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. pH <2.54> Dissolve 1.0 g of Nicotinamide in 20 mL of water: the pH of this solution is between 6.0 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Nicotinamide in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Take 0.5 g of Nicotinamide, and perform the test. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Take 1.0 g of Nicotinamide, and per- form the test. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Heavy metals <1.07>—Proceed with 1.0 g of Nicotin-amide according to Method 1, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Readily carbonizable substances <1.15>—Take 0.20 g of Nicotinamide, and perform the test. The solution has no more color than Matching Fluid A. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 25 mg each of Nicotinamide and Nicotinamide RS, both previously dried, dissolve sepa-rately in 3 mL of water, and add the mobile phase to make exactly 100 mL. Pipet 8 mL each of these solutions, and add the mobile phase to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard so-lution, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of nicotinamide to that of the internal stand-ard. Amount (g) of nicotinamide (C6H6N2O) ＝ MS × QT/QS MS: Amount (mg) of dried Nicotinamide RS taken Internal standard solution—A solution of nicotinic acid (1 in 25,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in water to make 1000 mL. To 700 mL of this solution add 300 mL of methanol. Flow rate: Adjust so that the retention time of nicotinamide is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and nicotinamide are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nicotinamide to that of the internal stand-ard is not more than 1.0z.
Nicotinic Acid, when dried, contains not less than 99.
Nicotinic Acid occurs as white, crystals or crystalline powder. It is odorless, and has a slightly acid taste. It is sparingly soluble in water, slightly soluble in ethanol (95), and very slightly soluble in diethyl ether. It dissolves in sodium hydroxide TS and in sodium car-bonate TS.
(1) Triturate 5 mg of Nicotinic Acid with 0.01 g of 1-chloro-2,4-dinitrobenzene, and fuse the mixture by gentle heating for 5 to 6 seconds. Cool, and add 4 mL of potassium hydroxide-ethanol TS: a dark red color is pro-duced. (2) Dissolve 0.02 g of Nicotinic Acid in water to make 1000 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Nicotinic Acid RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. pH <2.54> Dissolve 0.20 g of Nicotinic Acid in 20 mL of water: the pH of this solution is between 3.0 and 4.0.
(1) Clarity and color of solution—Dissolve 0.20 g of Nicotinic Acid in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 0.5 g of Nico- tinic Acid. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Dissolve 1.0 g of Nicotinic Acid in 3 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS and 3 mL of dilute hydrochloric acid, and dilute ‌with water to make 50 mL (not more than 0.019z). Nitro compounds—Dissolve 1.0 g of Nicotinic Acid in 8 mL of sodium hydroxide TS, and add water to make 20 mL: the solution has no more color than Matching Fluid A. Heavy metals <1.07>—Proceed with 1.0 g of Nicotinic Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Nicotinic Acid, pre-viously dried, dissolve in 50 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 5 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 12.31 mg of C H NO Assay Measure exactly a volume of Nicotinic Acid Injec-tion, equivalent to about 0.1 g of nicotinic acid (C6H5NO2), and add the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal stand-ard solution, then add the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Nicotinic Acid RS, previ-ously dried at 1059C for 1 hour, and dissolve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nicotinic acid to that of the internal standard. Amount (mg) of nicotinic acid (C6H5NO2) ＝ MS × QT/QS 6 5 2
Nifedipine occurs as a yellow crystalline pow-der. It is odorless and tasteless. It is freely soluble in acetone and in dichloromethane, sparingly soluble in methanol, in ethanol (95) and in acetic acid (100), slightly soluble in diethyl ether, and practically insoluble in water. It is affected by light.
(1) Dissolve 0.05 g of Nifedipine in 5 mL of ethanol (95), and add 5 mL of hydrochloric acid and 2 g of zinc powder. Allow to stand for 5 minutes, and filter: the filtrate responds to Qualitative Tests <1.09> for primary aro-matic amines, developing a red-purple color. Determine the absorption spectrum of a solution of Nifedipine in methanol (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Nifedipine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Nifedipine in 5 mL of acetone: the solution is clear and yellow. Chloride <1.03>—To 2.5 g of Nifedipine add 12 mL of dilute acetic acid and 13 mL of water, and heat to boil. After cooling, filter, and discard the first 10 mL of the fil-trate. To 5 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—To 4 mL of the filtrate obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.45 mL of 0.005 mol/L sulfuric acid VS (not more than 0.054z). Heavy metals <1.07>—Proceed with 2.0 g of Nifedi- pine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of cyclohexane and ethyl acetate (3:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution, cor-responding to that from the standard solution, is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
The procedure should be performed under protec-tion from light in light-resistant vessels. Weigh accurately about 0.12 g of Nifedipine, and dissolve in methanol to make exactly 200 mL. Measure exactly 5 mL of this solution, and add methanol to make exactly 100 mL. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 350 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of nifedipine (C17H18N2O6) ＝ A/142.3 × 40,000
Nilvadipine occurs as a yellow crystalline pow-der. It is freely soluble in acetonitrile, soluble in methanol, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Nilvadipine in acetonitrile (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Nilvadipine in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Nilvadipine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nil-vadipine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of
(1) Heavy metals <1.07>—Proceed with 2.0 g of Nilvadipine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ‌(2) Related substances—Dissolve 20 mg of Nilvadipine in 20 mL of acetonitrile, and use this solution as the sample so-lution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of each related substance is not more than 0.3z, and the total of them is not more than 0.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of phosphate buffer solution (pH 7.4), methanol and acetonitrile (32:27:18). Flow rate: Adjust so that the retention time of nilvadipine is about 12 minutes. Time span of measurement: About 2.5 times as long as the retention time of nilvadipine, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add acetonitrile to make 100 mL, and use this solu-tion as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add acetonitrile to make exactly 10 mL. Confirm that the peak area of nil-vadipine obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suita-bility test. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above op-erating conditions, the number of theoretical plates and the symmetry factor of the peak of nilvadipine is not less than 3300 and not more than 1.3, respectively. System repeatability: Pipet 1 mL of the solution for sys-tem suitability test, and add acetonitrile to make exactly 10 mL. When the test is repeated 6 times with 5 mL of this solu-tion under the above operating conditions, the relative stand-ard deviation of the peak area of nilvadipine is not more than 1.5z. Loss on drying <2.41> Not more than 0.1z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). MS: Amount (mg) of Nilvadipine RS taken Internal standard solution—A solution of acenaphthene in methanol (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.5 g of diammonium hydrogen phosphate in 1000 mL of water, add 10 mL of tetrabutylam-monium hydroxide TS, adjust the pH to 7.0 with diluted phosphoric acid (1 in 10), and add 900 mL of acetonitrile. Flow rate: Adjust so that the retention time of nilvadipine is about 12 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, nilvadipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of nilvadipine to that of the internal standard is not more than 1.0z.
Weigh accurately about 25 mg each of Nilvadipine and Nilvadipine RS, dissolve in methanol to make exactly 25 mL. Pipet 10 mL each of these solutions, add exactly 20 mL of the internal standard solution, 20 mL of water and metha-nol to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solu-tion as directed under the Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nilvadipine to that of the internal standard. Nilvadipine RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Amount (mg) of nilvadipine (C19H19N3O6) ＝ M S × QT/QS Melting point <2.60> 167 – 1719C Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Nilvadipine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). ‌(2) Related substances—Dissolve 20 mg of Nilvadipine in 20 mL of acetonitrile, and use this solution as the sample so-lution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of each related substance is not more than 0.3z, and the total of them is not more than 0.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of phosphate buffer solution (pH 7.4), methanol and acetonitrile (32:27:18). Flow rate: Adjust so that the retention time of nilvadipine is about 12 minutes. Time span of measurement: About 2.5 times as long as the retention time of nilvadipine, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add acetonitrile to make 100 mL, and use this solu-tion as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add acetonitrile to make exactly 10 mL. Confirm that the peak area of nil-vadipine obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suita-bility test. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above op-erating conditions, the number of theoretical plates and the symmetry factor of the peak of nilvadipine is not less than 3300 and not more than 1.3, respectively. System repeatability: Pipet 1 mL of the solution for sys-tem suitability test, and add acetonitrile to make exactly 10 mL. When the test is repeated 6 times with 5 mL of this solu-tion under the above operating conditions, the relative stand-ard deviation of the peak area of nilvadipine is not more than 1.5z. Loss on drying <2.41> Not more than 0.1z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). MS: Amount (mg) of Nilvadipine RS taken Internal standard solution—A solution of acenaphthene in methanol (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.5 g of diammonium hydrogen phosphate in 1000 mL of water, add 10 mL of tetrabutylam-monium hydroxide TS, adjust the pH to 7.0 with diluted phosphoric acid (1 in 10), and add 900 mL of acetonitrile. Flow rate: Adjust so that the retention time of nilvadipine is about 12 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, nilvadipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of nilvadipine to that of the internal standard is not more than 1.0z.
Nitrazepam occurs as white to light yellow, crystals or crystalline powder. It is odorless. It is freely soluble in acetic acid (100), soluble in acetone and in chloroform, slightly soluble in methanol, in ethanol (95) and in ethanol (99.5), very slightly soluble in diethyl ether, and practically insoluble in water. Melting point: about 2279C (with decomposition).
(1) To 3 mL of a solution of Nitrazepam in methanol (1 in 500) add 0.1 mL of sodium hydroxide TS: a yellow color is produced. To 0.02 g of Nitrazepam add 15 mL of dilute hydro-chloric acid, boil for 5 minutes, cool, and filter: the filtrate responds to Qualitative Tests <1.09> for primary aromatic amines. Neutralize 0.5 mL of the filtrate obtained in (2) with sodium hydroxide TS, add 2 mL of ninhydrin TS, and heat on a water bath: a purple color is produced. Determine the absorption spectrum of a solution of Nitrazepam in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths.
(1) Clarity and color of solution—Dissolve 0.10 g of Nitrazepam in 20 mL of acetone: the solution is clear and pale yellow to light yellow in color. Heavy metals <1.07>—Proceed with 1.0 g of Nitra- zepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Nitrazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Nitrazepam in a 10 mL of mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and chlo-roform (1:1) to make exactly 20 mL, pipet 2 mL of this solu-tion, add a mixture of methanol and chloroform (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of nitromethane and ethyl acetate (17:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: Nitrendipine
Weigh accurately about 0.4 g of Nitrazepam, previ-ously dried, and dissolve in 40 mL of acetic acid (100). Titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.13 mg of C15H11N3O3
Nitrendipine, when dried, contains not less than 98.
Nitrendipine occurs as a yellow crystalline powder. It is soluble in acetonitrile, sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. It is gradually colored to brownish yellow by light. A solution of Nitrendipine in acetonitrile (1 in 50) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Nitrendipine in methanol (1 in 80,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nitrendipine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Nitrendipine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Conduct this procedure rapidly using light-resistant vessels. Dissolve 40 mg of Nitrendipine ‌in 5 mL of acetonitrile, add the mobile phase to make 25 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test immediately with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area by the automatic integra-tion method, and calculate the amount of related substances by the following equation: the amount of the related sub-stance, having the relative retention time of about 0.8 to nitrendipine, is not more than 1.0z, the related substance, having the relative retention time of about 1.3, is not more than 0.25z, and other related substances are not more than 0.2z, respectively. The total amount of the related sub-stances other than nitrendipine is not more than 2.0z. Amount (z) of related substance ＝ AT/AS AT: Each peak area other than nitrendipine obtained from the sample solution AS: Peak area of nitrendipine obtained from the standard solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, tetrahydrofuran and acetonitrile (14:6:5). Flow rate: Adjust so that the retention time of nitrendi-pine is about 12 minutes. Time span of measurement: About 2.5 times as long as the retention time of nitrendipine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the Each mL of 0.1 mol/L serium (IV) tetraammonium sulfate VS ＝ 18.02 mg of C18H20N2O6
Nitrogen is the nitrogen produced by the air lique-faction separation method. It contains not less than 99.
Nitrogen is a colorless gas at room tempera-ture and under atmospheric pressure, and is odorless. 1 mL of Nitrogen dissolves in 65 mL of water and in 9 mL of ethanol (95) at 209C and at a pressure of 101.3 kPa. 1000 mL of Nitrogen at 09C and at a pressure of 101.3 kPa weighs 1.251 g.
Introduce 1 mL each of Nitrogen and nitro-gen into a gas-measuring tube or syringe for gas chromatog-raphy from a cylinder with a pressure-reducing valve, through a directly connected polyvinyl chloride or stainless steel tube. Perform the test with these gases as directed under Gas Chromatography <2.02> according to the following con-ditions: the principal peak in the chromatogram obtained form Nitrogen has the same retention time as that from nitrogen. Operating conditions— Proceed as directed in the operating conditions in the Assay.
Oxygen—The peak area of oxygen obtained from Nitrogen in the Assay is not larger than 1/2 times that from the standard gas mixture.
Introduce 1.0 mL of Nitrogen into a gas-measuring Amount (mg) of nitrendipine (C H N O ) tube or syringe for gas chromatography from a cylinder with ＝ MS × QT/QS × 1/50 18 20 2 6 a pressure-reducing valve, through a directly connected poly-vinyl chloride or stainless steel tube. Perform the test with MS: Amount (mg) of nitrendipine for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (4 in 5) (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter this gas as directed under Gas Chromatography <2.02> ac- cording to the following conditions. Measure the peak area AT of oxygen. Separately, introduce 1.0 mL of oxygen into the gas mixer, add carrier gas to make exactly 100 mL, mix thoroughly, and use this as the standard gas mixture. Pro-ceed with 1.0 mL of this mixture in the same manner under Nitrogen, and measure the peak area AS of oxygen. ‌Amount (volz) of nitrogen (N2) ＝ 100 － AT/AS Operating conditions— Detector: A thermal-conductivity detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with zeolite for gas chromatography (250 to 355 mm in particle diameter; 0.5 nm in pore size). Column temperature: A constant temperature of about 509C. Carrier gas: Hydrogen or helium. Flow rate: Adjust so that the retention time of oxygen is about 3 minutes. System suitability— System performance: Introduce 1.0 mL of oxygen into the gas mixer, add Nitrogen to make 100 mL, and mix thor-oughly. When the procedure is run with 1.0 mL of this mix-ture under the above operating conditions, oxygen and nitro-gen are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 1.0 mL of the standard gas mixture under the above conditions, the relative standard deviation of the peak area of oxygen is not more than 2.0z. Containers and storage Containers—Pressure-resistant cylinders. Storage—Not exceeding 409C. Nitroglycerin Tablets
Nitrous Oxide contains not less than 97.
Nitrous Oxide is a colorless gas at room tem-perature and at atmospheric pressure, and is odorless. 1 mL of Nitrous Oxide dissolves in 1.5 mL of water and in 0.4 mL of ethanol (95) at 209C and at a pressure of 101.3 kPa. It is soluble in diethyl ether and in fatty oils. 1000 mL of Nitrous Oxide at 09C and at a pressure of 101.3 kPa weighs about 1.96 g.
(1) A glowing splinter of wood held in Nitrous Oxide: it bursts into flame immediately. (2) Transfer 1 mL each of Nitrous Oxide and nitrous oxide directly from metal cylinders with a pressure-reducing valve to gas measuring tubes or syringes for gas chromatog-raphy, using a polyvinyl chloride induction tube. Perform the test with these gases as directed under Gas Chromatogra-phy <2.02> according to the operating conditions of the Assay: the retention time of the main peak in the chromato-gram obtained with Nitrous Oxide coincides with that in the chromatogram obtained with nitrous oxide.
Maintain the containers of Nitrous Oxide between 189C and 229C for more than 6 hours before the test, and correct the volume at 209C and at a pressure of 101.3 kPa. Acidity or alkalinity—To 400 mL of freshly boiled and cooled water add 0.3 mL of methyl red TS and 0.3 mL of bromothymol blue TS, and boil for 5 minutes. Transfer 50 mL of this solution to each of three Nessler tubes marked A, B and C. Add 0.10 mL of 0.01 mol/L hydrochloric acid VS to tube A, 0.20 mL of 0.01 mol/L hydrochloric acid VS to tube B, stopper each of the tubes, and cool. Pass 1000 mL of Nitrous Oxide through the solution in tube A for 15 minutes, employing delivery tube with an orifice approxi-mately 1 mm in diameter and extending to within 2 mm of the bottom of the Nessler tube: the color of the solution in tube A is not deeper orange-red than that of the solution in tube B and not deeper yellow-green than that of the solution in tube C. Carbon dioxide—Pass 1000 mL of Nitrous Oxide through 50 mL of barium hydroxide TS in a Nessler tube, in the same manner as directed in (1): any turbidity produced does not exceed that produced in the following control solu- tion. Control solution: To 50 mL of barium hydroxide TS in a Nessler tube add 1 mL of a solution of 0.1 g of sodium hydrogen carbonate in 100 mL of freshly boiled and cooled water. Oxidizing substances—Transfer 15 mL of potassium iodide-starch TS to each of two Nessler tubes marked A and B, add 1 drop of acetic acid (100) to each of the tubes, shake, and use these as solution A and solution B, respec-tively. Pass 2000 mL of Nitrous Oxide through solution A for 30 minutes in the same manner as directed in (1): the color of solution A is the same as that of the stoppered, un-treated solution B. Potassium permanganate-reducing substance—Pour 50 mL of water into each of two Nessler tubes marked A and B, add 0.10 mL of 0.02 mol/L potassium permanganate VS to each of the tubes, and use these as solution A and solution B, respectively. Pass 1000 mL of Nitrous Oxide through so-lution A in the manner as directed in (1): the color of solu-tion A is the same as that of solution B. Chloride—Pour 50 mL of water into each of two Nessler tubes marked A and B, add 0.5 mL of silver nitrate TS to each of the tubes, shake, and use these as solution A and solution B, respectively. Pass 1000 mL of Nitrous Oxide through solution A in the same manner as directed in (1): the turbidity of solution A is the same as that of solution B. Carbon monoxide—Introduce 5.0 mL of Nitrous Oxide into a gas-cylinder or a syringe for gas chromatogra-phy from a metal cylinder holding gas under pressure and fitted with a pressure-reducing valve, through a directly con-nected polyvinyl tube. Perform the test with this under Gas Chromatography <2.02> according to the following condi-tions: no peak is observed at the same retention time as that of carbon monoxide. Operating conditions— Detector: A thermal conductivity detector. Column: A column about 3 mm in inside diameter and about 3 m in length, packed with 300 to 500 mm zeolite for gas chromatography (0.5 nm in pore size). Column temperature: A constant temperature of about 509C. Carrier gas: Hydrogen or helium. Flow rate: Adjust so that the retention time of carbon monoxide is about 20 minutes. Selection of column: To 0.1 mL each of carbon monoxide and air in a gas mixer add carrier gas to make 100 mL, and mix well. Proceed with 5.0 mL of the mixed gas under the above operating conditions. Use a column giving well-resolved peaks of oxygen, nitrogen and carbon monoxide in this order. Detection sensitivity: Adjust the sensitivity so that the peak height of carbon monoxide obtained from 5.0 mL of the mixed gas used in the selection of column is about 10 cm.
Withdraw Nitrous Oxide as directed in the Purity. Introduce 1.0 mL of Nitrous Oxide into a gas-measuring tube or syringe for gas chromatography from a metal cylin-der under pressure through a pressure-reducing valve and a directly connected polyvinyl tube. Perform the test with this solution as directed under Gas Chromatography <2.02> ac-cording to the following conditions, and determine the peak area AT of air. Separately, introduce 3.0 mL of nitrogen into a gas mixer, add carrier gas to make exactly 100 mL, mix thoroughly, and use this as the standard mixed gas. Proceed with 1.0 mL of this mixture as directed in the case of Nitrous Oxide, and determine the peak area AS of nitrogen in the same manner. ‌Amount (volz) of nitrous oxide (N2O) ＝ 100 － 3 × AT/AS Operating conditions— Detector: A thermal conductivity detector. Column: A column about 3 mm in inside diameter and about 3 m in length, packed with silica gel for gas chroma-tography (300 to 500 mm in particle diameter). Column temperature: A constant temperature of about 509C. Carrier gas: Hydrogen or helium. Flow rate: Adjust so that the retention time of nitrogen is about 2 minutes. Selection of column: To 3.0 mL of nitrogen in a gas mixer add Nitrous Oxide to make 100 mL, and mix well. Proceed with 1.0 mL of the mixed gas under the above operating con-ditions. Use a column giving well-resolved peaks of nitrogen and nitrous oxide in this order. System repeatability: Repeat the test five times with the standard mixed gas under the above operating conditions: the relative standard deviation of the peak area of nitrogen is not more than 2.0z.
Containers—Metal cylinders. Storage—Not exceeding 409C.
Nizatidine, when dried, contains not less than 98.
Nizatidine occurs as a white to pale yellow-white crystalline powder, and has a characteristic odor. It is soluble in methanol, sparingly soluble in water, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Nizatidine in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Nizatidine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nizatidine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of dried Nizatidine RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Nizatidine according to Method 4, and perform the test using 3 mL of sulfuric acid. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Nizatidine in 10 mL of a mixture of the mobile phase A and mobile phase B (19:6), and use this solution as the sample solution. Pipet 3 mL of the sample solution, add the mixture of the mobile phase A and mobile phase B (19:6) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area from both solutions by the automatic integration method: the area of the peaks other than nizatidine obtained from the sample solution is not larger than 1/5 times the nizatidine from the standard solution. Furthermore, the total of the areas of peaks other than the nizatidine from the sample solution is not larger than the peak area of nizatidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 5.9 g of ammonium acetate in 760 mL of water, add 1 mL of diethylamine, and adjust to pH 7.5 with acetic acid (100). Mobile phase B: Methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 3 3 – 20 76 → 50 24 → 50 20 – 45 Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of nizatidine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of the mobile phase A and mo-bile phase B (19:6) to make exactly 25 mL. Confirm that the peak area of nizatidine obtained with 50 mL of this solution is equivalent to 15 to 25z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nizatidine are not less than 20,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nizatidine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (2 g, 1009C, 1 hour). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 15 mg each of Nizatidine and Nizatidine RS, both previously dried, dissolve each in the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under proceed as directed in the Assay. Amount (mg) of nizatidine (C ＝ MS × QT/QS × V/10 12H21N5O2S2) Liquid Chromatography <2.01> according to the following ‌conditions. Determine the peak area, AT and AS, of nizati-dine in each solution. Amount (mg) of nizatidine (C12H21N5O2S2) ＝ MS × AT/AS MS: Amount (mg) of Nizatidine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 5.9 g of ammonium acetate in 760 mL of water, add 1 mL of diethylamine, and adjust to pH 7.5 with acetic acid (100). To this solution add 240 mL of methanol. Flow rate: Adjust so that the retention time of nizatidine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nizatidine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nizatidine is not more than 1.0z.
Noradrenaline occurs as a white to light brown or slightly reddish brown crystalline powder. It is freely soluble in acetic acid (100), very slightly soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid. It gradually changes to brown by air and by light.
(1) Determine the absorption spectrum of a solution of Noradrenaline in 0.1 mol/L hydrochloric acid TS (3 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Noradrenaline, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.10 g of Noradrenaline in 10 mL of 0.1 mol/L hydrochloric acid TS, and add water to make 100 mL: the solution is clear and colorless. Arterenone—Dissolve 50 mg of Noradrenaline in 0.01 mol/L hydrochloric acid TS to make exactly 100 mL. Determine the absorbance of the solution at 310 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.1. Adrenaline—Dissolve 10.0 mg of Noradrenaline in 2.0 mL of diluted acetic acid (100) (1 in 2). Pipet 1 mL of this solution, add water to make 10 mL, then mix with 0.3 mL of a solution of sodium nitrite (1 in 100), and observe after 1 minute: the solution has no more color than the fol-lowing control solution. Control solution: Dissolve 2.0 mg of Adrenaline Bitartrate RS for Purity and 90 mg of Noradrenaline Bitartrate RS in water to make exactly 10 mL. Pipet 1 mL of this solution, add 1.0 mL of diluted acetic acid (100) (1 in 2) and water to make 10 mL, and proceed in the same manner. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, silica gel, 18 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Noradrenaline, pre-viously dried, dissolve in 50 mL of acetic acid for nona-queous titration by warming, if necessary, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solu-tion changes from blue-purple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.92 mg of C8H11NO3
Norethisterone, when dried, contains not less than 97.
Norethisterone occurs as a white to pale yel-low-white crystalline powder. It has no odor. It is sparingly soluble in ethanol (95), in acetone, and in tetrahydrofuran, slightly soluble in diethyl ether, and very
Norfloxacin, when dried, contains not less than 99.
Norfloxacin occurs as a white to pale yellow crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in ethanol (99.5) and in acetone, very slightly soluble in metha-nol, and practically insoluble in water. It dissolves in dilute hydrochloric acid TS and in sodium hydroxide TS. It is hygroscopic. It is gradually colored by light.
(1) Dissolve 0.01 g of Norfloxacin in a so-lution of sodium hydroxide (1 in 250) to make 100 mL. To 5 mL of this solution add a solution of sodium hydroxide (1 in 250) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro- ‌photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve a suitable amount of Norfloxacin in a suita-ble amount of acetone, evaporate the acetone under reduced pressure, and dry the residue. Determine the infrared ab-sorption spectrum of the residue so obtained as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—Dissolve 1.0 g of Norfloxacin in 7 mL of 0.5 mol/L sodium hydroxide TS and 23 mL of water, and add 1 drop of phenolphthalein TS. Add gradually diluted hydrochloric acid (1 in 3) to this solution until the red color disappears, then add 0.5 mL of dilute hydrochloric acid, and cool in ice for 30 minutes. Filter through a glass filter (G4), and wash the residue with 10 mL of water. Com-bine the filtrate and the washing, and add 1 mL of dilute hy-drochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows. To 0.50 mL of 0.005 mol/L sulfuric acid VS add 7 mL of 0.5 mol/L sodium hydroxide TS and 1 drop of phenolphthalein TS, add diluted hydrochloric acid (1 in 3) until the red color disappears, then add 1.5 mL of dilute hy-drochloric acid, 1 or 2 drops of bromophenol blue TS and water to make 50 mL (not more than 0.024z). Heavy metals <1.07>—Proceed with 2.0 g of Norflox- acin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Norfloxacin according to Method 3, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.10 g of Norfloxacin in 50 mL of a mixture of methanol and acetone (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of metha-nol and acetone (1:1) to make exactly 100 mL. Pipet 2 mL of this solution, add a mixture of methanol and acetone (1:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy (5 to 7 mm in particle diameter). Develop with a mix-ture of methanol, chloroform, toluene, diethylamine and water (20:20:10:7:4) to a distance of about 9 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm and 366 nm): the number of the spot other than the principal spot obtained from the sample solution is not more than 2 and they are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Norfloxacin, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.93 mg of C16H18FN3O3
Norgestrel, when dried, contains not less than 98.
Norgestrel occurs as white, crystals or crystal-line powder. It is soluble in tetrahydrofuran and in chloroform, spar-ingly soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water.
(1) Dissolve 1 mg of Norgestrel in 2 mL of ethanol (95), and add 1 mL of sulfuric acid: a red-purple color develops. With this solution, examine under ultraviolet light (main wavelength: 365 nm): the solution shows a red-orange fluorescence. (2) Determine the infrared absorption spectrum of Nor-gestrel, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Take 1.0 g of Norges-trel, heat gently to carbonize, cool, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and ignite the ethanol to burn. After cooling, add 1 mL of sulfuric acid, proceed with this solution according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 30 mg of Norgestrel in 5 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of dichloromethane and ethyl acetate (2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Weigh accurately about 0.2 g of Norgestrel, previ- ‌ously dried, dissolve in 40 mL of tetrahydrofuran, add 10 mL of a solution of silver nitrate (1 in 20), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titra-tion). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS quantities of Norgestrel RS and of Ethinylestradiol RS, equivalent to 100 times each of the labeled amounts, dissolve in diluted methanol (7 in 10) to make exactly 200 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and ＝ 31.25 mg of C 21H 28O2 standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the
Nortriptyline Hydrochloride, when dried, contains not less than 98.
Nortriptyline Hydrochloride occurs as a white to yellowish white crystalline powder. It is odorless, or has a faint, characteristic odor. It is freely soluble in acetic acid (100) and in chloroform, soluble in ethanol (95), sparingly soluble in water, and prac-tically insoluble in diethyl ether. The pH of a solution of 1.0 g of Nortriptyline Hydrochlo-ride in 100 mL of water is about 5.5. Melting point: 215 – 2209C
(1) To 5 mL of a solution of Nortriptyline Hydrochloride (1 in 100) add 1 mL of bromine TS: the color of the test solution disappears. To 5 mL of a solution of Nortriptyline Hydrochlo- Amount (mg) of ethinylestradiol (C ＝ MSb × QTb/QSb × 1/50 20H24O2) ride (1 in 100) add 1 to 2 drops of a solution of quinhydrone in methanol (1 in 40): a red color gradually develops. Determine the absorption spectrum of a solution of MSa: Amount (mg) of Norgestrel RS taken MSb: Amount (mg) of Ethinylestradiol RS taken Internal standard solution—A solution of diphenyl in diluted methanol (7 in 10) (1 in 50,000). Operating conditions— Detector: Norgestrel—An ultraviolet absorption photome-ter (wavelength: 241 nm). Ethinylestradiol—A fluorophotometer (excitation wave-length: 281 nm, fluorescence wavelength: 305 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and water (11:9). Flow rate: Adjust so that the retention time of norgestrel is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, ethinylestradiol, norgestrel and the internal standard are eluted in this order, and the resolution between the peaks of norgestrel and the internal standard is not less than 8. System repeatability: When the test is repeated 6 times Nortriptyline Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Nor-triptyline Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Nortriptyline Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 0.10 g of Nortriptyline Hydrochloride in 10 mL of water: the solu-tion is clear and colorless to very light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Nortrip- tyline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Nortriptyline Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Nortriptyline ‌Hydrochloride in 20 mL of chloroform, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add chloroform to make exactly 100 mL. Pipet 5 mL of this solution, add chloroform to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 4 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of cyclohexane, methanol and diethylamine (8:1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Nortriptyline Hydrochloride Tablets add a suitable volume of 0.1 mol/L hydrochloric acid TS, disperse the ˆne particles by sonicating, add a suitable volume of 0.1 mol/L hydrochloric acid TS, sonicate, and extract for 15 minutes while occasional shaking. Shake for 15 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 0.5 mg of nortriptyline (C19H21N). Centrifuge this solution, pipet 2 mL of the super-natant liquid, add water to make exactly 100 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. sample solution are not more intense than the spot from the standard solution. Amount (mg) of nortriptyline (C 19H 21N) Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Nortriptyline Hy-drochloride, previously dried, dissolve in 5 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ MS × AT/AS × V/50 × 0.878 MS: Amount (mg) of nortriptyline hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 30 minutes of a 10-mg tablet and a 25-mg tablet are not less than 70z and not less than 80z, respectively. Start the test with 1 tablet of Nortriptyline Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the speciˆed minute after starting the test, and ˆlter through a ＝ 29.98 mg of C19H N.HCl membrane ˆlter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the ˆrst ˆltrate, pipet V mL of
Noscapine Hydrochloride Hydrate, when dried, contains not less than 98.
Noscapine Hydrochloride Hydrate occurs as colorless or white, crystals or crystalline powder. It is odor-less, and has a bitter taste. It is freely soluble in water, in acetic anhydride, and in acetic acid (100), soluble in ethanol (95), and practically in-soluble in diethyl ether.
(1) To 1 mg of Noscapine Hydrochloride Hydrate add 1 drop of formaldehyde-sulfuric acid TS: a pur- ple color, changing to yellow-brown, is produced. To 1 mg of Noscapine Hydrochloride Hydrate add 1 drop of a solution of ammonium vanadate (V) in sulfuric acid (1 in 200): an orange color is produced. Dissolve 0.02 g of Noscapine Hydrochloride Hydrate in 1 mL of water, and add 3 drops of sodium acetate TS: a white, flocculent precipitate is produced. Dissolve 1 mg of Noscapine Hydrochloride Hydrate in 1 mL of diluted sulfuric acid (1 in 35), shake with 5 drops of a solution of disodium chromotropate dihydrate (1 in 50), and add 2 mL of sulfuric acid dropwise: a purple color is produced. Dissolve 0.1 g of Noscapine Hydrochloride Hydrate in 10 mL of water, make the solution alkaline with ammonia TS, and shake with 10 mL of chloroform. Separate the chlo-roform layer, wash with 5 mL of water, and filter. Distil most of the filtrate on a water bath, add 1 mL of ethanol (99.5), and evaporate to dryness. Dry the residue at 1059C for 4 hours: the residue so obtained melts <2.60> between 1749C and 1779C. ‌Make a solution of Noscapine Hydrochloride Hy-drate (1 in 50) alkaline with ammonia TS, and filter the pre-cipitate. Acidify the filtrate with dilute nitric acid: the solu-tion responds to Qualitative Tests <1.09> (2) for chloride.
Morphine—Dissolve 10 mg of Noscapine Hydro-chloride Hydrate in 1 mL of water, add 5 mL of 1-nitroso-2-naphthol TS and 2 mL of a solution of potassium nitrate (1 in 10), and warm at 409C for 2 minutes. Add 1 mL of a solu-tion of sodium nitrite (1 in 5000), and warm at 409C for 5 minutes. After cooling, shake the mixture with 10 mL of chloroform, centrifuge, and separate the aqueous layer: the solution so obtained has no more color than a pale red color. Loss on drying <2.41> Not more than 9.0z (0.5 g, 1209C, 4 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
Weigh accurately about 0.5 g of Noscapine Hydro-chloride Hydrate, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.99 mg of C22H23NO7.HCl
Noscapine occurs as white, crystals or crystal-line powder. It is odorless and tasteless. It is very soluble in acetic acid (100), slightly soluble in ethanol (95) and in diethyl ether, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Noscapine in methanol (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Noscapine, previously dried, as directed in the potassium bromide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋42 – ＋489(after drying, 0.5 g, 0.1 mol/L hydrochloric acid TS, 25 mL, 100 nm).
(1) Chloride <1.03>—Dissolve 0.7 g of Noscapine in 20 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test with this solution. Prepare the control solution as follows: To 0.4 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than After cooling, shake the solution with 10 mL of chloroform, centrifuge, and collect the aqueous layer: the solution so ob-tained has no more color than a pale red. (4) Related substances—Dissolve 0.7 g of Noscapine in 50 mL of acetone, and use this solution as the sample solu-tion. Pipet 5 mL of the sample solution, add acetone to make exactly 50 mL. Pipet 5 mL of this solution, add ace-tone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, toluene, ethanol (99.5) and ammonia solution (28) (60:60:9:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute bismuth subnitrate-potassium iodide TS for spraying on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.8 g of Noscapine, previ-ously dried, dissolve in 30 mL of acetic acid (100) and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 41.34 mg of C22H23NO7
Nux Vomica Extract contains not less than 6.
Nux Vomica Extract occurs as yellow-brown to brown powder. It has a slight characteristic odor, and an extremely bitter taste.
Extract 0.5 g of Nux Vomica Extract with 0.5 mL of ammonia TS and 10 mL of chloroform with occa-sional shaking. Filter the chloroform extract, evaporate the filtrate on a water bath until most of the chloroform is re-moved, and proceed as directed in the Identification under Nux Vomica.
Heavy metals <1.07>—Prepare the test solution with 1.0 g of Nux Vomica Extract as directed in the Extracts (4), and perform the test (not more than 30 ppm). Amount (mg) of strychnine ＝ MS × QT/QS × 1/5 × 0.841
Weigh accurately about 0.2 g of Nux Vomica Ex-tract, place in a glass-stoppered centrifuge tube, add 15 mL MS: Amount (mg) of strychnine nitrate for assay taken, calculated on the dried basis Internal standard solution—A solution of barbital sodium in the mobile phase (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: Room temperature. Mobile phase: Dissolve 6.8 g of potassium dihydrogen-phosphate in water to make 1000 mL, and mix with aceto-nitrile and triethylamine (45:5:1), and adjust the mixture with phosphoric acid to pH 3.0. Flow rate: Adjust so that the retention time of Strychnine is about 17 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and strychnine are eluted in this order with the resolution between these peaks being not less than 1.5.
Nux Vomica Tincture contains not less than 0.
Nux Vomica Tincture is a yellow-brown liquid. It has an extremely bitter taste. Specific gravity d 20: about 0.90
Heat 20 mL of Nux Vomica Tincture on a water bath to remove ethanol, cool, transfer to a separator, add 2 mL of ammonia TS and 20 mL of chloroform, and shake well for 2 to 3 minutes. Filter the chloroform layer through a pledget of absorbent cotton, warm the filtrate on a water bath to remove most of chloroform, and proceed as directed in the Identification under Nux Vomica. Alcohol number <1.01> Not less than 6.7 (Method 2).
Pipet 3 mL of Nux Vomica Tincture into a glass-stoppered centrifuge tube, add 10 mL of ammonia TS and 20 mL of diethyl ether, stopper tightly, shake for 15 minutes, centrifuge to separate the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, proceed in the same man-ner, and repeat this procedure twice. Combine the extracts, ‌JP XVIII Crude Drugs and Related Drugs / Ophiopogon Root 2081‌‌ and evaporate the solvent on a water bath. Dissolve the residue with 10 mL of the mobile phase, add exactly 5 mL of the internal standard solution, and add the mobile phase to make 50 mL. Filter the solution through a membrane filter with a pore size not exceeding 0.8-mm, discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 75 mg of strychnine nitrate for assay (separately determine the loss on drying), and dissolve in the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Proceed with the sample solution and the standard solution as di-rected in the Assay under Nux Vomica. using the former separator. Combine all the filtrates, distil the solvent on a water bath, passing nitrogen. Dissolve the residue in acetone to make exactly 20 mL, and use this solu-tion as the sample solution. Separately, dissolve 0.067 g of methyl behenate in acetone to make exactly 50 mL. Pipet 2 mL of this solution, add acetone to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> ac-cording to the following conditions. Measure the peak heights, HT and HS, of methyl behenate of respective solu-tions: HT is not higher than HS. Operating conditions— Detector: A hydrogen flame-ionization detector. Amount (mg) of strychnine ＝ MS × QT/QS × 1/20 × 0.841 Column: A glass column about 3 mm in inside diameter and about 2 m in length, packed with silanized siliceous MS: Amount (mg) of strychnine nitrate for assay taken, calculated on the dried basis Internal standard solution—A solution of barbital sodium in the mobile phase (1 in 500).
Nystatin is a mixture of polyene macrolide sub-stances having antifungal activity produced by the growth of Streptomyces noursei. It contains not less than 4600 units (potency) per mg, calculated on the dried basis.
Nystatin occurs as a white to light yellow-brown powder. It is soluble in formamide, sparingly soluble in methanol, slightly soluble in ethanol (95), and very slightly soluble in water. It dissolves in sodium hydroxide TS.
(1) Dissolve 1 mg of Nystatin in 5 mL of water and 1 mL of sodium hydroxide TS, heat for 2 minutes, and cool. To this solution add 3 mL of a solution of 4-aminoacetophenone in methanol (1 in 200) and 1 mL of hy-drochloric acid: a red-purple color develops. To 10 mg of Nystatin add 50.25 mL of a mixture of diluted methanol (4 in 5) and sodium hydroxide TS (200:1), heat at not exceeding 509C to dissolve, then add diluted methanol (4 in 5) to make 500 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Nystatin RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths.
Heavy metals <1.07>—Proceed with 1.0 g of Nysta-tin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 5.0z (0.3 g, in vacu-um, 609C, 3 hours).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Saccharomyces cerevisiae ATCC 9763 Culture medium—Use the medium 2) under (1) Agar media for seed and base layer. Standard solutions—Use a light-resistant container. Weigh accurately an amount of Nystatin RS equivalent to about 60,000 units, previously dried at 409C for 2 hours in vacuum (not more than 0.67 kPa), dissolve in formamide to make a solution of 3000 units per mL, and use this solution as the standard stock solution. Keep the standard stock solu-tion at 59C or below and use within 3 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 300 units and 150 units, and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Use a light-resistant container. Weigh accurately an amount of Nystatin equivalent to about 60,000 units, dissolve in formamide to make a solution of 3000 units per mL, and use this solution as the sample stock solution. Take exactly a suitable amount of the sample stock solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 300 units and 150 units, and use these solutions as the high concentration sample so-lution and the low concentration sample solution, respec-tively.
Ofloxacin occurs as pale yellowish white to light yellow-white, crystals or crystalline powder. It is freely soluble in acetic acid (100), slightly soluble in water, and very slightly soluble in acetonitrile and in ethanol (99.5). A soluton of Ofloxacin in sodium hydroxide TS (1 in 20) does not show optical rotation. It is changed in color by light. Melting point: about 2659C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ofloxacin in 0.1 mol/L hydrochloric acid TS (1 in 150,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. ‌(2) Determine the infrared absorption spectrum of Ofloxacin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ofloxacin according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Conduct this procedure without exposure to light. Dissolve 10 mg of Ofloxacin in 50 mL of a mixture of water and acetonitrile (6:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (6:1) to make exactly 20 mL. Pipet 1 mL of this solution, add a mixture of water and acetonitrile (6:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ofloxacin obtained from the sample so-lution is not larger than 2/5 times the peak area of ofloxacin from the standard solution, and the total area of the peaks other than ofloxacin is not larger than the peak area from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 294 nm). Column: A stainless steel column 4.6 mm in inside diame- ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 7.0 g of sodium perchlorate mono-hydrate and 4.0 g of ammonium acetate in 1300 mL of water, adjust the pH to 2.2 with phosphoric acid, and add 240 mL of acetonitrile. Flow rate: Adjust so that the retention time of ofloxacin is about 20 minutes. Time span of measurement: About 1.8 times as long as the retention time of ofloxacin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and acetonitrile (6:1) to make exactly 20 mL. Confirm that the peak area of ofloxa-cin obtained with 10 mL of this solution is equivalent to 4 to 6z of that with 10 mL of the standard solution. System performance: To 0.5 mL of the sample solution add 1 mL of a solution of ofloxacin demethyl substance in a mixture of water and acetonitrile (6:1) (1 in 20,000) and a mixture of water and acetonitrile (6:1) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ofloxacin demethyl sub-stance and ofloxacin are eluted in this order with the resolu-tion between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ofloxacin is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Ofloxacin, previ-ously dried, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.14 mg of C18H20FN3O4
Olmesartan Medoxomil contains not less than 98.
Olmesartan Medoxomil occurs as a white to pale yellow-white crystalline powder. It is slightly soluble in acetonitrile and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Olmesartan Medoxomil in acetonitrile (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Olmesartan Medoxomil RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Ol-mesartan Medoxomil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Olmesartan Medoxomil RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Olmesartan Medoxomil according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Olmesartan Medoxomil in 20 mL of acetonitrile, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the areas of the peaks, having the relative retention times of about 0.2 and about 1.6 to ol-mesartan medoxomil, obtained from the sample solution are not larger than 2/5 times and 3/10 times the peak area of ol-mesartan medoxomil from the standard solution, respec-tively, the area of the peaks other than olmesartan medox-omil and the peaks mentioned above from the sample solu-tion is not larger than 1/10 times the peak area of olmesar- tan medoxomil from the standard solution, and the total area of these peaks is not larger than 3/10 times the peak area of olmesartan medoxomil from the standard solution. In addition, the total area of the peaks other than olmesar-tan medoxomil from the sample solution is not larger than 4/5 times the peak area of olmesartan medoxomil from the standard solution. For the areas of the peaks, having the rel-ative retention times of about 0.7 and about 3.4 to olmesartan medoxomil, multiply their correction factors 0.65 and 1.39, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 3.5 with a solution prepared by dissolving 1.73 g of phosphoric acid in water to make 1000 mL. To 400 mL of this solution add 100 mL of acetonitrile. Mobile phase B: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 3.5 with a solution prepared by dissolving 1.73 g of phosphoric acid in water to make 1000 mL. To 100 mL of this solution add 400 mL of acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 10 – 35 75 → 0 25 → 100 35 – 45 Flow rate: 1.0 mL per minute. Time span of measurement: For 45 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add acetonitrile to make exactly 20 mL. Confirm that the peak area of olmesartan medoxomil obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of olmesartan medoxomil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olmesartan medoxomil is not more than 2.0z. Water <2.48> Not more than 0.5z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Olmesartan Medoxomil and Olmesartan Medoxomil RS (separately de-termine the water <2.48> and the residual solvent in the same manners as Olmesartan Medoxomil), dissolve them sepa-rately in a mixture of acetonitrile and water (4:1) to make ex-actly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, add a mixture of water and acetonitrile (3:2) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of olmesartan medoxomil to that of the internal standard. Amount (mg) of olmesartan medoxomil (C29H30N6O6) ＝ MS × QT/QS ‌MS: Amount (mg) of Olmesartan Medoxomil RS taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of isobutyl para-hydroxybenzoate in a mixture of water and acetonitrile (3:2) (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 3.4 with a solution prepared by dissolving 1.73 g of phosphoric acid in water to make 1000 mL. To 330 mL of this solution add 170 mL of acetonitrile. Flow rate: Adjust so that the retention time of olmesartan medoxomil is about 16 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, olmesartan medoxomil and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of olmesartan medoxomil to that of the inter-nal standard is not more than 0.5z.
Olopatadine Hydrochloride, when dried, contains not less than 99.
Olopatadine Hydrochloride occurs as white, ‌crystals or crystalline powder. It is very soluble in formic acid, sparingly soluble in water, and very slightly soluble in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. The pH of a solution obtained by dissolving 1.0 g of Olopatadine Hydrochloride in 100 mL of water is 2.3 to 3.3. Melting point: about 2509C (with decomposition).
(1) Determine the absorption spectrum of a solution of Olopatadine Hydrochloride in 0.01 mol/L hy-drochloric acid TS (1 in 40,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Olopatadine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Olopatadine Hydrochloride (1 in 100) add 1 mL of dilute nitric acid: this solution re-sponds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Olopatadine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Olopatadine Hydrochloride in 100 mL of a mixture of 0.05 mol/L phos-phate buffer solution (pH 3.5) and acetonitrile (3:2), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than olopatadine obtained from the sample solution is not larger than 1/10 times the peak area of olopatadine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 299 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.3 g of sodium lauryl sulfate in a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (11:9) to make 1000 mL. Flow rate: Adjust so that the retention time of olopatadine is about 11 minutes. Time span of measurement: About 2 times as long as the retention time of olopatadine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add a mixture of 0.05 mol/L phosphate buffer so-lution (pH 3.5) and acetonitrile (3:2) to make exactly 20 mL. Confirm that the peak area of olopatadine obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of olopatadine are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olopatadine is not more than 2.0z. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Olopatadine Hy-drochloride, previously dried, dissolve in 3 mL of formic acid, add 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 37.39 mg of C21H23NO3.HCl
Omeprazole, when dried, contains not less than 99.
Omeprazole occurs as a white to yellowish white crystalline powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in ethanol (99.5), and practically insoluble in water. A solution of Omeprazole in N,N-dimethylformamide (1 in 25) shows no optical rotation. It gradually turns yellowish white on exposure to light. ‌Melting point: about 1509C (with decomposition).
(1) Add phosphate buffer solution (pH 7.4) to 1 mL of a solution of Omeprazole in ethanol (99.5) (1 in 1000) to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Omeprazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Omeprazole in 25 mL of N,N-dimethylformamide: the solution is clear and colorless or light yellow. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 420 nm is not more than 0.3. Heavy metals <1.07>—Proceed with 2.0 g of Omepra-zole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct the procedure soon after preparation of the sample solution. Dissolve 50 mg of Omeprazole in 50 mL of the mobile phase, and use this solu-tion as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each of the peak areas of the sample solution by the auto-matic integration method, and calculate the amounts of them by the area percentage method: the amount of the peaks other than omeprazole is not more than 0.1z, and the total amount of the peaks other than omeprazole is not more than 0.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.83 g of disodium hydrogen phos-phate dodecahydrate and 0.21 g of sodium dihydrogen phos-phate dihydrate in water to make 1000 mL. If necessary, adjust the pH to 7.6 with diluted phosphoric acid (1 in 100). Add 11 volumes of acetonitrile to 29 volumes of this solu-tion. Flow rate: Adjust so that the retention time of omeprazole is about 8 minutes. Time span of measurement: About 10 times as long as the retention time of omeprazole, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the solution for sys-tem suitability test. Pipet 5 mL of the solution for system suitability test, and add the mobile phase to make exactly 25 mL. Confirm that the peak area of omeprazole obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the solution for system suitability test. System performance: Dissolve 10 mg of Omeprazole and 25 mg of 1,2-dinitrobenzene in 5 mL of sodium borate solu-tion (19 in 5000) and 95 mL of ethanol (99.5). When the procedure is run with 10 mL of this solution under the above conditions, omeprazole and 1,2-dinitrobenzene are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of omeprazole is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, in vacu-um, phosphorus (V) oxide, 509C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Omeprazole, previ-ously dried, dissolve in 70 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L tetramethylammonium hy-droxide VS (potentiometric titration). Separately, perform a blank determination in the same manner on a solution con-sisting of 70 mL of N,N-dimethylformamide and 12 mL of water, and make any necessary correction. Each mL of 0.1 mol/L tetramethylammonium hydroxide VS ＝ 34.54 mg of C17H19N3O3S
Containers—Tight containers. Storage—Light-resistant, in a cold place.
Opium Alkaloids Hydrochlorides consist of the hy-drochlorides of some of the main alkaloids obtained from opium. It contains not less than 47.
Opium Alkaloids Hydrochlorides occur as a white to light brown powder. It is soluble in water, and slightly soluble in ethanol (99.5). It is colored by light.
(1) Dissolve 0.1 g of Opium Alkaloids Hy-drochlorides in 10 mL of diluted ethanol (1 in 2), and use this solution as the sample solution. Separately, dissolve 60 mg of Morphine Hydrochloride Hydrate, 40 mg of Nosca-pine Hydrochloride Hydrate, 10 mg of Codein Phosphate Hydrate and 10 mg of Papaverine Hydrochloride in 10 mL each of diluted ethanol (1 in 2), and use these solutions as the standard solutions (1), (2), (3) and (4), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solu-tion and standard solutions on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of acetone, toluene, ethanol (99.5) and ammonia solution (28) (20:20:3:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): each spot obtained from the sample solution is the same in color tone and Rf value with the corresponding spot from the standard solutions (1), (2), (3) and (4) (morphine, noscapine, codeine and papaverine). (2) A solution of Opium Alkaloids Hydrochlorides (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Opium Alkaloids Hydrochlo-rides in 50 mL of water: the pH of the solution is between ‌3.0 and 4.0.
(1) Clarity and color of solution—Dissolve 0.5 g of Opium Alkaloids Hydrochlorides in 10 mL of water: the solution is clear, and its absorbance <2.24> at 420 nm is not more than 0.20. (2) Meconic acid—Dissolve 0.1 g of Opium Alkaloids Hydrochlorides in 2 mL of water, and pour into a polyethy-lene column 1 cm in inside diameter, packed with about 0.36 g of aminopropylsilanized silica gel for pretreatment (55 – 105 mm in particle diameter) and previously washed through with 5 mL of water. Then, wash the column with 5 mL of water, 5 mL of methanol and 10 mL of 0.1 mol/L hy-drochloric acid in this order, then elute with 2 mL of 1 mol/L hydrochloric acid, and use the eluate as the test solu-tion. To the test solution add 2 mL of dilute sodium hydrox-ide TS and 1 drop of iron (III) chloride TS: no red color de-velops. Loss on drying <2.41> Not more than 6.0z (0.5 g, 1209C, 8 hours). Residue on ignition <2.44> Not more than 0.5z (0.5 g).
Weigh accurately about 0.1 g of Opium Alkaloids Hydrochlorides, and dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 60 mg of morphine hydrochloride hydrate for assay, dissolve in water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: Dissolve 60 mg of Morphine Hydro-chloride Hydrate, 10 mg of Codeine Phosphate Hydrate, 10 mg of Papaverine Hydrochloride and 40 mg of Noscapine Hydrochloride Hydrate in water to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, morphine, codeine, papaverine and noscapine are eluted in this order with the complete separa-tion between these peaks and with the resolution between the peaks of morphine and codeine being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of morphine is not more than 1.0z.
Powdered Opium occurs as a yellow-brown to dark brown powder.
(1) To 0.1 g of Powdered Opium add 5 mL of diluted ethanol (7 in 10), dissolve by sonicating for 10 minutes, and add diluted ethanol (7 in 10) to make 10 mL. Filter this solution, and use the filtrate as the sample solu-tion. Separately, dissolve 25 mg of Morphine Hydrochloride Hydrate, 12 mg of Codeine Phosphate Hydrate, 2 mg of perature of 59C to 109C. Decant the diethyl ether layer and filter first, and then the aqueous layer through filter paper 7 cm in diameter. Wash the adhering crystals in the flask with three 5-mL portions of water saturated with diethyl ether, and wash the crystals on the filter paper with each of these washings. Wash the top of the glass-stoppered flask and the upper part of the filter paper with final 5 mL of water satu-rated with diethyl ether. Transfer the crystals and the filter paper to a beaker. Dissolve the crystals remaining in the glass-stoppered flask with the aid of 15 mL of 0.05 mol/L sulfuric acid VS, accurately measured, and pour the solution into the beaker. Wash the glass-stoppered flask with four 5-mL portions of water, and add the washings to the solu-tion in the beaker. Titrate <2.50> the excess sulfuric acid with 0.1 mol/L sodium hydroxide VS (indicator: 4 drops of methyl red-methylene blue TS). Each mL of 0.05 mol/L sulfuric acid VS ＝ 28.53 mg of C17H19NO3
Opium Tincture contains not less than 0.
Opium Tincture is a dark red-brown liquid. It is affected by light.
(1) To 1 mL of Opium Tincture add di-luted ethanol (7 in 10) to make 10 mL, filter, and use the fil-trate as the sample solution. Proceed as directed in the Iden-tification (1) under Powdered Opium. (2) Evaporate 1 mL of Opium Tincture to dryness on a water bath, and proceed with the residue as directed in the Identification (2) under Powdered Opium. Alcohol number <1.01> Not less than 3.5 (Method 1).
Evaporate 50 mL of Opium Tincture, accurately measured, on a water bath to dryness. Add 10 mL of ethanol (99.5) to the residue, evaporate to dryness again, cool, and triturate with exactly 10 mL of water. Proceed with this solu-tion as directed in the Assay under Powdered Opium. Each mL of 0.05 mol/L sulfuric acid VS ＝ 28.53 mg of C17H19NO3
Orange Peel Tincture is a yellowish brown liq-uid. It has a characteristic odor, and a bitter taste. Specific gravity d 20: about 0.90
To 5.0 mL of Orange Peel Tincture add 5 mL of ethanol (95), filter if necessary, and use the filtrate as the sample solution. Proceed as directed in the Identification under Bitter Orange Peel. Alcohol number <1.01> Not less than 6.6 (Method 2).
Orciprenaline Sulfate occurs as white, crystals or crystalline powder. It is freely soluble in water, slightly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether. A solution of Orciprenaline Sulfate (1 in 20) shows no op-tical rotation. Melting point: about 2209C (with decomposition).
(1) Determine the absorption spectrum of a solution of Orciprenaline Sulfate in 0.01 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Orci-prenaline Sulfate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>: it exhibits absorption at the wave numbers of about 1607 cm－1, 1153 cm－1, 1131 cm－1 and 1110 cm－1. A solution of Orciprenaline Sulfate (1 in 100) re-sponds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Orciprenaline Sulfate in 10 mL of water: the pH of this solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Orciprenaline Sulfate in 10 mL of water: the solution is clear, and has no more color than the following control solu-tion. Control solution: To 3 mL of Matching Fluid T add 1 mL of diluted hydrochloric acid (1 in 40). Heavy metals <1.07>—Proceed with 2.0 g of Orci- prenaline Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Orciprenalone—Dissolve 0.200 g of Orciprenaline Sulfate in 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absor-bance at 328 nm is not more than 0.075. Loss on drying <2.41> Not more than 1.5z (1 g, in vacu-um, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Orciprenaline Sul-fate, dissolve in 100 mL of acetic acid (100) by warming on a water bath, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 52.06 mg of (C11H17NO3)2.H2SO4
Orengedokuto Extract contains not less than 20 mg and not more than 80 mg of berberine [as berberine chloride (C20H18ClNO4: 371.
Orengedokuto Extract occurs as a yellow-brown to red-brown powder or blackish brown viscous ex-tract. It has a characteristic odor and a very bitter taste.
(1) Shake 0.5 g of the dry extract (or 1.5 g of the viscous extract) with 10 mL of methanol, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 1 mg of coptisine chloride for thin-layer chromatography in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ammonia so-lution (28) and methanol (15:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Coptis Rhizome). Shake 0.5 g of the dry extract (or 1.5 g of the viscous extract) with 5 mL of water, then add 25 mL of ethyl acetate, and shake. Separate the ethyl acetate layer, evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of limonin for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate and hexane (5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the pur- JP XVIII Crude Drugs and Related Drugs / Orengedokuto Extract 2085 ple spot from the standard solution (Phellodendron Bark). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Scutellaria Root). 5 hours). Total ash <5.01> Not more than 12.0z, calculated on the dried basis.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). Lead—Take 5.0 g of the dry extract (or an amount of the viscous extract, equivalent to 5.0 g of the dried sub-stance) in a platinum, quartz or porcelain crucible, heat gently, and then incinerate by ignition at 450 to 5509C. After cooling, add a small amount of 2 mol/L nitric acid TS, filter if necessary, and wash the crucible and filter several times with small portions of 2 mol/L nitric acid TS. Combine the washings and the filtrate, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sample so-lution. Separately, to 2.5 mL of Standard Lead Solution add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and the standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions: the absorbance of the sample solu-tion is not more than that of the standard solution (not more than 5 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: A lead hollow-cathode lamp. Wavelength: 283.3 nm. Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 7.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: 1.0 mL per minute (the retention time of ber-berine is about 8 minutes). System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, palmatine and berberine are eluted in this order with the resolution be-tween these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, add diluted methanol (7 in 10) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titra-tion, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis ‌2086 Oriental Bezoar / Crude Drugs and Related Drugs Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Geniposide—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.2 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of geniposide for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. Amount (mg) of geniposide ＝ MS × AT/AS × 1/2 MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (900:100:1). Flow rate: 1.0 mL per minute (the retention time of geniposide is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of geniposide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z.
(1) Berberine—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.2 g of dried substance), add exactly 50 mL of the mobile phase, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of berberine in each solution. Shake 0.5 g of the dry extract (or 1.5 g of the viscous extract) with 10 mL of methanol, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, dissolve Amount (mg) of berberine chloride (C ＝ MS × AT/AS × 1/2 20H 18ClNO4) 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solu-tion (Gardenia Fruit). Purity (1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). Lead—Take 5.0 g of the dry extract (or an amount of the viscous extract, equivalent to 5.0 g of the dried sub-stance) in a platinum, quartz or porcelain crucible, heat gently, and then incinerate by ignition at 450 to 5509C. After cooling, add a small amount of 2 mol/L nitric acid TS, filter if necessary, and wash the crucible and filter several times with small portions of 2 mol/L nitric acid TS. Combine the washings and the filtrate, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sample so-lution. Separately, to 2.5 mL of Standard Lead Solution add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and the standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions: the absorbance of the sample solu-tion is not more than that of the standard solution (not more than 5 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: A lead hollow-cathode lamp. Wavelength: 283.3 nm. Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 7.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: 1.0 mL per minute (the retention time of ber-berine is about 8 minutes). System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, palmatine and berberine are eluted in this order with the resolution be-tween these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, add diluted methanol (7 in 10) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titra-tion, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis ‌2086 Oriental Bezoar / Crude Drugs and Related Drugs Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Geniposide—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.2 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of geniposide for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. Amount (mg) of geniposide ＝ MS × AT/AS × 1/2 MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (900:100:1). Flow rate: 1.0 mL per minute (the retention time of geniposide is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of geniposide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z.
Powdered Oyster Shell occurs as a grayish white powder. It is almost odorless and tasteless.
(1) Dissolve 1 g of Powdered Oyster Shell in 10 mL of dilute hydrochloric acid by heating: it evolves a gas, and forms a very slightly red, turbid solution. Pass the gas evolved through calcium hydroxide TS: a white precipi-tate is produced. The solution obtained in (1) has a slight, characteris-tic odor. Filter this solution, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> for cal-cium salt. Ignite 1 g of Powdered Oyster Shell: it turns black-brown in color at first evolving a characteristic odor. Ignite it further: it becomes almost white.
(1) Water-soluble substances—Shake 3.0 g of Powdered Oyster Shell with 50 mL of freshly boiled and cooled water for 5 minutes, filter, and evaporate 25 mL of the filtrate to dryness. Dry the residue at 1059C for 1 hour, cool, and weigh: the mass of the residue does not exceed 15 mg. Acid-insoluble substances—To 5.0 g of Powdered Oyster Shell add 100 mL of water, and add hydrochloric acid in small portions with stirring until the solution becomes acid. Boil the acidic mixture with additional 1 mL of hydro-chloric acid. After cooling, collect the insoluble substance by filtration, and wash it with hot water until the last washing no longer gives any reaction in Qualitative Tests <1.09> (2) for chloride. Ignite the residue and weigh: the mass of the residue does not exceed 25 mg. Barium—Dissolve 1 g of Powdered Oyster Shell in 10 mL of dilute hydrochloric acid: the solution does not re-spond to Qualitative Tests <1.09> (1) for barium salt. Loss on drying <2.41> Not more than 4.0z (1 g, 1809C, 4 hours).
Irregularly curved, foliaceous or lamellated broken pieces. The unbroken oyster shell forms a bivalve 6 – 10 cm in length and 2 – 5 cm in width. The upper valve is flat and the lower one is somewhat hollow. Both the upper and lower edges of the valve are irregularly curved and bite with each other. The surface of the valve is externally light green-gray-brown and internally milky in color. Almost odorless and tasteless.
(1) Dissolve 1 g of sample pieces of Oyster Shell in 10 mL of dilute hydrochloric acid by heating: it evolves a gas, and forms a very slightly red, turbid solution in which a transparent, thin suspended matter remains. Pass the evolved gas through calcium hydroxide TS: a white pre-cipitate is produced. The solution obtained in (1) has a slight, characteris-tic odor. Filter this solution and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> for cal-cium salt. Ignite 1 g of pulverized Oyster Shell: it turns black-brown in color at first, and evolves a characteristic odor. Ignite it further: it becomes almost white.
Barium—Dissolve 1 g of pulverized Oyster Shell in 10 mL of dilute hydrochloric acid: the solution does not respond to Qualitative Tests (1) <1.09> for barium salt.
Otsujito Extract contains not less than 1.
Otsujito Extract occurs as light brown to brown powder or black-brown viscous extract, having a slightly order, and a hot and slight sweet taste. Identiˆcation (1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 10 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, add 10 mL of sodium hydroxide TS, shake, cen-trifuge, and use the diethyl ether layer as the sample solu-tion. Separately, use (Z)-ligustilide TS for thin-layer chro-matography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of butyl acetate and hexane (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Japanese Angelica Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromato-graphy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes. Examine under ul-traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown to grayish brown spot from the standard solution (Scutellaria Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Otsujito Extract / Crude Drugs and Related Drugs Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Use (E)-isoferulic acid-(E)-ferulic acid TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetone and water (20:12:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the light yellow-white fluorescent spot from the standard solution (Cimicifu-ga Rhizome). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of rhein for thin-layer chro-matography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the orange fluorescent spot from the standard solution (Rhubarb).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.5z, calculated on the dried basis.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, shake for 10 minutes, and centrifuge. After removing the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as de- scribed above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, separate the supernatant liquid, com-bine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of saikosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 CS: Concentration (mg/mL) of saikosaponin b2 in saiko-saponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of sai-kosaponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). JP XVIII Crude Drugs and Related Drugs / Otsujito Extract 2089 Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, shake for 10 minutes, and centrifuge. After removing the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as de-scribed above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Sennoside A—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and centri-fuge. Pipet 10 mL of the supernatant liquid, pour it into a column about 10 mm in inside diameter (previously prepared by packing 0.36 g of strongly basic ion-exchange resin for column chromatography, and washing with 10 mL of metha-nol and 10 mL of diluted methanol (1 in 2)) to flow out, wash out the column with 10 mL of diluted methanol (1 in 2), then flow out with a mixture of water, methanol and for-mic acid (25:25:1) to obtain exactly 5 mL of the outflow liq-uid, and use this liquid as the sample solution. Separately, weigh accurately about 5 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of sennoside A in each solution. centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these Amount (mg) of sennoside A (C ＝ MS × AT/AS × 1/8 42H 38O 20) supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (2460:540:1). Flow rate: 1.0 mL per minute (the retention time of senno-side A is about 14 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sennoside A are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z. Rhein—Weigh accurately about 0.5 g of the dry ex-tract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 80 mL of water, shake, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add 20 mL of iron (III) chloride TS, heat under a reflux condenser for 30 minutes, add 3 mL of hydrochloric acid, and heat in addition under a reflux condenser for 30 minutes. After cooling, extract three times with 25 mL each of diethyl ether, combine all the diethyl ether layers, evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in methanol to make exactly 10 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 5 mg of rhein for assay, and dissolve in acetone to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the ‌2090‌ Oyster Shell / Crude Drugs and Related Drugs standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of rhein in each solution. Amount (mg) of rhein ＝ MS × AT/AS × 2/5 MS: Amount (mg) of rhein for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. ‌Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (650:350:1).‌ Flow rate: 1.0 mL per minute (the retention time of rhein is about 17 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rhein are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rhein is not more than 1.5z.
Oxapium Iodide, when dried, contains not less than 98.
Oxapium Iodide occurs as a white crystalline powder. It is soluble in acetonitrile, in methanol and in ethanol (95), slightly soluble in water, in acetic anhydride and in ace-tic acid (100), and practically insoluble in diethyl ether. A solution of Oxapium Iodide in methanol (1 in 100) does not show optical rotation.
(1) Determine the infrared absorption spec-trum of Oxapium Iodide, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Dissolve 0.1 g of Oxapium Iodide in 10 mL of metha-nol, and add 2 mL of dilute nitric acid and 2 mL of silver nitrate TS: a greenish yellow precipitate is formed.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Oxapium Iodide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.05 g of Oxapium Iodide in 100 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the total area of the peaks other than oxapium obtained from the sample solution is not larger than the area of the peak of oxapium from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside di- ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of 209C to 309C. Mobile phase: To 57 mL of acetic acid (100) and 139 mL of triethylamine add water to make 1000 mL. To 50 mL of this solution add 500 mL of acetonitril, 10 mL of dilute ace-tic acid and 440 mL of water. Flow rate: Adjust so that the retention time of oxapium is about 4 minutes. ‌Selection of column: Dissolve 0.05 g of Oxapium Iodide and 3 mg of benzophenone in 100 mL of the mobile phase. Proceed with 20 mL of this solution under the above operat-ing conditions, and calculate the resolution. Use a column giving elution of oxapium and benzophenone in this order with the resolution between these peaks being not less than 5. Detection sensitivity: Adjust the detection sensitivity so that the peak height of oxapium obtained from 50 mL of the standard solution composes 5 to 15z of the full scale. Time span of measurement: About 6 times as long as the retention time of oxapium, beginning after the peak of iodide ion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Oxapium Iodide, previously dried, dissolve in 50 mL of a mixture of acetic an-hydride and acetic acid (100) (9:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration, platinum electrode). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 47.14 mg of C22H34INO2
Oxaprozin, when dried, contains not less than 98.
Oxaprozin occurs as a white to yellowish white crystalline powder. It is sparingly soluble in methanol and in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. It is gradually affected by light.
Determine the infrared absorption spectrum of Oxaprozin, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec-trum: both spectra exhibit similar intensities of absorption at the same wave numbers. 1 cm Absorbance <2.24> E1z (285 nm): 455 – 495 (after drying, 10 mg, methanol, 1000 mL).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Oxaprozin according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Oxaprozin according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.10 g of Oxaprozin in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 5 mL, 3 mL and 1 mL of the standard so-lution (1), add methanol to each to make exactly 10 mL, and use these solutions as the standard solutions (2), (3) and (4), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1), (2), (3) and (4) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (99:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the total intensity of the spots other than the principal spot obtained from the sample solution is not more than 1.0z calculated on the basis of in-tensities of the spots from the standard solutions (1), (2), (3) and (4). Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.5 g of Oxaprozin, previ-ously dried, dissolve in 50 mL of ethanol (95), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 29.33 mg of C18H15NO3
Oxazolam occurs as white, crystals or crystal-line powder. It is odorless and tasteless. It is freely soluble in acetic acid (100), soluble in 1,4-dioxane and in dichloromethane, slightly soluble in ethanol (95) and in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid. It gradually changes in color by light. Melting point: about 1879C (with decomposition).
(1) Dissolve 0.01 g of Oxazolam in 10 mL of ethanol (95) by heating, and add 1 drop of hydrochloric acid: a light yellow color develops, and the solution shows a yellow-green fluorescence under ultraviolet light (main wave-length: 365 nm). Add 1 mL of sodium hydroxide TS to this solution: the color and fluorescence of this solution disap-pear immediately. Dissolve 0.01 g of Oxazolam in 5 mL of dilute hydro-chloric acid by heating in a water bath for 10 minutes. After cooling, 1 mL of this solution responds to Qualitative Tests <1.09> for primary aromatic amines. Place 2 g of Oxazolam in a 200-mL flask, add 50 mL of ethanol (95) and 25 mL of 6 mol/L hydrochloric acid TS, and boil under a reflux condenser for 5 hours. After cooling, neutralize with a solution of sodium hydroxide (1 in 4), and extract with 30 mL of dichloromethane. Dehydrate with 3 g of anhydrous sodium sulfate, filter, and evaporate the dichloromethane of the filtrate. Dissolve the residue in 20 mL of methanol by heating on a water bath, and cool imme-diately in an ice bath. Collect the crystals, and dry in vacuum at 609C for 1 hour: the crystals melt <2.60> between 969C and 1009C. Determine the absorption spectrum of a solution of Oxazolam in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Proceed with Oxazolam as directed under Flame Coloration Test <1.04> (2), and perform the test: a green color appears. using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Oxazol- am according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Place 1.0 g of Oxazolam in a Kjel- dahl flask, add 5 mL of sulfuric acid and 5 mL of nitric acid, and heat gently. Repeat the addition of 2 to 3 mL of nitric acid at times, and continue to heat until a colorless to light yellow solution is obtained. After cooling, add 15 mL of saturated ammonium oxalate monohydrate solution, heat the solution until dense white fumes are evolved, and evapo-rate to a volume of 2 to 3 mL. After cooling, dilute with water to 10 mL, and perform the test with this solution as the test solution (not more than 2 ppm). Related substances—Dissolve 0.05 g of Oxazolam in 10 mL of dichloromethane, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dichloromethane to make exactly 200 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Immediately air-dry, develop the plate with a mixture of toluene and acetone (8:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Chloride <1.03>—To 1.0 g of Oxazolam add 50 mL of water, allow to stand for 1 hour with occasional shak-ing, and filter. To 25 mL of this filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test ‌Oxethazaine‌ Oxetacaine
Weigh accurately about 0.65 g of Oxazolam, previ-ously dried, dissolve in 100 mL of a mixture of acetic acid (100) and 1,4-dioxane (1:1). Titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.88 mg of C18H17ClN2O2
Oxethazaine occurs as a white to pale yellow-white crystalline powder. It is very soluble in acetic acid (100), freely soluble in methanol and in ethanol (95), sparingly soluble in diethyl ether, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Oxethazaine in ethanol (95) (1 in 2500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Oxethazaine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibits similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Oxetha-zaine in 20 mL of ethanol (95), add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of ethanol (95), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.011z). Heavy metals <1.07>—Proceed with 2.0 g of Oxetha- zaine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.40 g of Oxethazaine in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of isopropyl-ether, tetrahydrofuran, methanol and ammonia solution (28) (24:10:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. 2-Aminoethanol—To 1.0 g of Oxethazaine add
Weigh accurately about 0.9 g of Oxethazaine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 46.76 mg of C H N O 28 41 3 3 Description Oxethazaine occurs as a white to pale yellow-white crystalline powder. It is very soluble in acetic acid (100), freely soluble in methanol and in ethanol (95), sparingly soluble in diethyl ether, and practically insoluble in water. Identification (1) Determine the absorption spectrum of a solution of Oxethazaine in ethanol (95) (1 in 2500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Oxethazaine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibits similar intensities of absorption at the same wave numbers. Melting point <2.60> 101 – 1049C Purity (1) Chloride <1.03>—Dissolve 1.0 g of Oxetha-zaine in 20 mL of ethanol (95), add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of ethanol (95), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.011z). Heavy metals <1.07>—Proceed with 2.0 g of Oxetha- zaine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.40 g of Oxethazaine in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of isopropyl-ether, tetrahydrofuran, methanol and ammonia solution (28) (24:10:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. 2-Aminoethanol—To 1.0 g of Oxethazaine add
Oxprenolol Hydrochloride, when dried, contains not less than 98.
Oxprenolol Hydrochloride occurs as a white crystalline powder. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), slightly soluble in acetic anhydride, and practically insoluble in diethyl ether.
(1) To 2 mL of a solution of Oxprenolol Hydrochloride (1 in 100) add 1 drop of copper (II) sulfate TS and 2 mL of sodium hydroxide TS: a blue-purple color de-velops. To this solution add 1 mL of diethyl ether, shake well, and allow to stand: a red-purple color develops in the diethyl ether layer, and a blue-purple color develops in the water layer. To 3 mL of a solution of Oxprenolol Hydrochloride (1 in 150) add 3 drops of Reinecke salt TS: a light red pre-cipitate is formed. Determine the infrared absorption spectrum of Ox-prenolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Oxprenolol Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Oxprenolol Hydrochloride in 10 mL of water: the pH of this solution is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Oxprenolol Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Ox- prenolol Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g ‌of Oxprenolol Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Oxprenolol Hydrochloride in 10 mL of water, and use this solution as the sample solution. Pipet 4 mL of the sample solution, and add water to make exactly 100 mL. Pipet 5 mL of this solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate in a devel-oping chamber saturated with ammonia vapor with a mix-ture of chloroform and methanol (9:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Oxprenolol Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.18 mg of C15H23NO3.HCl
Oxycodone Hydrochloride Hydrate contains not less than 98.
Oxycodone Hydrochloride Hydrate occurs as a white crystalline powder. larger than 3/5 times the peak area of oxycodone from the standard solution. For the area of the peak, having the rela-tive retention time of about 1.8 to oxycodone, multiply the correction factor 0.17. Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 280 nm). Column: A stainless steel column 4.6 mm in inside di-ameter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle di-ameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust to pH 3.0 with sodium hydroxide TS. To 4 volumes of this so-lution add 1 volume of tetrahydrofuran for liquid chro-matography. Mobile phase B: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust to pH 3.0 with sodium hydroxide TS. To 1 volume of this solu-tion add 1 volume of tetrahydrofuran for liquid chro-matography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. It is freely soluble in water, in methanol and in acetic acid (100), sparingly soluble in ethanol (95), slightly soluble in acetic anhydride. The pH of a solution dissolved 1.0 g of Oxycodone Hy-drochloride Hydrate in 10 mL of water is between 3.8 and 5.8. Time after injection 0 – 30 30 – 70 100 → 0 0 → 100 of sample (min) Mobile phase A (volz) Mobile phase B (volz) It is affected by light.
(1) Determine the absorption spectrum of a solution of Oxycodone Hydrochloride Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Oxyc-odone Hydrochloride Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Oxycodone Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －140 – －1499(0.5 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Oxycodone Hydrochloride Hydrate in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve 26 mg of Oxycodone Hydrochloride Hydrate in 20 mL of the mobile phase A, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than oxycodone obtained from the sample so-lution is not larger than 1/5 times the peak area of oxyco-done from the standard solution, and the total area of the peaks other than oxycodone from the sample solution is not Flow rate: 1.0 mL per minute. Time span of measurement: About 5 times as long as the retention time of oxycodone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2.5 mL of the stan-dard solution, add the mobile phase A to make exactly 50 mL. Confirm that the peak area of oxycodone obtained with 50 mL of this solution is equivalent to 3.5 to 6.5z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of oxycodone are not less than 3000 and between 0.7 and 1.3, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of oxycodone is not more than 2.0z. Water <2.48> 12 – 15z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.5 g of Oxycodone Hydro-chloride Hydrate, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 35.18 mg of C18H21NO4.HCl
Oxydol occurs as a clear, colorless liquid. It is odorless or has an odor resembling that of ozone. It gradually decomposes upon standing or upon vigorous agitation. It rapidly decomposes when in contact with oxidizing sub-stances as well as reducing substances. It, when alkalized, decomposes with effervescence. It is affected by light. pH: 3.0 – 5.0 Specific gravity d 20: about 1.01
1 mL of Oxydol responds to Qualitative Tests <1.09> for peroxide.
(1) Acidity—To 25.0 mL of Oxydol add 2 drops of phenolphthalein TS and 2.5 mL of 0.1 mol/L sodium hy-droxide VS: a red color develops. Heavy metals <1.07>—To 5.0 mL of Oxydol add 20 mL of water and 2 mL of ammonia TS, evaporate on a water bath to dryness, dissolve the residue in 2 mL of dilute acetic acid by heating, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 2.5 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 5 ppm). Arsenic <1.11>—To 1.0 mL of Oxydol add 1 mL of ammonia TS, evaporate on a water bath to dryness, take the residue, prepare the test solution according to Method 1, and perform the test (not more than 2 ppm). Organic stabilizer—Extract 100 mL of Oxydol with 50-mL, 25-mL and 25-mL portions of a mixture of chlo-roform and diethyl ether (3:2) successively, combine the ex-tracts in a tared vessel, and evaporate the combined extract on a water bath. Dry the residue over silica gel to constant mass: the mass of the residue is not more than 50 mg. Nonvolatile residue—Evaporate 20.0 mL of Oxydol on a water bath to dryness, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 20 mg.
Pipet 1.0 mL of Oxydol, transfer it to a flask con-taining 10 mL of water and 10 mL of dilute sulfuric acid, and titrate <2.50> with 0.02 mol/L potassium permanganate VS. Each mL of 0.02 mol/L potassium permanganate VS ＝ 1.701 mg of H2O2
Oxygen is oxygen produced by the air liquification separation method. It contains not less than 99.
Oxygen is a colorless gas under atmospheric pressure, and is odorless. 1 mL of Oxygen dissolves in 32 mL of water, and in 7 mL of ethanol (95) at 209C and at a pressure of 101.3 kPa. 1000 mL of Oxygen at 09C and at a pressure of 101.3 kPa weighs 1.429 g.
Transfer 1 mL each of Oxygen and oxygen directly from cylinders with a pressure-reducing valve to gas-measuring tubes or syringes for gas chromatography, using a polyvinyl chloride induction tube. Perform the test with these gases as directed under Gas Chromatography <2.02> according to the following conditions: the retention time of principal peak in the chromatogram obtained from Oxygen is the same as that of the peak in the chromatogram from oxygen. Operating conditions— Proceed as directed in the operating conditions in the Purity.
Nitrogen—Transfer 1.0 mL of Oxygen directly from cylinder with a pressure-reducing valve to gas-measuring tube or syringe for gas chromatography, using a polyvinyl chloride induction tube. Perform the test with this gas as directed under Gas Chromatography <2.02> according to the following conditions, and determine the peak area AT of nitrogen. Introduce 0.50 mL of nitrogen into the gas mixer, draw carrier gas into the mixer to make exactly 100 mL, allow to mix thoroughly and use this gas as the standard mixed gas. Perform the test in the same manner with 1.0 mL of this mixture as directed above, and determine the peak area AS of nitrogen: AT is not larger than AS. Operating conditions— Detector: A thermal conductivity detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with zeolite for gas chromatography 250- to 355-mm in particle diameter (a porosity of 0.5 nm). Column temperature: A constant temperature of about 509C. Carrier gas: Hydrogen or helium. Flow rate: Adjust so that the retention time of nitrogen is about 5 minutes. System suitability— System performance: Introduce 0.5 mL of nitrogen into a gas mixer, add Oxygen to make 100 mL, and mix thor-oughly. When the test is run with 1.0 mL of the mixture under the above operating conditions, oxygen and nitrogen are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 1.0 mL of the standard mixed gas under the above oper-ating conditions, the relative standard deviation of the peak area of nitrogen is not more than 2.0z.
Perform the assay by using the magnetic analysis. Apparatus—Oxygen zero gas for assay and oxygen span gas for assay are delivered individually into an appa- ratus via a sample gas-introducing system and a switching valve. An apparatus with a pressure detector of magnetic force method has an introducing system not only for the above gases but also for oxygen reference gas for assay. Each gas is delivered at the flow rate specified for the appa-ratus under control of a flow meter and a manometer. ‌Procedure—Inject oxygen zero gas for assay into the apparatus at the set flow rate, and set the zero after reaching a stable indication. Then inject oxygen span gas for assay at the specified flow rate, and set the span after reaching a sta-ble indication. Confirm that both these indicated values after the settings are within the specification of the apparatus to confirm the suitability of the apparatus. Stop injecting each calibration gas, inject the sample gas at the specified flow rate, and read the indication V (volz). Volume (volz) of oxygen ＝ V (volz) When the apparatus is controlled by periodic calibration, determine appropriately the frequency of calibration accord-ing to the apparatus manufacturer's recommendations, past control records, or statistical methods, and maintain the en-vironment for use and the conditions of injection of the sam-ple gas within the range recommended by the apparatus manufacturer.
Oxymetholone, when dried, contains not less than 97.
Oxymetholone occurs as a white to pale yel-low-white crystalline powder. It is odorless. It is freely soluble in chloroform, soluble in 1,4-dioxane, sparingly soluble in methanol, in ethanol (95) and in ace-tone, slightly soluble in diethyl ether, and practically insolu-ble in water. It is gradually colored and decomposed by light.
(1) Dissolve 2 mg of Oxymetholone in 1 mL of ethanol (95), and add 1 drop of iron (III) chloride TS: a purple color develops. Dissolve 0.01 g of Oxymetholone in methanol to make 50 mL. To 5 mL of the solution add 5 mL of sodium hydroxide-methanol TS and methanol to make 50 mL. De-termine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Oxymetholone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋34 – ＋389(after drying, 0.2 g, 1,4-dioxane, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Oxymetholone in 25 mL of 1,4-dioxane: the solution is clear, and shows a colorless to pale yellow color. (2) Related substances—Dissolve 50 mg of Oxymetho-lone in 5 mL of chloroform, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add chlo-roform to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, and air-dry the spot. Develop immediately the plate with a mixture of toluene and ethanol (99.5) (49:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid TS on the plate, and heat the plate at 1009C for 3 to 5 minutes: any spot other than the principal spot and starting point obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 40 mg of Oxymetholone, previously dried, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, and add methanol to make exactly 50 mL. To exactly measured 5 mL of this solu-tion add 5 mL of sodium hydroxide-methanol TS and methanol to make exactly 50 mL. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 315 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using a solution, prepared by adding methanol to 5 mL of sodium hydroxide-methanol TS to make 50 mL, as the blank. Amount (mg) of oxymetholone (C21H32O3) ＝ A/541 × 50,000
Oxytetracycline Hydrochloride occurs as yel-low, crystals or crystalline powder. It is freely soluble in water, and slightly soluble in ethanol (99.5). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Oxytetracycline Hydrochloride in 0.1 mol/L hy-drochloric acid TS (1 in 50,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Oxytetracycline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Oxyt-etracycline Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of Oxytetracycline Hydro-chloride RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. If any difference ap-pears between the spectra, dissolve the sample and the Refer-ence Standard separately in methanol, evaporate the solvent, and perform the test with the residues. Dissolve 20 mg of Oxytetracycline Hydrochloride in 3 mL of water, and add 1 drop of silver nitrate TS: a white turbidity is produced. D Optical rotation <2.49> [a]20: －188 – －2009(0.25 g calcu-lated on the dried basis, 0.1 mol/L hydrochloric acid, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 0.5 g of Oxytetracycline Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 50 ppm). (2) Related substances—Dissolve 20 mg of Oxytetracy-cline Hydrochloride in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the sample so-lution. Separately, dissolve 20 mg of 4-epioxytetracycline in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as 4-epioxytetracycline stock solution. Separately, dissolve 20 mg of tetracycline hydrochloride in mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as tetracycline hydrochloride stock so-lution. Separately, dissolve 8 mg of b-apooxytetracycline in 5 mL of 0.01 mol/L sodium hydroxide TS, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as b-apooxytetracycline stock solution. Pipet 1 mL of 4-epioxytetracycline stock solution, 4 mL of tetracycline hydrochloride stock solution and 40 mL of b-apooxytetracy-cline stock solution, add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic inte-gration method: the peak areas of 4-epioxytetracycline and tetracycline obtained from the sample solution are not larger than each of the peak area from the standard solution, and the total area of the peaks, a-apooxytetracycline having the relative retention time of about 2.1 to oxytetracycline, b-apooxytetracycline and the peaks, which appear between a-apooxytetracycline and b-apooxytetracycline, is not larger than the peak area of b-apooxytetracycline from the stand-ard solution. The peak area of 2-acetyl-2-decarboxamide oxytetracycline, which appears after the principal peak, from the sample solution is not larger than 4 times the peak area of 4-epioxytetracycline from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (8 mm in particle di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase A: Mix 60 mL of 0.33 mol/L potassium dihydrogen phosphate TS, 100 mL of a solution of tetrabut-ylammonium hydrogensulfate (1 in 100), 10 mL of a solution of disodium dihydrogen ethylenediamine tetraacetate dihy-drate (1 in 2500) and 200 mL of water, and adjust the pH to 7.5 with 2 mol/L sodium hydroxide TS. To this solution add 30 g of t-butyl alcohol and water to make 1000 mL. Mobile phase B: Mix 60 mL of 0.33 mol/L potassium dihydrogen phosphate TS, 50 mL of a solution of tetrabut-ylammonium hydrogensulfate (1 in 100), 10 mL of a solution of disodium dihydrogen ethylenediamine tetraacetate dihy-drate (1 in 2500) and 200 mL of water, and adjust the pH to 7.5 with 2 mol/L sodium hydroxide TS. To this solution add 100 g of t-butyl alcohol and water to make 1000 mL. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection Mobile phase A Mobile phase B of sample (min) (volz) (volz) 0 – 20 70 → 10 30 → 90 20 – 35 10 → 20 90 → 80 Flow rate: 1.0 mL per minute. Time span of measurement: About 3.5 times as long as the retention time of oxytetracycline, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of 4-epiox- ‌ytetracycline stock solution, and add 0.01 mol/L hydrochlo-ric acid TS to make exactly 200 mL. Pipet 4 mL of this solu-tion, and add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Confirm that the peak area of 4-epiox-ytetracycline obtained with 20 mL of this solution is equiva-lent to 14 to 26z of that with 20 mL of the standard solu-tion. System performance: Dissolve 8 mg of a-apooxytetracy-cline in 5 mL of 0.01 mol/L sodium hydroxide TS, add 0.01 mol/L hydrochloric acid TS to make 100 mL, and use this solution as a-apooxytetracycline stock solution. Mix 3 mL of the sample solution, 2 mL of 4-epioxytetracycline stock solu-tion, 6 mL of tetracycline hydrochloride stock solution, 6 mL of b-apooxytetracycline stock solution and 6 mL of a-apooxytetracycline stock solution, and add 0.01 mol/L hy-drochloric acid TS to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-epioxytetracycline, oxytetracycline, tetracy-cline, a-apooxytetracycline and b-apooxytetracycline are eluted in this order with the resolutions between the peaks, 4-epioxytetracycline and oxytetracycline, oxytetracycline and cline is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of the theoretical plates and the sym-metry factor of the peak of oxytetracycline are not less than 1000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of oxytetracycline is not more than 1.0z.
Oxytocin is synthetic human oxytocin, and is a pep-tide consisting of 9 amino acid residues. It contains not less than 540 oxytocin Units and not more than 600 oxytocin Units per mg, calculated on the anhydrous and residual acetic acid-free basis.
Oxytocin occurs as a white powder. It is very soluble in water, and freely soluble in ethanol (99.5). It dissolves in hydrochloric acid TS. The pH of a solution prepared by dissolving 0.10 g of Oxytocin in 10 mL of freshly boiled and cooled water is be-tween 4.0 and 6.0. It is hygroscopic.
Determine the absorption spectrum of a so-lution of Oxytocin (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Constituent amino acids Put about 1 mg of Oxytocin in a test tube for hydrolysis, add 6 mol/L hydrochloric acid TS to dissolve, replace the air in the tube with Nitrogen, seal the tube under reduced pressure, and heat at 110 to 1159C for 16 hours. After cooling, open the tube, evaporate the hydrolyzate to dryness under reduced pressure, add 2 mL of mol/L hydrochloric acid TS to dissolve the residue, and use this solution as the sample solution. Separately, weigh accurately about 27 mg of L-aspartic acid, about 24 mg of L-threonine, about 21 mg of L-serine, about 29 mg of L- glutamic acid, about 23 mg of L-proline, about 15 mg of glycine, about 18 mg of L-alanine, about 23 mg of L-valine, about 48 mg of L-cystine, about 30 mg of methionine, about 26 mg of L-isoleucine, about 26 mg of L-leucine, about 36 mg of L-tyrosine, about 33 mg of phenylalanine, about 37 mg of L-lysine hydrochloride, about 42 mg of L-histidine hy-drochloride monohydrate and about 42 mg of L-arginine hy-drochloride, dissolve them in 10 mL of 1 mol/L hydrochlo-ric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the re-spective molar ratios with respect to leucine: 0.95 – 1.05 for aspartic acid, 0.95 – 1.05 for glutamic acid, 0.95 – 1.05 for proline, 0.95 – 1.05 for glycine, 0.80 – 1.10 for isoleucine, 0.80 – 1.05 for tyrosine and 0.80 – 1.05 for cystine, and not more than 0.01 each for others. Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 nm and 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (Na type) com-posed with a sulfonated polystyrene copolymer (3 mm in par-ticle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B and C accord-ing to the following table. hydrate Mobile phase A B C Citric acid mono- 19.80 g 22.00 g 6.10 g ditions, aspartic acid, threonine, serine, glutamic acid, pro-line, glycine, alanine, valine, cystine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and argi-nine are eluted in this order with the resolutions between the peaks of threonine and serine, glycine and alanine, and isoleucine and leucine being not less than 1.5, 1.4 and 1.2, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of aspartic acid, proline, valine and arginine are not more than 2.0z, respectively.
(1) Acetic acid—Weigh accurately about 15 mg of Oxytocin, dissolve in the internal standard solution to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 1 g of acetic acid (100), add the internal standard solution to make exactly 100 mL. Pipet 2 mL of this solution, add the internal standard solu-tion to make exactly 200 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of acetic acid to that of the internal standard: the amount of acetic acid is not less than 6.0z and not more than 10.0z. Trisodium citrate dihydrate 6.19 g 7.74 g 26.67 g Amount (z) of acetic acid (C2H4O2) ＝ MS/MT × QT/QS × 1/10 Sodium chloride 5.66 g 7.07 g 54.35 g Ethanol (99.5) 260.0 mL 20.0 mL — Benzyl alcohol — — 5.0 mL Thiodiglycol 5.0 mL 5.0 mL — MS: Amount (mg) of acetic acid (100) taken MT: Amount (mg) of Oxytocin taken Internal standard solution—A solution of propionic acid in Lauromacrogol solution (1 in 4) 4.0 mL 4.0 mL 4.0 mL the mobile phase (1 in 10,000). Capryric acid 0.1 mL 0.1 mL 0.1 mL Water a sufficient a sufficient a sufficient amount amount amount Total amount 2000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B and C as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) 0 – 9 9 – 25 25 – 61 100 → 0 0 → 100 61 – 80 Reaction reagent: Mix 407 g of lithium acetate dihydrate, 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for more than 10 minutes while passing Nitrogen, and use this solution as So-lution A. Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for more than 30 minutes while passing Nitrogen, and use this solution as Solution B. Mix Solution A and Solution B before use. Flow rate of mobile phase: About 0.26 mL per minute. Flow rate of reaction reagent: About 0.3 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 0.7 mL of phosphoric acid add 900 mL of water, adjust the pH to 3.0 with 8 mol/L sodium hydrox-ide TS, and add water to make 1000 mL. To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of acetic acid is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, acetic acid and propionic acid are eluted in this order with the resolution between these peaks being not less than 14. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of acetic acid to that of the internal standard is not more than 2.0z. (2) Related substances—Dissolve 25 mg of Oxytocin in 100 mL of the mobile phase A, and use this solution as the sample solution. Perform the test with 50 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of each peak other than Oxytocin is not more than 1.5z, and the total of them is not more than 5.0z. ‌Operating conditions— Detector, column, column temperature, mobile phase, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of oxytocin. System suitability— Test for required detectability: Measure exactly 1 mL of the sample solution, add the mobile phase A to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suita-bility test, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of oxytocin obtained with 50 mL of this solution is equivalent to 5 to 15z of that with 50 mL of the solution for system suitability test. System performance: Dissolve an adequate amount of oxytocin and vasopressin in the mobile phase A, so that each mL contains about 0.1 mg each of them. When the proce-dure is run with 50 mL of this solution under the above oper-ating conditions, vasopressin and oxytocin are eluted in this order with the resolution between these peaks being not less than 14, and the symmetry factor of the peak of oxytocin is not more than 1.5. System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of oxytocin is not more than 2.0z. Water <2.48> Not more than 5.0z (50 mg, coulometric titration).
Weigh accurately an amount of Oxytetracycline Hy-drochloride and Oxytetracycline Hydrochloride RS, equiva-lent to about 50 mg (potency), and dissolve each in diluted hydrochloric acid (1 in 100) to make exactly 50 mL. Pipet 5 mL each of these solutions, add diluted methanol (3 in 20) to make exactly 50 mL, and use these solutions as the sample solution and the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of oxytetracycline in each solution. Amount [ mg (potency)] of oxytetracycline (C22H24N2O9) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Oxytetracycline Hydro-chloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with strongly acidic ion ex-change resin for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.402 g of potassium dihydrogen phosphate and 9.306 g of disodium dihydrogen ethylene-diamine tetraacetate dihydrate in 700 mL of water, add 300 mL of methanol, and adjust the pH to 4.5 with dilute hydro-chloric acid. Flow rate: Adjust so that the retention time of oxytetracy- [50-56-6] Oxytocin is synthetic human oxytocin, and is a pep-tide consisting of 9 amino acid residues. It contains not less than 540 oxytocin Units and not more than 600 oxytocin Units per mg, calculated on the anhydrous and residual acetic acid-free basis. Description Oxytocin occurs as a white powder. It is very soluble in water, and freely soluble in ethanol (99.5). It dissolves in hydrochloric acid TS. The pH of a solution prepared by dissolving 0.10 g of Oxytocin in 10 mL of freshly boiled and cooled water is be-tween 4.0 and 6.0. It is hygroscopic. Identification Determine the absorption spectrum of a so-lution of Oxytocin (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Constituent amino acids Put about 1 mg of Oxytocin in a test tube for hydrolysis, add 6 mol/L hydrochloric acid TS to dissolve, replace the air in the tube with Nitrogen, seal the tube under reduced pressure, and heat at 110 to 1159C for 16 hours. After cooling, open the tube, evaporate the hydrolyzate to dryness under reduced pressure, add 2 mL of mol/L hydrochloric acid TS to dissolve the residue, and use this solution as the sample solution. Separately, weigh accurately about 27 mg of L-aspartic acid, about 24 mg of L-threonine, about 21 mg of L-serine, about 29 mg of L- glutamic acid, about 23 mg of L-proline, about 15 mg of glycine, about 18 mg of L-alanine, about 23 mg of L-valine, about 48 mg of L-cystine, about 30 mg of methionine, about 26 mg of L-isoleucine, about 26 mg of L-leucine, about 36 mg of L-tyrosine, about 33 mg of phenylalanine, about 37 mg of L-lysine hydrochloride, about 42 mg of L-histidine hy-drochloride monohydrate and about 42 mg of L-arginine hy-drochloride, dissolve them in 10 mL of 1 mol/L hydrochlo-ric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the re-spective molar ratios with respect to leucine: 0.95 – 1.05 for aspartic acid, 0.95 – 1.05 for glutamic acid, 0.95 – 1.05 for proline, 0.95 – 1.05 for glycine, 0.80 – 1.10 for isoleucine, 0.80 – 1.05 for tyrosine and 0.80 – 1.05 for cystine, and not more than 0.01 each for others. Operating conditions— Detector: A visible spectrophotometer (wavelength: 440 nm and 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 8 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography (Na type) com-posed with a sulfonated polystyrene copolymer (3 mm in par-ticle diameter). Column temperature: A constant temperature of about 579C. Chemical reaction bath temperature: A constant tempera-ture of about 1309C. Color developing time: About 1 minute. Mobile phase: Prepare mobile phases A, B and C accord-ing to the following table. hydrate Mobile phase A B C Citric acid mono- 19.80 g 22.00 g 6.10 g ditions, aspartic acid, threonine, serine, glutamic acid, pro-line, glycine, alanine, valine, cystine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, histidine and argi-nine are eluted in this order with the resolutions between the peaks of threonine and serine, glycine and alanine, and isoleucine and leucine being not less than 1.5, 1.4 and 1.2, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of aspartic acid, proline, valine and arginine are not more than 2.0z, respectively. Purity (1) Acetic acid—Weigh accurately about 15 mg of Oxytocin, dissolve in the internal standard solution to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 1 g of acetic acid (100), add the internal standard solution to make exactly 100 mL. Pipet 2 mL of this solution, add the internal standard solu-tion to make exactly 200 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of acetic acid to that of the internal standard: the amount of acetic acid is not less than 6.0z and not more than 10.0z. Trisodium citrate dihydrate 6.19 g 7.74 g 26.67 g Amount (z) of acetic acid (C2H4O2) ＝ MS/MT × QT/QS × 1/10 Sodium chloride 5.66 g 7.07 g 54.35 g Ethanol (99.5) 260.0 mL 20.0 mL — Benzyl alcohol — — 5.0 mL Thiodiglycol 5.0 mL 5.0 mL — MS: Amount (mg) of acetic acid (100) taken MT: Amount (mg) of Oxytocin taken Internal standard solution—A solution of propionic acid in Lauromacrogol solution (1 in 4) 4.0 mL 4.0 mL 4.0 mL the mobile phase (1 in 10,000). Capryric acid 0.1 mL 0.1 mL 0.1 mL Water a sufficient a sufficient a sufficient amount amount amount Total amount 2000 mL 1000 mL 1000 mL Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B and C as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz) 0 – 9 9 – 25 25 – 61 100 → 0 0 → 100 61 – 80 Reaction reagent: Mix 407 g of lithium acetate dihydrate, 245 mL of acetic acid (100) and 801 mL of 1-methoxy-2-propanol, add water to make 2000 mL, stir for more than 10 minutes while passing Nitrogen, and use this solution as So-lution A. Separately, to 1957 mL of 1-methoxy-2-propanol add 77 g of ninhydrin and 0.134 g of sodium borohydride, stir for more than 30 minutes while passing Nitrogen, and use this solution as Solution B. Mix Solution A and Solution B before use. Flow rate of mobile phase: About 0.26 mL per minute. Flow rate of reaction reagent: About 0.3 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 0.7 mL of phosphoric acid add 900 mL of water, adjust the pH to 3.0 with 8 mol/L sodium hydrox-ide TS, and add water to make 1000 mL. To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of acetic acid is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, acetic acid and propionic acid are eluted in this order with the resolution between these peaks being not less than 14. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of acetic acid to that of the internal standard is not more than 2.0z. (2) Related substances—Dissolve 25 mg of Oxytocin in 100 mL of the mobile phase A, and use this solution as the sample solution. Perform the test with 50 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of each peak other than Oxytocin is not more than 1.5z, and the total of them is not more than 5.0z. ‌Operating conditions— Detector, column, column temperature, mobile phase, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of oxytocin. System suitability— Test for required detectability: Measure exactly 1 mL of the sample solution, add the mobile phase A to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suita-bility test, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of oxytocin obtained with 50 mL of this solution is equivalent to 5 to 15z of that with 50 mL of the solution for system suitability test. System performance: Dissolve an adequate amount of oxytocin and vasopressin in the mobile phase A, so that each mL contains about 0.1 mg each of them. When the proce-dure is run with 50 mL of this solution under the above oper-ating conditions, vasopressin and oxytocin are eluted in this order with the resolution between these peaks being not less than 14, and the symmetry factor of the peak of oxytocin is not more than 1.5. System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of oxytocin is not more than 2.0z. Water <2.48> Not more than 5.0z (50 mg, coulometric titration). Assay Weigh accurately an amount of Oxytocin, equiva-lent to about 13,000 Units, dissolve in the mobile phase A to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 1 bottle of the Oxytocin RS in the mobile phase A to make a known concentration solution containing each mL contains about 130 Units, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of oxytocin in each solution. Units per mg of Oxytocin, calculated on the anhydrous and residual acetic acid-free basis ＝ MS/MT × AT/AS × 100 MS: Units per mL of the standard solution MT: Amount (mg) of Oxytocin taken, calculated on the anhydrous and residual acetic acid-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 15.6 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water. Mobile phase B: A mixture of water and acetonitrile (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 30 70 → 40 30 → 60 30 – 30.1 40 → 70 60 → 30 30.1 – 45 Flow rate: 1.0 mL per minute. System suitability— System performance: Dissolve an adequate amount of oxytocin and vasopressin in the mobile phase A, so that each mL contains about 0.1 mg each of them. When the proce-dure is run with 25 mL of this solution under the above oper-ating conditions, vasopressin and oxytocin are eluted in this order with the resolution between these peaks being not less than 14, and the symmetry factor of the peak of oxytocin is not more than 1.5. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of oxytocin is not more than 1.0z.
Containers—Tight containers. Storage—At 2 to 89C.
Pancuronium Bromide occurs as a white crys-talline powder. It is very soluble in water, and freely soluble in ethanol (95) and in acetic anhydride. It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Pancuronium Bromide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of Pancuronium Bromide (1 in 100) re-sponds to Qualitative Tests <1.09> (1) for bromide. D Optical rotation <2.49> [a]20: ＋38 – ＋429(0.75 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution of Pancuronium Bromide (1 in 100) is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Pancuronium Bromide in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve 50 mg of Pancuronium Bromide in 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution (1). Separately, weigh exactly 5 mg of dacuronium bromide for thin-layer chromatography, add ethanol (95) to make exactly 25 mL, and use this solution as the standard solution (2). Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of 2-propanol, aceto-nitrile and a solution of sodium iodide (1 in 5) (17:2:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly a solution of sodium nitrite in methanol (1 in 100) on the plate, allow to stand for 2 minutes, and spray evenly potas-sium bismuth iodide TS on the plate: a spot obtained from the sample solution, corresponding to that from the standard solution (2), has no more color than that from the standard solution (2), and the spots other than the principal spot and the above mentioned spot from the sample solution have no more color than the spot from the standard solution (1). Water <2.48> Not more than 8.0z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Pancuronium Bromide, dissolve in 50 mL of acetic anhydride by warming, and titrate <2.50> with 0.1 mol/L perchloric acid VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.63 mg of C35H60Br2N2O4
Panipenem contains not less than 900 mg (potency) and not more than 1010 mg (potency) per mg, calcu-lated on the anhydrous and residual solvent-free basis.
Panipenem occurs as a white to light yellow, crystalline powder or mass. It is very soluble in water, freely soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in diethyl ether. It is hygroscopic. It deliquesces in the presence of moisture.
(1) Dissolve 20 mg of Panipenem in 2 mL of water, add 1 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 3 minutes, add 1 mL of acidic ammo-nium iron (III) sulfate TS, and shake: a red-brown color de-velops. Determine the absorption spectrum of a solution of Panipenem in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Panipenem as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋55 – ＋659(0.1 g calculated on the anhydrous and residual solvent-free basis, 0.1 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0), 10 mL, 100 mm). pH <2.54> Dissolve 0.5 g of Panipenem in 10 mL of water: the pH of the solution is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.30 g of Panipenem in 40 mL of water, and observe immediately: the solution is clear and its absorbance at 400 nm determined as directed under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.4. Heavy metals <1.07>—Proceed with 1.0 g of Panipen-em according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Keep the sample solution at 59C or below. Dissolve 50 mg of Panipenem in 50 mL of water, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area per-centage method: the amount of the peak other than panipen-em is not more than 2.0z, and the total amount of the peaks other than panipenem is not more than 6.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized porous glass for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 3.12 g of sodium dihydrogen phosphate dihydrate in 700 mL of water, adjust to pH 8.0 with dilute sodium hydroxide TS, then add water to make 1000 mL, and add 20 mL of acetonitrile. Mobile phase B: Dissolve 3.12 g of sodium dihydrogen phosphate dihydrate in 700 mL of water, adjust to pH 8.0 with dilute sodium hydroxide TS, then add water to make 1000 mL. To 750 mL of this solution add 250 mL of aceto-nitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 15 – 50 100 → 0 0 → 100 Flow rate: 1.0 mL per minute (the retention time of panipenem is about 16 minutes). Time span of measurement: For 50 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Use a solution of Panipen-em (1 in 100,000) as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add water to make exactly 10 mL. Confirm that the peak area of panipenem obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above conditions, the number of theoretical plates and the symme-try factor of the peak of panipenem are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above conditions, the relative standard deviation of the peak area of panipenem is not more than 2.0z. Water Weigh accurately about 0.5 g of Panipenem, trans-fer to a 15-mL narrow-mouthed cylindrical glass bottle, add exactly 2 mL of the internal standard solution to dissolve, seal tightly a rubber stopper with aluminum cap, and use this solution as the sample solution. Separately, weigh accurately 2 g of water, and add the internal standard solution to make exactly 100 mL. Pipet 5 mL and 10 mL of this solution, add the internal standard solution to make exactly 20 mL, and use these solutions as the standard solution (1) and the stand-ard solution (2). Perform the test with 1 mL of the sample so-lution and standard solutions (1) and (2) as directed under Gas Chromatography <2.02> according to the following con-dition, and calculate the ratios, QT, QS1 and QS2 of the peak area of water to that of the internal standard. Calculate the amount of water by the following formula: water is not more than 5.0z. Amount of water (z) ＝ MS/MT × (QT ＋ QS2 － 2QS1)/2(QS2 － QS1) × 1/100 × 100 MS: Amount (g) of water taken MT: Amount (g) of Panipenem taken Internal standard solution—A solution of acetonitrile in methanol (1 in 100). Operating conditions— Detector: A thermal conductivity detector. Column: A glass column 3 mm in inside diameter and 2 m in length, packed with porous ethyl vinylbenzene-divinylben-zene copolymer for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature of about 1259C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of acetonitrile is about 8 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution (2) under the above operating condi-tions, water, methanol, and the internal standard are eluted in this order with the resolution between the peaks of water and internal standard being not less than 10. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution (2) under the above oper-ating conditions, the relative standard deviation of the ratios of the peak area of water to that of the internal standard is not more than 5.0z. Residue on ignition <2.44> Not more than 0.5z (1 g).
Conduct this procedure within 30 minutes after preparation of the sample and standard solutions. Weigh accurately an amount of Panipenem and Panipenem RS, equivalent to about 0.1 g (potency), dissolve them separately in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 100 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL of the sample solu-tion and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of panipenem to that of the internal standard. (III) sulfate TS, and shake: a red-brown color develops (panipenem). Amount [mg (potency)] of panipenem (C ＝ MS × QT/QS × 1000 15H21N3O4S) (2) Powder Panipenem and Betamipron for Injection. Dissolve a portion of the powder, equivalent to 50 mg of Be-tamipron, in 4 mL of diluted methanol (1 in 2), and use this MS: Amount [mg (potency)] of Panipenem RS taken ‌Internal standard solution—A solution of sodium p-styrenesulfonate in 0.02 mol/L 3-(N-morpholino)pro-panesulfonic acid buffer solution (pH 7.0) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized sili-cone polymer coated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.02 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 8.0) and acetonitrile (50:1). Flow rate: Adjust so that the retention time of the internal standard is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, panipenem and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of panipenem to that of the internal stand-ard is not more than 2.0z.
Containers—Tight containers. Storage—At a temperature not exceeding －109C.
Pantethine is an aqueous solution containing 80z of pantethine. Pantethine contains not less than 98.
Pantethine is a clear, colorless to pale yellow viscous liquid. It is miscible with water, with methanol and with ethanol (95). It is decomposed by light.
(1) To 0.7 g of Pantethine add 5 mL of so-dium hydroxide TS, shake, and add 1 to 2 drops of copper (II) sulfate TS: a blue-purple color develops. To 0.7 g of Pantethine add 3 mL of water, shake, add 0.1 g of zinc powder and 2 mL of acetic acid (100), and boil for 2 to 3 minutes. After cooling, add 1 to 2 drops of so-dium pentacyanonitrosylferrate (III) TS: a red-purple color develops. To 1.0 g of Pantethine add 500 mL of water, and shake. To 5 mL of this solution add 3 mL of 1 mol/L hydro-chloric acid TS, and heat on a water bath for 30 minutes. After cooling, add 7 mL of a solution of hydroxylammo-nium chloride in sodium hydroxide TS (3 in 140), and allow to stand for 5 minutes. Add 3 drops of 2,4-dinitrophenol TS, and add 1 mol/L hydrochloric acid TS dropwise until the so-lution has no color, and then add 1 mL of iron (III) chloride TS: a red-purple color develops. D Optical rotation <2.49> [a]20: ＋15.0 – ＋18.09(1 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Pantethine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Pantethine according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.6 g of Pantethine in 10 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with 2-buta-none saturated with water to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand for about 10 minutes in iodide vapor: the spots other than the principal spot obtained from the sample solution are not more intense Each mL of 0.05 mol/L bromine VS ＝ 5.547 mg of C22H42N4O8S2
Weigh accurately about 0.3 g of Pantethine, add water to make exactly 20 mL. Transfer exactly 5 mL of this solution in an iodine bottle, and add exactly 25 mL of 0.05 mol/L bromine VS and 100 mL of water. Add 5 mL of di-luted sulfuric acid (1 in 5) rapidly, stopper tightly immedi-ately, and warm at 40 to 509C for 15 minutes with occa-sional shaking. After cooling, carefully add 5 mL of a solu-tion of potassium iodide (2 in 5), then immediately stopper tightly, shake, add 100 mL of water and titrate <2.50> the lib-erated iodine with 0.1 mol/L sodium thiosulfate VS (indica- tethine (C basis. 22H42N4O8S2), calculated on the anhydrous tor: 2 mL of starch TS). Perform a blank determination in the same manner. Description Pantethine is a clear, colorless to pale yellow viscous liquid. It is miscible with water, with methanol and with ethanol (95). It is decomposed by light. Identification (1) To 0.7 g of Pantethine add 5 mL of so-dium hydroxide TS, shake, and add 1 to 2 drops of copper (II) sulfate TS: a blue-purple color develops. To 0.7 g of Pantethine add 3 mL of water, shake, add 0.1 g of zinc powder and 2 mL of acetic acid (100), and boil for 2 to 3 minutes. After cooling, add 1 to 2 drops of so-dium pentacyanonitrosylferrate (III) TS: a red-purple color develops. To 1.0 g of Pantethine add 500 mL of water, and shake. To 5 mL of this solution add 3 mL of 1 mol/L hydro-chloric acid TS, and heat on a water bath for 30 minutes. After cooling, add 7 mL of a solution of hydroxylammo-nium chloride in sodium hydroxide TS (3 in 140), and allow to stand for 5 minutes. Add 3 drops of 2,4-dinitrophenol TS, and add 1 mol/L hydrochloric acid TS dropwise until the so-lution has no color, and then add 1 mL of iron (III) chloride TS: a red-purple color develops. D Optical rotation <2.49> [a]20: ＋15.0 – ＋18.09(1 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Pantethine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Pantethine according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.6 g of Pantethine in 10 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with 2-buta-none saturated with water to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand for about 10 minutes in iodide vapor: the spots other than the principal spot obtained from the sample solution are not more intense Each mL of 0.05 mol/L bromine VS ＝ 5.547 mg of C22H42N4O8S2
Papaverine Hydrochloride, when dried, contains not less than 98.
Papaverine Hydrochloride occurs as white crystals or crystalline powder. It is sparingly soluble in water and in acetic acid (100), slightly soluble in ethanol (95), and practically insoluble in acetic anhydride and in diethyl ether. The pH of a solution of 1.0 g of Papaverine Hydrochlo-ride in 50 mL of water is between 3.0 and 4.0.
(1) To 1 mg of Papaverine Hydrochloride add 1 drops of formaldehyde-sulfuric acid TS: a colorless to light yellow-green color is produced, and it gradually changes to deep red, then to brown. Dissolve 0.02 g of Papaverine Hydrochloride in 1 mL of water, and add 3 drops of sodium acetate TS: a white pre-cipitate is produced. Dissolve 1 mg of Papaverine Hydrochloride in 3 mL of acetic anhydride and 5 drops of sulfuric acid, heat in a water bath for 1 minute, and examine under ultraviolet light (main wavelength: 365 nm): the solution shows a yellow-green fluorescence. ‌Dissolve 0.1 g of Papaverine Hydrochloride in 10 mL of water, make alkaline with ammonia TS, and shake with 10 mL of diethyl ether. Draw off the diethyl ether layer, wash with 5 mL of water, and filter. Evaporate the filtrate on a water bath, and dry the residue at 1059C for 3 hours: the residue so obtained melts <2.60> between 1459C and 1489C.‌ Alkalify a solution of Papaverine Hydrochloride (1 in 50) with ammonia TS, and filter the precipitate. Acidify the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 0.10 g of Papaverine Hydrochloride in 10 mL of water: the solution is clear and colorless. Morphine—Dissolve 10 mg of Papaverine Hydro-chloride in 1 mL of water, add 5 mL of 1-nitroso-2-naphthol TS and 2 mL of a solution of potassium nitrate (1 in 10), and warm at 409C for 2 minutes. Add 1 mL of a solution of sodi-um nitrate (1 in 5000), and warm at 409C for 5 minutes. After cooling, shake the mixture with 10 mL of chloroform, centrifuge, and separate the aqueous layer: the solution so obtained has no more color than a pale red color. Readily carbonizable substances <1.15>—Perform the test with 0.12 g of Papaverine Hydrochloride: the solution has no more color than Matching Fluid S or P. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Papaverine Hydro-chloride, previously dried, dissolve in 100 mL of a mixture of acetic anhydride and acetic acid (100) (7:3) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. water to 10 mL, render the solution alkaline with ammonia TS, and shake with 10 mL of diethyl ether. Draw off the diethyl ether layer, wash with 5 mL of water, and filter. Evaporate the filtrate on a water bath to dryness, and dry the residue at 1059C for 3 hours: the residue so obtained melts <2.60> between 1459C and 1489C. Proceed with 1 mg each of the residue obtained in (2) as directed in the Identification (1) and (3) under Papaverine Hydrochloride. Alkalify 2 mL of Papaverine Hydrochloride Injection with ammonia TS, filter the precipitate off, and acidity the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chloride. Bacterial endotoxins <4.01> Less than 6.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Dilute an exactly measured volume of Papaverine Hydrochloride Injection, equivalent to about 0.2 g of papaverine hydrochloride (C20H21NO4.HCl), with water to 10 mL, render the solution alkaline with ammonia TS, and extract with 20-mL, 15-mL, 10-mL and 10-mL portions of chloroform. Combine the extracts, wash with 10 mL of water, and re-extract the washings with two 5-mL portions of chloroform. Combine all the chloroform extracts, and distil the chloroform on a water bath. Dissolve the residue in 30 mL of acetic acid (100), and titrate <2.50> with 0.05 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 18.79 mg of C H NO .HCl 20 21 4 Each mL of 0.1 mol/L perchloric acid VS ＝ 37.59 mg of C20H21NO4.HCl
Acetaminophen occurs as white, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (95), spar-ingly soluble in water, and very slightly, soluble in diethyl ether. It dissolves in sodium hydroxide TS.
Determine the infrared absorption spectra of Acetaminophen, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Acetaminophen RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 4.0 g of Acetamino-phen in 100 mL of water by heating, cool with shaking in ice water, allow to stand until ordinary temperature is attained, add water to make 100 mL, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Sulfate <1.14>—To 25 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Heavy metals <1.07>—Proceed with 2.0 g of Acet-aminophen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Acetaminophen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Acetamino-phen in 1 mL of methanol, add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area of both solutions by the automatic integration method: the total area of all peaks other than acetaminophen obtained from the sample solution is not larger than the peak area of acetaminophen from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogenphosphate (pH 4.7) and methanol (4:1). Flow rate: Adjust so that the retention time of acetamino-phen is about 5 minutes. Selection of column: Dissolve 0.01 g each of Acetamino-phen and 4-aminophenol hydrochloride in 1 mL of metha-nol, add the mobile phase to make 50 mL, to 1 mL of this solution add the mobile phase to make 10 mL. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of 4-aminophenol and acetaminophen in this order with the resolution between these peaks being not less than 7. Detection sensitivity: Adjust the detection sensitivity so that the peak height of acetaminophen obtained from 10 mL ‌JP XVIII Official Monographs / Acetic Acid 403 of the standard solution is about 15z of the full scale. ‌Time span of measurement: About 6 times as long as the retention time of acetaminophen, beginning after the solvent peak. Loss on drying <2.41> Not more than 0.3z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Acetamino-phen and Acetaminophen RS, previously dried, dissolve in 2 mL of methanol, and add water to make exactly 100 mL. Pipet 3 mL each of these solutions, add water to make ex-actly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absor-bances, AT and AS, of the sample solution and standard solution at the wavelength of maximum absorption at about 244 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank. Amount (mg) of acetaminophen (C8H9NO2) ＝ MS × AT/AS MS: Amount (mg) of Acetaminophen RS taken
Paraffin occurs as a colorless or white, more or less transparent, crystalline mass. It is odorless and taste-less. It is sparingly soluble in diethyl ether and practically in-soluble in water, in ethanol (95) and in ethanol (99.5). Specific gravity d 20: about 0.92 (proceed as directed in 4.2. in 4. Specific gravity under Fats and Fatty Oils Test <1.13>).
(1) Heat Paraffin strongly in a porcelain dish, and ignite: it burns with a bright flame and the odor of paraffin vapor is perceptible. (2) Heat 0.5 g of Paraffin with 0.5 g of sulfur with shak-ing carefully: the odor of hydrogen sulfide is perceptible.
(1) Acidity or alkalinity—Boil 10.0 g of Paraffin with 10 mL of hot water and 1 drop of phenolphthalein TS in a water bath for 5 minutes, and shake vigorously: a red color is not produced. Add 0.20 mL of 0.02 mol/L sodium hydroxide VS to this solution, and shake: a red color is pro-duced. Heavy metals <1.07>—Ignite 2.0 g of Paraffin in a ‌crucible, first moderately until charred, then between 4509C and 5509C to ash. Cool, add 2 mL of hydrochloric acid, and evaporate on a water bath to dryness. To the residue add 2 mL of dilute acetic acid and water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Paraffin according to Method 3, and perform the test (not more than 2 ppm). Sulfur compounds—To 4.0 g of Paraffin add 2 mL of ethanol (99.5), further add 2 drops of a clear saturated so-lution of lead (II) oxide in a solution of sodium hydroxide (1 in 5), and heat for 10 minutes at 709C with occasional shak-ing: no dark brown color develops in the aqueous layer. Readily carbonizable substances—Melt 5.0 g of Paraffin placed in a Nessler tube at a temperature near the melting point. Add 5 mL of sulfuric acid for readily car-bonizable substances, and warm at 709C for 5 minutes in a water bath. Remove the tube from the water bath, immedi-ately shake vigorously and vertically for 3 seconds, and warm for 1 minute in a water bath at 709C. Repeat this procedure 5 times: the color of the sulfuric acid layer is not darker than that of the following control solution. Control solution: Add 1.5 mL of Cobalt (II) Chloride CS, 0.5 mL of Copper (II) Sulfate CS and 5 mL of liquid paraffin to 3.0 mL of Iron (III) Chloride CS, and shake vigorously.
Paraformaldehyde contains not less than 95.
Paraformaldehyde occurs as a white powder. It has a slight odor of formaldehyde, but a very strong irritating odor is perceptible when it is heated. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in hot water, in hot dilute hydrochloric acid, in sodium hydroxide TS and in ammonia TS. It sublimes at about 1009C. ‌Identification (1) Dissolve 0.1 g of Paraformaldehyde in 5 mL of ammonia TS, add 5 mL of silver nitrate TS, shake, and add 3 mL of a solution of sodium hydroxide (1 in 10): a mirror of metallic silver is immediately formed on the sides of the container. (2) Add a solution of 0.04 g of salicylic acid in 5 mL of sulfuric acid to 0.02 g of Paraformaldehyde, and warm slowly: a persistent, dark red color is produced.
(1) Clarity and color of solution—Dissolve 0.20 g of Paraformaldehyde in 10 mL of ammonia TS: the solution is clear and colorless. Acidity or alkalinity—To 0.5 g of Paraformaldehyde add 10 mL of water, shake vigorously for 1 minute, and filter: the filtrate is neutral. Chloride <1.03>—Dissolve 1.5 g of Paraformalde- hyde in 75 mL of water and 7.5 mL of sodium carbonate TS, evaporate on a water bath to dryness, and ignite at about 5009C. Dissolve the residue in 15 mL of water, filter, if nec-essary, neutralize with diluted nitric acid (3 in 10), and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydro-chloric acid VS add 7.5 mL of sodium carbonate TS, a volume of diluted nitric acid (3 in 10) required for neutraliza-tion of the sample, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.006z). Sulfate <1.14>—Dissolve 1.5 g of Paraformaldehyde in 45 mL of water and 4.5 mL of sodium carbonate TS, evaporate on a water bath to dryness, and ignite at about 5009C. Dissolve the residue in 15 mL of water, filter, if nec-essary, neutralize the diluted hydrochloric acid (3 in 5), and boil for 5 minutes. After cooling, add 1 mL of dilute hydro-chloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 4.5 mL of sodium carbonate TS add an equal volume of diluted hydrochloric acid (3 in 5) for the neutralization of the sample and 15 mL of water, and boil for 5 minutes. After cooling, add 0.35 mL of 0.005 mol/L sulfuric acid VS, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.011z). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 50 mg of Paraformaldehyde, accu-rately weighed, in 10 mL of potassium hydroxide TS in an iodine flask. Add 40 mL of water and an exactly measured 50 mL of 0.05 mol/L iodine VS, stopper, and allow to stand for 5 minutes. Then add 5 mL of dilute hydrochloric acid, stopper immediately, allow to stand for 15 minutes, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thio-sulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 1.501 mg of CH2O
Paroxetine Hydrochloride Hydrate contains not less than 98.
Paroxetine Hydrochloride Hydrate occurs as a white crystalline powder. It is freely soluble in methanol, soluble in ethanol (99.5), and slightly soluble in water. D Optical rotation [a]20: －83 – －939(0.1 g calculated on the anhydrous basis, ethanol (99.5), 20 mL, 100 mm). Melting point: about 1409C (with decomposition).
(1) Determine the absorption spectrum of a solution of Paroxetine Hydrochloride Hydrate in ethanol (99.5) (1 in 20,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Paroxe-tine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Paroxetine Hydrochloride Hydrate as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Paroxetine Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Paroxetine Hydrochloride Hydrate (1 in 500) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Paroxetine Hydrochloride Hydrate according to Method 4, and perform the test. Use a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 30). Prepare the control so-lution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). 4-(4-Fluorophenyl)-1-methyl-1,2,3,6-tetrahydropyri-dine—Dissolve 0.42 g of Paroxetine Hydrochloride Hydrate in 10 mL of a mixture of water and acetonitrile (4:1), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add a mixture of water and acetonitrile (4:1) to make exactly 100 mL. Pipet 1 mL of this solution, and add a mixture of water and acetonitrile (4:1) to make ex-actly 100 mL. Pipet 2 mL of this solution, add a mixture of water and acetonitrile (4:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 75 mL each of the sample solution and standard solu- tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.8 to paroxetine, obtained from the sample solution is not larger than the peak area of paroxetine from the standard solution. For the area of the peak, having the relative retention time of about 0.8 to paroxetine, multiply the correction factor 0.86. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: Dissolve 30 g of sodium perchlorate monohydrate in 900 mL of water, add 3.5 mL of phosphoric acid, 2.4 mL of triethylamine and water to make 1000 mL, and then adjust to pH 2.0 with phosphoric acid or triethyla-mine. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 20 85 → 80 15 → 20 20 – 27 80 → 55 20 → 45 27 – 36 Flow rate: 1.5 mL per minute. System suitability— System performance: When the procedure is run with 75 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of paroxetine are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 75 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 5.0z. Related substances—Dissolve 20 mg of Paroxetine Hydrochloride Hydrate in 20 mL of a mixture of water and tetrahydrofuran (9:1), and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add a mixture of water and tetrahydrofuran (9:1) to make exactly 100 mL. Pipet 1 mL of this solution, and add a mixture of water and tetrahydrofuran (9:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than paroxetine obtained from the sample solu-tion is not larger than the peak area of paroxetine from the standard solution. For the areas of the peaks, having the rel-ative retention time of about 0.29, about 0.66, about 0.73, about 0.85, about 0.91, about 1.14, about 1.51, and about 1.84 to paroxetine, multiply their correction factors 0.46, 0.82, 1.10, 0.95, 0.93, 0.82, 1.55, and 1.54, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, tetrahydrofuran and trifluoroacetic acid (180:20:1). Mobile phase B: A mixture of acetonitrile, tetrahyrofuran and trifluoroacetic acid (180:20:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. factor of the peak of paroxetine are not less than 500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 2.0z. Water <2.48> 2.0 – 3.0z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 50 mg each of Paroxetine Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Hydrochloride Hydrate and Paroxetine Hydrochloride RS (separately determine the water <2.48> in the same manner as Paroxetine Hydrochloride Hydrate), dissolve them sepa- 0 – 30 80 20 30 – 50 80 → 20 20 → 80 50 – 60 20 80 Flow rate: 1.0 mL per minute. Time span of measurement: For 60 minutes after injec-tion, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of paroxetine are not less than 5000 and not more than 2.0, respectively. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 2.0z. Enantiomer—Dissolve 0.10 g of Paroxetine Hydro-chloride Hydrate in 20 mL of methanol, add a solution of sodium chloride (29 in 1000) to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add 10 mL of methanol, and add a solution of so-dium chloride (29 in 1000) to make exactly 50 mL. Pipet 2 mL of this solution, add 4 mL of methanol, add a solution of sodium chloride (29 in 1000) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak of the enantiomer, having the relative retention time of about 0.4 to paroxetine, obtained from the sample solution is not larger than the peak area of paroxetine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4 mm in inside diameter and 10 cm in length, packed with a1-acid glycoprotein-bind-ing silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 189C. Mobile phase: A mixture of sodium chloride solution (29 in 1000) and methanol (4:1). Flow rate: Adjust so that the retention time of paroxetine is about 22 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry rately in water to make exactly 100 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of paroxe-tine in each solution. Amount (mg) of paroxetine hydrochloride (C19H20FNO3.HCl) ＝ MS × AT/AS MS: Amount (mg) of Paroxetine Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with trimethylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 1000 mL of water, and adjust to pH 4.5 with acetic acid (100). To 600 mL of this solution, add 400 mL of acetonitrile and 10 mL of triethylamine, then adjust to pH5.5 with acetic acid (100). Flow rate: Adjust so that the retention time of paroxetine is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of paroxetine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 1.0z.
Pazufloxacin Mesilate occurs as a white to light yellow crystalline powder. It is freely soluble in water, and slightly soluble in ethanol (99.5). It dissolves in sodium hydroxide TS. The pH of a solution prepared by dissolving 0.4 g of Pazufloxacin Mesilate in 10 mL of water is between 3.0 and 4.0. Melting point: about 2589C (with decomposition). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Pazufloxacin Mesilate in a mixture of methanol and 1 mol/L hydrochloric acid TS (49:1) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pazufloxacin Mesilate RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Pazufloxacin Mesilate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Pazufloxacin Mesilate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Pazufloxacin Mesilate responds to Qualitative Tests <1.09> for mesilate. D Optical rotation <2.49> [a]20: －61 – －659(after drying, 0.2 g, sodium hydroxide TS, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Pazufloxacin Mesilate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 26 mg of Pazufloxacin Mesilate in 100 mL of the mobile phase, and use this solu-tion as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the amount of the peak other than pazufloxacin is not more than 0.10z. For the area of the peak, having the relative retention time of about 2.7 to pazufloxacin, multiply the correction factor, 1.6. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.08 g of sodium 1-octanesul-fonate in 1000 mL of a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (39:11). Flow rate: Adjust so that the retention time of pazufloxa-cin is about 8 minutes. Time span of measurement: About 6 times as long as the retention time of pazufloxacin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of pazufloxacin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pazufloxacin are not less than 2500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of pazufloxacin is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.2 g, platinum crucible).
Weigh accurately about 26 mg each of Pazufloxacin Mesilate and Pazufloxacin Mesilate RS, both previously dried, dissolve each in water to make exactly 100 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of pazufloxacin to that of the internal standard. Amount (mg) of pazufloxacin mesilate (C16H15FN2O4.CH4O3S) ＝ MS × QT/QS MS: Amount (mg) of Pazufloxacin Mesilate RS taken Internal standard solution—A solution of acetanilide in the mobile phase (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 200 mL of water add gradually 30 mL ‌of methanesulfonic acid while ice-cooling, then add 30 mL of triethylamine in the same manner, and add water to make 300 mL. To 50 mL of this solution add 150 mL of aceto-nitrile, 35 mL of 1 mol/L dipotassium hydrogen phosphate TS for buffer solution and water to make 1000 mL.‌ Flow rate: Adjust so that the retention time of pazufloxa-cin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pazufloxacin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pazufloxacin to that of the internal standard is not more than 1.0z.
Penbutolol Sulfate, when dried, contains not less than 98.
Penbutolol Sulfate occurs as a white crystalline powder. It is very soluble in acetic acid (100), freely soluble in methanol, sparingly soluble in ethanol (95), slightly soluble in water, and practically insoluble in acetic anhydride and in diethyl ether.
(1) Determine the absorption spectrum of a solution of Penbutolol Sulfate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pen-butolol Sulfate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra propanol, ethanol (95) and ammonia solution (28) (85:12:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.8 g of Penbutolol Sulfate, previously dried, dissolve in 50 mL of a mixture of acetic an-hydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 68.09 mg of (C18H29NO2)2.H2SO4
Pentazocine occurs as a white to pale yellow-white, crystalline powder. It is odorless. It is freely soluble in acetic acid (100) and in chloroform, soluble in ethanol (95), sparingly soluble in diethyl ether and practically insoluble in water.
(1) To 1 mg of Pentazocine add 0.5 mL of formaldehyde-sulfuric acid TS: a deep red color is produced, and it changes to grayish brown immediately. Dissolve 5 mg of Pentazocine in 5 mL of sulfuric acid, add 1 drop of iron (III) chloride TS, and heat in a water bath for 2 minutes: the color of the solution changes from light yellow to deep yellow. Shake the solution with 1 drop of nitric acid: the solution remains yellow in color. Determine the absorption spectrum of a solution of Pentazocine in 0.01 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.
Weigh accurately about 0.5 g of Pentazocine, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS (2RS,6RS,11RS )-6,11-Dimethyl- 3-(3-methylbut-2-en-1-yl)-1,2,3,4,5,6-hexahydro- ＝ 28.54 mg of C 19H 27NO 2,6-methano-3-benzoazocin-8-ol [359-83-1] Pentazocine, when dried, contains not less than 99.0z of pentazocine (C19H27NO). Description Pentazocine occurs as a white to pale yellow-white, crystalline powder. It is odorless. It is freely soluble in acetic acid (100) and in chloroform, soluble in ethanol (95), sparingly soluble in diethyl ether and practically insoluble in water. Identification (1) To 1 mg of Pentazocine add 0.5 mL of formaldehyde-sulfuric acid TS: a deep red color is produced, and it changes to grayish brown immediately. Dissolve 5 mg of Pentazocine in 5 mL of sulfuric acid, add 1 drop of iron (III) chloride TS, and heat in a water bath for 2 minutes: the color of the solution changes from light yellow to deep yellow. Shake the solution with 1 drop of nitric acid: the solution remains yellow in color. Determine the absorption spectrum of a solution of Pentazocine in 0.01 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.
Pentoxyverine Citrate, when dried, contains not less than 98.
Pentoxyverine Citrate occurs as a white crys-talline powder. It is very soluble in acetic acid (100), freely soluble in water and in ethanol (95), and practically insoluble in diethyl ether.
(1) Dissolve 0.1 g of Pentoxyverine Citrate in 10 mL of water, and add 10 mL of Reinecke salt TS: a light red precipitate is formed. Determine the infrared absorption spectrum of Pen- ‌toxyverine Citrate, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Pentoxyverine Citrate (1 in 10) re-sponds to Qualitative Tests <1.09> (1) and (2) for citrate.
(1) Clarity and color of solution—Dissolve 1.0 g of Pentoxyverine Citrate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Pentox- yverine Citrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pentoxyverine Citrate according to Method 3, and per-form the test (not more than 2 ppm). Peplomycin Sulfate
Peplomycin Sulfate is the sulfate of a substance having antitumor activity produced by the growth of Streptomyces verticillus. It contains not less than 865 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the dried basis.
Peplomycin Sulfate occurs as a white to light yellow-white powder. It is freely soluble in water, and practically insoluble in ethanol (95). It is hygroscopic.
(1) To 4 mg of Peplomycin Sulfate add 5 mL of copper (II) sulfate TS, and dissolve in water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Peplomycin Sulfate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. ＝ 52.56 mg of C20H31 NO3.C6H8O7 (2) Determine the infrared absorption spectrum of Peplomycin Sulfate as directed in the paste method under In-
(1) Clarity and color of solution—Dissolve 80 mg of Peplomycin Sulfate in 4 mL of water: the solution is clear and colorless. Copper—Dissolve exactly 75 mg of Peplomycin Sul-fate in 10 mL of diluted nitric acid (1 in 100), and use this so-lution as the sample solution. Separately, to 5.0 mL of Standard Copper Stock Solution add diluted nitric acid (1 in 100) to make exactly 100 mL. To 3.0 mL of this solution add diluted nitric acid (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions: the absorbance of the sample solu-tion is not more than that of the standard solution (not more than 200 ppm). Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Copper hollow cathode lamp. Wavelength: 324.8 nm. Related substances—Dissolve about 10 mg of Peplomycin Sulfate in 6 mL of water, add 0.5 mL of a solu-tion of copper (II) sulfate pentahydrate (1 in 125), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine the areas of the peaks, appeared after the peak of copper sulfate, by the automatic integration method, and calculate the amounts of them by the area percentage method: the total amount of the peaks other than peplomy-cin is not more than 7.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase stock solution: Dissolve 0.96 g of sodium 1-pentanesulfonate and 1.86 g of disodium dihydrogen ethyl-enediamine tetraacetate dihydrate in 1000 mL of water and 5 mL of acetic acid (100), and adjust the pH to 4.3 with am-monia TS. Mobile phase A: A mixture of mobile phase stock solution and methanol (9:1). Mobile phase B: A mixture of mobile phase stock solution and methanol (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: 1.2 mL per minute. Time span of measurement: As long as 20 minutes after elution of peplomycin, beginning after the peak of copper sulfate. System suitability— Test for required detectability: Measure exactly 1 mL of the sample solution, add water to make exactly 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add water to make exactly 10 mL. Confirm that the peak area of peplomycin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the sample solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of peplomycin are not less than 30,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of peplomycin is not more than 2.0z. Loss on drying <2.41> Not more than 3.0z (60 mg, in vacuum, phosphorus (V) oxide, 609C, 3 hours). Handle the sample avoiding absorption of moisture.
Weigh accurately about 0.5 g of Pentoxyverine Citrate, previously dried, dissolve in 30 mL of acetic acid (100), add 30 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L of perchloric acid VS until the color of the solu-tion changes from purple through blue-green to green (indi-cator: 3 drops of crystal violet TS). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS Description Peplomycin Sulfate occurs as a white to light yellow-white powder. It is freely soluble in water, and practically insoluble in ethanol (95). It is hygroscopic. Identification (1) To 4 mg of Peplomycin Sulfate add 5 mL of copper (II) sulfate TS, and dissolve in water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Peplomycin Sulfate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. ＝ 52.56 mg of C20H31 NO3.C6H8O7 (2) Determine the infrared absorption spectrum of Peplomycin Sulfate as directed in the paste method under In-
Perphenazine Maleate, when dried, contains not less than 98.
Perphenazine Maleate occurs as a white to light yellow powder. It is odorless. It is sparingly soluble in acetic acid (100), slightly soluble in water and in ethanol (95), and practically insoluble in chloroform. It dissolves in dilute hydrochloric acid. It is gradually colored by light. Melting point: about 1759C (with decomposition).
(1) Dissolve 8 mg of Perphenazine Maleate in 5 mL of sulfuric acid: a red color is produced, which becomes deep red-purple on warming. Dissolve 0.3 g of Perphenazine Maleate in 3 mL of dilute hydrochloric acid, add 2 mL of water and 3 mL of am-monia solution (28), shake, and extract with three 10-mL portions of chloroform. [Reserve the aqueous layer, and use for test (5)]. Evaporate the combined chloroform extracts on a water bath to dryness, dissolve the residue in 20 mL of methanol, and pour into 10 mL of a warm solution of 2,4,6-trinitrophenol in methanol (1 in 25). Allow to stand for 4 hours, collect the crystals, wash with a small amount of methanol, and dry at 1059C for 1 hour: the crystals melt <2.60> between 2379C and 2449C (with decomposition). Determine the absorption spectrum of a solution of Perphenazine Maleate (1 in 20,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Separately, to 10 mL of the solution add 30 mL of water. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. Perform the test with Perphenazine Maleate as di-rected under Flame Coloration Test <1.04> (2): a green color appears. Evaporate the aqueous layer reserved in (2) to dry-ness. To the residue add 1 mL of dilute sulfuric acid and 5 mL of water, and extract with four 25-mL portions of diethyl ether. Combine the diethyl ether extracts, and evapo-rate in a water bath at about 359C with the aid of a current of air: the residue melts <2.60> between 1289C and 1369C.
(1) Heavy metals <1.07>—Proceed with 2.0 g of perphenazine maleate according to Method 2, and perform ‌the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of Perphenazine Maleate according to Method 3, and per-form the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Perphenazine Male-ate, previously dried, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.81 mg of C H ClN OS.2C H O fades immediately. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Disintegrate 1 tablet of Perphenazine Maleate Tablets by shaking with 15 mL of 0.1 mol/L hydrochloric acid TS, shake vigorously with 50 mL of methanol, add water to make exactly 100 mL, and centrifuge. Pipet V mL of the supernatant liquid, add water to make exactly V? mL of a solution containing about 6 mg of perphenazine maleate (C21H26ClN3OS.2C4H4O4) in each ml, and use this solution as the sample solution. Separately, weigh accurately 30 mg of perphenazine maleate for assay, previously dried at 1059C for 3 hours, dissolve in 15 mL of 0.1 mol/L hydrochloric acid TS and 50 mL of methanol, and add water to make ex-actly 100 mL. Pipet 5 mL of this solution, add 3 mL of 0.1 mol/L hydrochloric acid TS, 10 mL of methanol and water to make exactly 250 mL, and use this solution as the stand- 21 26 3 4 4 4 ard solution. Determine the absorbances, AT and AS, of the
Perphenazine, when dried, contains not less than 98.
Perphenazine occurs as white to light yellow, crystals or crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in methanol and in ethanol (95), soluble in acetic acid (100), sparingly soluble in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid. It is gradually colored by light.
(1) Dissolve 5 mg of Perphenazine in 5 mL of sulfuric acid: a red color, changing to deep red-purple upon warming, is produced. Dissolve 0.2 g of Perphenazine in 2 mL of methanol, add this solution to 10 mL of a warm solution of 2,4,6-trinitrophenol in methanol (1 in 25), and allow to stand for 4 hours. Collect the crystals, wash with a small volume of methanol, and dry at 1059C for 1 hour: the crystals so ob-tained melt <2.60> between 2379C and 2449C (with decom-position). Determine the absorption spectrum of a solution of Perphenazine in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum 1 or the spectrum of a solution of Perphena-zine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, to 10 mL of the solution add 10 mL of water. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum 2 or the spectrum of a solution of Perphena-zine RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Perform the test with Perphenazine as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Perphenazine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Perform the test in the current of nitrogen in light-resistant containers under the protection from light. Dissolve 0.10 g of Perphenazine in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add ethanol (95) to make exactly 10 mL. Pipet 1 mL of this solution, add ethanol (95) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol and 1 mol/L ammonia TS (5:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): any spot other than the principal spot obtained from the sample solution is not more intense than that from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 659C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Perphenazine, pre-viously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue-pur-ple to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.20 mg of C21H26ClN3OS
Pethidine Hydrochloride occurs as a white crystalline powder. It is very soluble in water and in acetic acid (100), freely soluble in ethanol (95), sparingly soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution dissolved 1.0 g of Pethidine Hydro-chloride in 20 mL of water is between 3.8 and 5.8.
(1) Clarity and color of solution—Dissolve 1.0 g of Pethidine Hydrochloride in 10 mL of water: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.20 g of Pethidine Hydrochloride. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.240z). Related substances—Dissolve 0.05 g of Pethidine Hy-drochloride in 20 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area obtained from both solutions by the automatic integra-tion method: the total area of the peaks other than pethidine obtained from the sample solution is not larger than the peak area of perthidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 257 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.0 g of sodium lauryl sulfate in 1000 mL of diluted phosphoric acid (1 in 1000), adjust the pH to 3.0 with sodium hydroxide TS, and to 550 mL of this solution add 450 mL of acetonitrile. Flow rate: Adjust so that the retention time of pethidine is about 7 minutes. Time span of measurement: About 2 times as long as the retention time of pethidine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of pethidine obtained with 20 mL of this solution is equivalent to 5 to 15z of that with 20 mL of the standard solution. System performance: To 2 mL each of the sample solution and a solution of isoamyl parahydroxybenzoate in the mo-bile phase (1 in 50,000) add the mobile phase to make 10 mL. When the procedure is run with 20 mL of this solution ac-cording to the above operating conditions, pethidine and isoamyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌area of pethidine is not more than 2.0z.‌ Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.5 g of Pethidine Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.38 mg of C15H21NO2.HCl
Petroleum Benzin occurs as a colorless, clear, volatile liquid. It shows no fluorescence. It has a chracteris-tic odor. It is miscible with ethanol (99.5) and with diethyl ether. It is practically insoluble in water. It is very flammable. Specific gravity d 20: 0.65 – 0.71
(1) Acid—Shake vigorously 10 mL of Petroleum Benzin with 5 mL of water for 2 minutes, and allow to stand: the separated aqueous layer does not change moistened blue litmus paper to red. Sulfur compounds and reducing substances—To 10 mL of Petroleum Benzin add 2.5 mL of ammonia-ethanol TS and 2 to 3 drops of silver nitrate TS, and warm the mix-ture at about 509C for 5 minutes, protected from light: no brown color develops. Fatty oil and sulfur compounds—Drop and evapo-rate 10 mL of Petroleum Benzin in small portions on odor-less filter paper spread on a previously warmed glass plate: no spot or no foreign odor is perceptible. Benzene—Warm 5 drops of Petroleum Benzin with 2 mL of sulfuric acid and 0.5 mL of nitric acid for about 10 minutes, allow to stand for 30 minutes, transfer the mixture to a porcelain dish, and dilute with water: no odor of nitrobenzene is perceptible. Residue on evaporation—Evaporate 140 mL of Petroleum Benzin on a water bath to dryness, and heat the residue at 1059C to constant mass: the mass is not more than 1 mg. Readily carbonizable substances—Shake vigorously 5 mL of Petroleum Benzin with 5 mL of sulfuric acid for rea-dily carbonizable substances for 5 minutes in a Nessler tube, and allow to stand: the sulfuric acid layer has no more color than Matching Fluid A. Distilling range <2.57> 50 – 809C, not less than 90 volz.
Containers—Tight containers. Storage—Remote from fire, and not exceeding 309C.
Antipyrine occurs as colorless or white crys-tals, or a white, crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in water, freely soluble in ethanol (95), and sparingly soluble in diethyl ether. A solution of Antipyrine (1 in 10) is neutral.
(1) To 5 mL of a solution of Antipyrine (1 in 100) add 2 drops of sodium nitrite TS and 1 mL of dilute sulfuric acid: a deep green color develops. To 2 mL of a solution of Antipyrine (1 in 100) add 4 drops of dilute iron (III) chloride TS: a yellow-red color de-velops. Then add 10 drops of dilute sulfuric acid: the color changes to light yellow. To 5 mL of a solution of Antipyrine (1 in 100) add 2 to 3 drops of tannic acid TS: a white precipitate is produced. To 0.1 g of Antipyrine add 0.1 g of vanillin, 5 mL of water and 2 mL of sulfuric acid, boil the mixture, and cool: a yellow-red precipitate is produced.
(1) Chloride <1.03>—Perform the test with 1.0 g of Antipyrine. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Antipy-rine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Readily carbonizable substances<1.15>—Perform the test with 0.5 g of Antipyrine: the solution remains colorless. Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Dissolve about 0.2 g of Antipyrine, previously dried and accurately weighed, in 20 mL of sodium acetate TS, add exactly 30 mL of 0.05 mol/L iodine VS, and allow to stand for 20 minutes with occasional shaking. Dissolve the precipi-tate in 10 mL of chloroform, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 9.412 mg of C11H12N2O
Phenobarbital, when dried, contains not less than 99.
Phenobarbital occurs as white, crystals or crys-talline powder. It is very soluble in N,N-dimethylformamide, freely solu-ble in ethanol (95) and in acetone, sparingly soluble in aceto-nitrile, and very slightly soluble in water. It dissolves in sodium hydroxide TS. The pH of a saturated solution of Phenobarbital is be-tween 5.0 and 6.0.
(1) Determine the absorption spectrum of a solution of Phenobarbital in boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.6) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Phenobarbital as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Phenobarbital in 5 mL of sodium hydroxide TS: the solu-tion is clear and colorless. Chloride <1.03>—Dissolve 0.30 g of Phenobarbital in 20 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: Amount (unit) of phenethicillin potassium (C ＝ MS × VT/VS 17H19KN2O5S) take 0.30 mL of 0.01 mol/L hydrochloric acid VS, 20 mL of acetone and 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.035z). MS: Amount (unit) of Phenethicillin Potassium RS taken
Weigh accurately about 0.5 g of Phenobarbital, pre-viously dried, dissolve in 50 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until the color of the solution change from yel-low to yellow-green (indicator: 1 mL of alizarin yellow GG-thymolphthalein TS). Perform a blank determination using a mixture of 50 mL of N,N-dimethylformamide and 22 mL of ethanol (95) in the same manner, and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 23.22 mg of C12H12N2O3
Phenol for Disinfection contains not less than 95.
Phenol for Disinfection occurs as colorless to slightly, red crystals, crystalline masses, or liquid containing ‌these crystals. It has a characteristic odor.‌‌ It is very soluble in ethanol (95) and in diethyl ether, and freely soluble in water. Phenol for Disinfection (10 g) is liquefied by addition of 1 mL of water. It cauterizes the skin, turning it white. Congealing point: about 309C.
(1) To 10 mL of a solution of Phenol for Disinfection (1 in 100) add 1 drop of iron (III) chloride TS: a blue-purple color is produced. (2) To 5 mL of a solution of Phenol for Disinfection (1 in 10,000) add bromine TS dropwise: a white precipitate is formed, and it dissolves at first upon shaking but becomes permanent as excess of the reagent is added.
(1) Clarity of solution—Dissolve 1.0 g of Phenol for Disinfection in 15 mL of water: the solution is clear. (2) Residue on evaporation—Weigh accurately about 5 g of Phenol for Disinfection, evaporate on a water bath, and stopper immediately, shake for 30 minutes, and allow to stand for 15 minutes. Add 7 mL of potassium iodide TS, stopper tightly at once, shake well, and titrate <2.50> the lib-erated iodine with 0.1 mol/L sodium thiosulfate VS (indica-tor: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 1.569 mg of C6H6O
Phenol [108-95-2] Phenol contains not less than 98.
Phenol occurs as colorless to slightly red, crys-tals or crystalline masses. It has a characteristic odor. It is very soluble in ethanol (95) and in diethyl ether, and soluble in water. Phenol (10 g) is liquefied by addition of 1 mL of water. The color changes gradually through red to dark red by light or air. It cauterizes the skin, turning it white. Congealing point: about 409C
(1) Add 1 drop of iron (III) chloride TS to 10 mL of a solution of Phenol (1 in 100): a blue-purple color develops. (2) Add bromine TS dropwise to 5 mL of a solution of Phenol (1 in 10,000): a white precipitate is produced, which at first dissolves with shaking, but becomes permanent as excess of the reagent is added.
(1) Clarity and color of solution and acidity or alkalinity—Dissolve 1.0 g of Phenol in 15 mL of water: the solution is clear, and neutral or only faintly acid. Add 2 drops of methyl orange TS: no red color develops. (2) Residue on evaporation—Weigh accurately about 5 g of Phenol, evaporate on a water bath, and dry the residue at 1059C for 1 hour: the mass is not more than 0.05z of the mass of the sample.
Dissolve about 1.5 g of Phenol, accurately weighed, in water to make exactly 1000 mL. Transfer exactly 25 mL of this solution to an iodine flask, add exactly 30 mL of 0.05 mol/L bromine VS, then 5 mL of hydrochloric acid, and im- Liquefied Phenol
Phenolated Water for Disinfection contains not less than 2.
Dissolve about 1 g of Phenol for Disinfection, accu-rately weighed, in water to make exactly 1000 mL. Pipet 25 mL of the solution into an iodine flask, add exactly 30 mL of 0.05 mol/L bromine VS and 5 mL of hydrochloric acid, stopper immediately, shake for 30 minutes and allow to stand for 15 minutes. Add 7 mL of potassium iodide TS, stopper immediately, shake well, and titrate <2.50> the liber-ated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 1.569 mg of C6H6O
Phenolated Water contains not less than 1.
Phenolated Water is a colorless, clear liquid, having the odor of phenol.
(1) Add 1 drop of iron (III) chloride TS to 10 mL of Phenolated Water: a blue-purple color develops. (2) To 5 mL of a solution of Phenolated Water (1 in 200) add bromine TS dropwise: a white precipitate is formed, and it dissolves at first upon shaking but becomes permanent as excess of the reagent is added.
Take exactly 2 mL of Phenolated Water into an iodine flask, add 25 mL of water, then add exactly 40 mL of 0.05 mol/L bromine VS and 5 mL of hydrochloric acid, Method of preparation Phenol for Disinfection 31 g Water, Purified Water or Purified Water in Containers a sufficient quantity To make 1000 mL Mix the above ingredients. Description Phenolated Water for Disinfection is a clear, colorless liquid, having the odor of phenol. Identification (1) Add 1 drop of iron (III) chloride TS to 10 mL of Phenolated Water for Disinfection: a blue-purple color develops. (2) Proceed with 5 mL of a solution of Phenolated Water for Disinfection (1 in 200) as directed in the Identifi-cation (2) under Phenol for Disinfection. Assay Take exactly 5 mL of Phenolated Water for Disin-fection, add water to make exactly 100 mL, then pipet 25 mL of the solution into an iodine flask, and proceed as directed in the Assay under Phenol for Disinfection. Each mL of 0.05 mol/L bromine VS ＝ 1.569 mg of C6H6O
Phenolsulfonphthalein, when dried, contains not less than 98.
Phenolsulfonphthalein occurs as a vivid red to dark red crystalline powder. It is very slightly soluble in water and in ethanol (95). It dissolves in sodium hydroxide TS.
(1) Dissolve 5 mg of Phenolsulfonphtha-lein in 2 to 3 drops of sodium hydroxide TS, add 2 mL of 0.05 mol/L bromine VS and 1 mL of dilute sulfuric acid, shake well, and allow to stand for 5 minutes. Render the so-lution alkaline with sodium hydroxide TS: a deep blue-pur-ple color develops. (2) Dissolve 0.01 g of Phenolsulfonphthalein in diluted sodium carbonate TS (1 in 10) to make 200 mL. To 5 mL of this solution add diluted sodium carbonate TS (1 in 10) to make 100 mL. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Insoluble substances—To about 1 g of Phenol-sulfonphthalein, accurately weighed, add 20 mL of a solu-tion of sodium hydrogen carbonate (1 in 40). Allow the mix-ture to stand for 1 hour with frequent shaking, dilute with water to 100 mL, and allow to stand for 24 hours. Collect the insoluble substances using a tared glass filter (G4), wash with 25 mL of a solution of sodium hydrogen carbonate (1 in 100) and with five 5-mL portions of water, and dry at 1059C for 1 hour: the mass of the residue is not more than 0.2z. (2) Related substances—Dissolve 0.10 g of Phenolsul-fonphthalein in 5 mL of dilute sodium hydroxide TS, and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of t-amyl alcohol, acetic acid (100) and water (4:1:1) to a distance of about 15 cm, and air-dry the plate. After allowing the plate to stand in an ammonia vapor, examine under ultraviolet light (main wave-length: 254 nm): the spots other than the principal spot ob-tained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). ‌Assay Weigh accurately about 0.15 g of Phenolsul-fonphthalein, previously dried, transfer to an iodine flask, dissolve in 30 mL of a solution of sodium hydroxide (1 in 250), and add water to make 200 mL. Add exactly measured 50 mL of 0.05 mol/L bromine VS, add 10 mL of hydrochlo-ric acid to the solution quickly, and stopper immediately. Allow the mixture to stand for 5 minutes with occasional shaking, add 7 mL of potassium iodide TS, stopper again immediately, and shake gently for 1 minute. Titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determina-tion in the same manner.‌ Each mL of 0.05 mol/L bromine VS ＝ 4.430 mg of C H O S add a solution of anhydrous sodium carbonate (1 in 100) to make exactly 250 mL. Pipet 5 mL of this solution, add a so-lution of anhydrous sodium carbonate (1 in 100) to make ex-actly 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of phenolsul-fonphthalein for assay, previously dried in a desiccator (sili-ca gel) for 4 hours, and dissolve in a solution of anhydrous sodium carbonate (1 in 100) to make exactly 250 mL. Pipet 5 mL of this solution, add a solution of anhydrous sodium carbonate (1 in 100) to make exactly 200 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 559 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. 19 14 5
Phenylbutazone, when dried, contains not less than 99.
Phenylbutazone occurs as a white to pale yel-low-white crystalline powder. It is odorless, and is at first tasteless but leaves a slightly bitter aftertaste. It is freely soluble in acetone, soluble in ethanol (95) and in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) To 0.1 g of Phenylbutazone add 1 mL of acetic acid (100) and 1 mL of hydrochloric acid, and heat on a water bath under a reflux condenser for 30 minutes. Add 10 mL of water, and cool with ice water. Filter, and to the filtrate add 3 to 4 drops of sodium nitrite TS. To 1 mL of this solution add 1 mL of 2-naphthol TS and 3 mL of chlo-roform, and shake: a deep red color develops in the chlo-roform layer. (2) Dissolve 1 mg of Phenylbutazone in 10 mL of dilute sodium hydroxide TS, and dilute with water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Clarity of solution—Dissolve 1.0 g of Phenyl-butazone in 20 mL of sodium hydroxide solution (2 in 25), and allow to stand at 25 ± 19C for 3 hours: the solution is clear. Determine the absorbance of this solution at 420 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.05. Heavy metals <1.07>—Proceed with 2.0 g of Phenyl-butazone according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of phenylbutazone, according to Method 3, and perform the test (not more than 2 ppm). Readily carbonizable substances—Dissolve 1.0 g of Phenylbutazone in 20 mL of sulfuric acid, and allow to stand at 25 ± 19C for exactly 30 minutes: the solution is clear. Determine the absorbance of this solution at 420 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.10. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Phenylbutazone, previously dried, dissolve in 25 mL of acetone, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the solu- tion shows a blue color which persists for 15 seconds (indica-tor: 5 drops of bromothymol blue TS). Perform a blank de-termination with a mixture of 25 mL of acetone and 16 mL of water in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 30.84 mg of C19H20N2O2
Phenylephrine Hydrochloride occurs as white crystals or crystalline powder. It is odorless, and has a bitter taste. It is very soluble in water, freely soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Phenylephrine Hydro-chloride in 100 mL of water is 4.5 to 5.5.
(1) To 1 mL of a solution of Phenylephrine Hydrochloride (1 in 100) add 1 drop of copper (II) sulfate TS and 1 mL of a solution of sodium hydroxide (1 in 5): a blue color is produced. To the solution so obtained add 1 mL of diethyl ether, and shake vigorously: no blue color develops in the diethyl ether layer. To 1 mL of a solution of Phenylephrine Hydrochlo-ride (1 in 100) add 1 drop of iron (III) chloride TS: a persis-tent purple color is produced. Dissolve 0.3 g of Phenylephrine Hydrochloride in 3 mL of water, add 1 mL of ammonia TS, and rub the inner side of the test tube with a glass rod: a precipitate is pro-duced. Collect the precipitate, wash with a few drops of ice-cold water, and dry at 1059C for 2 hours: it melts <2.60> be-tween 1709C and 1779C. A solution of Phenylephrine Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. D Optical rotation <2.49> [a]20: －42.0 – －47.59(after dry-ing, 0.5 g, water, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Phenylephrine Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Take 0.5 g of Phenylephrine Hydro- chloride, and perform the test. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Ketone—Dissolve 0.20 g of Phenylephrine Hydro-chloride in 1 mL of water, and add 2 drops of sodium penta-cyanonitrosylferrate (III) TS, 1 mL of sodium hydroxide TS and then 0.6 mL of acetic acid (100): the solution has no more color than the following control solution. Control solution: Prepare as directed above without Phenylephrine Hydrochloride. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.1 g of Phenylephrine Hy-drochloride, previously dried, dissolve in 40 mL of water contained in an iodine flask, add exactly measured 50 mL of 0.05 mol/L bromine VS, then add 5 mL of hydrochloric acid, and immediately stopper tightly. Shake the mixture, and allow to stand for 15 minutes. To this solution add 10 mL of potassium iodide TS carefully, stopper tightly imme-diately, shake thoroughly, allow to stand for 5 minutes, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (indica-tor: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 3.395 mg of C9H13NO2.HCl
Phenytoin, when dried, contains not less than 99.
Phenytoin occurs as a white, crystalline pow-der or granules. It is odorless and tasteless. It is sparingly soluble in ethanol (95) and in acetone, slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS. Melting point: about 2969C (with decomposition).
(1) Dissolve 0.02 g of Phenytoin in 2 mL of ammonia TS, and add 5 mL of silver nitrate TS: a white pre-cipitate is produced. Boil a mixture of 0.01 g of Phenytoin, 1 mL of am-monia TS and 1 mL of water, and add dropwise 2 mL of a mixture prepared from 50 mL of a solution of copper (II) sulfate pentahydrate (1 in 20) and 10 mL of ammonia TS: a red, crystalline precipitate is produced. Heat 0.1 g of Phenytoin with 0.2 g of sodium hydrox-ide, and fuse: the gas evolved turns moistened red litmus paper blue. Add 3 mL of chlorinated lime TS to 0.1 g of Phenytoin, shake for 5 minutes, and dissolve the oily pre-cipitate in 15 mL of hot water. After cooling, add 1 mL of dilute hydrochloric acid dropwise, then add 4 mL of water. Filter the white precipitate thus obtained, wash with water, and press it with dry filter paper to remove the accom-panying water. Dissolve the precipitate with 1 mL of chlo-roform, add 5 mL of diluted ethanol (9 in 10), and rub the inner surface of the flask to produce a white, crystalline pre-cipitate. Collect the precipitate, wash with ethanol (95), and dry: the melting point <2.60> is between 1659C and 1699C.
(1) Clarity and color of solution—Dissolve 0.20 g of Phenytoin in 10 mL of 0.2 mol/L sodium hydroxide VS: the solution is clear and colorless. Then heat the solution: no ‌turbidity is produced. Cool, and mix the solution with 5 mL of acetone: the solution is clear and colorless.‌ Acidity or alkalinity—Shake 2.0 g of Phenytoin with 40 mL of water for 1 minute, filter, and perform the follow-ing tests using this filtrate as the sample solution. To 10 mL of the sample solution add 2 drops of phe-nolphthalein TS: no color develops. Then add 0.15 mL of 0.01 mol/L sodium hydroxide VS: a red color develops. To 10 mL of the sample solution add 0.30 mL of 0.01 mol/L hydrochloric acid VS and 5 drops of methyl red TS: a red to orange color develops. standard solution, and use this solution as the sample solu-tion. Separately, weigh accurately about 25 mg of phenytoin for assay, previously dried at 1059C for 2 hours, and dis-solve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of phenytoin to that of the inter-nal standard. Chloride <1.03>—Dissolve 0.30 g of Phenytoin in 30 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the Amount (mg) of phenytoin (C ＝ MS × QT/QS × 2 15H12N2O2) test solution. Prepare the control solution from 0.60 mL of 0.01 mol/L hydrochloric acid VS, 30 mL of acetone and 6 mL of dilute nitric acid, and add water to 50 mL (not more than 0.071z). Heavy metals <1.07>—Proceed with 1.0 g of Phenytoin according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Phenytoin, previ-ously dried, dissolve in 40 mL of ethanol (95) with the aid of gentle heating, add 0.5 mL of thymolphthalein TS immedi-ately, and titrate with 0.1 mol/L sodium hydroxide VS until a light blue color develops. Then add 1 mL of pyridine, 5 drops of phenolphthalein TS and 25 mL of silver nitrate TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until a light red color, which persists for 1 minute, develops. Each mL of 0.1 mol/L sodium hydroxide VS MS: Amount (mg) of phenytoin for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 25,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 258 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and 0.02 mol/L phosphate buffer solution (pH 3.5) (11:9). Flow rate: Adjust so that the retention time of phenytoin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, phenytoin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. ＝ 25.23 mg of C 15H 12N2O2 System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-
Pilocarpine Hydrochloride, when dried, contains not less than 99.
Pilocarpine Hydrochloride occurs as colorless crystals or white powder. It is odorless, and has a slightly bitter taste. It is very soluble in acetic acid (100), freely soluble in water, in methanol and in ethanol (95), soluble in acetic an-hydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Pilocarpine Hydrochlo- tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia TS (85:14:2) to a distance of about 13 cm, and dry the plate at 1059C for 10 minutes. Cool, and spray evenly bismuth potassium iodide TS on the plate: the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Readily carbonizable substances <1.15>—Take 0.25 g of Pilocarpine Hydrochloride, and perform the test: the so-lution has no more color than Matching Fluid B. Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.5z (0.1 g).
Weigh accurately about 0.5 g of Pilocarpine Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.47 mg of C11H16N2O2.HCl
Pilsicainide Hydrochloride Hydrate contains not less than 99.
Pilsicainide Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is very soluble in acetic acid (100), and freely soluble in water, in methanol and in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS.
(1) Determine the absorption spectrum of a solution of Pilsicainide Hydrochloride Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 2000) as directed under Ultravio- with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilsicainide is not more than 2.0z. Water <2.48> 2.5 – 3.3z (50 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Pilsicainide Hydrochloride Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 40 mg of Pilsicainide Hydrochloride Hydrate in 20 mL of water, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add water to make exactly 20 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks other than pilsicainide obtained from the sample solution is not larger than the peak area of pilsicainide from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 750 mL of water add 5 mL of triethyla-mine, adjust to pH 4.0 with phosphoric acid, and add water to make 1000 mL. To this solution add 200 mL of aceto-nitrile for liquid chromatography. Flow rate: Adjust so that the retention time of pilsicainide is about 5 minutes. Time span of measurement: About 5 times as long as the retention time of pilsicainide, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry sicainide hydrochloride hydrate (C 1/2 H2O). 17H24 N2O.HCl. factor of the peak of pilsicainide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times Description Pilsicainide Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is very soluble in acetic acid (100), and freely soluble in water, in methanol and in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. Identification (1) Determine the absorption spectrum of a solution of Pilsicainide Hydrochloride Hydrate in 0.1 mol/L hydrochloric acid TS (1 in 2000) as directed under Ultravio- with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilsicainide is not more than 2.0z. Water <2.48> 2.5 – 3.3z (50 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Pilsicainide Hydro-chloride Hydrate, dissolve it in 10 mL of acetic acid (100), ‌add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 31.79 mg of C17H24N2O.HCl.1/2 H2O
Pimozide occurs as a white to pale yellow-white powder. It is freely soluble in acetic acid (100), slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Pimozide in methanol (1 in 25,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pimozide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Pimozide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution by using 5 mL of sulfuric acid (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pimozide according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Pimozide in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq- than 1.5 times of the peak area of pimozide from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 2.5 g of ammonium acetate and 8.5 g of tetrabutylammonium hydrogensulfate in water to make 1000 mL. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 80 → 70 20 → 30 10 – 15 Flow rate: 2.0 mL per minute. Time span of measurement: 1.5 times as long as the reten-tion time of pimozide. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add methanol to make exactly 10 mL. Confirm that the peak area of pimozide obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 mL of the standard solution. System performance: Dissolve 5 mg of Pimozide and 2 mg of mebendazole in methanol to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, mebendazole and pimozide are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pimozide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 70 mg of Pimozide, previ-ously dried, dissolve in 25 mL of acetic acid for nonaqueous titration, and titrate <2.50> with 0.02 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L perchloric acid VS ＝ 9.231 mg of C28H29F2N3O
Pindolol occurs as a white crystalline powder. It has a slight, characteristic odor. It is sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in water and in diethyl ether. It dissolves in dilute sulfuric acid and in acetic acid (100).
(1) To 1 mL of a solution of Pindolol in methanol (1 in 10,000) add 1 mL of a solution of 1-(4- pyridyl)-pyridinium chloride hydrochloride (1 in 1000) and 1 mL of sodium hydroxide TS, then add 1 mL of hydrochloric acid: a blue to blue-purple color, changing to red-purple, is produced. Dissolve 0.05 g of Pindolol in 1 mL of dilute sulfuric acid, and add 1 mL of Reinecke salt TS: a light red precipi-tate is produced. Determine the absorption spectrum of a solution of Pindolol in methanol (1 in 50,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pin-dolol, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetone and isopropylamine (5:4:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly diluted sulfuric acid (3 in 5) and a sodium nitrite solution (1 in 50) on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Pindolol, previ-ously dried, dissolve in 80 mL of methanol, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L hydrochloric acid VS ＝ 24.83 mg of C14H20N2O2
Pioglitazone Hydrochloride contains not less than 1 cm Absorbance <2.
Pioglitazone Hydrochloride occurs as white, crystals or crystalline powder. It is soluble in N,N-dimethylformamide and in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. A solution of Pioglitazone Hydrochloride in N,N-dimethylformamide (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Pioglitazone Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pioglitazone Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pioglitazone Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, ‌and compare the spectrum with the Reference Spectrum or the spectrum of Pioglitazone Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. Dissolve 50 mg of Pioglitazone Hydrochloride in 1 mL of nitric acid, and add 4 mL of dilute nitric acid: the so-lution responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 0.5 g of Pindolol in 10 mL of acetic acid (100), and observe imme-diately: the solution is clear, and has no more color than the following control solution. Control solution: Measure accurately 4 mL of Matching Fluid A, add exactly 6 mL of water, and mix. Heavy metals <1.07>—Proceed with 1.0 g of Pindolol according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pindolol according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Pindolol in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 2 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the Description Pioglitazone Hydrochloride occurs as white, crystals or crystalline powder. It is soluble in N,N-dimethylformamide and in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. A solution of Pioglitazone Hydrochloride in N,N-dimethylformamide (1 in 20) shows no optical rotation. Identification (1) Determine the absorption spectrum of a solution of Pioglitazone Hydrochloride in 0.1 mol/L hydro-chloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pioglitazone Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pioglitazone Hydrochloride as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, ‌and compare the spectrum with the Reference Spectrum or the spectrum of Pioglitazone Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. Dissolve 50 mg of Pioglitazone Hydrochloride in 1 mL of nitric acid, and add 4 mL of dilute nitric acid: the so-lution responds to Qualitative Tests <1.09> (2) for chloride. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Pioglitazone Hydrochloride according to Method 4, and per-form the test. After incineration, use 3 mL of hydrobromic acid instead of 3 mL of hydrochloric acid. Prepare the con-trol solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Pioglitazone Hydrochloride in 20 mL of methanol, add the mobile phase to make 100 mL, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of the peaks, having the relative retention times of about 0.7, about 1.4 and about 3.0 to pioglitazone obtained from the sample solution, is not larger than 2/5 times the peak area of pioglitazone from the standard solution, and the area of each peak other than pioglitazone and those peaks men-tioned above from the sample solution is smaller than 1/5 times the peak area of pioglitazone from the standard solu-tion. Furthermore, the total area of the peaks other than pioglitazone from the sample solution is not larger than the peak area of pioglitazone from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of pioglitazone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of pioglitazone obtained with 40 mL of this solution is equivalent to 7 to 13z of that with 40 mL of the standard solution. System performance: Dissolve 50 mg of Pioglitazone Hy-drochloride in 10 mL of a solution of benzophenone in methanol (1 in 750), and add methanol to make 100 mL. To 1 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 40 mL of this solution under the above operating conditions, pioglitazone and benzophe-none are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pioglitazone is not more than 2.0z. Water <2.48> Not more than 0.2z (0.5 g, coulometric titration). For anolyte solution, use anode solution A for water determination. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Pioglitazone Hydrochloride and Pioglitazone Hydrochloride RS (sepa-rately, determine the water <2.48> in the same manner as Pioglitazone Hydrochloride), add exactly 10 mL of the inter-nal standard solution and methanol to make 100 mL. Pipet 2 mL each of these solutions, add the mobile phase to make 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of pioglitazone to that of the internal stand-ard. Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × QT/QS MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of benzophenone in methanol (1 in 750). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 269 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of ammonium acetate solution (77 in 10,000), acetonitrile and acetic acid (100) (25:25:1). Flow rate: Adjust so that the retention time of pioglita-zone is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, pioglitazone and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pioglitazone is not more than 1.0z.
Pipemidic Acid Hydrate contains not less than 98.
Pipemidic Acid Hydrate occurs as a pale yel-low crystalline powder. It is freely soluble in acetic acid (100), very slightly soluble in water, and practically insoluble in methanol and in ethanol (99.5). It dissolves in sodium hydroxide TS. It is gradually colored on exposure to light. Melting point: about 2509C (with decomposition).
(1) Dissolve 0.1 g of Pipemidic Acid Hy-drate in 20 mL of sodium hydroxide TS, and dilute with water to make 200 mL. To 1 mL of the solution add water to make 100 mL. Determine the absorption spectrum of the so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Pipemidic Acid Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Pipemidic Acid Hydrate in 35 mL of water and 10 mL of sodium hy-droxide TS, then add 15 mL of dilute nitric acid, shake well, and filter through a glass filter (G3). To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 5 mL of sodium hydroxide TS, 13.5 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate <1.14>—Dissolve 1.0 g of Pipemidic Acid Hy- drate in 35 mL of water and 10 mL of sodium hydroxide TS, then add 15 mL of dilute hydrochloric acid, shake well, and filter through a glass filter (G3). To 30 mL of the filtrate add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.50 mL of 0.005 mol/L sulfuric acid VS add 5 mL of sodi- um hydroxide TS, 7.5 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Pipe- midic Acid Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pipemidic Acid Hydrate according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Pipemidic Acid Hydrate in 10 mL of diluted acetic acid (100) (1 in 20), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted acetic acid (100) (1 in 20) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform, metha-nol, formic acid and triethylamine (25:15:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 14.5 – 16.0z (20 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Pipemidic Acid Hydrate, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.33 mg of C14H17N5O3
Piperacillin Hydrate contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Piperacillin Hydrate occurs as a white crystal-line powder. It is freely soluble in methanol, soluble in ethanol (99.5) and in dimethylsulfoxide, and very slightly soluble in water.
(1) Determine the infrared absorption spec-trum of Piperacillin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Piperacillin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Determine the 1H spectrum of a solution of Piperacil-lin Hydrate in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 3) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tet-ramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a triplet signal A at about d 1.1 ppm, a singlet signal B at about d 4.2 ppm, and a multiplet signal C at about d 7.4 ppm, and the ratio of the integrated intensity of each signal, A:B:C, is about 3:1:5. D Optical rotation <2.49> [a]20: ＋162 – ＋1729(0.2 g, metha-nol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Piperacillin Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Related substances 1—Conduct this procedure rapid-ly after the preparation of the sample solution and standard solution. Dissolve 20 mg of Piperacillin Hydrate in 20 mL of the mobile phase, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution (1). Pipet 2 mL of the standard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution and the stand-ard solutions (1) and (2) as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks, having the relative reten-tion time of about 0.38 and about 0.50 to piperacillin, ob-tained from the sample solution is not larger than 2 times the peak area of piperacillin from the standard solution (2), the total area of the peaks, having the relative retention time of about 0.82 and about 0.86, from the sample solution is not larger than the peak area of piperacillin from the standard solution (2), and the area of the peak other than piperacillin and the peaks having the relative retention time of about 0.38, about 0.50, about 0.82 and about 0.86, from the sam-ple solution, is not larger than the peak area of piperacillin from the standard solution (2). Furthermore, the total area of the peaks other than piperacillin from the sample solution is not larger than the peak area of piperacillin from the standard solution (1). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of piperacillin, beginning after the solvent peak. System suitability— Test for required detectability: Confirm that the peak area of piperacillin obtained with 20 mL of the standard solution (2) is equivalent to 15 to 25z of that with 20 mL of the standard solution (1). System performance: When the procedure is run with 20 mL of the standard solution (1) under the above operating conditions, the number of theoretical plates and the symme- try factor of the peak of piperacillin are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (2) under the above op-erating conditions, the relative standard deviation of the peak area of piperacillin is not more than 3.0z. Related substances 2—Dissolve 20 mg of Piperacillin Hydrate in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution (1). Pipet 2 mL of the standard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solu-tion, and the standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 6.6 to piperacillin, obtained from the sample solution is not larger than 3 times the peak area of piperacillin from the standard solution (2), and the area of the peaks other than piperacillin and the peak mentioned above from the sample solution are not larger than 1.4 times the peak area of piperacillin from the standard solution (2). Furthermore, the total area of the peaks other than the peak of piperacillin from the sample solution is not larger than the area of the peak of piperacillin from the standard solution (1). For the area of the peak, having the relative retention time of about 6.6, multiply the correction factor, 2.0. Operating conditions— Detector, column and column temperature: Proceed as di-rected in the operating conditions in the Assay. Mobile phase: Take 60.1 g of acetic acid (100) and 101.0 g of triethylamine, add water to make 1000 mL. To 25 mL of this solution add 300 mL of acetonitrile and 25 mL of dilute acetic acid, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of piperacillin is about 1.2 minutes. Time span of measurement: About 8 times as long as the retention time of piperacillin, beginning after the piperacillin peak. System suitability— Test for required detectability: Confirm that the peak area of piperacillin obtained with 20 mL of the standard solution (2) is equivalent to 15 to 25z of that with 20 mL of the standard solution (1). System performance: When the procedure is run with 20 mL of the standard solution (1) under the above operating conditions, the number of theoretical plates and the symme-try factor of the peak of piperacillin are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (2) under the above op-erating conditions, the relative standard deviation of the peak area of piperacillin is not more than 4.0z. Residual solvents <2.46>—Transfer exactly 10 mg of Piperacillin Hydrate to an about 3 mL-vial, add exactly 1 mL of saturated sodium hydrogen carbonate solution to dis-solve and stop the vial tightly. After heating this at 909C for 10 minutes, use the gas inside the container as the sample gas. Separately, measure exactly 1 mL of ethyl acetate, dis-solve in water to make exactly 200 mL. Pipet 10 mL of this solution, add water to make exactly 20 mL. Pipet 2 mL of this solution in an about 3-mL vial containing exactly 1 mL of saturated sodium hydrogen carbonate solution, and stop the vial tightly. Run the procedure similarly to the sample, and use the gas as the standard gas. Perform the test with ex- ‌actly 0.5 mL each of the sample gas and standard gas as di-rected under Gas Chromatography <2.02> according to the following conditions, and determine the peak area of ethyl acetate by the automatic integration method: the peak area of ethyl acetate obtained from the sample gas is not larger than that from the standard gas. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with porous stylene-divinyl benzene co-polymer for gas chromatography (average pore diameter of 0.0085 mm, 300 – 400 m2/g) with the particle size of 125 to 150 mm. Column temperature: A constant temperature of about 1459C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethyl ace-tate is about 4 minutes. System suitability— System performance: Take 1 mL of saturated sodium hydrogen carbonate solution in an about 3 mL-vial, add 2 mL each of ethyl acetate solution (1 in 400) and acetone solu-tion (1 in 400), and stop the vial tightly. When the procedure is run under the above operating conditions, acetone and ethyl acetate are eluted in this order with the resolution be-tween these peaks being not less than 2.0. System repeatability: Take 1 mL of saturated sodium hydrogen carbonate solution in an about 3 mL-vial, add 2 mL of ethyl acetate solution (1 in 400), stop the vial tightly, and perform the test under the above operating conditions. When the procedure is repeated 6 times, the relative standard deviation of the peak area of ethyl acetate is not more than 10z. Water <2.48> Not less than 3.2z and not more than 3.8z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Bacterial endotoxins <4.01> Less than 0.07 EU/mg (po-tency).
Weigh accurately an amount of Piperacillin Hydrate and Piperacillin RS, equivalent to about 50 mg (potency), dissolve each in the mobile phase to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, HT and HS, of the peak height of piperacillin to that of the internal standard. Amount [ mg (potency)] of piperacillin (C23H27N5O7S) ＝ MS × HT/HS × 1000 MS: Amount [mg (potency)] of Piperacillin RS taken Internal standard solution—A solution of acetanilide in the mobile phase (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Take 60.1 g of acetic acid (100) and 101.0 g of triethylamine, add water to make 1000 mL. To 25 mL of this solution add 210 mL of acetonitrile and 25 mL of dilute acetic acid, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of piperacillin is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and piperacillin are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak height of piperacillin to that of the internal standard is not more than 1.0z.
Piperazine Adipate, when dried, contains not less than 98.
Piperazine Adipate occurs as a white crystal-line powder. It is odorless, and has a slightly acid taste. It is soluble in water and in acetic acid (100), and practi-cally insoluble in ethanol (95), in acetone and in diethyl ether. Melting point: about 2509C (with decomposition).
(1) Dissolve 0.5 g of Piperazine Adipate in 10 mL of water, add 1 mL of hydrochloric acid, and extract with two 20-mL portions of diethyl ether. Combine the diethyl ether extracts, evaporate to dryness on a water bath, and dry the residue at 1059C for 1 hour: the melting point <2.60> is between 1529C and 1559C. To 3 mL of a solution of Piperazine Adipate (1 in 100) add 3 drops of Reinecke salt TS: a light red precipitate is formed. Determine the infrared absorption spectrum of Piper-azine Adipate, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> The pH of a solution of 1.0 g of Piperazine Adi-pate in 20 mL of water is between 5.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Piperazine Adipate in 30 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Proceed with 2.0 g of Pipera- zine Adipate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Piperazine Adipate, previously dried, dissolve in a mixture of 20 mL of acetic acid for nonaqueous titration and 40 mL of acetone for nonaqueous titration, and titrate <2.50> with 0.1 mol/L per-chloric acid VS until the red-purple color of the solution changes to blue-purple (indicator: 6 drops of bromocresol green-methylrosaniline chloride TS). Perform a blank deter-mination, and make any necessary correction in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.61 mg of C4H10N2.C6H10O4
Piperazine Phosphate Hydrate occurs as white, crystals or crystalline powder. It is odorless, and has a slightly acid taste. It is soluble in formic acid, sparingly soluble in water, very slightly soluble in acetic acid (100), and practically insoluble in methanol, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid. Melting point: about 2229C (with decomposition).
(1) To 3 mL of a solution of Piperazine Phosphate Hydrate (1 in 100) add 3 drops of Reinecke salt TS:a light red precipitate is formed. Determine the infrared absorption spectrum of Piperazine Phosphate Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Piperazine Phosphate Hydrate (1 in 100) responds to Qualitative Tests <1.09> (1) and (3) for phosphate. pH <2.54> Dissolve 1.0 g of Piperazine Phosphate Hydrate in 100 mL of water: the pH of the solution is between 6.0 and 6.5.
(1) Chloride <1.03>—To 0.5 g of Piperazine Phos-phate Hydrate add 6 mL of dilute nitric acid and water to make 50 mL. Use this solution as the test solution, and per-form the test. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Heavy metals <1.07>—To 2.0 g of Piperazine Phos-phate Hydrate add 5 mL of dilute hydrochloric acid, 30 mL of water and 2 mL of dilute acetic acid, and dissolve. Add sodium hydroxide TS, adjust the pH of the solution to 3.3, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Dissolve 2.0 g of Piperazine Phos- phate Hydrate in 5 mL of dilute hydrochloric acid, and use this solution as the test solution. Perform the test (not more than 1 ppm). Related substances—Dissolve 50 mg of Piperazine Phosphate Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28), acetone and ethanol (99.5) (8:3:3:2) to a distance of about 13 cm, and air-dry the plate. Spray evenly 4-dimethylaminocinnamaldehyde TS, and allow to stand for 15 minutes: the spots other than the principal spot and the spot on the starting line obtained from the sample solution are not more intense than the spot from the standard solu-tion. Water <2.48> 8.0 – 9.5z (0.3 g, volumetric titration, direct titration).
Weigh accurately about 0.15 g of Piperazine Phos-phate Hydrate, dissolve in 10 mL of formic acid, add 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 9.207 mg of C4H10N2.H3PO4
Pirarubicin occurs as a red-orange crystalline powder. It is soluble in chloroform, very slightly soluble in aceto-nitrile, in methanol and in ethanol (99.5), and practically in-soluble in water.
(1) Dissolve 10 mg of Pirarubicin in 80 mL of methanol and 6 mL of diluted hydrochloric acid (1 in 5000), and add water to make 100 mL. To 10 mL of this so-lution add diluted methanol (4 in 5) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pirarubicin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Dissolve 5 mg each of Pirarubicin and Pirarubicin RS in 5 mL of chloroform, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform and methanol (5:1) to a distance of about 10 cm, and air-dry the plate. Examine the spots: the principal spot obtained from the sample solution and the spot from the standard solution show a red-orange color and the same Rf value. D Optical rotation <2.49> [a]20: ＋195 – ＋2159(10 mg, chlo-roform, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 10 mg of Pirarubicin in 10 mL of 0.01 mol/L hydrochloric acid TS: the solution is clear and red. Heavy metals <1.07>—Proceed with 1.0 g of Pirarubi- cin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 10 mg of Pirarubicin in 20 mL of the mobile phase, and use this solution as the sam- ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of doxorubicin, having the relative retention time of about 0.45 to pirarubi-cin, and the area of the peak, having the relative retention time of about 1.2, obtained from the sample solution are not larger than the peak area of pirarubicin from the standard solution, respectively, and the sum of the areas of the peaks, having the relative retention times of about 1.9 and about 2.0, from the sample solution is not larger than 5 times the peak area of pirarubicin from the standard solution. For the peak area for doxorubicin, multiply the correction factor 0.94 and the area for the two peaks, having the relative retention times of about 1.9 and about 2.0, multiply their correction factors, 1.09, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of pirarubicin. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mobile phase to make ex-actly 10 mL. Confirm that the peak area of pirarubicin ob-tained with 20 mL of this solution is equivalent to 14 to 26z of that with 20 mL of the standard solution. Water <2.48> Not more than 2.0z (0.1 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Pirarubicin and Pirarubicin RS, equivalent to about 10 mg (potency), and dissolve in the mobile phase to make exactly 10 mL. Pipet 5 mL of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample so-lution and standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of pirarubicin to that of the internal standard. Amount [ mg (potency)] of pirarubicin (C32H37NO12) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Pirarubicin RS taken Internal standard solution—A solution of 2-naphthol in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L ammonium for-mate buffer solution (pH 4.0) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of pirarubicin is about 7 minutes. System suitability— System performance: When the procedure is run with 20 ‌mL of the standard solution under the above operating con-ditions, pirarubicin and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of pirarubicin to that of the internal stand-ard is not more than 1.0z.
Pirenoxine, when dried, contains not less than 98.
Pirenoxine occurs as a yellow-brown powder. It is odorless, and has a slightly bitter taste. It is very slightly soluble in dimethylsulfoxide, and practi-cally insoluble in water, in acetonitrile, in ethanol (95), in tetrahydrofuran and in diethyl ether. Melting point: about 2509C (with decomposition).
(1) Dissolve 2 mg of Pirenoxine in 10 mL of phosphate buffer solution (pH 6.5), add 5 mL of a solu-tion of L-ascorbic acid (1 in 50), and shake vigorously: a dark purple precipitate is formed. Determine the absorption spectrum of a solution of Pirenoxine in phosphate buffer solution (pH 6.5) (1 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Pirenoxine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Pirenoxine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 10 mg of Pirenoxine in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the total area of the peaks other than pirenoxine obtained from the sample solution is not larger than the peak area of pirenoxine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.39 g of tetra n-butylammonium chloride and 4.5 g of disodium hydrogen phosphate dodeca-hydrate in 1000 mL of water, and adjust the pH to 6.5 with phosphoric acid. To 700 mL of this solution add 200 mL of acetonitrile and 30 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of pirenoxine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of pirenoxine. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 30 mL. Confirm that the peak area of pirenoxine obtained with 5 mL of this solution is equivalent to 5 to 8z of that with 5 mL of the standard solution. System performance: Dissolve 3 mg of Pirenoxine and 16 mg of methyl parahydroxybenzoate in 100 mL of the mobile phase. When the procedure is run with 5 mL of this solution under the above operating conditions, pirenoxine and methyl parahydroxybenzoate are eluted in this order with the resolu-tion between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of pirenoxine is not more than 1.0z. Loss on drying <2.41> Not more than 1.5z (0.5 g, in vacu-um, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Pirenoxine, previ-ously dried, dissolve in 140 mL of dimethylsulfoxide by heat-ing on a water bath. After cooling, add 30 mL of water, and titrate <2.50> immediately with 0.02 mol/L sodium hydrox-ide VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.02 mol/L sodium hydroxide VS ＝ 6.165 mg of C16H8N2O5
Pirenzepine Hydrochloride Hydrate occurs as a white to pale yellow crystalline powder. It is freely soluble in water and in formic acid, slightly soluble in methanol, and very slightly soluble in ethanol (99.5). mL of this solution, add 5 mL of methanol and the mobile phase A to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than pirenzepine obtained from the sample solution is not larger than 3/10 times the peak area of pirenzepine from the stand-ard solution, and the total area of the peaks other than pirenzepine from the sample solution is not larger than 3/5 times the peak area of pirenzepine from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2 g of sodium lauryl sulfate in 900 mL of water, adjust the pH to 3.2 with acetic acid (100), and add water to make 1000 mL. Mobile phase B: Methanol. Mobile phase C: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A, B and C as directed in the following table. The pH of a solution obtained by dissolving 1 g of Piren-zepine Hydrochloride Hydrate in 10 mL of water is between 1.0 and 2.0. Melting point: about 2459C (with decomposition). It is gradually colored by light. Time after injection 0 – 15 55 → 25 15 → 45 15 – of sample (min) Mobile phase A (volz) Mobile phase B (volz) Mobile phase C (volz)
(1) Determine the absorption spectrum of a solution of Pirenzepine Hydrochloride Hydrate (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Piren-zepine Hydrochloride Hydrate as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Pirenzepine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Pirenzepine Hydrochloride Hy-drate in 10 mL of water is clear and not more color than that of the following control solution. Control solution: To 1.2 mL of Matching Fluid F add 8.8 mL of diluted hydrochloric acid (1 in 40). Heavy metals <1.07>—Proceed with 2.0 g of Pirenze- pine Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.3 g of Pirenzepine Hydrochloride Hydrate in 10 mL of water. To 1 mL of this solution add 5 mL of methanol and the mobile phase A to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add 5 mL of metha-nol and the mobile phase A to make exactly 10 mL. Pipet 1 Flow rate: Adjust so that the retention time of pirenzepine is about 8 minutes. Time span of measurement: About 2 times as long as the retention time of pirenzepine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 5 mL of methanol and the mobile phase A to make exactly 10 mL. Confirm that the peak area of piren-zepine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 0.1 g of 1-phenylpiperazine hydrochloride in 10 mL of methanol. Mix 1 mL of this solu-tion and 1 mL of the sample solution, and add 5 mL of methanol and the mobile phase A to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pirenzepine and phenylpiperazine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pirenzepine is not more than 2.0z. Water <2.48> Not less than 3.5z and not more than 5.0z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Pirenzepine Hydro-chloride Hydrate, dissolve in 2 mL of formic acid, add 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L ‌perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction.‌ Each mL of 0.1 mol/L perchloric acid VS ＝ 14.14 mg of C19H21N5O2.2HCl
Piroxicam occurs as a white to light yellow crystalline powder. It is slightly soluble in acetonitrile and in ethanol (99.5), and practically insoluble in water. Melting point: about 2009C (with decomposition). It shows crystal polymorphism.
(1) Dissolve 0.1 g of Piroxicam in a mix-ture of methanol and 0.5 mol/L hydrochloric acid TS (490:1) to make 200 mL. To 1 mL of this solution add the mixture of methanol and 0.5 mol/L hydrochloric acid TS (490:1) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Piroxicam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve the Piroxicam with dichloromethane, evaporate the solvent, dry the residue on a water bath, and perform the test.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Piroxicam according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Dissolve 75 mg of Piroxicam in 50 mL of acetonitrile for liquid chromatography, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add acetonitrile for liquid chromatography to make exactly 10 mL. Pipet 1 mL of this solution, add aceto-nitrile for liquid chromatography to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of piroxicam are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of piroxicam is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.25 g of Piroxicam, dis-solve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 33.14 mg of C15H13N3O4S
Pivmecillinam Hydrochloride contains not less than 630 mg (potency) and not more than 710 mg (potency) per mg, calculated on the anhydrous basis.
Pivmecillinam Hydrochloride occurs as a white to yellowish white crystalline powder. It is very soluble in methanol and in acetic acid (100), freely soluble in water and in ethanol (99.5), and soluble in acetonitrile.
(1) Determine the infrared absorption spec-trum of Pivmecillinam Hydrochloride as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Pivmecillinam Hy-drochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.5 g of Pivmecillinam Hydrochloride in 10 mL of water, and add 1 mL of dilute nitric acid and 1 drop of silver nitrate TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋200 – ＋2209(1 g calculated on the anhydrous basis, water, 100 mL, 100 mm).
(1) Heavy metals <1.07>—To 1.0 g of Pivmecillin-am Hydrochloride in a crucible add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), fire the ethanol to burn, and heat gradually to incinerate. If a carbonized substance remains, moisten with a small amount of nitric acid, and ignite to incinerate. Cool, add 3 mL of hydrochloric acid to the residue, dissolve by warming on a water bath, and heat to dryness. To the residue add 10 mL of water, and dissolve by warming on a water bath. After cool-ing, adjust the pH to 3 to 4 with ammonia TS, add 2 mL of dilute acetic acid, filter if necessary, and wash the crucible and the filter with 10 mL of water. Put the filtrate and the washings to a Nessler tube, add water to make 50 mL, and use this solution as the test solution. Prepare the control so-lution in the same manner as the test solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pivmecillinam Hydrochloride according to Method 4, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Pivmecillin-am Hydrochloride in 4.0 mL of a mixture of acetonitrile and acetic acid (100) (97:3), and use this solution as the sample solution. Separately, dissolve 2.0 mg of Pivmecillinam Hy-drochloride RS in 4.0 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography, allow to stand for 30 minutes, then spot 2 mL of the sample solution on the plate. Immediately, develop the plate with a mixture of acetone, water and acetic acid (100) (10:1:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand for 10 minutes in iodine vapor: the spot obtained from the sample solution appeared at the position corresponding to the spot from the standard solution is not larger and not more intense than the spot from the standard solution, and any spot other than the principal spot and the above spot from the sample solution is not observable. Water <2.48> Not more than 1.0z (0.25 g, coulometric titration).
Weigh accurately an amount of Pivmecillinam Hy-drochloride and Pivmecillinam Hydrochloride RS, equiva-lent to about 20 mg (potency), dissolve in a suitable amount of the mobile phase, add exactly 10 mL of the internal stand-ard solution and the mobile phase to make 100 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pivmecillinam to that of the internal standard. Amount [ mg (potency)] of mecillinam (C15H23N3O3S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Pivmecillinam Hydrochlo-ride RS taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 12,500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.771 g of ammonium acetate in about 900 mL of water, adjust the pH to 3.5 with acetic acid (100), and add water to make 1000 mL. To 400 mL of this solution add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of pivmecillin-am is about 6.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pivmecillinam and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the ratios of the peak area of pivmecillinam to that of the internal standard is not more than 1.0z.‌
Platycodon Fluidextract is a red-brown liquid. It is miscible with water, producing slight turbidity. It has a mild taste at first, followed by an acrid and bitter taste. Identiˆcation To 2 mL of Platycodon Fluidextract add 20 mL of water and 5 mL of 1-butanol, mix, shake for 10 minutes, centrifuge, and use the 1-butanol layer as the sam-ple solution. Separately, dissolve 1 mg of platycodin D for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromato-graphy <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (5:3:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Alcohol number <1.01> Apply to Platycodon Fluidextract prepared by the Method of preparation 2). 2.0 – 3.0 (Method 1).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Platycodon Fluidextract as directed in the Fluidextracts (4), and perform the test (not more than 30 ppm). (2) Starch—Mix 1 mL of Platycodon Fluidextract with 4 mL of water, and add 1 drop of dilute iodine TS: no purple or blue color develops. Content of the active principle Transfer exactly 5 mL of Platycodon Fluidextract to a tared beaker or porcelain dish, evaporate to dryness on a water bath, and dry at 1059C for 5 hours: the mass of the residue is not less than 0.50 g.
Macrogol 1500 occurs as a white, smooth petrolatum-like solid. It is odorless or has a faint, character-istic odor. It is very soluble in water, in pyridine and in diphenyl ether, freely soluble in methanol, sparingly soluble in ethanol (95), very slightly soluble in ethanol (99.5), and prac-tically insoluble in diethyl ether. Congealing point: 37 – 419C
Dissolve 50 mg of Macrogol 1500 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, and filter, if necessary. To the filtrate add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10): a yellow-green precipitate is formed. pH <2.54> Dissolve 1.0 g of Macrogol 1500 in 20 mL of water: the pH of the solution is between 4.0 and 7.0.
(1) Clarity and color of solution—Dissolve 5.0 g of Macrogol 1500 in 50 mL of water: the solution is clear and colorless. Acidity—Dissolve 5.0 g of Macrogol 1500 in 20 mL of neutralized ethanol, and add 2 drops of phenolphthalein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: the so-lution is red in color. ‌Ethylene glycol and diethylene glycol—Place 50.0 g of Macrogol 1500 in a distilling flask, add 75 mL of diphenyl ether, warm to dissolve if necessary, distil slowly under a reduced pressure of 0.13 to 0.27 kPa and take 25 mL of the distillate in a 100-mL container with 1-mL graduation. To the distillate add exactly 20 mL of water, shake vigorously, cool in ice water, congeal the diphenyl ether, and filtrate into a 25-mL volumetric flask. Wash the residue with 5.0 mL of ice-cold water, combine the washings with the filtrate, warm to room temperature, and add water to make 25 mL. Trans-fer this solution to a glass-stoppered flask, shake with 25.0 mL of freshly distilled acetonitrile, and use this solution as the sample solution. Separately, to 62.5 mg of diethylene glycol add a mixture of water and freshly distilled aceto-nitrile (1:1) to make exactly 25 mL, and use this solution as the standard solution. Take exactly 10 mL each of the sam-ple solution and the standard solution, and add to each exactly 15 mL of cerium (IV) diammonium nitrate TS. Per-form the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24> within 2 to 5 minutes: the absorbance of the solution obtained from the sample solu-tion at the wavelength of maximum absorption at about 450 nm is not larger than the absorbance of the solution obtained from the standard solution.‌ Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Macrogol 20000 occurs as white, paraffin-like flakes or powder. It is odorless or has a faint, characteristic odor. It is freely soluble in water and in pyridine, and practically insoluble in methanol, in ethanol (95), in petroleum benzine and in macrogol 400. Congealing point: 56 – 649C
Dissolve 50 mg of Macrogol 20000 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, and filter, if necessary. To the filtrate add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10): a yellow-green precipitate is formed. pH <2.54> Dissolve 1.0 g of Macrogol 20000 in 20 mL of water: the pH of this solution is between 4.5 and 7.5.
(1) Clarity and color of solution—Dissolve 5.0 g of Macrogol 20000 in 50 mL of water: the solution is clear and colorless. (2) Acidity—Dissolve 5.0 g of Macrogol 20000 in 20 mL of neutralized ethanol by warming, cool, and add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol-phthalein TS: the color of the solution is red. Average molecular mass Weigh accurately about 15 g of Macrogol 20000, transfer to an about 200-mL glass-stoppered pressure bottle, add about 25 mL of pyridine, dissolve by warming, and allow to cool. Separately, pipet 300 mL of freshly distilled pyridine into a 1-L light-resistant glass-stoppered bottle, add 42 g of phthalic anhydride, dis-solve with vigorous shaking, and allow to stand for 16 hours or more. Pipet 25 mL of this solution, transfer to the former pressure bottle, stopper the bottle tightly, wrap it securely with strong cloth, and immerse in a water bath, having a temperature of 98 ± 29C, to the same depth as the mixture in the bottle. Maintain the temperature of the bath at 98 ± 29C for 60 minutes. Remove the bottle from the bath, and allow to cool in air to room temperature. Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 drops of a solution of phenolphthalein in pyridine (1 in 100). Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a light red color remains for not less than 15 seconds. Perform a blank deter-mination in the same manner. Average molecular mass ＝ (M × 4000)/(a － b) M: Amount (g) of Macrogol 20000 taken a: Volume (mL) of 0.5 mol/L sodium hydroxide VS used in the blank determination b: Volume (mL) of 0.5 mol/L sodium hydroxide VS used in the test of the sample Average molecular mass is between 15000 and 25000. Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Macrogol 400 occurs as a clear, colorless and viscous liquid. It has no odor or a slight, characteristic odor. It is miscible with water, with methanol, with ethanol (95) and with pyridine. It is soluble in diethyl ether. It is slightly hygroscopic. Congealing point: 4 – 89C Specific gravity d 20: 1.110 – 1.140
Dissolve 50 mg of Macrogol 400 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, and filter, if necessary. To the filtrate add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10): a yellow-green precipitate is formed. pH <2.54> Dissolve 1.0 g of Macrogol 400 in 20 mL of water: the pH of this solution is between 4.0 and 7.0.
(1) Acidity—Dissolve 5.0 g of Macrogol 400 in 20 mL of neutralized ethanol, and add 2 drops of phenol-phthalein TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: the solution is red in color. (2) Ethylene glycol and diethylene glycol—Dissolve 4.0 g of Macrogol 400 in water to make exactly 10 mL, and use this solution as the sample solution. Weigh accurately about 50 mg each of ethylene glycol and diethylene glycol, dissolve in water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con-ditions. Determine the peak heights, HTa and HSa, of ethyl-ene glycol of each solution, and the peak heights, HTb and HSb, of diethylene glycol, and calculate the amount of ethyl-ene glycol and diethylene glycol: the sum of the contents of ethylene glycol and diethylene glycol is not more than 0.25z. Amount (mg) of ethylene glycol ＝ MSa × HTa/HSa × 1/10 Amount (mg) of diethylene glycol solution under the above operating conditions, and calculate the resolution. Use a column clearly dividing peaks of ethyl-ene glycol and diethylene glycol in this order. Detection sensitivity: Adjust the detection sensitivity so that the peak height of diethylene glycol obtained from 2 mL of the standard solution composes about 80z of the full scale. Average molecular mass Add 42 g of phthalic anhydride to 300 mL of freshly distilled pyridine, exactly measured, in a 1-L light-resistant glass-stoppered bottle. Shake the bottle vigorously to dissolved the solid, and allow to stand for 16 hours or more. Pipet 25 mL of this solution into an about 200-mL glass-stoppered pressure bottle. Add about 1.5 g of Macrogol 400, accurately weighed, stopper the bottle, wrap it securely with strong cloth, and immerse in a water bath, having a temperature of 98 ± 29C, to the level so that the mixture in the bottle soaks completely in water. Maintain the temperature of the bath at 98 ± 29C for 30 minutes. Re-move the bottle from the bath, and allow to cool in air to room temperature. Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 drops of a solution of phenolphthalein in pyridine (1 in 100). Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a light red color remains for not less than 15 seconds. Perform a blank determination in the same man-ner. Average molecular mass ＝ (M × 4000)/(a － b ) M: Amount (g) of Macrogol 400 taken a: Volume (mL) of 0.5 mol/L sodium hydroxide VS used in the blank determination b: Volume (mL) of 0.5 mol/L sodium hydroxide VS used in the test of the sample Average molecular mass is between 380 and 420. Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Macrogol 4000 is a white, paraffin-like solid, occurring as flakes or powder. It is odorless or has a faint, characteristic odor. It is very soluble in water, freely soluble in methanol and in pyridine, and practically insoluble in ethanol (99.5) and in diethyl ether. Congealing point: 53 – 579C
Dissolve 50 mg of Macrogol 4000 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, and filter, if necessary. To the filtrate add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10): a yel-low-green precipitate is formed. pH <2.54> Dissolve 1.0 g of Macrogol 4000 in 20 mL of water: the pH of this solution is between 4.0 and 7.5.
(1) Clarity and color of solution—A solution of 5.0 g of Macrogol 4000 in 50 mL of water is clear and color-less. (2) Acidity—Dissolve 5.0 g of Macrogol 4000 in 20 mL of neutralized ethanol by warming, cool, and add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol-phthelein TS: the color of the solution is red. Average molecular mass Weigh accurately about 12.5 g of Macrogol 4000, transfer to an about 200-mL glass-stoppered pressure bottle, add about 25 mL of pyridine, dissolve by warming, and allow to cool. Separately, pipet 300 mL of freshly distilled pyridine into a 1-L light-resistant, glass-stoppered bottle, add 42 g of phthalic anhydride, dissolve with vigorous shaking, and allow to stand for 16 hours or more. Pipet 25 mL of this solution, transfer to the former pressure bottle, stopper the bottle tightly, wrap it securely with strong cloth, and immerse in a water bath, previously heated at 98 ± 29C, to the level so that the mixture in the bottle soaks completely in water. Maintain the temperature of the bath at 98 ± 29C for 30 minutes. Remove the bottle from the bath, and allow to cool in air to room temperature. Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 drops of a solution of phenolphthalein in pyridine (1 in 100). Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a light red color remains for not less than 15 seconds. Perform a blank determination in the same manner. Average molecular mass ＝ (M × 4000)/(a － b) M: Amount (g) of Macrogol 4000 taken a: Volume (mL) of 0.5 mol/L sodium hydroxide VS con-sumed in the blank determination b: Volume (mL) of 0.5 mol/L sodium hydroxide VS con-sumed in the test of the sample Average molecular mass is between 2600 and 3800. Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Macrogol 6000 is a white, paraffin-like solid, occurring as flakes or powder. It is odorless or has a faint, characteristic odor. It is very soluble in water, freely soluble in pyridine, and practically insoluble in methanol, in ethanol (95), in ethanol (99.5) and in diethyl ether. Congealing point: 56 – 619C
Dissolve 50 mg of Macrogol 6000 in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, and filter, if necessary. To the filtrate add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10): a yellow-green precipitate is formed. pH <2.54> Dissolve 1.0 g of Macrogol 6000 in 20 mL of water: the pH of this solution is between 4.5 and 7.5.
(1) Clarity and color of solution—Dissolve 5.0 g of Macrogol 6000 in 50 mL of water: the solution is clear and colorless. ‌(2) Acidity—Dissolve 5.0 g of Macrogol 6000 in 20 mL of neutralized ethanol by warming, cool, and add 0.20 mL of 0.1 mol/L sodium hydroxide VS and 1 drop of phenol-phthalein TS: the color of the solution is red.‌ Average molecular mass Weigh accurately about 12.5 g of Macrogol 6000, transfer to an about 200-mL glass-stoppered pressure bottle, add about 25 mL of pyridine, dissolve by warming, and allow to cool. Separately, pipet 300 mL of freshly distilled pyridine into a 1-L light-resistant, glass-stoppered bottle, add 42 g of phthalic anhydride, dissolve with vigorous shaking, and allow to stand for 16 hours or more. Pipet 25 mL of this solution, transfer to the former pressure bottle, stopper the bottle tightly, wrap it securely with strong cloth, and immerse in a water bath, previously heated at 98 ± 29C, to the level so that the mixture in the bottle soaks completely in water. Maintain the temperature of the bath at 98 ± 29C for 30 minutes. Remove the bottle from the bath, and allow to cool in air to room temperature. Add exactly 50 mL of 0.5 mol/L sodium hydroxide VS and 5 drops of a solution of phenolphthalein in pyridine (1 in 100). Titrate <2.50> with 0.5 mol/L sodium hydroxide VS until a light red color remains for not less than 15 seconds. Perform a blank determination in the same manner. Average molecular mass ＝ (M × 4000)/(a － b) M: Amount (g) of Macrogol 6000 taken a: Volume (mL) of 0.5 mol/L sodium hydroxide VS con-sumed in the blank determination b: Volume (mL) of 0.5 mol/L sodium hydroxide VS con-sumed in the test of the sample Average molecular mass is between 7300 and 9300. Water <2.48> Not more than 1.0z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Polymixin B Sulfate occurs as a white powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) To 5 mL of a solution of Polymixin B Sulfate (1 in 10) add 5 mL of a solution of sodium hydroxide (1 in 10), add 5 drops of a solution of copper (II) sulfate pen-tahydrate (1 in 100) while shaking: a purple color develops. (2) Transfer 5 mg each of Polymixin B Sulfate and Poly-mixin B Sulfate RS separately into two glass stoppered test tubes, add 1 mL of diluted hydrochloric acid (1 in 2), stopper the tube, heat at 1359C for 5 hours, then heat to dryness on a water bath, and keep the heating until no more hydrochloric acid odor is evolved. Dissolve the residue in 0.5 mL of water, and use these solutions as the sample solution and standard solution (1). Separately, dissolve 20 mg each of L-leucine, L-threonine, phenylalanine and L-serine separately in 10 mL of water, and use these solutions as the standard solutions (2), (3), (4) and (5), respectively. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 3 mL each of the sample solution, the standard solutions (1), (2), (3), (4) and (5) on a plate of silica gel for thin-layer chromatography, and expose the plate to a saturated vapor of the developing solvent for 15 hours. De-velop the plate with a mixture of phenol and water (3:1) to a distance of about 13 cm while without exposure to light, and dry the plate at 1109C for 5 minutes. Spray evenly nin-hydrin-acetic acid TS on the plate, and heat at 1109C for 5 minutes: Rf value of each spot obtained from the sample so-lution is the same with Rf value of the corresponding spots from the standard solution (1). Each of the spots from the sample solution appears at the position corresponding to each of the spots from the standard solutions (2), (3) and (4), but not appears at the position corresponding to the spot from the standard solution (5). A solution of Polymixin B Sulfate (1 in 20) responds to Qualitative Tests <1.09> for sulfate. D Optical rotation <2.49> [a]20: －78 – －909(0.5 g calculated on the dried basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Polymixin B Sulfate in 50 mL of water is between 5.0 and 7.0. Phenylalanine Weigh accurately about 0.375 g of Polymix-in B Sulfate, dissolve in 0.1 mol/L hydrochloric acid VS to make exactly 100 mL. Determine absorbances, A1, A2, A3, A4 and A5, of this solution at 252 nm, at 258 nm, at 264 nm, at 280 nm and at 300 nm, respectively, as directed under Ultraviolet-visible Spectrophotometry <2.24>, and calculate the amount of phenylalanine by the following equation: the amount of phenylalanine is not less than 9.0z and not more than 12.0z. Amount (z) of phenylalanine ＝ (A2 － 0.5A1 ＋ 0.5A3 － 1.8A4 ＋ 0.8A5)/MT × 9.4787 MT: Amount (g) of Polymixin B Sulfate taken, calculated on the dried basis
Heavy metals <1.07>—Proceed with 1.0 g of Poly-mixin B Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 6.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.75z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Escherichia coli NIHJ Agar media for seed and base layer Peptone 10.0 g Meat extract 3.0 g Sodium chloride 30.0 g Agar 20.0 g Water 1000 mL Mix all the ingredients, and sterilize. Adjust the pH <2.54> of the solution so that it will be 6.5 to 6.6 after sterilization. Standard solutions—Weigh accurately an amount of Polymixin B Sulfate RS, equivalent to about 200,000 units, dissolve in phosphate buffer solution (pH 6.0) to make ex-actly 20 mL, and use this solution as the standard stock solu-tion. Keep the standard stock solution at not exceeding 59C and use within 14 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 4000 units and 1000 units, and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Polymixin B Sulfate, equivalent to about 200,000 units, and dissolve in phosphate buffer solution (pH 6.0) to make ex-actly 20 mL. Take exactly a suitable amount of this solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 4000 units and 1000 units, and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Polyoxyl 40 Stearate occurs as a white to light yellow, waxy solid or powder. It is odorless or has a faint fat-like odor. It is soluble in water, in ethanol (95) and in diethyl ether. Congealing point <2.42> 39.0 – 44.09C Congealing point of the fatty acid <1.13> Not below 539C. Acid value <1.13> Not more than 1. Saponification value <1.13> 25 – 35
(1) Clarity and color of solution—Dissolve 1.0 g of Polyoxyl 40 Stearate in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Polyoxyl 40 Stearate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of Polyoxyl 40 Stearate, according to Method 3, and per-form the test (not more than 3 ppm). Residue on ignition <2.44> Not more than 0.1z (1 g).
Powdered Polyporus Sclerotium occurs as a light grayish brown to light brown powder. Almost odorless and tasteless. Under a microscope <5.01>, Powdered Polyporus Scleroti- um reveals hypha, 1 to 2 mm, rarely up to 13 mm in diameter, and colorless transparent; granule strongly refracting light; and a few mucilage plates; sometimes fragments of false tis-sue consisting of them; somewhat brown false tissues; and solitary crystal of calcium oxalate. Solitary crystal is 10 to 40 mm in diameter, sometimes 100 mm in diameter.
Warm, while shaking, 0.5 g of Powdered Polyporus Sclerotium with 5 mL of acetone on a water bath for 2 minutes, filter and evaporate the filtrate to dryness. Dissolve the residue in 5 drops of acetic anhydride, and add 1 drop of sulfuric acid: a red-purple color develops, and im-mediately changes to dark green. ‌JP XVIII Crude Drugs and Related Drugs / Prepared Glycyrrhiza 2105‌‌
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Polyporus Sclerotium according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Polyporus Sclerotium according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 16.0z. Acid-insoluble ash <5.01> Not more than 4.0z.
Irregularly shaped mass, usually 5 – 15 cm in length; externally black-brown to grayish brown, with numerous dents and coarse wrinkles; breakable; fractured surface rather soft and cork-like, and almost white to light brown in color, and a white speckled pattern on the inner region; light in texture. Practically odorless and tasteless.
Warm, while shaking, 0.5 g of pulverized Polyporus Sclerotium with 5 mL of acetone on a water bath for 2 minutes, filter, and evaporate the filtrate to dryness. Dissolve the residue in 5 drops of acetic anhydride, and add 1 drop of sulfuric acid: a red-purple color develops, and im-mediately changes to dark green.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Polyporus Sclerotium according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Polyporus Sclerotium according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 16.0z. Acid-insoluble ash <5.01> Not more than 4.0z.
Polysorbate 80 is a mixture of partial esters of fatty acids, mainly oleic acid, with sorbitol and its anhy-drides ethoxylated with approximately 20 moles of eth-ylene oxide for each mole of sorbitol and sorbitol an-hydrides. Description Polysorbate 80 is a colorless or brownish yel-low, clear or slightly opalescent, oily liquid. It is miscible with water, with methanol, with ethanol (99.5) and with ethyl acetate. It is practically insoluble in fatty oils and in liquid paraffin. Viscosity: about 400 mPa･s (259C). Specific gravity d 20: about 1.10◆ Identification It meets the requirements of the Composi-tion of fatty acids. Composition of fatty acids Dissolve 0.10 g of Polysorbate 80 in 2 mL of a solution of sodium hydroxide in methanol (1 this solution as the sample solution. Perform the test with 1 mL each of the sample solution and fatty acid methyl esters mixture TS as directed under Gas Chromatography <2.02> according to the following conditions. Identify each peak obtained with the sample solution using the chromatogram with fatty acid methyl esters mixture TS. Determine each peak area with the sample solution by the automatic integra-tion method, and calculate the composition of fatty acids by the area percentage method: myristic acid is not more than 5.0z, palmitic acid is not more than 16.0z, palmitoleic acid is not more than 8.0z, stearic acid is not more than 6.0z, oleic acid is not less than 58.0z, linoleic acid is not more than 18.0z and linolenic acid is not more than 4.0z. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.32 mm in inside diameter and 30 m in length, coated with polyethylene glycol 20 M for gas chromatography 0.5 mm in thickness. Column temperature: Inject at a constant temperature of about 809C, raise the temperature to 2209C at a rate of 109C per minute, and maintain at 2209C for 40 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: 50 cm per second. Split ratio: 1: 50. System suitability— Test for required detectability: Dissolve 0.50 g of the mix-ture of fatty acid methyl esters described in the following table in heptane to make exactly 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test add heptane to make ex-actly 10 mL. When the procedure is run with 1 mL of this so-lution under the above operating conditions, the SN ratio of methyl myristate is not less than 5. Mixture of fatty acid methyl esters Composition (z) Methyl myristate for gas chromatography 5 Methyl palmitate for gas chromatography 10 Methyl stearate for gas chromatography 15 Methyl arachidate for gas chromatography 20 Methyl oleate for gas chromatography 20 Methyl eicosenoate for gas chromatography 10 Methyl behenate 10 Methyl lignocerate for gas chro- matography 10 System performance: When the procedure is run with 1 mL of the solution for system suitability test under the above op-erating conditions, ◆methyl stearate and methyl oleate are eluted in this order,◆ the resolution between these peaks is not less than 1.8, and the number of theoretical plates of the peak of methyl stearate is not less than 30,000. Acid value <1.13> Not more than 2.0 (using ethanol (95) instead).◆ Saponification value Introduce about 4 g of Polysorbate 80, accurately weighed, into a 250-mL borosilicate glass flask. Add exactly 30 mL of 0.5 mol/L potassium hydrox-ide-ethanol VS and a few glass beads. Attach a reflux con-denser, and heat for 60 minutes. Add 1 mL of phenolphthal-ein TS and 50 mL of ethanol (99.5), and titrate <2.50> imme-diately with 0.5 mol/L hydrochloric acid VS. Perform a blank determination in the same manner. Calculate the saponification value by the following equation: 45 – 55. Saponification value ＝ (a － b) × 28.05/M M: Amount (g) of Polysorbate 80 taken a: Volume (mL) of 0.5 mol/L hydrochloric acid VS re-quired for blank determination b: Volume (mL) of 0.5 mol/L hydrochloric acid VS re-quired for sample determination Hydroxyl value Introduce about 2 g of Polysorbate 80, ac-curately weighed, into a 150-mL round bottom flask, add ex-actly 5 mL of acetic anhydride-pyridine TS, and attach an air condenser. Heat the flask in a water bath for 1 hour keeping the level of the water about 2.5 cm above the level of the liq-uid in the flask. Withdraw the flask and allow to cool. Add 5 mL of water through the condenser. If a cloudiness appears add sufficient pyridine to clear it, noting the volume added. Shake the flask, and heat in the water bath for 10 minutes. Withdraw the flask and allow to cool. Rinse the condenser and the walls of the flask with 5 mL of neutralized ethanol, and titrate <2.50> with 0.5 mol/L potassium hydroxide-ethanol VS (indicator: 0.2 mL of phenolphthalein TS). Per-form a blank determination in the same manner. Calculate the hydroxyl value by the following equation: 65 – 80. Hydroxyl value ＝ (a － b) × 28.05/M ＋ acid value M: Amount (g) of Polysorbate 80 taken a: Volume (mL) of 0.5 mol/L potassium hydroxide-ethanol VS required for blank determination b: Volume (mL) of 0.5 mol/L potassium hydroxide-ethanol VS required for sample determination Purity ◆(1) Heavy metals <1.07>—Proceed with 1.0 g of Polysorbate 80 according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ Ethylene oxide and 1,4-dioxane—Transfer exactly 1.00 g of Polysorbate 80 into a 10-mL headspace vial, add exactly 2 mL of water, seal the vial immediately with a sep-tum of silicon rubber coated the surface with fluororesin and an aluminum cap. Mix carefully, and use the content as the sample solution. Separately, pipet 0.5 mL of a solution, pre-pared by dissolving ethylene oxide in dichloromethane so that each mL contains 50 mg, and add water to make exactly 50 mL.
Polysorbate 80 is a colorless or brownish yel-low, clear or slightly opalescent, oily liquid. It is miscible with water, with methanol, with ethanol (99.5) and with ethyl acetate. It is practically insoluble in fatty oils and in liquid paraffin. Viscosity: about 400 mPa･s (259C). Specific gravity d 20: about 1.10◆
It meets the requirements of the Composi-tion of fatty acids. Composition of fatty acids Dissolve 0.10 g of Polysorbate 80 in 2 mL of a solution of sodium hydroxide in methanol (1 this solution as the sample solution. Perform the test with 1 mL each of the sample solution and fatty acid methyl esters mixture TS as directed under Gas Chromatography <2.02> according to the following conditions. Identify each peak obtained with the sample solution using the chromatogram with fatty acid methyl esters mixture TS. Determine each peak area with the sample solution by the automatic integra-tion method, and calculate the composition of fatty acids by the area percentage method: myristic acid is not more than 5.0z, palmitic acid is not more than 16.0z, palmitoleic acid is not more than 8.0z, stearic acid is not more than 6.0z, oleic acid is not less than 58.0z, linoleic acid is not more than 18.0z and linolenic acid is not more than 4.0z. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.32 mm in inside diameter and 30 m in length, coated with polyethylene glycol 20 M for gas chromatography 0.5 mm in thickness. Column temperature: Inject at a constant temperature of about 809C, raise the temperature to 2209C at a rate of 109C per minute, and maintain at 2209C for 40 minutes. Injection port temperature: A constant temperature of about 2509C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: 50 cm per second. Split ratio: 1: 50. System suitability— Test for required detectability: Dissolve 0.50 g of the mix-ture of fatty acid methyl esters described in the following table in heptane to make exactly 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test add heptane to make ex-actly 10 mL. When the procedure is run with 1 mL of this so-lution under the above operating conditions, the SN ratio of methyl myristate is not less than 5. Mixture of fatty acid methyl esters Composition (z) Methyl myristate for gas chromatography 5 Methyl palmitate for gas chromatography 10 Methyl stearate for gas chromatography 15 Methyl arachidate for gas chromatography 20 Methyl oleate for gas chromatography 20 Methyl eicosenoate for gas chromatography 10 Methyl behenate 10 Methyl lignocerate for gas chro- matography 10 System performance: When the procedure is run with 1 mL of the solution for system suitability test under the above op-erating conditions, ◆methyl stearate and methyl oleate are eluted in this order,◆ the resolution between these peaks is not less than 1.8, and the number of theoretical plates of the peak of methyl stearate is not less than 30,000. Acid value <1.13> Not more than 2.0 (using ethanol (95) instead).◆ Saponification value Introduce about 4 g of Polysorbate 80, accurately weighed, into a 250-mL borosilicate glass flask. Add exactly 30 mL of 0.5 mol/L potassium hydrox-ide-ethanol VS and a few glass beads. Attach a reflux con-denser, and heat for 60 minutes. Add 1 mL of phenolphthal-ein TS and 50 mL of ethanol (99.5), and titrate <2.50> imme-diately with 0.5 mol/L hydrochloric acid VS. Perform a blank determination in the same manner. Calculate the saponification value by the following equation: 45 – 55. Saponification value ＝ (a － b) × 28.05/M M: Amount (g) of Polysorbate 80 taken a: Volume (mL) of 0.5 mol/L hydrochloric acid VS re-quired for blank determination b: Volume (mL) of 0.5 mol/L hydrochloric acid VS re-quired for sample determination Hydroxyl value Introduce about 2 g of Polysorbate 80, ac-curately weighed, into a 150-mL round bottom flask, add ex-actly 5 mL of acetic anhydride-pyridine TS, and attach an air condenser. Heat the flask in a water bath for 1 hour keeping the level of the water about 2.5 cm above the level of the liq-uid in the flask. Withdraw the flask and allow to cool. Add 5 mL of water through the condenser. If a cloudiness appears add sufficient pyridine to clear it, noting the volume added. Shake the flask, and heat in the water bath for 10 minutes. Withdraw the flask and allow to cool. Rinse the condenser and the walls of the flask with 5 mL of neutralized ethanol, and titrate <2.50> with 0.5 mol/L potassium hydroxide-ethanol VS (indicator: 0.2 mL of phenolphthalein TS). Per-form a blank determination in the same manner. Calculate the hydroxyl value by the following equation: 65 – 80. Hydroxyl value ＝ (a － b) × 28.05/M ＋ acid value M: Amount (g) of Polysorbate 80 taken a: Volume (mL) of 0.5 mol/L potassium hydroxide-ethanol VS required for blank determination b: Volume (mL) of 0.5 mol/L potassium hydroxide-ethanol VS required for sample determination
◆(1) Heavy metals <1.07>—Proceed with 1.0 g of Polysorbate 80 according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ Ethylene oxide and 1,4-dioxane—Transfer exactly 1.00 g of Polysorbate 80 into a 10-mL headspace vial, add exactly 2 mL of water, seal the vial immediately with a sep-tum of silicon rubber coated the surface with fluororesin and an aluminum cap. Mix carefully, and use the content as the sample solution. Separately, pipet 0.5 mL of a solution, pre-pared by dissolving ethylene oxide in dichloromethane so that each mL contains 50 mg, and add water to make exactly 50 mL. Allow to stand to reach room temperature. Pipet 1 mL of this solution, add water to make exactly 250 mL, and use this solution as ethylene oxide stock solution. Separately, pipet 1 mL of 1,4-dioxane, add water to make exactly 200 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as 1,4-dioxane stock solution. To exact 6 mL of ethylene oxide stock solution and exact 2.5 mL of 1,4-dioxane stock solution add water to make ex-actly 25 mL, and use this solution as ethylene oxide-1,4-dioxane standard stock solution. Separately, transfer exactly 1.00 g of Polysorbate 80 into a 10-mL headspace vial, add exactly 2 mL of ethylene oxide-1,4-dioxane standard stock solution, seal the vial immediately with a septum of silicon rubber coated the surface with fluororesin and an aluminum cap. Mix carefully, and use the content as the standard solu-tion. Perform the test with the sample solution and standard solution as directed in the head-space method under Gas Chromatography <2.02> according to the following condi-tions. The amounts of ethylene oxide and 1,4-dioxane, calcu-lated by the following equations, are not more than 1 ppm and not more than 10 ppm, respectively. Amount (ppm) of ethylene oxide ＝ 2 × CEO × Aa/(Ab － Aa) CEO: Concentration (mg/mL) of added ethylene oxide in the standard solution Aa: Peak area of ethylene oxide obtained with the sample solution Ab: Peak area of ethylene oxide obtained with the stand-ard solution Amount (ppm) of 1,4-dioxane ＝ 2 × 1.03 × CD × A?a × 1000/(A?b － A?a) CD: Concentration (mL/mL) of added 1,4-dioxane in the standard solution 1.03: Density (g/mL) of 1,4-dioxane A?a: Peak area of 1,4-dioxane obtained with the sample so-lution A?b: Peak area of 1,4-dioxane obtained with the standard solution Head-space injection conditions— Equilibration temperature in vial: A constant temperature of about 809C. Equilibration time in vial: 30 minutes. Carrier gas: Helium. Injection volume of sample: 1.0 mL. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.53 mm in inside diameter and 50 m in length, coated the inside surface with 5z diphenyl-95z dimethylpolysiloxane for gas chromatography 5 mm in thickness. Column temperature: Inject at a constant temperature of about 709C, raise the temperature to 2509C at a rate of 109C per minute, and maintain at 2509C for 5 minutes. Injection port temperature: A constant temperature of about 859C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Helium. Flow rate: 4.0 mL per minute. Split ratio: 1:3.5. System suitability— System performance: Introduce 0.100 g of acetaldehyde in a 100-mL volumetric flask, and add water to make 100 mL. To exact 1 mL of this solution add water to make exactly 100 mL. Transfer exactly 2 mL of this solution and exactly 2 mL of ethylene oxide stock solution into a 10-mL headspace vial, seal the vial immediately with a fluororesin coated silicon septum and an aluminum cap. Mix carefully, and use the content as the solution for system suitability test. When per-form the test with ◆the standard solution and◆ the solution for system suitability test under the above conditions, acetal-dehyde, ethylene oxide and 1,4-dioxane are eluted in this order, and the resolution between the peaks of acetaldehyde and ethylene oxide is not less than 2.0. Peroxide value—Introduce about 10 g of Polysorbate 80, accurately weighed, into a 100-mL beaker, dissolve in 20 mL of acetic acid (100). Add 1 mL of saturated potassium ‌iodide solution and allow to stand for 1 minute. Add 50 mL of fleshly boiled and cooled water, and titrate <2.50> with‌ 0.01 mol/L sodium thiosulfate VS, while stirring with a magnetic stirrer (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Calculate peroxide value by the following equa-tion: not more than 10.0. Peroxide value ＝ (a － b) × 10/M M: Amount (g) of Polysorbate 80 taken a: Volume (mL) of 0.01 mol/L sodium thiosulfate VS re-quired for sample determination b: Volume (mL) of 0.01 mol/L sodium thiosulfate VS re-quired for blank determination Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration). Residue on ignition Heat a quartz or platinum crucible to redness for 30 minutes, allow to cool in a desiccator (silica gel or other appropriate desiccants), and weigh accurately. Evenly distribute 2.00 g of Polysorbate 80 in the crucible, dry at 100 – 1059C for 1 hour, ◆and gradually heat with as lower temperature as possible to carbonize completely.◆ Then after igniting to constant mass in an electric furnace at 600 ± 259C, allow the crucible to cool in a desiccator, and weigh the mass accurately. Flames should not be produced at any time during the procedure. If after prolonged ignition the ash still contains black particles, take up the ash with hot water, filter through a filter paper for quantitative analysis, and ignite the residue and the filter paper. Combine the fil-trate with the ash, carefully evaporate to dryness, and ignite to constant mass: not more than 0.25z.
Powdered Poria Sclerotium occurs as a white to grayish white powder. It is almost odorless and almost tasteless, but is slightly mucous. Under a microscope <5.01>, Powdered Poria Sclerotium reveals colorless and transparent hyphae strongly refracting material. Description Usually cut; external surface dark brown to dark red-brown and with longitudinal wrinkles; cut surface brown to light yellow-brown; in case periderm fallen off, ex-ternal surface brown to light yellow-brown and fibrous; on transversely cut surface cortex and xylem almost distinctly defined, and exhibits radial structure; sometimes radial cleft observed. Odor, fragrant; taste sweet, followed by slight bitterness.
To 2.0 g of pulverized Prepared Glycyrrhiza add 10 mL of ethyl acetate, shake for 15 minutes, centrifuge, and separate the ethyl acetate layer. Shake the residue with 5 mL of ethyl acetate and 5 mL of 0.1 mol/L hydrochloric acid TS for 15 minutes, centrifuge, and use the ethyl acetate layer as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (7:2:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, heat the plate at 1059C for 3 minutes, and allow to cool: a red-purple spot is observed at an Rf value of about 0.6.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Prepared Glycyrrhiza according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g ‌2106 Processed Aconite Root / Crude Drugs and Related Drugs of pulverized Prepared Glycyrrhiza according to Method 4, and perform the test (not more than 5 ppm). Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 8.0z (6 hours). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
Weigh accurately about 0.5 g of pulverized Prepared Glycyrrhiza in a glass-stoppered centrifuge tube, add 70 mL of dilute ethanol, shake for 15 minutes, centrifuge, and sepa-rate the supernatant liquid. To the residue add 25 mL of dilute ethanol, and proceed in the same manner. Combine all the extracts, add dilute ethanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in dilute ethanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: Adjust so that the retention time of glycyrrhizic acid is about 15 minutes. System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak with the relative retention time of about 0.9 to glycyrrhizic acid, and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Mass, about 10 – 30 cm in diameter, up to 0.1 – 2 kg in mass; usually it appears as broken or chipped pieces; white or slightly reddish white; sclerotium with remaining outer layer is dark brown to dark red-brown in color, coarse, which fissures; hard in texture, but brittle. Almost odorless, almost tasteless, and slightly mucous.
(1) Warm 1 g of pulverized Poria Scleroti-um with 5 mL of acetone on a water bath for 2 minutes with shaking, and filter. Evaporate the filtrate to dryness, dis-solve the residue in 0.5 mL of acetic anhydride, and add 1 drop of sulfuric acid: a light red color develops, which light, and fragments of false tissue consisting of granules and mucilage plates. Thin hyphae, 2 – 4 mm in diameter; thick ones, usually 10 – 20 mm, up to 30 mm. Identification (1) Warm 1 g of Powdered Poria Scleroti-um with 5 mL of acetone on a water bath for 2 minutes with shaking, and filter. Evaporate the filtrate to dryness, dis-solve the residue in 0.5 mL of acetic anhydride, and add 1 drop of sulfuric acid: a light red color develops, which changes immediately to dark green. (2) To Powdered Poria Sclerotium add 1 drop of iodine TS: a deep red-brown color is produced.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Poria Sclerotium according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Poria Sclerotium according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Poria Sclerotium does not show starch grains. Total ash <5.01> Not more than 1.0z.
Potash Soap occurs as a yellow-brown, trans-parent, unctuous, soft mass, having a characteristic odor. It is freely soluble in water and in ethanol (95).
Silicic acid and alkalinity—Dissolve 10 g of Potash Soap in 30 mL of ethanol (95), and add 0.50 mL of 1 mol/L hydrochloric acid VS: no turbidity is produced. Add 1 drop of phenolphthalein TS to this solution: no red color devel- ops.
Weigh accurately about 5 g of Potash Soap, dissolve in 100 mL of hot water, and transfer to a separator. Acidify the mixture with dilute sulfuric acid, and cool. Extract the solution with 50-mL, 40-mL, and 30-mL portions of diethyl ether. Wash the combined diethyl ether extracts with 10-mL portions of water until the washing contains no acid. Trans-fer the diethyl ether solution to a tared flask, evaporate diethyl ether on a water bath at a temperature as low as pos-sible. Dry the residue at 809C to constant mass, and weigh as fatty acids.
Potato Starch occurs as a white powder. It is practically insoluble in water and in ethanol (99.5).◆
(1) Examined under a microscope <5.01> using a mixture of water and glycerin (1:1), Potato Starch presents granules, either irregularly shaped, ovoid or pear-shaped, usually 30 – 100 mm in size but occasionally exceeding 100 mm, or rounded, 10 – 35 mm in size. There are occasional compound granules having two to four compo-nents. The ovoid and pear-shaped granules have an eccentric hilum and the rounded granules acentric or slightly eccentric hilum. All granules show clearly visible concentric striations. Between orthogonally oriented polarizing plates or prisms, the granules show a distinct black cross intersecting at the hilum. To 1 g of Potato Starch add 50 mL of water, boil for 1 minute, and allow to cool: a thick, opalescent mucilage is formed. To 1 mL of the mucilage obtained in (2) add 0.05 mL of diluted iodine TS (1 in 10): an orange-red to deep blue color is formed, and the color disappears by heating. pH <2.54> Put 5.0 g of Potato Starch in a non-metal vessel, add 25.0 mL of freshly boiled and cooled water, mix gently for 1 minute, and allow to stand for 15 minutes: the pH of the solution is between 5.0 and 8.0.
(1) Iron—To 1.5 g of Potato Starch add 15 mL of 2 mol/L hydrochloric acid TS, mix, filter, and use the fil-trate as the test solution. To 2.0 mL of Standard Iron Solu-tion add water to make 20 mL, and use this solution as the control solution. Put 10 mL each of the test solution and the ‌control solution in test tubes, add 2 mL of a solution of citric acid (1 in 5) and 0.1 mL of mercapto acetic acid, and mix. Alkalize with ammonia solution (28) to litmus paper, add water to make 20 mL, and mix. Transfer 10 mL each of these solutions into test tubes, allow to stand for 5 minutes, and compare the color of these solutions against a white background: the color of the test solution is not more intense than that of the control solution (not more than 10 ppm). Oxidizing substances—To 4.0 g of Potato Starch add 50.0 mL of water, shake for 5 minutes, and centrifuge. To 30.0 mL of the supernatant liquid add 1 mL of acetic acid (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow to stand for 25 to 30 minutes at a dark place. Add 1 mL of starch TS, and titrate <2.50> with 0.002 mol/L sodium thio-sulfate VS until the color of the solution disappears. Per-form a blank determination in the same manner, and make any necessary correction: the volume of 0.002 mol/L sodium thiosulfate VS consumed is not more than 1.4 mL (not more than 20 ppm, calculated as hydrogen peroxide). Sulfur dioxide— Apparatus Use as shown in the figure. A: Three-necked round-bottom flask (500 mL) B: Cylindrical dropping funnel (100 mL) C: Condenser D: Test tube E: Tap Procedure Introduce 150 mL of water into the three-necked round-bottom flask, close the tap of the cylindrical dropping funnel, and pass carbon dioxide through the whole system at a rate of 100 ± 5 mL per minute. Pass cooling water through the condenser, and place 10 mL of hydrogen peroxide-sodium hydroxide TS in the test tube. After 15 minutes, remove the funnel without interrupt-ing the stream of carbon dioxide, and introduce through the opening into the flask about 25 g of Potato Starch, accu-rately weighed, with the aid of 100 mL of water. Apply tap grease to the outside of the connection part of the funnel, and load the funnel. Close the tap of the funnel, pour 80 mL of 2 mol/L hydrochloric acid TS into the funnel, open the tap to introduce the hydrochloric acid into the flask, and close the tap while several mL of the hydrochloric acid remains, in order to avoid losing sulfur dioxide. Place the flask in a water bath, and heat the mixture for 1 hour. Transfer the contents of the test tube with the aid of a little water to a wide-necked conical flask. Heat in a water bath for 15 minutes, and cool. Add 0.1 mL of bromophenol blue TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the amount of sulfur dioxide by applying the following for-mula: it is not more than 50 ppm. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 M: Amount (g) of Potato Starch taken V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed (4) Foreign matter—Under a microscope <5.01>, Potato Starch does not contain starch granules of any other origin. It may contain a minute quantity, if any, of frag-ments of the tissue of the original plant.◆ Loss on drying <2.41> Not more than 20.0z (1 g, 1309C, 90 minutes). Residue on ignition <2.44> Not more than 0.6z (1 g).
Povidone-Iodine is a complex of iodine with 1-vinyl- K ＝ ＋ 0.15 ＋ 0.003 c 0.15 c ＋ 0.003 c2 pyrrolidone polymer. c: Mass (g) of Povidone in 100 mL of the solution, calcu-lated on the anhydrous basis It contains not less than 9.
Povidone-Iodine occurs as a dark red-brown powder. It has a faint, characteristic odor. Nominal K-value calculated on anhydrous basis It is freely soluble in water and in ethanol (99.5). The pH of a solution obtained by dissolving 1.0 g of Povidone-Iodine in 100 mL of water is between 1.5 and 3.5. Not more than 18 5.00 More than 18 and not more than 95 1.00 More than 95 0.10 The K-value is not less than 85z and not more than 115.0z of the nominal K-value when the nominal K-value is not more than 15, and the K-value is not less than 90.0z and not more than 108.0z of the nominal K-value when the nominal K-value is more than 15.
(1) To 10 mL of diluted starch TS (1 in 10) add 1 drop of a solution of Povidone-Iodine (1 in 10): a deep blue color develops. (2) To 1 mL of a solution of Povidone-Iodine (1 in 100) add 1 mL of sodium thiosulfate TS, and add 1 mL of ammo-nium thiocyanate-cobalt (II) nitrate TS and 2 drops of 1 mol/L hydrochloric acid TS: a blue color develops, and a blue precipitate is gradually formed.
(1) Clarity and color of solution—Dissolve 0.30 g of Povidone-Iodine in 100 mL of water: the solution is clear and brown. Heavy metals <1.07>—Proceed with 1.0 g of Povidone-Iodine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Povidone-Iodine according to Method 4, and perform the test (not more than 2 ppm). Iodide ion—Weigh accurately about 0.5 g of Povidone-Iodine, dissolve in 100 mL of water, and add sodi-um hydrogensulfite TS until the color of iodine completely disappears. To this solution add exactly 25 mL of 0.1 mol/L silver nitrate VS, shake thoroughly with 10 mL of nitric acid, titrate <2.50> the excess silver nitrate with 0.1 mol/L ammo-nium thiocyanate VS until the solution develops a red-brown color, and calculate the total amount of iodine (indicator: 1 mL of ammonium iron (III) sulfate TS). Perform a blank de-termination in the same manner. Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 12.69 mg of I Obtain the amount of iodide ion, calculated on the dried basis, by deducting the amount (z) of available iodine from the total amount (z) of iodine: not more than 6.6z. Loss on drying <2.41> Not more than 8.0z (1 g, 1009C, 3 hours). Residue on ignition <2.44> Not more than 0.05z (5 g). ‌Assay (1) Available iodine—Weigh accurately about 0.5 g of Povidone-Iodine, dissolve in 30 mL of water, and titrate <2.50> with 0.02 mol/L sodium thiosulfate VS (indica-tor: 2 mL of starch TS). Each mL of 0.02 mol/L sodium thiosulfate VS ＝ 2.538 mg of I (2) Nitrogen—Weigh accurately about 20 mg of Povidone-Iodine, and perform the test as directed under Nitrogen Determination <1.08>.
Weigh accurately about 0.1 g of Povidone, and place in a Kjeldahl flask. Add 5 g of a decomposition ac-celerator (a powdered mixture of 33 g of potassium sulfate, 1 g of copper (II) sulfate pentahydrate and 1 g of titanium (IV) oxide), and wash down any adhering sample from the neck of the flask with a small amount of water. Add 7 mL of sul-furic acid allowing to flow down the inside wall of the flask. Heat the flask gradually over a free flame until the solution has a clear, yellow-green color and the inside wall of the flask is free from a carbonaceous material, and then heat for further 45 minutes. After cooling, add cautiously 20 mL of water, and connect the flask to the distillation apparatus pre-viously washed by passing steam through it. To the absorp-tion flask add 30 mL of a solution of boric acid (1 in 25), 3 drops of bromocresol green-methyl red TS and sufficient water to immerse the lower end of the condenser tube. Add 30 mL of a solution of sodium hydroxide (2 in 5) through the funnel, rinse cautiously the funnel with 10 ml of water, im-mediately close the clamp attached to the rubber tube, then start the distillation with steam to get 80 to 100 mL of the distillate. Remove the absorption flask from the lower end of the condenser tube, rinsing the end part with a small quantity of water, and titrate <2.50> the distillate with 0.025 mol/L sulfuric acid VS until the color of the solution changes from green through pale grayish blue to pale grayish red-purple. Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.025 mol/L sulfuric acid VS ＝ 0.700 mg of N Identification (1) To 10 mL of diluted starch TS (1 in 10) add 1 drop of a solution of Povidone-Iodine (1 in 10): a deep blue color develops. (2) To 1 mL of a solution of Povidone-Iodine (1 in 100) add 1 mL of sodium thiosulfate TS, and add 1 mL of ammo-nium thiocyanate-cobalt (II) nitrate TS and 2 drops of 1 mol/L hydrochloric acid TS: a blue color develops, and a blue precipitate is gradually formed. Purity (1) Clarity and color of solution—Dissolve 0.30 g of Povidone-Iodine in 100 mL of water: the solution is clear and brown. Heavy metals <1.07>—Proceed with 1.0 g of Povidone-Iodine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Povidone-Iodine according to Method 4, and perform the test (not more than 2 ppm). Iodide ion—Weigh accurately about 0.5 g of Povidone-Iodine, dissolve in 100 mL of water, and add sodi-um hydrogensulfite TS until the color of iodine completely disappears. To this solution add exactly 25 mL of 0.1 mol/L silver nitrate VS, shake thoroughly with 10 mL of nitric acid, titrate <2.50> the excess silver nitrate with 0.1 mol/L ammo-nium thiocyanate VS until the solution develops a red-brown color, and calculate the total amount of iodine (indicator: 1 mL of ammonium iron (III) sulfate TS). Perform a blank de-termination in the same manner. Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 12.69 mg of I Obtain the amount of iodide ion, calculated on the dried basis, by deducting the amount (z) of available iodine from the total amount (z) of iodine: not more than 6.6z. Loss on drying <2.41> Not more than 8.0z (1 g, 1009C, 3 hours). Residue on ignition <2.44> Not more than 0.05z (5 g). ‌Assay (1) Available iodine—Weigh accurately about 0.5 g of Povidone-Iodine, dissolve in 30 mL of water, and titrate <2.50> with 0.02 mol/L sodium thiosulfate VS (indica-tor: 2 mL of starch TS). Each mL of 0.02 mol/L sodium thiosulfate VS ＝ 2.538 mg of I (2) Nitrogen—Weigh accurately about 20 mg of Povidone-Iodine, and perform the test as directed under Nitrogen Determination <1.08>.
Povidone occurs as a white to slightly yellow-ish fine powder. It is odorless or has a faint, characteristic odor. It is freely soluble in water, in methanol and in ethanol (99.5). It is hygroscopic.◆
(1) To 0.5 g of Povidone add 10 mL of water, and shake: it dissolves. Determine the infrared absorption spectrum of Povi-done, previously dried at 1059C for 6 hours, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Povidone for Iden-tification RS (previously dried at 1059C for 6 hours): both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 1.0 g of Povidone in 20 mL of water: the pH of this solution is between 3.0 and 5.0 for Povidone having the nominal K-value of 30 or less, and between 4.0 and 7.0 for Povidone having the nominal K-value exceeding 30.
◆(1) Clarity and color of solution—Dissolve 1.0 g of Povidone in 20 mL of water: the solution is clear and col-orless to pale yellow, or pale red.◆ (2) Heavy metals <1.07>—Proceed with 2.0 g of Povi- done according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm).◆ Aldehydes—Weigh accurately about 1 g of Povidone and dissolve in 0.05 mol/L pyrophosphate buffer solution (pH 9.0) to make exactly 100 mL. Stopper, heat at 609C for 60 minutes, allow to cool to room temperature, and use this solution as the sample solution. Separately, dissolve 0.140 g of acetaldehyde ammonia trimer trihydrate in water to make exactly 200 mL. Pipet 1 mL of this solution, add 0.05 mol/L pyrophosphate buffer solution (pH 9.0) to make exactly 100 mL, and use this solution as the standard solution. Measure exactly 0.5 mL each of the sample solution, standard solu-tion and water, transfer to separate 1-cm cells, add 2.5 mL of 0.05 mol/L pyrophosphate buffer solution (pH 9.0) and 0.2 mL of b-nicotinamide adenine dinucleotide TS to each of these cells, mix and stopper tightly. Allow to stand for 2 to 3 minutes at 22 ± 29C, and perform the test with these solu-tions as directed under Ultraviolet-visible Spectrophotome-try <2.24> using water as the control solution. Determine the absorbances, AT1, AS1 and AB1 of the subsequent solutions of the sample solution, the standard solution and water at 340 nm. Add 0.05 mL of aldehyde dehydrogenase TS to each of the cells, mix and stopper tightly. Allow to stand for 5 minutes at 22 ± 29C. Determine the absorbances, AT2, AS2 and AB2 of these solutions in the same manner as above, and calculate the content of aldehydes by the follwing equation: the content of aldehydes is not more than 500 ppm. Content (ppm) of aldehydes [as acetaldehyde (CH3CHO)] ＝ C/M ×{(AT2 － AT1) － (AB2 － AB1)}/{(AS2 － AS1) － (AB2 － AB1)}× 100,000 M: Amount (g) of Povidone taken, calculated on the an-hydrous basis C: Concentration (mg/mL) of acetaldehyde in the stand-ard solution, using 0.72 as conversion factor for acetal-dehyde ammonia trimer trihydrate to acetaldehyde 1-Vinyl-2-pyrrolidone—Weigh accurately about 0.25 g of Povidone, dissolve in a mixture of water and acetonitrile (9:1) to make exactly 10 mL, and use this solu-tion as the sample solution. Separately, dissolve 50 mg of 1-vinyl-2-pyrrolidone in a mixture of water and acetonitrile (9:1) to make exactly 100 mL. Pipet 1 mL of this solution and add a mixture of water and acetonitrile (9:1) to make ex-actly 100 mL. Pipet 5 mL of this solution, add a mixture of water and acetonitrile (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, determine the peak areas, AT and AS, of 1-vinyl-2-pyrrolidone in each solution, and calculate the content of 1-vinyl-2-pyrrolidone by the fol-lowing equation: it is not more than 10 ppm. Content (ppm) of 1-vinyl-2-pyrrolidone ＝ 1/M × AT/AS × 2.5 M: Amount (g) of Povidone taken, calculated on the an-hydrous basis Operating conditions— Detector: An ultraviolet spectrophotometer (detection wavelength: 235 nm). Column: Stainless steel columns 4.0 mm in inside diameter and 10 mm in length, and 4.6 mm in inside diameter and 150 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter), and use them as a guard column and a separation column, respec-tively. Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (9:1). Flow rate: 1.0 mL per minutes. System suitability— System performance: Dissolve 10 mg of 1-vinyl-2-pyrroli-done and 0.5 g of vinyl acetate in 100 mL of methanol. To 1 mL of this solution add a mixture of water and acetonitrile (9:1) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 1-vinyl-2-pyrrolidone and vinyl acetate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 1-vinyl-2-pyrrolidone is not more than 2.0z. Peroxides—Weigh exactly an amount of Povidone, equivalent to 4.0 g calculated on the anhydrous basis, dis-solve in water to make exactly 100 mL, and use this solution as the sample solution. To 25 mL of the sample solution add 2 mL of titanium (III) chloride-sulfuric acid TS, and mix. Allow to stand for 30 minutes, and perform the test with this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared by adding 2 mL of diluted sulfuric acid (13 in 100) to 25 mL of the sample solu-tion as a blank: the absorbance of the solution at 405 nm is not more than 0.35 (not more than 400 ppm, expressed as hydrogen peroxide). Hydrazine—Weigh exactly an amount of Povidone equivalent to 2.5 g calculated on the anhydrous basis, trans-fer to a 50-mL centrifuge tube, add 25 mL of water, and stir to dissolve. Add 500 mL of a solution of salicylaldehyde in methanol (1 in 20), stir and warm at 609C for 15 minutes in a water bath. Allow to cool, add 2.0 mL of toluene, stopper tightly, shake vigorously for 2 minutes, centrifuge, and use the upper layer of the mixture as the sample solution. Sepa-rately, dissolve 90 mg of salicylaldazine in toluene to make exactly 100 mL. Pipet 1 mL of this solution, add toluene to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of dimethylsilanized silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol and water (2:1) to a distance of about three-fourths of the length of the plate, and air-dry the plate. Exa-mine under ultraviolet light (main wavelength: 365 nm): the fluorescence of the spot obtained from the sample solution Content of formic acid(z) ＝ MS/MT × AT/AS MS: Amount (g) of formic acid taken MT: Amount (g) of Povidone taken, calculated on the an- hydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 7.8 mm in inside di-ameter and 300 mm in length, packed with strongly acidic ion exchange resin for liquid chromatography (9 mm in parti-cle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Diluted perchloric acid (1 in 700). Flow rate: 1.0 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of formic acid are not less han 1000 and 0.5 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of formic acid is not more than 2.0z. (8) 2-Pyrrolidone—Weigh accurately about 0.5 g of Povidone, dissolve in a mixture of water and methanol for liquid chromatography (19:1) to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 0.150 g of 2-pyrrolidone in a mixture of water and methanol for liquid chromatography (19:1) to make exactly 100 mL. Pipet 2 mL of this solution, add a mixture of water and methanol for liquid chromatography (19:1) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of 2-pyrrolidone in each solution. Calculate the content of 2-pyrrolidone by the following equation: not more than 3.0z. corresponding to the spot having a Rf value of about 0.3 from the standard solution is not more intense than that of Content (z) of 2-pyrrolidone ＝ 1/M × AT/AS × 0.3 the spot from the standard solution (not more than 1 ppm). (7) Formic acid—Weigh accurately about 2 g of Pivo-done, dissolve in water to make exactly 100 mL, and use this solution as the sample stock solution. Transfer a strongly acidic ion exchange resin (H type) for column chromato-graphy previously suspended in water to a column of about 8 mm in inside diameter to give a packing depth of about 20 mm in length, and keep the resin layer constantly immersed in water. Pour 5 mL of water to the column, and adjust the flow rate about 1 mL per minute. When the level of the water comes down to near the top of the resin layer, put the sample stock solution into the column, discard the first 2 mL of the eluent, take 1.5 mL of the subsequent eluent, and use this solution as the sample solution. Separately, weigh ac-curately about 0.1 g of formic acid, dissolve in water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the stan-dard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of for-mic acid in each solution. Calculate the content of formic acid by the following equation: it is not more than 0.5z. M: The amount (g) of Povidone taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: Stainless steel columns 4.0 mm in inside diameter and 10 mm in length, and 4.6 mm in inside diameter and 150 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter), and use them as a guard column and a separation column, respec-tively. Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol for liquid chromatography (19 : 1). Flow rate: 0.8 mL per min. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of 2-pyrrolidone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times ‌with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 2-pyrrolidone is not more than 2.0z. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). K-value Weigh accurately an amount of Povidone, calcu-lated on the anhydrous basis, specified in the table below ac-cording to the nominal K-value, dissolve in water to make exactly 100 mL, allow to stand for 60 minutes, and use this solution as the sample solution. Perform the test with the sample solution and with water at 259C as directed in Method 1 under Viscosity Determination <2.53>, and calcu-late the K-value by the following formula. 300 c log nrel. ＋ (c ＋ 1.5 c log nrel.)2 1.5 log nrel. － 1 ◆Containers and storage Containers—Tight containers.◆ Povidone-Iodine
Pranlukast Hydrate contains not less than 98.
Pranlukast Hydrate occurs as a white to light yellow crystalline powder. It is very slightly soluble in ethanol (99.5), and practically insoluble in water. Melting point: about 2339C (with decomposition).
(1) Determine the absorption spectrum of a solution of Pranlukast Hydrate in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Pranlukast RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pran-lukast Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Pranlukast RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Suspend 1.0 g of Pran-lukast Hydrate in 10 mL of N,N-dimethylformamide, proceed according to Method 4, and perform the test. Pre-pare the control solution with 10 mL of N,N-dimethylfor-mamide in the same manner as preparation of the test solu-tion, and add 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Suspend 1.0 g of Pranlukast Hy- drate in 10 mL of N,N-dimethylformamide, then proceed according to Method 4, and perform the test (not more than 2 ppm). Related substances—Dissolve 20 mg of Pranlukast Hydrate in 50 mL of a mixture of acetonitrile and dimethyl-sulfoxide (3:1), and use this solution as the sample solution. not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pranlukast is not more than 2.0z. Water <2.48> 1.5 – 2.2z (50 mg, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 20 mg each of Pranlukast Hydrate and Pranlukast RS (separately determine the water <2.48> in the same manner as Pranlukast Hydrate), dissolve them separately in a mixture of acetonitrile and dimethylsul-foxide (3:1) to make exactly 50 mL. To exactly 5 mL each of these solutions add exactly 5 mL of the internal standard so-lution, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 4 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of pranlukast to that of the internal standard. Amount (mg) of pranlukast (C27H23N5O4) ＝ MS × QT/QS MS: Amount (mg) of Pranlukast RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isoamyl para-hydroxybenzoate in a mixture of acetonitrile and dimethyl-sulfoxide (3:1) (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for ‌liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.02 mol/L potassium dihy-drogen phosphate TS, acetonitrile and methanol (5:5:1). Flow rate: Adjust so that the retention time of pranlukast is about 10 minutes. System suitability— System performance: When the procedure is run with 4 mL of the standard solution under the above operating condi-tions, pranlukast and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 4 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of pranlukast to that of the internal standard is not more than 1.0z.
Pranoprofen, when dried, contains not less than 98.
Pranoprofen occurs as a white to pale yellow-white crystalline powder. It is freely soluble in N,N-dimethylformamide, soluble in acetic acid (100), sparingly soluble in methanol, slightly solu-ble in acetonitrile, in ethanol (95) and in acetic anhydride, very slightly soluble in diethyl ether, and practically insolu-ble in water. A solution of Pranoprofen in N,N-dimethylformamide (1 in 30) shows no optical rotation.
(1) Dissolve 0.02 g of Pranoprofen in 1 mol/L hydrochloric acid TS to make 100 mL, and dilute 10 mL of the solution with water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. (2) Determine the infrared absorption spectrum of Pranoprofen as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 0.5 g of Prano-profen in 40 mL of methanol, and 6 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows. To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of methanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Heavy metals <1.07>—Proceed with 2.0 g of Prano- profen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of the Standard Lead Solu-tion (not more than 10 ppm). Related Substances—Dissolve 50 mg of Pranoprofen in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area from both so-lutions by the automatic integration method: the each area of the peaks other than pranoprofen obtained from the sam-ple solution is not larger than the peak area of pranoprofen from the standard solution, and the total peak area of them is not larger than 2 times the peak area of pranoprofen from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column about 6 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.02 g of sodium perchlorate monohydrate in 1000 mL of water, and adjust the pH to 2.5 with perchloric acid. To 2 volumes of this solution add 1 volume of acetonitrile. Flow rate: Adjust so that the retention time of pranoprofen is about 10 minutes. Selection of column: Dissolve 4 mg each of Pranoprofen and ethyl parahydroxybenzoate in 200 mL of the mobile phase. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of pranoprofen and ethyl parahydrox-ybenzoate in this order with the resolution between these peaks being not less than 2.1. Detection sensitivity: Adjust the detection sensitivity so that the peak height of pranoprofen obtained from 10 mL of the standard solution is between 10 mm and 20 mm. Time span of measurement: About three times as long as the retention time of pranoprofen. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Pranoprofen, previ-ously dried, dissolve in 70 mL of a mixture of acetic anhy-dride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 25.53 mg of C15H13NO3
Prazepam occurs as white to light yellow crys-tals or crystalline powder. It is odorless. It is freely soluble in acetone, soluble in acetic anhydride, sparingly soluble in ethanol (99.5) and in diethyl ether, and practically insoluble in water.
(1) Dissolve 0.01 g of Prazepam in 3 mL of sulfuric acid, and observe under ultraviolet light (main wave-length: 365 nm): the solution shows a grayish blue fluores-cence. Dissolve 0.01 g of Prazepam in 1000 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the absorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Prazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform Flame Coloration Tests <1.04> (2) with Prazepam: a green color appears.
(1) Chloride <1.03>—To 1.0 g of Prazepam add 50 mL of water, allow to stand for 1 hour with occasional shak-ing, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Sulfate <1.14>—To 20 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Praze- pam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Prazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.40 g of Prazepam in 10 mL of acetone, and use this solution as the sample solu- ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.48 mg of C19H17ClN2O
Weigh accurately about 0.4 g of Prazepam, previ- of prazepam (C 19H17 ClN2O). ously dried, dissolve in 60 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti- Description Prazepam occurs as white to light yellow crys-tals or crystalline powder. It is odorless. It is freely soluble in acetone, soluble in acetic anhydride, sparingly soluble in ethanol (99.5) and in diethyl ether, and practically insoluble in water. Identification (1) Dissolve 0.01 g of Prazepam in 3 mL of sulfuric acid, and observe under ultraviolet light (main wave-length: 365 nm): the solution shows a grayish blue fluores-cence. Dissolve 0.01 g of Prazepam in 1000 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the absorption spectrum of the solution as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Prazepam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform Flame Coloration Tests <1.04> (2) with Prazepam: a green color appears. Melting point <2.60> 145 – 1489C Purity (1) Chloride <1.03>—To 1.0 g of Prazepam add 50 mL of water, allow to stand for 1 hour with occasional shak-ing, and filter. To 20 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Sulfate <1.14>—To 20 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Praze- pam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Prazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.40 g of Prazepam in 10 mL of acetone, and use this solution as the sample solu- ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.48 mg of C19H17ClN2O
Prazosin Hydrochloride, when dried, contains not less than 97.
Prazosin Hydrochloride occurs as a white crys-talline powder. It is slightly soluble in methanol, very slightly soluble in ethanol (99.5) and practically insoluble in water. It gradually turns pale yellow-white on exposure to light. Melting point: about 2709C (with decomposition).
(1) Determine the absorption spectrum of a solution of Prazosin Hydrochloride in 0.01 mol/L hydro-chloric acid-methanol TS (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum (100): the solution responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Prazosin Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Prazosin Hy-drochloride in 20 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area of both solutions by the automatic integration method: the area of each peak other than prazo-sin obtained from the sample solution is not larger than 2 times the peak area of prazosin from the standard solution, and the total area of the peaks other than prazosin from the sample solution is not larger than 5 times the peak area of prazosin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.484 g of sodium 1-pentane sul-fonate and 18 mL of tetramethylammonium hydroxide in 900 mL of water, adjust the pH to 5.0 with acetic acid (100), and add water to make 1000 mL. To this solution add 1000 mL of methanol. Flow rate: Adjust so that the retention time of prazosin is about 9 minutes. Time span of measurement: About 6 times as long as the retention time of prazosin. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of prazosin obtained with 20 mL of this solution is equivalent to 35 to 65z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of prazosin are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌area of prazosin is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 25 mg each of Prazosin Hy-drochloride and Prazosin Hydrochloride RS, previously dried, and dissolve each in methanol to make exactly 50 mL. Pipet 3 mL each of these solutions, and add a mixture of methanol and water (7:3) to make exactly 100 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of prazosin in each solution. Description Prednisolone occurs as a white crystalline pow-der. It is soluble in methanol and in ethanol (95), slightly solu-ble in ethyl acetate, and very slightly soluble in water. Melting point: about 2359C (with decomposition). It shows crystal polymorphism. Identification (1) To 2 mg of Prednisolone add 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluorescence, develops. To this solution add 10 mL of water cautiously: the color disappears and a gray, flocculent precipitate is formed. (2) Determine the infrared absorption spectrum of Pred-nisolone, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Prednisolone RS: both Amount (mg) of prazosin hydrochloride (C ＝ MS × AT/AS 19H21N5O4.HCl) spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spec-tra, dissolve Prednisolone and Prednisolone RS in ethyl MS: Amount (mg) of Prazosin Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, water, acetic acid (100) and diethylamine (3500:1500:50:1). Flow rate: Adjust so that the retention time of prazosin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of prazosin are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of prazosin is not more than 1.0z.
Prednisolone Acetate, when dried, contains not less than 96.
Prednisolone Acetate occurs as a white crystal-line powder. It is slightly soluble in methanol and in ethanol (99.5), and practically insoluble in water. Melting point: about 2359C (with decomposition). It shows crystal polymorphism.
(1) To 2 mg of Prednisolone Acetate add 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluorescence, develops. To this solu-tion add 10 mL of water cautiously: the color disappears and a gray, flocculent precipitate is formed. (2) Determine the infrared absorption spectra of Pred-nisolone Acetate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum in a range between 4000 cm－1 and 650 cm－1 with the Infrared Reference Spec-trum or the spectrum of previously dried Prednisolone Acetate RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. If any difference ap-pears, dissolve the sample and the RS in ethanol (99.5), respectively, evaporate to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋128 – ＋1379(after drying, 70 mg, methanol, 20 mL, 100 mm).
Related substances—Dissolve 0.20 g of Predniso-lone Acetate in exactly 10 mL of a mixture of chloroform and methanol (9:1), and use this solution as the sample solu- plate. Examine under ultraviolet light (wavelength: 254 mm): the spots obtained from the sample solution corresponding to those from the standard solution are not more intense than the spots from the standard solution, and any spot from the sample solution other than the principal spot and the spots from prednisolone, cortisone acetate and hydrocor-tisone acetate does not appear. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Dissolve about 10 mg each of Prednisolone Acetate and Prednisolone Acetate RS, previously dried and accu-rately weighed, in 60 mL each of methanol, add exactly 2 mL each of the internal standard solution, then add metha-nol to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak height of prednisolone acetate to that of the inter-nal standard. Amount (mg) of prednisolone acetate (C23H30O6) ＝ MS × QT/QS MS: Amount (mg) of Prednisolone Acetate RS taken Internal standard solution—A solution of butyl parahydrox-ybenzoate in methanol (3 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (3:2). Flow rate: Adjust so that the retention time of predniso- lone acetate is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, prednisolone acetate and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak height of prednisolone acetate to that of the in-ternal standard is not more than 1.0z.
プレドニゾロンコハク酸エステル
Prednisolone Succinate, when dried, contains not less than 97.
Prednisolone Succinate occurs as a white, fine, crystalline powder. It is odorless. It is freely soluble in methanol, soluble in ethanol (95), and very slightly soluble in water and in diethyl ether. Melting point: about 2059C (with decomposition).
(1) To 2 mg of Prednisolone Succinate add 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluorescence, develops. To this so-lution add 10 mL of water cautiously: the color disappears and a gray, flocculent precipitate is formed. (2) Determine the infrared absorption spectrum of Pred- nisolone Succinate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Prednisolone Succinate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋114 – ＋1209(after drying, 67 mg, methanol, 10 mL, 100 mm).
Related substances—Dissolve 0.10 g of Predniso-lone Succinate in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 30 mg of prednisolone in methanol to make exactly 10 mL. Pipet 1 mL of the solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and ethanol (95) (2:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 10 mg each of Prednisolone Succinate and Prednisolone Succinate RS, previously dried, and dissolve each in methanol to make exactly 100 mL. Pipet 5 mL each of these solutions, add methanol to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of prednisolone succinate (C25H32O8) ＝ MS × AT/AS MS: Amount (mg) of Prednisolone Succinate RS taken
Prednisolone, when dried, contains not less than 97.
(1) Selenium—To 0.10 g of Prednisolone add 0.5 mL of a mixture of perchloric acid and sulfuric acid (1:1) and 2 mL of nitric acid, and heat on a water bath until no more brown gas evolves and the solution becomes to be a light yellow clear solution. After cooling, add 4 mL of nitric acid to this solution, then add water to make exactly 50 mL, and use this solution as the sample solution. Separately, pipet 3 mL of Standard Selenium Solution, add 0.5 mL of a mixture of perchloric acid and sulfuric acid (1:1) and 6 mL of nitric acid, then add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions, and determine constant ab-sorbances, AT and AS, obtained on a recorder after rapid increasing of the absorption: AT is smaller than AS (not more than 30 ppm). Perform the test by using a hydride generating system and a thermal absorption cell. Lamp: A selenium hollow cathode lamp. Wavelength: 196.0 nm. Temperature of sample atomizer: When an electric fur-nace is used, about 10009C. Carrier gas: Nitrogen or argon. (2) Related substances—Dissolve 20 mg of Prednisolone in exactly 2 mL of a mixture of methanol and chloroform (1:1), and use this solution as the sample solution. Sepa-rately, dissolve 20 mg of hydrocortisone and 10 mg of pred-nisolone acetate each in a mixture of methanol and chlo-roform (1:1) to make exactly 100 mL, and use these solutions as the standard solution (1) and standard solution (2). Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, toluene and diethylamine (55:45:2) to a distance of about 15 cm, and air-dry the plate (do not dip the filter paper in the developing vessel). Spray evenly alkaline blue tetrazolium TS on the plate: the spots obtained from the sample solution corresponding to those from the standard solutions (1) and (2) are not more intense than the spots from the standard solutions (1) and (2), and ‌no spots other than the principal spot, hydrocortisone and prednisolone acetate appear from the sample solution. Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Dissolve about 25 mg each of Prednisolone and Prednisolone RS, previously dried, and accurately weighed, in 50 mL of methanol, add exactly 25 mL of the internal standard solution to each, and add methanol to make 100 mL. To 1 mL each of these solutions add the mobile phase to make 10 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of prednisolone to that of the internal standard. (2) Determine the infrared absorption spectra of the residue obtained in (1) and Prednisolone RS, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: both spectra ex-hibit similar intensities of absorption at the same wave num-bers. If any difference appears, dissolve the sample and the RS in ethyl acetate, evaporate to dryness, and repeat the test on the residues. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Prednisolone Tablets to a volumetric flask, and shake with 10 mL of water until the tablet is disin-tegrated. Add 50 mL of methanol, shake for 30 minutes, and add methanol to make exactly 100 mL. Centrifuge this solu-tion, pipet V mL of the supernatant liquid, and add metha-nol to make exactly V? mL to provide a solution that con- Amount (mg) of prednisolone (C 21H28O5) ＝ M S × QT/QS tains about 10 mg of prednisolone (C21H28O5) per ml, and use this solution as the sample solution. Separately, weigh accu- MS: Amount (mg) of Prednisolne RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 247 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with fluorosilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol (13:7). Flow rate: Adjust so that the retention time of predniso- lone is about 15 minutes. System suitability— System performance: Dissolve 25 mg of Prednisolone and 25 mg of hydrocortisone in 100 mL of methanol. To 1 mL of this solution add the mobile phase to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrocortisone and prednisolone are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of prednisolone to that of the internal standard is not more than 1.0z.
Primidone occurs as a white, crystalline pow-der or granules. It is odorless and has a slightly bitter taste. It is soluble in N,N-dimethylformamide, sparingly soluble in pyridine, slightly soluble in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether.
(1) Heat 0.5 g of Primidone with 5 mL of diluted sulfuric acid (1 in 2): the odor of formaldehyde is perceptible. (2) Mix 0.2 g of Primidone with 0.2 g of anhydrous sodi-um carbonate, and heat: the gas evolved changes moistened red litmus paper to blue.
(1) Clarity and color of solution—Dissolve 0.10 g of Primidone in 10 mL of N,N-dimethylformamide: the so-lution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Primi- done according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). 2-Ethyl-2-phenylmalonediamide—Dissolve 0.10 g of Primidone in 2 mL of pyridine, add exactly 2 mL of the internal standard solution, then add 1 mL of bis-trimethyl silyl acetamide, shake well, and heat at 1009C for 5 minutes. Cool, add pyridine to make 10 mL, and use this solution as the sample solution. Separately, dissolve 50 mg of 2-ethyl-2-phenylmalonediamide in pyridine to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, proceed in the same manner as Primi-done, and use this solution as the standard solution. Per-form the test with 2 mL of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of 2-ethyl-2-phenyl-malonediamide to that of the internal standard: QT is not more than QS. Internal standard solution—A solution of stearylalcohol in pyridine (1 in 2000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 150 cm in length, packed with siliceous earth for gas chromatog-raphy (125 to 150 mm in particle diameter) coated with 50z phenyl-methyl silicon polymer for gas chromatography at the ratio of 3z. Column temperature: A constant temperature of about 1959C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of stearylalco- hol is about 10 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tion, 2-ethyl-2-phenylmalonediamide and the internal stand-ard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 5 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of 2-ethyl-2-phenylmalonediamide to that of the internal standard is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 20 mg each of Primidone and Primidone RS, previously dried, dissolve each in 20 mL of ethanol (95) by warming, and after cooling, add ethanol (95) to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absorbance, A1, of the sample solution and standard solution at the wavelength of maximum absorption at about 257 nm, and the absorbances, A2 and A3, at the wavelength of minimum absorption at about 254 nm and at about 261 nm, as directed under Ultraviolet-visible Spectro-photometry <2.24>, using ethanol (95) as the blank. Amount (mg) of primidone (C12H14N2O2) ＝ MS × (2A1 － A2 － A3)T/(2A1 － A2 － A3)S MS: Amount (mg) of Primidone RS taken where, (2A1 － A2 － A3)T is the value from the sample solu-tion, and (2A1 － A2 － A3)S is from the standard solution.
Probenecid, when dried, contains not less than 98.
Probenecid occurs as white, crystals or crystal-line powder. It is odorless, and has a slightly bitter taste, fol-lowed by unpleasant bitter. It is sparingly soluble in ethanol (99.5), and practically in-soluble in water. It dissolves in sodium hydroxide TS and in ammonia TS. Melting point: 198 – 2009C
(1) Heat Probenecid strongly: the odor of sulfur dioxide is perceptible. (2) Determine the absorption spectrum of a solution of Probenecid in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Probenecid RS prepared in the same manner ‌as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths.
(1) Acidity—To 2.0 g of Probenecid add 100 mL of water, heat on a water bath with occasional shaking for 30 minutes, cool, and filter. To the filtrate add 1 drop of phenolphthalein TS and 0.50 mL of 0.1 mol/L sodium hy-droxide VS: a red color develops. Chloride <1.03>—To 1.0 g of Probenecid add 100 mL of water and 1 mL of nitric acid, and heat on a water bath with occasional shaking for 30 minutes. After cooling, add, if necessary, water to make 100 mL, and filter. Perform the test using 50 mL of the filtrate as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochlo-ric acid VS (not more than 0.021z). Sulfate <1.14>—To 1.0 g of Probenecid add 100 mL of water and 1 mL of hydrochloric acid, and heat on a water bath with occasional shaking for 30 minutes. After cooling, add, if necessary, water to make 100 mL, and filter. Perform the test using 50 mL of the filtrate as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 2.0 g of Probenecid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Probenecid according to Method 3, and perform the test (not more than 2 ppm). tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Probenecid RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Probenecid Tablets add 30 mL of water and 2 mL of 1 mol/L hydrochloric acid TS, sonicate with occa-sional shaking to disintegrate the tablet completely, and add ethanol (99.5) to make exactly 100 mL. Centrifuge this solu-tion, pipet 3 mL of the supernatant liquid, and add 1 mL of 1 mol/L hydrochloric acid TS and ethanol (99.5) to make ex-actly 50 mL. Pipet 5 mL of this solution, and add ethanol (99.5) to make exactly V mL so that each mL contains about 15 mg of probenecid (C13H19NO4S), and use this solution as the sample solution. Separately, weigh accurately about 0.125 g of Probenecid RS, previously dried at 1059C for 4 hours, dissolve in 15 mL of water, 1 mL of 1 mol/L hydro-chloric acid TS and ethanol (99.5) to make exactly 50 mL. Pipet 3 mL of this solution, and add 1 mL of 1 mol/L hy-drochloric acid TS and ethanol (99.5) to make exactly 50 mL. Pipet 5 mL of this solution, add ethanol (99.5) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by adding ethanol (99.5) to 1 mL of 0.1 mol/L hydrochloric acid TS to make exactly Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Probenecid, previ-ously dried, and dissolve in 50 mL of neutralized ethanol. Titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indica- 50 mL, as the blank, and determine the absorbances, AT AS, at 248 nm. Amount (mg) of probenecid (C13H19NO4S) ＝ MS × AT/AS × V/25 MS: Amount (mg) of Probenecid RS taken and tor: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS Dissolution <6.10> When the test is performed at 50 revolu- tions per minute according to the Paddle method, using 900 ＝ 28.54 mg of C 13H 19NO4S mL of the 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Probenecid
Probucol, when dried, contains not less than 98.
Probucol occurs as a white crystalline powder. It is very soluble in tetrahydrofuran, freely soluble in ethanol (99.5), soluble in methanol, and practically insoluble in water. It gradually turns light yellow on exposure to light.
(1) Determine the absorption spectrum of a solution of Probucol in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Probucol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Probucol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Probucol RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Probucol according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.40 g of Probucol in 5 mL of ethanol (99.5), add the mobile phase to make 20 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of both solutions by the au-tomatic integration method: the area of the peak having the relative retention time of about 0.9 to probucol obtained from the sample solution is not larger than the peak area of probucol from the standard solution, and the area of peak having the relative retention time of about 1.9 from the sam-ple solution is not larger than 25 times the peak area of probucol from the standard solution, and the area of each peak other than probucol and the peaks mentioned above from the sample solution is not larger than 5 times the peak area of probucol from the standard solution. Furthermore, the total area of the peaks other than probucol from the sample solution is not larger than 50 times the peak area of probucol from the standard solution. For the areas of the peaks, having the relative retention times of about 0.9 and about 1.9 to probucol, multiply their correction factors, 1.2 and 1.4, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of probucol, beginning after the solvent peak, excluding the peak having the relative retention time of about 0.5 to probucol. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of probucol obtained with 5 mL of this solution is equivalent to 14 to 26z of that with 5 mL of the standard solution. System performance: To 1 mL of the sample solution add the mobile phase to make 50 mL. To 1 mL of this solution add 1 mL of a solution of phthalic acid bis(cis-3,3,5-trimethylcyclohexyl) in the mobile phase (1 in 1000), 5 mL of ethanol (99.5), and the mobile phase to make 20 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, phthalic acid bis(cis-3,3,5-trimethylcyclohexyl) and probucol are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of probucol is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 809C, 1 hour). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 60 mg each of Probucol and Probucol RS, previously dried, dissolve each in 5 mL of tetrahydrofuran, and add the mobile phase to make exactly ‌50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution and the mobile phase to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of probucol to that of the internal standard. Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS MS: Amount (mg) of Probucol RS taken Internal standard solution—Dissolve 0.2 g of bis(cis-3,3,5-trimethylcyclohexyl) phthalate in 1 mL of tetrahydrofuran, and add the mobile phase to make 50 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and water (93:7). Flow rate: Adjust so that the retention time of probucol is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and probucol are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of probucol to that of the internal standard is not more than 1.0z.
Procainamide Hydrochloride, when dried, contains not less than 98.
Procainamide Hydrochloride occurs as a white to light yellow crystalline powder. It is very soluble in water and soluble in ethanol (99.5). It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Procainamide Hydrochloride, previously dried, as directed in the potassium chloride disk method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. A solution of Procainamide Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Procainamide Hydrochloride in 10 mL of water: the pH of this solution is between 5.0 and 6.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Procainamide Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Procain- amide Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Procainamide Hydrochloride according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Procainamide Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so- ‌lution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area of both solutions by the automatic integration method: the total area of the peaks other than procainamide obtained from the sample solution is not larger than the peak area of procainamide from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.02 mol/L phosphate buffer solution (pH 3.0) and methanol (9:1). Flow rate: Adjust so that the retention time of procainamide is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of procainamide. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of procainamide obtained with 10 mL of this solution is equivalent to 40 to 60z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of procainamide are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of procainamide is not more than 2.0z. Loss on drying <2.41> Not more than 0.3z (2 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (2 g).
Weigh accurately about 0.5 g of Procainamide Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 27.18 mg of C13H21N3O.HCl
Procaine Hydrochloride occurs as white, crys-tals or crystalline powder. It is very soluble in water, soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Procaine Hydrochloride (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Procaine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Procaine Hydrochloride (1 in 10) re-sponds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution prepared by dissoluing 1.0 g of Procaine Hydrochloride in 20 mL of water is be-tween 5.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Procaine Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Procaine Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—To 1.0 g of Procaine Hydro-chloride add 5 mL of ethanol (95), dissolve by mixing well, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of 4-aminobenzoic acid in ethanol (95) to make exactly 20 mL, then pipet 1 mL of this solution, add 4 mL of ethanol (95) and water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of dibutyl ether, n-hexane and acetic acid (100) (20:4:1) to a distance of about 10 cm, and air-dry the plate. After drying the plate more at 1059C for 10 minutes, examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. The principal spot from the sample solution stays at the origin. Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Procaine Hydro-chloride, previously dried, dissolve in 5 mL of hydrochloric acid and 60 mL of water, add 10 mL of a solution of potas-sium bromide (3 in 10), cool to below 159C, and titrate <2.50> with 0.1 mol/L sodium nitrite VS (potentiometric titration or amperometric titration). Each mL of 0.1 mol/L sodium nitrite VS ＝ 27.28 mg of C13H20N2O2.HCl
Procarbazine Hydrochloride, when dried, contains not less than 98.
Procarbazine Hydrochloride occurs as white to light yellowish white, crystals or crystalline powder. It is freely soluble in water, and slightly soluble in ethanol (99.5). It dissolves in dilute hydrochloric acid. Melting point: about 2239C (with decomposition).
(1) Dissolve 0.01 g of Procarbazine Hydro-chloride in 1 mL of diluted copper (II) sulfate TS (1 in 10), and add 4 drops of sodium hydroxide TS: a green precipitate is formed immediately, and the color changes from green through yellow to orange. Determine the absorption spectrum of a solution of Procarbazine Hydrochloride in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Procarbazine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Procarbazine Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 0.10 g of Procarbazine Hydrochloride in 10 mL of water: the pH of this solution is between 3.0 and 5.0.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Procarbazine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Procarbazine Hydrochloride in 5.0 mL of a solution of L-cysteine hydro-chloride monohydrate in diluted methanol (7 in 10) (1 in 200), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a solution of L-cysteine hydrochloride monohydrate in diluted methanol (7 in 10) (1 in 200) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Immerse slowly, by inclining, a plate of silica gel with fluorescent indicator for thin-layer chromatography in a solution of L-cysteine hydrochloride monohydrate in diluted methanol (7 in 10) (1 in 200), allow to stand for 1 minute, lift the plate from the solution, dry it in cold wind for 10 minutes, then dry in warm wind for 5 minutes, and then dry at 609C for 5 minutes. After cooling, spot 5 mL each of the sample solu-tion and standard solution on the plate. Develop the plate with a mixture of methanol and ethyl acetate (1:1) to a dis-tance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): not more than 1 spot other than the principal spot and the spot of the starting point obtained from the sample solution appears, and is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.15 g of Procarbazine Hy-drochloride, previously dried, place in a glass-stoppered flask, dissolve in 25 mL of water, add 25 mL of hydrochloric acid, and cool to room temperature. To this solution add 5 mL of chloroform, and titrate <2.50>, while shaking, with 0.05 mol/L potassium iodate VS until the purple color of the chloroform layer disappears. The end point is reached when the red-purple color of the chloroform layer no more reap-pears within 5 minutes after the purple color disappeared. ‌Each mL of 0.05 mol/L potassium iodate VS ＝ 8.592 mg of C12H19N3O.HCl
Procaterol Hydrochloride Hydrate occurs as white to pale yellow-white, crystals or crystalline powder. It is soluble in water, in formic acid and in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Procaterol Hydrochloride Hydrate in 100 mL of water is between 4.0 and 5.0. It is gradually colored by light. The solution of Procaterol Hydrochloride Hydrate (1 in 20) shows no optical rotation. Melting point: about 1959C (with decomposition).
(1) Determine the absorption spectrum of a solution of Procaterol Hydrochloride Hydrate (7 in 1,000,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Procaterol Hydrochloride Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A solution of Procaterol Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Procaterol Hydrochloride Hydrate in 30 mL of water: the solution is clear, and has no more color than the following control solution. Control solution: To 3.0 mL of Iron (III) Chloride CS add water to make 50 mL. Heavy metals <1.07>—Proceed with 2.0 g of Procaterol Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Procaterol Hydrochloride Hydrate in 100 mL of diluted methanol (1 in 2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of these solutions by the au-tomatic integration method: the total area of the peaks other than procaterol obtained from the sample solution is not larger than the peak area of procaterol from the standard so-lution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.87 g of sodium 1-pentanesul-fonate in 1000 mL of water. To 760 mL of this solution add 230 mL of methanol and 10 mL of acetic acid (100). Flow rate: Adjust so that the retention time of procaterol is about 15 minutes. Selection of column: Dissolve 20 mg each of Procaterol Hydrochloride Hydrate and threoprocaterol hydrochloride in 100 mL of diluted methanol (1 in 2). To 15 mL of this so-lution add diluted methanol (1 in 2) to make 100 mL. Pro-ceed with 2 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of procaterol and threoprocaterol in this order with the resolution of these peaks being not less than 3. Detection sensitivity: Adjust the detection sensitivity so that the peak height of procaterol obtained from 2 mL of the standard solution is not less than 10 mm. Time span of measurement: 2.5 times as long as the reten-tion time of procaterol, beginning after the solvent peak. Water <2.48> 2.5 – 3.3z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Procaterol Hydro-chloride Hydrate, add 2 mL of formic acid, dissolve by warming, and add exactly 15 mL of 0.1 mol/L perchloric acid VS. Add 1 mL of acetic anhydride, heat on a water bath for 30 minutes, cool, add 60 mL of acetic anhydride, and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodi-um acetate VS (potentiometric titration). Perform a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.68 mg of C16H22N2O3.HCl
プロクロルペラジンマレイン酸塩
Prochlorperazine Maleate occurs as a white to light yellow powder. It is odorless, and has a slightly bitter taste. It is slightly soluble in acetic acid (100), very slightly solu-ble in water and in ethanol (95), and practically insoluble in diethyl ether. It gradually acquires a red tint by light. Melting point: 195 – 2039C (with decomposition).
(1) Dissolve 5 mg of Prochlorperazine Maleate in 5 mL of sulfuric acid: a red color develops, which darkens slowly on standing. Warm a half of the solution: the color changes to red-purple. To the remainder add 1 drop of potassium dichromate TS: a green-brown color develops, which changes to brown on standing. Boil 0.5 g of Prochlorperazine Maleate with 10 mL of hydrobromic acid under a reflux condenser for 10 minutes. After cooling, add 100 mL of water, and filter through glass filter (G4). Wash the residue with three 10-mL portions of water, and dry at 1059C for 1 hour: it melts <2.60> between 1959C and 1989C (with decomposition). Dissolve 0.2 g of Prochlorperazine Maleate in 5 mL of a solution of sodium hydroxide (1 in 10), and extract with three 3-mL portions of diethyl ether [reserve the aqueous layer, and use for test (4)]. Evaporate the combined diethyl ether extracts on a water bath to dryness, dissolve the residue in 10 mL of methanol by warming, and pour into 30 mL of a solution of 2,4,6-trinitrophenol in methanol (1 in 75), previ-ously warmed to 509C. Allow to stand for 1 hour, collect the crystals, wash with a small amount of methanol, and dry at 1059C for 1 hour: the crystals melt <2.60> between 2529C and 2589C (with decomposition). To the aqueous layer reserved in (3) add boiling chips, and heat on a water bath for 10 minutes. Cool, add 2 mL of bromine TS, heat on a water bath for 10 minutes, and heat the solution to boil. After cooling, add 2 drops of this solution to 3 mL of a solution of resorcinol in sulfuric acid (1 in 300), and heat on a water bath for 15 minutes: a red-purple color is produced.
Heavy metals <1.07>—Proceed with 1.0 g of Pro-chlorperazine Maleate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Prochlorperazine Maleate, previously dried, dissolve in 60 mL of acetic acid (100) while stirring and warming. Cool, and titrate <2.50> with 0.05 mol/L perchloric acid VS until the color of the so-lution changes from orange to green (indicator: 0.5 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 15.15 mg of C20H24ClN3S.2C4H4O4
Progesterone, when dried, contains not less than 97.
Progesterone occurs as white, crystals or crys-talline powder. It is soluble in methanol and in ethanol (99.5), and practi-cally insoluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Progesterone in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Progesterone RS prepared in ‌the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Progesterone, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Progesterone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Progester-one and Progesterone RS in ethanol (95), respectively, then evaporate the ethanol to dryness, and repeat the test on the residues. D Optical rotation <2.49> [a]20: ＋184 – ＋1949(after drying, 0.2 g, ethanol (99.5), 10 mL, 100 mm).
Related substances—Dissolve 80 mg of Progester-one in 2 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of diethyl ether and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 10 mg each of Progesterone and Progesterone RS, previously dried, and dissolve each in ethanol (99.5) to make exactly 100 mL. Pipet 5 mL each of these solutions, add ethanol (99.5) to make exactly 50 mL, and use these solution as the sample solution and the stand-ard solution, respectively. Determine the absorbances, AT and AS, of the sample solution and standard solution at the wavelength of maximum absorption at about 241 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Identification To 1 mL of Progesterone Injection add 1 mL of diluted ethanol (9 in 10), shake well, take the ethanol layer, shake well with 1 mL of petroleum benzin, and use the ethanol layer as the sample solution. Separately, dissolve about 5 mg of Progesterone RS in 1 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether and diethylamine (19:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Measure the specific gravity of Progesterone Injec-tion. Weigh accurately the mass of Progesterone Injection, equivalent to about 1 mL, mix with 2 mL of tetrahydrofu-ran, and add ethanol (99.5) to make exactly V mL so that each mL contains about 0.5 mg of progesterone (C21H30O2). Pipet 2 mL of this solution, add exactly 10 mL of the inter-nal standard solution and ethanol (99.5) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Progesterone RS, previously dried in vacuum for 4 hours using phosphorus (V) oxide as the desiccant, dissolve in 2 mL of tetrahydrofuran, and add ethanol (99.5) to make exactly 20 mL. Pipet 2 mL of this so-lution, add exactly 10 mL of the internal standard solution and ethanol (99.5) to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of progesterone to that of the internal standard. Amount (mg) of progesterone (C 21H30O2) ＝ M S × AT/AS Amount (mg) of progesterone (C21H30O2) ＝ MS × QT/QS × V/20 MS: Amount (mg) of Progesterone RS taken
Proglumide, when dried, contains not less than 98.
Proglumide occurs as white, crystals or crystal-line powder. It is freely soluble in methanol, soluble in ethanol (95), sparingly soluble in diethyl ether, and very slightly soluble in water. A solution of Proglumide in methanol (1 in 10) shows no optical rotation.
(1) Put 0.5 g of Proglumide in a round bot-tom tube, add 5 mL of hydrochloric acid, seal the tube, and heat the tube carefully at 1209C for 3 hours. After cooling, open the tube, filter the content to collect crystals separated out, wash the crystals with 50 mL of cold water, and dry at 1009C for 1 hour: the melting point <2.60> of the crystals is between 1219C and 1249C. (2) Determine the infrared absorption spectrum of Proglumide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of cyclohexane, ethyl acetate, acetic acid (100) and methanol (50:18:5:4) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.10z (1 g, reduced pressure, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.16 g of Proglumide, previ-ously dried, dissolve in 40 mL of methanol, add 10 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 33.44 mg of C18H26N2O4
Promethazine Hydrochloride occurs as a white to light yellow powder. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), sparingly soluble in acetic anhy-dride, and practically insoluble in diethyl ether. It is gradually colored by light. A solution of Promethazine Hydrochloride (1 in 25) shows no optical rotation. Melting point: about 2239C (with decomposition).
(1) Determine the absorption spectrum of a solution of Promethazine Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Promethazine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.5 g of Promethazine Hydrochloride in 5 mL of water, add 2 mL of ammonia TS, and filter. To 5 mL of the filtrate add dilute nitric acid to make acidic: the solu-tion responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> The pH of a solution of Promethazine Hydro-chloride (1 in 10) is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Promethazine Hydrochloride in 10 mL of water, protect-ing from light: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Promethazine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Perform the test under the pro-tection from sunlight. Dissolve 0.10 g of Promethazine Hydrochloride in exactly 5 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution (1). Separately, dis-solve 20 mg of isopromethazine hydrochloride for thin-layer chromatography in ethanol (95) to make exactly 100 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu- tion and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of methanol and diethyla-mine (19:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots obtained from the sample solution cor-responding to the spots from the standard solution (2) are not more intense than the spot from the standard solution (2), and any spot other than the principal spot from the sam-ple solution is not more intense than the spot from the stand-ard solution (1). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Promethazine Hy-drochloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.09 mg of C17H20N2S.HCl
Propafenone Hydrochloride, when dried, contains not less than 98.
Propafenone Hydrochloride occurs as white crystals or a white crystalline powder. It is freely soluble in formic acid, sparingly soluble in methanol, and slightly soluble in water and in ethanol (99.5). A solution of Propafenone Hydrochloride in methanol (1 in 100) shows no optical rotation.
(1) Dissolve 0.1 g of Propafenone Hydro-chloride in 20 mL of water by warming. After cooling, to 3 mL of this solution add water to make 500 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Propafenone Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at ‌the same wave numbers. Dissolve 0.1 g of Propafenone Hydrochloride in 20 mL of water by warming. After cooling, to 10 mL of this so-lution add 1 mL of dilute nitric acid, and filter to separate formed precipitate: the filtrate responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Propafenone Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Propafenone Hydrochloride in 20 mL of the mobile phase in the operating conditions 1, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add the mobile phase in the operating conditions 1 to make exactly 50 mL. Pipet 2.5 mL of this solution, add 2.5 mL of a solution of diphenyl phthalate in methanol (1 in 2000), add the mobile phase in the operating conditions 1 to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions 1 and 2. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than the peak of propafenone obtained from the sample solution is not larger than the peak area of propafenone from the standard solu-tion. Operating conditions 1— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.6 g of sodium 1-nonanesulfonate and 2.3 g of phosphoric acid in water to make 1000 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 900 mL of the filtrate add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of diphenyl phthalate is about 39 minutes. Time span of measurement: Beginning after the solvent peak to the retention time of diphenyl phthalate. System suitability 1— System performance: Dissolve 12 mg of Propafenone Hy-drochloride and 50 mg of isopropyl benzoate in 100 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions 1, propafenone and isopropyl benzoate are eluted in this order with the reso-lution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions 1, the relative standard deviation of the peak area of propafenone is not more than 2.0z. Operating conditions 2— Detector, column and column temperature: Proceed as di-rected in the operating conditions 1. Mobile phase: Dissolve 7.33 g of sodium 1-decanesul-fonate and 2.3 g of phosphoric acid in water to make 1000 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 700 mL of the filtrate add 700 mL of acetonitrile. Flow rate: Adjust so that the retention time of diphenyl phthalate is about 11 minutes. Time span of measurement: About 2.5 times as long as the retention time of diphenyl phthalate, beginning after the retention time of diphenyl phthalate. System suitability 2— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions 2, propafenone and diphenyl phthalate are eluted in this order with the resolution between these peaks being not less than 21. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions 2, the relative standard deviation of the peak area of propafenone is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Propafenone Hy-drochloride, previously dried, dissolve in 2 mL of formic acid, add 50 mL of acetic anhydride, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 18.90 mg of C21H27NO3.HCl
Propantheline Bromide, when dried, contains not less than 98.
Propantheline Bromide occurs as a white to yellowish white crystalline powder. It is odorless and has a very bitter taste. It is very soluble in water, in ethanol (95), in acetic acid (100) and in chloroform, soluble in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Propantheline Bromide in 50 mL of water is between 5.0 and 6.0. Melting point: about 1619C (with decomposition, after drying).
(1) To 5 mL of a solution of Propantheline Bromide (1 in 20) add 10 mL of sodium hydroxide TS, heat to boil for 2 minutes. Cool to 609C, and add 5 mL of dilute hydrochloric acid. After cooling, collect the precipitates, and wash with water. Recrystallize from dilute ethanol, and dry at 1059C for 1 hour: the crystals melt <2.60> between 2179C and 2229C. Dissolve 0.01 g of the crystals obtained in (1) in 5 mL ‌of sulfuric acid: a vivid yellow to yellow-red color develops. To 5 mL of a solution of Propantheline Bromide (1 in 10) add 2 mL of dilute nitric acid: this solution responds to Qualitative Tests <1.09> (1) for bromide.
Xanthene-9-carboxylic acid and xanthone—Dis-solve 10 mg of Propantheline Bromide in exactly 2 mL of chloroform, and use this solution as the sample solution. Separately, dissolve 1.0 mg of xanthene-9-carboxylic acid and 1.0 mg of xanthone in exactly 40 mL of chloroform, and use this solution as the standard solution. Perform the test immediately with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, and air-dry the plate for 10 minutes. Develop the plate with a mixture of 1,2-dichloroethane, methanol, water and formic acid (56:24:1:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light: the spots obtained from the sample solution corresponding to the spots from the standard solution are not more intense than those from the standard solution. Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Propantheline Bromide, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.84 g of C23H30BrNO3
Propiverine Hydrochloride, when dried, contains not less than 98.
Propiverine Hydrochloride occurs as white, crystals or crystalline powder. It is soluble in water and in ethanol (99.5).
(1) Dissolve 50 mg of Propiverine Hydro-chloride in 20 mL of water, and add acetonitrile to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Propiverine Hydrochroride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Propiverine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of dried Propiverine Hy-drochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Propiverine Hydrochloride (1 in 100) add 6 mL of ethyl acetate, and add 3 drops of sil-ver nitrate TS: a white precipitate is formed, which does not dissolve on the addition of 0.5 mL of dilute nitric acid and shaking. The precipitate dissolves on the addition of 2 mL of ammonia TS and shaking.
(1) Sulfate <1.14>—Perform the test with 0.40 g of Propiverine Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Propiverine Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Propiverine Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having the relative retention time about 0.28 to propiverine, obtained from the sample solution is not larger than 3/10 times the peak area of propiverine from the stand-ard solution, the area of the peak other than propiverine and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of propiverine from the standard solution, and the total area of the peaks other than propiverine from the sample solution is not larger than 1/2 times the peak area of propiverine from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of propiverine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of propiverine obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propiverine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propiverine is not more than 2.0z. ‌Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 1 hour). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Propiverine Hydrochloride and Propiverine Hydrachloride RS, both pre-viously dried, and dissolve each in the mobile phase to make exactly 100 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 15 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of propiver-ine in each solution. Amount (mg) of propiverine hydrochloride (C23H29NO3.HCl) ＝ MS × AT/AS MS: Amount (mg) of Propiverine Hydrochloride RS taken System suitability— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.21 g of potassium dihydrogen phosphate and 1.51 g of sodium 1-octane sulfonate in 650 mL of water, adjust to pH 3.2 with phosphoric acid, and add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of propiverine is about 17 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propiverine are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propiverine is not more than 1.0z.
Propranolol Hydrochloride, when dried, contains not less than 99.
Propranolol Hydrochloride occurs as a white crystalline powder. It is freely soluble in methanol, soluble in water and in acetic acid (100), and sparingly soluble in ethanol (99.5). A solution of Propranolol Hydrochloride in methanol (1 in 40) shows no optical rotation. It is gradualy colored to yellowish white to light brown by light.
(1) Determine the absorption spectrum of a solution of Propranolol Hydrochloride in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Propranolol Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Propranolol Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> The pH of a solution prepared by dissolving 0.5 g of Propranolol Hydrochloride in 50 mL of water is 5.0 – 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Propranolol Hydrochloride in 20 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Propranolol Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Propranolol Hydrochloride in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample so-lution, and add the mobile phase to make exactly 100 mL. ‌Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than propranolol ob-tained from the sample solution is not larger than 1/2 times the peak area of propranolol from the standard solution, and the total area of the peaks other than the peak of propranolol from the sample solution is not larger than 2 times the peak area of propranolol from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 292 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.6 g of sodium lauryl sulfate and 0.31 g of tetrabutylammonium dihydrogen phosphate in 450 mL of water, add 1 mL of sulfuric acid and 550 mL of aceto-nitrile for liquid chromatography, and adjust to pH 3.3 with 2 mol/L sodium hydroxide TS. Flow rate: Adjust so that the retention time of propranolol is about 4 minutes. Time span of measurement: About 5 times as long as the retention time of propranolol. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add the mobile phase to make ex-actly 20 mL. Confirm that the peak area of propranolol ob-tained with 20 mL of this solution is equivalent to 17 to 33z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propranolol is not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propranolol is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Propranolol Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.58 mg of C16H21NO2.HCl
Propyl Parahydroxybenzoate contains not less than 98.
Propyl Parahydroxybenzoate occurs as col-orless crystals or a white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tone, and very slightly soluble in water.◆
Determine the infrared absorption spectrum of Propyl Parahydroxybenzoate as directed in the potassium flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of propyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. ‌Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of propyl parahydroxy-benzoate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Clarity and color of solution—Dissolve 1.0 g of Propyl Parahydroxybenzoate in ethanol (95) to make 10 mL: the solution is clear and not more intensely colored than the following control solution. Control solution: To 5.0 mL of Cobalt (II) Chloride CS, 12.0 mL of Iron (III) Chloride CS and 2.0 mL of Copper (II) Sulfate CS add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Acidity—To 2 mL of the solution of Propyl Parahy-droxybenzoate obtained in (1) add 3 mL of ethanol (95), add 5 mL of freshly boiled and cooled water and 0.1 mL of bromocresol green-sodium hydroxide-ethanol TS, then add 0.1 mol/L sodium hydroxide VS until the solution shows a blue color: the volume of 0.1 mol/L sodium hydroxide VS used does not exceed 0.1 mL. (3) Heavy metals <1.07>—Dissolve 1.0 g of Propyl Par- ahydroxybenzoate in 25 mL of acetone, add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm).◆ Related substances—Dissolve 50.0 mg of Propyl Par-ahydroxybenzoate in 2.5 mL of methanol, and add the mo-bile phase to make exactly 50 mL. Pipet 10 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the peak area of parahydroxybenzoic acid having a relative retention time of about 0.3 to propyl par-ahydroxybenzoate obtained from the sample solution is not larger than the peak area of propyl parahydroxybenzoate from the standard solution (0.5z). For the peak area of parahydroxybenzoic acid, multiply the correction factor, 1.4. Furthermore, the area of the peak other than propyl parahydroxybenzoate and parahydroxybenzoic acid from the sample solution is not larger than the peak area of propyl parahydroxybenzoate from the standard solution (0.5z), and the total area of the peaks other than propyl parahy-droxybenzoate from the sample solution is not larger than 2 times the peak area of propyl parahydroxybenzoate from the standard solution (1.0z). For this calculation the peak area not larger than 1/5 times the peak area of propyl parahy-droxybenzoate from the standard solution is excluded (0.1z). droxybenzoate (C 10H12 O3). Operating conditions— Detector, column, column temperature, mobile phase, and Description Propyl Parahydroxybenzoate occurs as col-orless crystals or a white crystalline powder. It is freely soluble in methanol, in ethanol (95) and in ace-tone, and very slightly soluble in water.◆ Identification Determine the infrared absorption spectrum of Propyl Parahydroxybenzoate as directed in the potassium flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of propyl parahydroxybenzoate. System suitability— System performance: Proceed as directed in the system suitability in the Assay. ‌Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of propyl parahydroxy-benzoate obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution.◆ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propyl parahydroxybenzoate is not more than 2.0z.◆ Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50.0 mg each of Propyl Par-ahydroxybenzoate and Propyl Parahydroxybenzoate RS, dissolve separately in 2.5 mL each of methanol, and add the mobile phase to make exactly 50 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of propyl parahydroxybenzoate in each solution. Propylthiouracil
Propylene Glycol is a clear, colorless, viscous liquid. It is odorless, and has a slightly bitter taste. It is miscible with water, with methanol, with ethanol (95) and with pyridine. It is freely soluble in diethyl ether. It is hygroscopic.
(1) Mix 2 to 3 drops of Propylene Glycol with 0.7 g of triphenylchloromethane, add 1 mL of pyridine, and heat under a reflux condenser on a water bath for 1 hour. After cooling, dissolve the mixture in 20 mL of ace-tone by warming, shake with 0.02 g of activated charcoal, and filter. Concentrate the filtrate to about 10 mL, and cool. Collect the separated crystals, and dry in a desiccator (silica gel) for 4 hours: the crystals melt <2.60> between 1749C and 1789C. (2) Heat gently 1 mL of Propylene Glycol with 0.5 g of potassium hydrogen sulfate: a characteristic odor is evolved. Specific gravity <2.56> d 20: 1.035 – 1.040
(1) Acidity—Mix 10.0 mL of Propylene Glycol with 50 mL of freshly boiled and cooled water, and add 5 drops of phenolphthalein TS and 0.30 mL of 0.1 mol/L so-dium hydroxide VS: the solution has a red color. Chloride <1.03>—Perform the test with 2.0 g of Propylene Glycol. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.007z). Sulfate <1.14>—Perform the test with 10.0 g of Propylene Glycol. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.002z). Heavy metals <1.07>—Perform the test with 5.0 g of Propylene Glycol according to Method 1. Prepare the con-trol solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Propylene Glycol according to Method 1, and perform the test (not more than 2 ppm). Glycerin—Heat 1.0 g of Propylene Glycol with 0.5 g of potassium hydrogen sulfate and evaporate to dryness: no odor of acrolein is perceptible. Ethylene glycol, diethylene glycol and related sub-stances—Weigh accurately about 5 g of Propylene Glycol, mix with methanol to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 0.1 g each of ethylene glycol and diethylene glycol, and mix with methanol to make exactly 100 mL. Pipet 5 mL of this solution, and transfer to a 100-mL volumetric flask. Separately, weigh 5.0 g of propylene glycol for gas chroma-tography, mix with a suitable amount of methanol and put in the 100-mL volumetric flask, dilute with methanol to volume, and use this solution as the standard solution. Per-form the test with exactly 1 mL each of the sample solution and standard solution as directed under Gas Chromatogra-phy <2.02> according to the following conditions, and deter-mine the peak areas, AT1 and AS1, of ethylene glycol and, AT2 and AS2, of diethylene glycol by the automatic integra-tion method. The amounts of ethylene glycol and diethylene glycol calculated by the following equations are not more than 0.1z, respectively. The amount of the peak other than propylene glycol, ethylene glycol and diethylene glycol ob-tained from the sample solution, calculated by the area per-centage method, is not more than 0.1z, and the total amount of the peaks other than propylene glycol is not more than 1.0z. Amount (z) of ethylene glycol ＝ MS1/MT × AT1/AS1 × 5 Amount (z) of diethylene glycol ＝ MS2/MT × AT2/AS2 × 5 MS1: Amount (g) of ethylene glycol taken MS2: Amount (g) of diethylene glycol taken MT: Amount (g) of Propylene Glycol taken Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica tube 0.32 mm in inside diameter and 30 m in length, coated the inside surface 1 mm in thick-ness with 14z cyanopropylphenyl-86z dimethyl silicone polymer for gas chromatography. Column temperature: Inject at a constant temperature of about 1009C, rise the temperature at the rate of 7.59C per minute to 2209C, and maintain at a constant temperature of about 2209C. Injection port temperature: A constant temperature of about 2209C. Detector temperature: A constant temperature of about 2509C. ‌Carrier gas: Helium. Flow rate: about 38 cm per second. Split ratio: 1:20. Time span of measurement: About 3 times as long as the retention time of propylene glycol, beginning after the sol-vent peak. System suitability— System performance: Mix 50 mg each of ethylene glycol, diethylene glycol and propylene glycol for gas chromatogra-phy with 100 mL of methanol. When the procedure is run with 1 mL of this mixture under the above operating condi-tions, ethylene glycol, propylene glycol and diethylene glycol are eluted in this order, and the resolution between the peaks of ethylene glycol and propylene glycol is not less than 5, and that between the peaks of propylene glycol and diethy-lene glycol is not less than 50. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ethylene glycol and diethylene glycol is not more than 10z. Water <2.48> Not more than 0.5z (2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Weigh accurately about 20 g of Propylene Glycol in a tared crucible, and heat to boiling. Stop heating, and immediately ignite to burn. Cool, moisten the residue with 0.2 mL of sulfuric acid, and heat strongly with care to constant mass: the mass of the residue is not more than 0.005z. Distilling range <2.57> 184 – 1899C, not less than 95 volz.
Propylthiouracil, when dried, contains not less than 98.
Propylthiouracil occurs as a white powder. It is odorless, and has a bitter taste. It is sparingly soluble in ethanol (95), and very slightly soluble in water and in diethyl ether. It dissolves in sodium hydroxide TS and in ammonia TS.
(1) Shake well 0.02 g of Propylthiouracil Amount (mg) of propyl parahydroxybenzoate (C ＝ MS × AT/AS 10H12O3) with 7 mL of bromine TS for 1 minute, and heat until the color of bromine TS disappears. Cool, filter, and add 10 mL of barium hydroxide TS to the filtrate: a white precipitate is MS: Amount (mg) of Propyl Parahydroxybenzoate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and potassium dihy-drogen phosphate solution (17 in 2500) (13:7). Flow rate: 1.3 mL per minute. System suitability— System performance: Dissolve 5 mg each of Propyl Par-ahydroxybenzoate, ethyl parahydroxybenzoate and para-hydroxybenzoic acid in the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, and add the mobile phase to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, parahydroxybenzoic acid, ethyl parahydroxybenzoate and propyl parahydroxybenzoate are eluted in this order, the rel-ative retention times of parahydroxybenzoic acid and ethyl parahydroxybenzoate to propyl parahydroxybenzoate are about 0.3 and about 0.7, respectively, and the resolution be-tween the peaks of ethyl parahydroxybenzoate and propyl parahydroxybenzoate is not less than 3.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propyl parahydroxybenzoate is not more than 0.85z.
(1) Sulfate <1.14>—Triturate Propylthiouracil finely in a mortar. To 0.75 g of the powder add 25 mL of water, heat for 10 minutes on a water bath, cool, filter, and wash the residue with water until the volume of the filtrate becomes 30 mL. To 10 mL of the filtrate add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.077z). (2) Thiourea—Dissolve 0.30 g of Propylthiouracil in 50 mL of water by heating under a reflux condenser for 5 minutes, cool, and filter. To 10 mL of the filtrate add 3 mL of ammonia TS, shake well, and add 2 mL of silver nitrate TS: the solution has no more color than the following con-trol solution. Control solution: Weigh exactly 60 mg of thiourea, and dissolve in water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and proceed with 10 mL of this solution in the same manner. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Propylthiouracil, previously dried, and add 30 mL of water. Add 30 mL of 0.1 mol/L sodium hydroxide VS from a burette, heat to boil, and dissolve by stirring. Wash down the solid adhering to the wall of the flask with a small amount of water, and add 50 mL of 0.1 mol/L silver nitrate VS with stirring. Boil gently for 5 minutes, add 1 to 2 mL of bromothymol blue TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until a persistent blue-green color develops. Determine the total volume of 0.1 mol/L sodium hydroxide VS consumed. ‌Each mL of 0.1 mol/L sodium hydroxide VS‌ ＝ 8.512 mg of C7H10N2OS
Isopropylantipyrine occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in acetic acid (100), freely soluble in ethanol (95) and in acetone, soluble in diethyl ether, and slightly soluble in water.
(1) To 2 mL of a solution of Isopropylan-tipyrine (1 in 500) add 1 drop of iron (III) chloride TS: a light red color develops. Further add 3 drops of sulfuric acid to this solution: the color changes to pale yellow. Add 5 mL of a solution of Isopropylantipyrine (1 in 500) to a mixture of 5 mL of potassium hexacyanoferrate (III) TS and 1 to 2 drops of iron (III) chloride TS: a dark green color gradually develops. To 2 mL of a solution of Isopropylantipyrine (1 in 500) add 2 to 3 drops of tannic acid TS: a white precipitate is produced.
(1) Chloride <1.03>—Dissolve 1.0 g of Isopropyl-antipyrine in 30 mL of dilute ethanol, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid, 30 mL of dilute ethanol and water to make 50 mL (not more than 0.014z). Sulfate <1.14>—Dissolve 1.0 g of Isopropylantipyrine in 30 mL of dilute ethanol, and add 1 mL of dilute hydro-chloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid and 30 mL of dilute ethanol, and dilute with water to make 50 mL (not more than 0.019z). Heavy metals <1.07>—Dissolve 1.0 g of Isopropylan- tipyrine in 25 mL of acetone, add 2 mL of dilute acetic acid ‌1204 and water to make 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid, 25 mL of acetone, and dilute with water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Isopropylantipyrine according to Method 3, and perform the test (not more than 2 ppm). Antipyrine—Dissolve 1.0 g of Isopropylantipyrine in 10 mL of dilute ethanol, and add 1 mL of sodium nitrite TS and 1 mL of dilute sulfuric acid: no green color develops. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Isopropylantipy-rine, previously dried, dissolve in 60 mL of a mixture of ace-tic acid (100) and acetic anhydride (2:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 23.03 mg of C14H18N2O
Protamine Sulfate occurs as a white powder. It is sparingly soluble in water.
(1) Dissolve 1 mg of Protamine Sulfate in 2 mL of water, add 0.4 mL of sodium hydroxide TS, and im-mediately add 5 drops of a solution prepared by dissolving 0.1 g of 1-naphthol in 100 mL of diluted ethanol (7 in 10) and 5 drops of sodium hypochlorite TS: a vivid red color de-velops. Dissolve 5 mg of Protamine Sulfate in 1 mL of water by warming, add 1 drop of a solution of sodium hydroxide (1 in 10) and 2 drops of copper (II) sulfate TS: a red-purple color develops. An aqueous solution of Protamine Sulfate (1 in 20) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Protamine Sulfate in 100 mL of water: the pH of this solution is between 6.5 and 7.5.
(1) Clarity and color of solution—Dissolve 0.10 g of Protamine Sulfate in 10 mL of water: the solution is clear and colorless. ‌Absorbance—Dissolve 0.10 g of Protamine Sulfate in 10 mL of water, and determine the absorption spectrum as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance between 260 nm and 280 nm is not more than 0.1. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 3 hours). Nitrogen content Weigh accurately about 10 mg of Prota-mine Sulfate, and perform the test as directed under Nitro-gen Determination <1.08>: the amount of nitrogen (N:14.01) is 22.5 – 25.5z, calculated on the dried basis. Heparin-binding capacity Sample solution (a)—Weigh accurately about 15 mg of Protamine Sulfate, and dissolve in water to make exactly 100 mL. Repeat this procedure 3 times, and use the solutions so obtained as the sample solutions (a1), (a2) and (a3). Sample solution (b)—Pipet 10 mL each of the sample solutions (a1), (a2) and (a3), add exactly 5 mL of water to them, and use these solutions as the sample solutions (b1), (b2) and (b3). Sample solution (c)—Pipet 10 mL each of the sam-ple solutions (a1), (a2) and (a3), add exactly 20 mL of water to them, and use these solutions as the sample solutions (c1), (c2) and (c3). Standard solution—Dissolve Heparin Sodium RS in water to make a solution containing exactly about 20 Units per mL. Procedure—Transfer exactly 2 mL of the sample solution to a cell for spectrophotometer, add the standard solution dropwise while mixing, and determine the transmit-tance at 500 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>. Continue the addition until a sharp change in the transmittance is observed, and note the volume, V mL, of the standard solution added. Repeat this procedure 2 times for each sample solution. Calculation—Calculate the amount of heparin bound with 1 mg of the sample by the following formula from the volume of titrant on each sample solution, and cal-culate the average of 18 results obtained. The assay is not valid unless each relative standard deviation of 6 results ob-tained from the sample solution (a), sample solution (b) and sample solution (c) is not more than 5z, respectively, and also unless each relative standard deviation of 6 results ob-tained from 3 sets, (a1, b1, c1), (a2, b2, c2) and (a3, b3, c3) is not more than 5z, respectively. Amount (heparin Unit) of heparin bound to 1 mg of Protamine Sulfate ＝ S × V × 50/MT × d S: Amount (heparin Unit) of heparin sodium in 1 mL of the standard solution MT: Amount (mg) of Protamine Sulfate taken, calculated on the dried basis d: Dilution factor for each sample solution from the sam-ple solution (a) Sulfate content Weigh accurately about 0.15 g of Prota-mine Sulfate, dissolve in 75 mL of water, add 5 mL of 3 mol/L hydrochloric acid TS, and heat to boil. Add gradually 10 mL of barium chloride TS while boiling, and allow to stand for 1 hour while heating. Filter the precipitate formed, wash the precipitate with warm water several times, and transfer the precipitate into a tared crucible. Dry the precipi-tate, and incinerate by ignition to constant mass: the amount of sulfate (SO4) is 16 – 22z, calculated on the dried basis, where 1 g of the residue is equivalent to 0.4117 g of SO4.
Prothionamide occurs as yellow, crystals or crystalline powder. It has a slight, characteristic odor. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid and in dilute sulfu-ric acid.
(1) Mix 0.05 g of Prothionamide with 0.1 g of 1-chloro-2,4-dinitrobenzene, transfer about 10 mg of this mixture to a test tube, and heat for several seconds over a small flame until the mixture is fused. Cool, and add 3 mL of potassium hydroxide-ethanol TS: a red to orange-red color develops. Place 0.5 g of Prothionamide in a 100-mL beaker, and dissolve in 20 mL of sodium hydroxide TS by heating while shaking occasionally: the gas evolved turns a moistened red litmus paper to blue. Boil gently, and evapo-rate the solution to 3 to 5 mL. After cooling, add gradually 20 mL of acetic acid (100), and heat on a water bath: the gas evolved darkens moistened lead (II) acetate paper. Evaporate the solution on a water bath to 3 to 5 mL with the aid of a current of air, cool, add 10 mL of water, and mix well. Filter the crystals by suction, recrystallize from water immediately, and dry in a desiccator (in vacuum, silica gel) for 6 hours: the crystals melt <2.60> between 1989C and 2039C (with decomposition).
(1) Clarity and color of solution—Dissolve 0.5 g of Prothionamide in 20 mL of ethanol (95): the solution is clear, and shows a yellow color. Acidity—Dissolve 3.0 g of Prothionamide in 20 mL of methanol with warming. Add 100 mL of water to the so-lution, cool in an ice water bath with agitation, and remove any precipitate by filtration. Allow 80 mL of the filtrate to cool to room temperature, and add 0.8 mL of cresol red TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Pro- thionamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.6 g of Prothionamide according to Method 3, and perform the test. To the test solution add 10 mL of a solution of magne-sium nitrate hexahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and ignite to burn (not more than 3.3 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Prothionamide, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from orange-red to dark orange-brown (indicator: 2 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.03 mg of C9H12N2S
Protirelin Tartrate Hydrate, calculated on the anhy-drous basis, contains not less than 98.
Protirelin Tartrate Hydrate occurs as white to pale yellowish white, crystals or crystalline powder. It is freely soluble in water, sparingly soluble in acetic acid (100), and practically insoluble in ethanol (95) and in diethyl ether. Melting point: about 1879C (with decomposition).
(1) To 1 mL of a solution of Protirelin Tartrate Hydrate (1 in 1000) add 2 mL of a solution of 4- nitrobenzene diazonium fluoroborate (1 in 2000) and 2 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.0): a red color develops. Dissolve 0.03 g of Protirelin Tartrate Hydrate in 5 mL of sodium hydroxide TS, add 1 drop of copper (II) sul-fate TS: a purple color develops. To 0.20 g of Protirelin Tartrate Hydrate add 5.0 mL of 6 mol/L hydrochloric acid TS, and boil for 7 hours under a reflux condenser. After cooling, evaporate 2.0 mL of this solution on a water bath to dryness, dissolve the residue in 2.0 mL of water and use this solution as the sample solution. Separately, dissolve 22 mg of L-glutamic acid, 32 mg of L-histidine hydrochloride monohydrate and 17 mg of L-proline in 2.0 mL of 0.1 mol/L hydrochloric acid TS by heating, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of 1-butanol, water, acetic acid (100) and pyridine (4:1:1:1) to a distance of about 12 cm, and dry at 1009C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the three spots obtained from the sample solution show, respectively, the same color and the same Rf value as the corresponding spot from the standard solution. A solution of Protirelin Tartrate Hydrate (1 in 40) responds to Qualitative Tests <1.09> for tartrate. D Optical rotation <2.49> [a]20: －50.0 – －53.09(0.5 g calcu-lated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Protirelin Tartrate Hydrate in 100 mL of water: the pH of this solution is between 3.0 and 4.0.
(1) Clarity and color of solution—Dissolve 0.10 g of Protirelin Tartrate Hydrate in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Protire- lin Tartrate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Take 1.0 g of Protirelin Tartrate Hy- drate in a porcelain crucible. Add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), ignite the ethanol, and heat gradually to incinerate. If a car-bonized material still remains in this method, moisten with a small quantity of nitric acid, and ignite to incinerate. After cooling, add 10 mL of dilute hydrochloric acid, heat on a water bath to dissolve the residue, use this solution as the test solution, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.60 g of Protirelin Tartrate Hydrate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate (1) of silica gel for thin-layer chromatography. Spot 5 ‌mL of the sample solution on a plate (2) of silica gel for thin-layer chromatography. Develop the plates with a mixture of chloroform, methanol and ammonia solution (28) (6:4:1) to a distance of about 10 cm, and dry at 1009C for 30 minutes. Spray evenly a mixture of a solution of sulfanilic acid in 1 mol/L hydrochloric acid TS (1 in 200) and a solution of so-dium nitrite (1 in 20) (1:1) on the plate (1), and air-dry the plate. Then, spray evenly a solution of sodium carbonate decahydrate (1 in 10) on the plate: the spots other than the principal spot obtained from the sample solution are not more intense than those from the standard solution in color. On the other hand, spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate (2), and heat the plate at 809C for 5 minutes: no colored spot is obtained.‌ Water <2.48> Not more than 4.5z (0.2 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Weigh accurately about 0.5 g of Protirelin Tartrate Hydrate, dissolve in 80 mL of acetic acid (100) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 51.25 mg of C16H22N6O4.C4H6O6
Protirelin occurs as a white powder. It is freely soluble in water, in methanol, in ethanol (95) and in acetic acid (100). It is hygroscopic.
(1) Take 0.01 g of Protirelin in a test tube made of hard glass, add 0.5 mL of 6 mol/L hydrochloric acid TS, seal the upper part of the tube, and heat carefully at 1109C for 5 hours. After cooling, open the seal, transfer the contents into a beaker, and evaporate on a water bath to dryness. Dissolve the residue in 1 mL of water, and use this solution as the sample solution. Separately, dissolve 0.08 g of L-glutamic acid, 0.12 g of L-histidine hydrochloride monohydrate and 0.06 g of L-proline in 20 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water, acetic acid (100) and pyridine (4:1:1:1) to a distance of about 12 cm, and dry the plate at 1009C for 30 minutes. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 809C for 5 minutes: the three spots obtained from the sample solution show the same color and the same Rf value as each corresponding spots from the standard solution. Determine the infrared absorption spectrum of Pro-tirelin, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －66.0 – －69.09(0.1 g, calcu- ‌lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 0.20 g of Protirelin in 10 mL of water: the pH of this solution is between 7.5 and 8.5.
(1) Clarity and color of solution—Dissolve 0.10 g of Protirelin in 10 mL of water: the solution is clear and col-orless. Heavy metals <1.07>—Proceed with 1.0 g of Protire- lin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.20 g of Protirelin in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate (1) of silica gel for thin-layer chromatography, and spot 5 mL of the sample so-lution on a plate (2) of silica gel for thin-layer chromatogra-phy. Develop the plates with a mixture of 1-butanol, water, pyridine and acetic acid (100) (4:2:1:1) to a distance of about 12 cm, and dry the plates at 1009C for 30 minutes. Spray evenly a mixture of a solution of sulfanilic acid in 1 mol/L hydrochloric acid TS (1 in 200) and a solution of sodium nitrite (1 in 20) (1:1) on the plate (1), and air-dry the plates. Successively spray evenly a solution of sodium carbonate decahydrate (1 in 10) on it: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Spray evenly a so-lution of ninhydrin in acetone (1 in 50) on the plate (2), and heat the plate at 809C for 5 minutes: no colored spot ap-pears. Water <2.48> Not more than 5.0z (0.1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.3z (0.2 g).
Weigh accurately about 70 mg of Protirelin dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.02 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L perchloric acid VS ＝ 7.248 mg of C16H22N6O4
Spikes in nearly cylindrical and wheat ear-like shape, 3 – 6 cm in length, 1 – 1.5 cm in diameter, externally grayish brown; spikes composed of a floral axis having numerous bracts and calyxes; corollas often remaining on the upper part; a calyx usually enclosing four mericarps; bract, cordate to eccentric, and exhibiting white hairs on the vein, as on the calyx; light in texture. Almost odorless and tasteless.
(1) Stem—When perform the test of foreign mat- ter <5.01>, the amount of the stems contained in Prunella Spike does not exceed 5.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than the stems contained in Prunella Spike does not exceed 1.0z. Total ash <5.01> Not more than 13.0z. Acid-insoluble ash <5.01> Not more than 5.0z.
Pullulan occurs as a white powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) Dissolve 10 g of Pullulan in 100 mL of water with stirring by adding in small portions: a viscous so-lution is produced. Mix 10 mL of the viscous solution obtained in (1) with 0.1 mL of pullulanase TS, and allow to stand: the solu-tion loses its viscosity. To 10 mL of a solution of Pullulan (1 in 50) add 2 mL of macrogol 600: a white precipitate is formed immedi-ately. Viscosity <2.53> Weigh exactly 10.0 g of Pullulan, previ-ously dried, dissolve in water to make exactly 100 g, and per- form the test at 30 ± 0.19C as directed in Method 1: the kinematic viscosity is between 100 mm2/s and 180 mm2/s. pH <2.54> Dissolve 1.0 g of Pullulan in 10 mL of freshly boiled and cooled water: the pH is between 4.5 and 6.5.
(1) Heavy metals <1.07>—Proceed with 4.0 g of Pullulan according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 5 ppm). Nitrogen—Weigh accurately about 3 g of Pullulan, previously dried, and perform the test as directed under Nitrogen Determination <1.08>: the amount of nitrogen (N: 14.01) is not more than 0.05z. Use 12 mL of sulfuric acid for the decomposition, and add 40 mL of a solution of sodi-um hydroxide (2 in 5). Monosaccharide and oligosaccharides—Dissolve 0.8 g of Pullulan, previously dried, in 100 mL of water, and designate this solution as the sample stock solution. To 1 mL of the sample stock solution add 0.1 mL of potassium chlo-ride saturated solution, and shake vigorously with 3 mL of methanol. Centrifuge, and use the supernatant liquid as the sample solution. Separately, pipet 1 mL of the sample stock solution, add water to make exactly 50 mL, and use this so-lution as the standard solution. Pipet 0.2 mL each of the sample solution, the standard solution and water, transfer them gently to each test tube containing 5 mL of a solution of anthrone in diluted sulfuric acid (3 in 4) (1 in 500) cooled in ice water, stir immediately, then heat at 909C for 10 minutes, and cool immediately. Perform the test with these solutions so obtained as directed under Ultraviolet-visible Spectrophtometry <2.24> using water as a blank, and deter-mine the absorbances at 620 nm, AT, AS and AB: the amount of monosaccharide and oligosaccharides is not more than 10.0z. Amount (z) of monosaccharide and oligosaccharides ＝ (AT － AB)/(AS － AB) × 8.2 Loss on drying <2.41> Not more than 6.0z (1 g, in vacu-um, 909C, 6 hours). Residue on ignition <2.44> Not more than 0.3z (2 g).
Purified Dehydrocholic Acid, when dried, contains odorless.
Puriˆed Dehydrocholic Acid occurs as a white crystalline powder. It is odorless, and has a bitter taste. ‌JP XVIII Official Monographs / Dehydrocholic Acid Injection 833 It is sparingly soluble in acetone, slightly soluble in ethanol (95), and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Dissolve 5 mg of Purified Dehydro-cholic Acid in 1 mL of sulfuric acid and 1 drop of formalde-hyde solution, and allow to stand for 5 minutes. Add 5 mL of water to the solution: the solution shows a yellow color and blue-green fluorescence. (2) To 0.02 g of Purified Dehydrocholic Acid add 1 mL of ethanol (95), shake, add 5 drops of 1,3-dinitrobenzene TS and 0.5 mL of a solution of sodium hydroxide (1 in 8), and allow to stand: a purple to red-purple color develops, and gradually changes to brown. D Optical rotation <2.49> [a]25: ＋20 – ＋269(after drying, 0.2 g, acetone, 10 mL, 100 mm).
(1) Odor—To 2.0 g of Purified Dehydrocholic Acid add 100 mL of water, and boil for 2 minutes: the solu-tion is odorless. Clarity and color of solution—Dissolve 0.10 g of Purified Dehydrocholic Acid, previously powdered in a mor-tar, in 30 mL of ethanol (95) by shaking for 10 minutes: the solution is clear and colorless. Chloride <1.03>—To 2.0 g of Purified Dehydrocholic Acid add 100 mL of water, shake for 5 minutes and filter, and use this filtrate as the sample solution. To 25 mL of the sample solution add 6 mL of dilute nitric acid, heat in a water bath for 6 minutes, filter after cooling, and collect the clear filtrate. Wash the residue with 10 mL of water, com-bine the washings and the filtrate, dilute with water to 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Add 1 mL of dilute hydrochloric acid to 25 mL of the sample solution obtained in (3), heat in a water bath for 6 minutes, filter after cooling, and collect the clear filtrate. Wash the residue with 10 mL of water, com-bine the washings and the filtrate, dilute with water to 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Purified Dehydrocholic Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Barium—To the solution obtained in (1) add 2 mL of hydrochloric acid, and boil for 2 minutes, cool, filter, and
Purified Gelatin occurs as colorless or white to light yellow-brown sheets, shreds, granules or powder. It is very soluble in hot water, and practically insoluble in ethanol (95). The gelling grade does not dissolve in water. It slowly swells and softens when immersed in water, and absorbs water 5 to 10 times its own mass. The non-gelling grade is freely soluble in water.
(1) To 5 mL of a solution of Purified Gela-tin (1 in 100) add 2,4,6-trinitrophenol TS dropwise: a pre-cipitate is formed. To 5 mL of a solution of Purified Gelatin (1 in 5000) add tannic acid TS dropwise: the solution becomes turbid. In a test tube about 15 mm in internal diameter, place 0.5 g of Purified Gelatin, add 10 mL of water, and allow to stand for 10 minutes. Heat at 609C for 15 minutes, then keep the tube upright at 2 to 89C for 6 hours, and invert the tube: the contents do not flow out immediately for the gell-ing grade. In case of the non-gelling grade the contents flow JP XVIII Official Monographs / Purified Gelatin 1063 out immediately. Gel strength (Bloom value) Apply to the gelling grade. De-termine the mass (g) necessary to produce the force which, applied to a plunger 12.7 mm in diameter, makes a depres-sion 4 mm deep in the surface of the gel having a concentra-tion of 6.67z and matured at 109C. Apparatus, instruments: Texture analyzer or gelome-ter with a cylindrical piston 12.7 ± 0.1 mm in diameter with a plane bottom and a sharp bottom edge, and with a cup 59 ± 1 mm in internal diameter and 85 mm high (jelly cup). Procedure: Place 7.5 g of Purified Gelatin in a jelly cup, add 105 mL of water, close the cup, and allow to stand for 1 to 4 hours. Heat in a water bath at 65 ± 29C for 15 minutes. While heating, stir gently with a glass rod. Incor-porate any condensed water on the inner wall of the cup into the solution, and ensure that the solution is uniform. Allow to cool at room temperature for 15 minutes and transfer the cup to a thermostatically controlled bath at 10.0 ± 0.19C, and fitted with a device to ensure that the platform on which the cup stands is perfectly horizontal. Close the cup, and allow to stand for 17 ± 1 hours. Remove the sample cup from the bath and quickly wipe the water from the exterior of the cup. Put the cup on the platform of the apparatus so that the tip of plunger contacts the sample as nearly at its midpoint as possible, and start the measurement with 4 mm depression distance and 0.5 mm per second test speed: 80 to 120z of the labeled nominal value. pH <2.54> Dissolve 1.00 g of Purified Gelatin in freshly boiled water and kept at about 559C, to make 100 mL. pH at 559C of this solution is 3.8 – 9.0.
(1) Heavy metals<1.07>—Proceed with 1.0 g of Purified Gelatin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Iron—To 5.00 g of Purified Gelatin, in a glass-stoppered flask, add 10 mL of hydrochloric acid, close the flask, and place in a water bath at 75 – 809C for 2 hours. If necessary for proper solubilization, the gelatin may be al-lowed to swell after addition of the acid and the heating time may be prolonged or a higher temperature may be used. After cooling, adjust the content of the flask to 100.0 g with water, and use this solution as the sample solution. Sepa-rately, place 5.00 g each of Purified Gelatin in three glass-stoppered flasks, proceed with them in the same manner as the sample solution, then add exactly 10 mL, 20 mL and 30 mL of Standard Iron Solution (2) for Atomic Absorption Spectrophotometry to each flask, respectively. Adjust the content of these flasks to 100.0 g each with water, and use these solutions as the standard solutions. The volume of Standard Iron Solution may be adjusted appropriately ac-cording to the sensitivity of the instrument to be used. Per-form the test with the sample solution and standard solu-tions as directed in the standard addition method under Atomic Absorption Spectrophotometry<2.23> according to the following conditions, and determine the content of iron: not more than 30 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Iron hollow cathode lamp. Wavelength: 248.3 nm. Chromium—Use the sample solution obtained in (2) as the sample solution. Separately, place 5.00 g each of Puri-fied Gelatin in three glass-stoppered flasks, proceed with them in the same manner as the sample solution, then add exactly 0.25 mL, 0.50 mL and 0.75 mL of Standard Chromi- um Solution for Atomic Absorption Spectrophotometry to each flask, respectively. Adjust the content of these flasks to 100.0 g each with water, and use these solutions as the stand-ard solutions. The volume of Standard Iron Solution may be adjusted appropriately according to the sensitivity of the instrument to be used. Perform the test with the sample solu-tion and standard solutions as directed in the standard addi-tion method under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and determine the content of chromium: not more than 10 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Chromium hollow cathode lamp. Wavelength: 357.9 nm. Zinc—Use the sample solution obtained in (2) as the sample solution. Separately, place 5.00 g each of Purified Gelatin in three glass-stoppered flasks, proceed with them in the same manner as the sample solution, then add exactly 7.5 mL, 15 mL and 22.5 mL of Standard Zinc Solution for Atomic Absorption Spectrophotometry to each flask, re-spectively. Adjust the content of these flasks to 100.0 g each with water, and use these solutions as the standard solutions. The volume of Standard Iron Solution may be adjusted ap-propriately according to the sensitivity of the instrument to be used. Perform the test with the sample solution and standard solutions as directed in the standard addition method under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and determine the content of zinc: not more than 30 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm. Arsenic<1.11>—Place 15.0 g of Purified Gelatin in a flask, add 60 mL of diluted hydrochloric acid (1 in 5), and dissolve by heating. Add 15 mL of bromine TS, heat until the excess of bromine is expelled, neutralize with ammonia TS, add 1.5 g of disodium hydrogen phosphate dodecahy-drate, and allow to cool. To this solution add 30 mL of mag-nesia TS, allow to stand for 1 hour, and collect the precipi-tates. Wash the precipitates with five 10-mL portions of diluted ammonia TS (1 in 4), and dissolve in diluted hydro-chloric acid (1 in 4) to make exactly 50 mL. Perform the test with 5 mL of this solution: the solution has no more color than the following color standard. Color Standard: Proceed with 12 mL of Standard Arsenic Solution, instead of Purified Gelatin, in the same manner (not more than 0.8 ppm). Peroxides— Enzyme reaction: Peroxidase transfers oxygen atom at on from peroxides to an organic redox indicator which is converted to a blue oxidized form. The intensity of the color obtained is proportional to the quantity of peroxide. The peroxide concentration can be determined by comparing it with the color scale provided with the test strips employing this reaction. Procedure: Weigh 20.0 ± 0.1 g of Purified Gelatin in a beaker, add 80.0 ± 0.2 mL of water, and stir to moisten all the gelatin. Allow to stand at room temperature for 1—3 hours. Cover the beaker with a watch-glass, and heat the beaker for 20 ± 5 minutes in a water bath at 65 ± 29C to dissolve the sample. Stir the contents of the beaker with a glass rod to achieve a homogeneous solution, and use this as the sample solution. Dip a peroxide test strip for 1 second into the sample solution, such that the reaction zone is prop-erly wetted. Remove the test strip, shake off excess liquid, and compare the reaction zone after 15 seconds with the ‌1064 color scale provided. Multiply the concentration read from the color scale by a factor of 5 to calculate the concentration of peroxide in the test substance: not more than 10 ppm. Sensitivity: To exactly 10 mL of Standard Hydrogen Peroxide Solution add water to make exactly 300 mL. Pipet exactly 2 mL of this solution, add water to make exactly 1000 mL (2 ppm). Dip a peroxide test strip for 1 second into this solution, such that the reaction zone is properly wetted. Remove the test strip, shake off excess liquid and compare the color of the reaction zone after 15 seconds with the color scale: the color of the zone is equivalent to 2 ppm of the color scale. Sulfur dioxide— Apparatus: Use as shown in the figure. Conductivity <2.51> Dissolve 1.00 g of Purified Gelatin in freshly boiled water and kept at about 559C, to make 100 mL. Perform the test at 30 ± 1.09C with this solution, with-out temperature compensation: not more than 1 mS･cm－1. Loss on drying <2.41> Not more than 15.0z (5 g, 1059C, 16 hours). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 103 CFU/g and 102 CFU/g, respectively. Es-cherichia coli and Salmonella are not observed.
Containers—Tight containers. Storage—Protect from heat and moisture.
Purified Glucose is D-glucopyranose derived from starch. It contains not less than 97.
Purified Glucose occurs as white, crystals or crystalline powder, and has a sweet taste. It is freely soluble in water, and slightly soluble in metha-nol and in ethanol (95).◆
◇(1) Add 2 to 3 drops of a solution of Purified Glucose (1 in 20) to 5 mL of boiling Fehling's TS: a red precipitate is produced.◇ Perform the test with 20 mL each of the sample solu-tion and standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions: the principal peak in the chromatogram obtained from the sample solution is similar in retention time and size to the principal peak in the chromatogram from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay.
(1) Clarity and color of solution—Dissolve 10.0 g of Purified Glucose in 15 mL of water by heating on a water bath, and allow to cool to room temperature, and use this solution as the test solution. Perform the test with the test solution as directed under Turbidity Measurement <2.61>: the solution is clear. Perform the test with the test solution according to Method 2 under Methods for Color Matching <2.65>: the solution is not more colored than Matching Fluid BY7. (2) Heavy metals <1.07>—Proceed with 5.0 g of Puri- ‌fied Glucose according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 4 ppm).◆ Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 250 mL, and use this solution as the standard solution (1). Pipet 25 mL of the standard solution (1), add water to make exactly 200 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution, the
Weigh accurately about 0.3 g each of Purified Glu-cose and ◆Glucose RS◆ (separately determine the water <2.48> in the same manner as Purified Glucose), dissolve separately in water to make exactly 10 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of glucose in each solution. standard solution (1) and the standard solution (2) as di-rected under Liquid Chromatography <2.01> according to Amount (g) of glucose (C6H12O6) ＝ M S × AT/AS the following conditions. Determine each peak area by the automatic integration method: the total area of the peaks of maltose and isomaltose, having the relative retention time of about 0.8 to glucose, obtained from the sample solution, is not larger than the peak area of glucose from the standard solution (1) (not more than 0.4z), and the peak area of mal-totriose, having the relative retention time of about 0.7 from the sample solution, is not larger than 1/2 times the peak area of glucose from the standard solution (1) (not more than 0.2z), and the peak area of fructose, having the rela-tive retention time of about 1.3 from the sample solution, is not larger than 3 times the peak area of glucose from the standard solution (2) (not more than 0.15z), and the area of the peak other than glucose and the peaks mentioned above from the sample solution is not larger than 2 times the peak area of glucose from the standard solution (2) (not more than 0.10z). Furthermore, the total area of the peaks other than glucose from the sample solution is not larger than 1.25 times the peak area of glucose from the standard solution (1) (not more than 0.5z). For these calculations the peak areas not larger than the peak area of glucose from the standard solution (2) are excluded (disregard limit: 0.05z). Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of glucose. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Confirm that the peak area of glucose obtained with 20 mL of the standard solution (2) is equivalent to 8.8 to 16.3z of that with 20 mL of the standard solution (1). System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviation of the peak area of glucose is not more than 1.0z.◇ Dextrin—To 1.0 g of powdered Purified Glucose add 20 mL of ethanol (95), and boil under a reflux condenser: the solution is clear. Soluble starch and sulfite—To 6.7 g of Purified Glu-cose add 15 mL of water, dissolve by heating on a water bath, cool, and add 25 mL of 0.05 mol/L iodine VS: a yellow color develops (not more than 15 ppm as SO3). Conductivity <2.51> Dissolve 20.0 g of Purified Glucose in a fleshly boiled and cooled distilled water to make 100 mL, and use this solution as the sample solution. Measure the conductivity of the sample solution at 25 ± 0.19C while gently stirring with a magnetic stirrer: not more than 20 mS･ cm－1. Water <2.48> Not more than 1.0z (0.5 g, volumetric titra-tion, direct titration). MS: Amount (g) of Glucose RS taken, calculated on the anhydrous basis Operating conditions— Detector: A differential refractometer maintained at a constant temperature (409C for example). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with strongly acidic ion-ex-change resin for liquid chromatography (Ca type) composed with a sulfonated polystyrene cross-linked with divinylben-zene (degree of cross-linkage: 8z) (9 mm in particle diame-ter). Column temperature: A constant temperature of about 859C. Mobile phase: Water. Flow rate: 0.3 mL per minute (the retention time of glu-cose is about 21 minutes). System suitability— System performance: Dissolve 5 mg of maltose, 5 mg of maltotriose and 5 mg of fructose in 50 mL of water, and use this solution as the solution for system suitability test. When the procedure is run with 20 mL each of the solution for sys-tem suitability test and the standard solution (2) in Purity (3) under the above operating conditions, maltotriose, maltose, glucose and fructose are eluted in this order, the relative retention times of maltotriose, maltose, isomaltose and fruc-tose to glucose are about 0.7, about 0.8, about 0.8 and about 1.3, respectively, and the resolution between the peaks of maltotriose and maltose is not less than 1.3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glucose is not more than 1.0z.◇ ◆Containers and storage Containers—Tight containers.◆ Glucose Injection
Purified Shellac occurs as light yellow-brown to brown, lustrous, hard, brittle scutella. It is odorless or has a faint, characteristic odor. It is freely soluble in ethanol (95) and in ethanol (99.5), and practically insoluble in water and in diethyl ether. It dissolves in sodium hydroxide TS. Acid value <1.13> 60 – 80 Weigh accurately about 1 g of Purified Shellac, add 40 mL of neutralized ethanol, and dissolve by warming. After cooling, titrate <2.50> with 0.1 mol/L potassium hydroxide VS (potentiometric titration).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Purified Shellac according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Purified Shellac according to Method 3, and perform the test. Add 10 mL of a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and fire to burn (not more than 5 ppm). Ethanol-insoluble substances—Dissolve about 5 g of Purified Shellac, accurately weighed, in 50 mL of ethanol (95) on a water bath while shaking. Pour the ethanol solu-tion into a tared extraction thimble, previously dried at 1059C for 2 hours, in a Soxhlet extractor, and extract with ethanol (95) for 3 hours. Dry the extraction thimble at 1059C for 3 hours: the mass of the residue is not more than 2.0z. Use a cylindrical weighing bottle for taring the extraction thimble. Rosin—To 2.0 g of Purified Shellac add 10 mL of ethanol (99.5), shake thoroughly to dissolve, and add gradu-ally 50 mL of hexane while shaking. Transfer the solution to a separator, wash with two 50-mL portions of water, filter the upper layer, and evaporate the filtrate on a water bath to dryness. To the residue add 5 mL of acetic anhydride, and heat on a water bath to dissolve, if necessary. Transfer this solution to a Nessler tube, and add 1 drop of sulfuric acid: the color of the solution does not change from red-purple through purple to reddish yellow. Wax—Dissolve 10.0 g of Purified Shellac in 150 mL of a solution of sodium carbonate decahydrate (9 in 200) with shaking on a water bath, and continue the heating for 2 hours. After cooling, collect the floating wax by filtration, wash the wax and the filter paper with water, transfer to a beaker, and dry at 659C until the water is almost evaporated. Transfer the wax together with the filter paper to an ex-traction thimble in a Soxhlet extractor. Dissolve the wax remaining in the beaker with a suitable quantity of chlo-roform by warming. Pour the solution into the thimble, and extract with chloroform for 2 hours. Evaporate the chlo-roform solution to dryness, and dry the residue at 1059C for 3 hours: the mass of the residue is not more than 20 mg. Loss on drying Not more than 2.0z. Weigh accurately about 1 g of moderately fine powder of Purified Shellac, and dry at 409C for 4 hours, then for 15 hours in a desiccator (calcium chloride for drying). Total ash <5.01> Not more than 1.0z (1 g).
Purified Water in Containers is a clear and colorless liquid, having no odor.
Potassium permanganate-reducing substances—To 100 mL of Purified Water in Containers add 10 mL of dilute sulfuric acid, boil, then add 0.10 mL of 0.02 mol/L potas-sium permanganate VS, and boil again for 10 minutes: the red color of the solution does not disappear. Conductivity <2.51> When the test is performed according to the following method, the conductivity (259C) is not more than 25 mS・cm－1 for containers with a nominal volume of 10 mL or less, and not more than 5 mS・cm－1 for containers with a nominal volume greater than 10 mL. Transfer a suitable amount of Purified Water in Contain-ers to a beaker, and stir the water specimen. Adjust the temperature to 25 ± 19C, and begin agitating the water specimen vigorously, while observing its conductivity periodically. When the change in conductivity becomes not greater than 0.1 mS・cm－1 per 5 minutes, adopt the observed value as the conductivity of the water specimen. Microbial limit <4.05> The acceptance criteria of TAMC is 102 CFU/mL. Perform the test using soybean-casein digest agar medium.
Purified Water is a clear and colorless liquid, having no odor.
Total organic carbon <2.59>—Not more than 0.50 mg/L. Conductivity <2.51> When the test is performed according to the following method, the conductivity (259C) is not more than 2.1 mS・cm－1. Transfer a suitable amount of Purified Water to a beaker, and stir the water specimen. Adjust the temperature to 25 ± 19C, and begin agitating the water specimen vigorously, while observing its conductivity periodically. When the change in conductivity becomes not greater than 0.1 mS・ cm－1 per 5 minutes, adopt the observed value as the conduc-tivity of the water specimen. Purified Water in Containers
Pyrantel Pamoate, when dried, contains not less than 98.
Pyrantel Pamoate occurs as a light yellow to ‌yellow crystalline powder. It is odorless and tasteless. It is sparingly soluble in N,N-dimethylformamide, very slightly soluble in methanol and in ethanol (95), and practi-cally insoluble in water, in ethyl acetate and in diethyl ether. Melting point: 256 – 2649C (with decomposition).
(1) To 0.05 g of Pyrantel Pamoate add 10 mL of methanol and 1 mL of a mixture of hydrochloric acid and methanol (1:1), and shake vigorously: a yellow precipi-tate is produced. Filter the solution, and use the filtrate as the sample solution. Use the precipitate for the test (2). To 0.5 mL of the sample solution add 1 mL of a solution of 2,3-indolinedione in sulfuric acid (1 in 1000): a red color devel-ops. Collect the precipitate obtained in the test (1), wash with methanol, and dry at 1059C for 1 hour. To 0.01 g of the dried precipitate add 10 mL of methanol, shake well, and filter. To 5 mL of the filtrate add 1 drop of iron (III) chlo-ride TS: a green color develops. Dissolve 0.1 g of Pyrantel Pamoate in 50 mL of N,N-dimethylformamide, and add methanol to make 200 mL. To 2 mL of the solution add a solution of hydrochloric acid in methanol (9 in 1000) to make 100 mL. Determine the absorp- ethyl acetate, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the spot of pyrantel and the spot of pamoic acid obtained from the sample solution are not more intense than the spot of pyrantel (Rf value: about 0.3) from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
(1) Chloride <1.03>—To 1.0 g of Pyrantel Pamo-ate add 10 mL of dilute nitric acid and 40 mL of water, and heat on a water bath with shaking for 5 minutes. After cool-ing, add water to make 50 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute nitric acid and water to make 50 mL. Proceed the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.036z). Sulfate <1.14>—To 0.75 g of Pyrantel Pamoate add 5 mL of dilute hydrochloric acid and water to make 100 mL, and heat on a water bath for 5 minutes with shaking. After cooling, add water to make 100 mL, and filter. To 20 mL of the filtrate add water to make 50 mL. Proceed the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.005 mol/L sulfuric acid VS (not more than 0.144z). Heavy metals <1.07>—Proceed with 1.0 g of Pyrantel Pamoate according to Method 2, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pyrantel Pamoate according to Method 3, and perform the test (not more than 2 ppm). Related substances—The procedure should be per-formed under protection from light in light-resistant vessels. Dissolve 0.10 g of Pyrantel Pamoate in 10 mL of N,N-dimethylformamide, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add N,N-dimeth-ylformamide to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of
Weigh accurately about 0.5 g of Pyrantel Pamoate, previously dried, add 25 mL of chloroform and 25 mL of so-dium hydroxide TS, shake for 15 minutes, and extract. Ex-tract further with two 25-mL portions of chloroform. Filter each extract through 5 g of anhydrous sodium sulfate on a pledget of absorbent cotton. Combine the chloroform ex-tracts, add 30 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 59.47 mg of C H N S.C H O tion spectrum of the solution as directed under Ultraviolet- 11 14 2 23 16 6 visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pyrantel Pamoate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. Purity (1) Chloride <1.03>—To 1.0 g of Pyrantel Pamo-ate add 10 mL of dilute nitric acid and 40 mL of water, and heat on a water bath with shaking for 5 minutes. After cool-ing, add water to make 50 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute nitric acid and water to make 50 mL. Proceed the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hy-drochloric acid VS (not more than 0.036z). Sulfate <1.14>—To 0.75 g of Pyrantel Pamoate add 5 mL of dilute hydrochloric acid and water to make 100 mL, and heat on a water bath for 5 minutes with shaking. After cooling, add water to make 100 mL, and filter. To 20 mL of the filtrate add water to make 50 mL. Proceed the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.005 mol/L sulfuric acid VS (not more than 0.144z). Heavy metals <1.07>—Proceed with 1.0 g of Pyrantel Pamoate according to Method 2, and perform the test. Pre-pare the control solution with 3.0 mL of Standard Lead So-lution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pyrantel Pamoate according to Method 3, and perform the test (not more than 2 ppm). Related substances—The procedure should be per-formed under protection from light in light-resistant vessels. Dissolve 0.10 g of Pyrantel Pamoate in 10 mL of N,N-dimethylformamide, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add N,N-dimeth-ylformamide to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of
Pyrazinamide, when dried, contains not less than 99.
Pyrazinamide occurs as white, crystals or crys-talline powder. It is sparingly soluble in water and in methanol, and slightly soluble in ethanol (99.5) and in acetic anhydride.
(1) Determine the absorption spectrum of a solution of Pyrazinamide in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pyrazinamide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Pyrazinamide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Pyrazinamide in 10 mL of methanol, and use this solution as the sample so- ‌lution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution.‌ Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Pyrazinamide, pre-viously dried, dissolve in 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 12.31 mg of C5H5N3O
Pyridostigmine Bromide occurs as a white crystalline powder. It is odorless or has a slightly characteris-tic odor. It is very soluble in water, freely soluble in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Pyridostigmine Bromide in 10 mL of water is between 4.0 and 6.0. It is deliquescent.
(1) Dissolve 0.02 g of Pyridostigmine Bromide in 10 mL of water, add 5 mL of Reinecke salt TS: a light red precipitate is produced. To 0.1 g of Pyridostigmine Bromide add 0.6 mL of sodium hydroxide TS: the unpleasant odor of dimethylamine is perceptible. Determine the absorption spectrum of a solution of Pyridostigmine Bromide in 0.1 mol/L hydrochloric acid TS (1 in 30,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Pyridostigmine Bromide (1 in 50) re-sponds to Qualitative Tests <1.09> for Bromide.
(1) Clarity and color of solution—Dissolve 1.0 g of Pyridostigmine Bromide in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Pyridostigmine Bromide according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pyridostigmine Bromide according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Pyridostig-mine Bromide in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add ethanol (95) to make exactly 10 mL. Pipet 1 mL of this solution, add ethanol (95) to make exactly 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of methanol, chloroform and ammonium chloride TS (5:4:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution in color. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, phosphorus (V) oxide, 1009C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Pyridostigmine Bromide, previously dried, dissolve in 10 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.11 mg of C9H13BrN2O2
ピリドキサールリン酸エステル水和物
Pyridoxal Phosphate Hydrate contains not less than 98.
Pyridoxal Phosphate Hydrate occurs as a pale yellow-white to light yellow crystalline powder. It is slightly soluble in water, and practically insoluble in ethanol (99.5). It dissolves in dilute hydrochloric acid and in sodium hy-droxide TS. The pH of a solution prepared by dissolving 0.1 g of Pyridoxal Phosphate Hydrate in 200 mL of water is between 3.0 and 3.5. Pyridoxal Phosphate Hydrate is colored to light red by light.
(1) Determine the absorption spectrum of a solution of Pyridoxal Phosphate Hydrate in phosphate buffer solution (pH 6.8) (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pyridoxal Phosphate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pyridoxal Phosphate Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Pyridoxal Phosphate RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 4.0 g of Pyridoxal Phosphate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 5 ppm). Arsenic <1.11>—Dissolve 1.0 g of Pyridoxal Phos- phate Hydrate in 5 mL of dilute hydrochloric acid. Use this solution as the test solution, and perform the test (not more than 2 ppm). Free phosphoric acid—Weigh accurately about 0.1 g of Pyridoxal Phosphate Hydrate, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Pipet 5 mL each of the sample solution and Standard Phos-phoric Acid Solution, to each add 2.5 mL of hexaammoni-um heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, and shake. Add water to make exactly 25 mL, and allow to stand at 20 ± 19C for 30 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of water in the same manner as the blank. Determine the absorbances, AT and AS, of each solution from the sample solution and Standard Phosphoric Acid Solution at 740 nm: the amount of free phosphoric acid is not more than 0.5z. Content (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Pyridoxal Phosphate Hydrate taken, calculated on the anhydrous basis Related substances—Dissolve 50 mg of Pyridoxal Phosphate Hydrate in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and stand-ard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than pyridoxal phosphate obtained from the sample solution is not larger than the peak area of pyridoxal phosphate from the standard solution, and the total area of the peaks other than pyridoxal phosphate from the sample solution is not larger than 2 times the peak area of pyridoxal phosphate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.63 g of potassium dihydrogen phosphate and 5.68 g of anhydrous disodium hydrogen phosphate in water to make 1000 mL. Flow rate: Adjust so that the retention time of pyridoxal phosphate is about 6 minutes. Time span of measurement: About 2.5 times as long as the retention time of pyridoxal phosphate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of pyridoxal phosphate obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of pyridoxal phosphate are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of pyridoxal phosphate is not more than 2.0z. Water <2.48> 6.0 – 9.0z (0.1 g, volumetric titration, direct titration. Use a solution prepared by dissolving 50 g of im-idazole for water determination in 100 mL of the dissolving solution instead of methanol for water determination). Dissolving solution: A solution containing 80z of 1-methoxy-2-propanol, 18z of ethanol (99.5), 1z of imida-zole and 1z of imidazole hydrobromide.
Weigh accurately about 45 mg each of Pyridoxal Phosphate Hydrate and Pyridoxal Phosphate RS (separately determine the water <2.48> in the same manner as Pyridoxal Phosphate Hydrate), and dissolve each in phosphate buffer solution (pH 6.8) to make exactly 250 mL. Pipet 10 mL each of these solutions, add phosphate buffer solution (pH 6.8) to make exactly 100 mL, and use these solutions as the sample solution and the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 388 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24> using phosphate buffer solution (pH 6.8) as the blank. Amount (mg) of pyridoxal phosphate (C8H10NO6P) ＝ MS × AT/AS MS: Amount (mg) of Pyridoxal Phosphate RS taken, cal-culated on the anhydrous basis
Pyroxylin occurs as a white cotton-like sub-stance or white flakes. It is freely soluble in acetone, and very slightly soluble in diethyl ether. Upon heating or exposure to light, it is decomposed with the evolution of nitrous acid vapors.
Ignite Pyroxylin: it burns very rapidly with a luminous flame.
(1) Clarity of solution—Dissolve 1.0 g of Pyroxy-lin, previously dried at 809C for 2 hours, in 25 mL of a mix-ture of diethyl ether and ethanol (95) (3:1): the solution is clear. Acidity—Shake 1.0 g of Pyroxylin, previously dried at 809C for 2 hours, with 20 mL of water for 10 minutes: the filtrate is neutral. Water-soluble substances—Evaporate 10 mL of the filtrate obtained in (2) on a water bath to dryness, and dry at 1059C for 1 hour: the mass of the residue is not more than 1.5 mg. Residue on ignition—Weigh accurately about 2 g of Pyroxylin, previously dried at 809C for 2 hours, and moisten with 10 mL of a solution of castor oil in acetone (1 in 20) to gelatinize the sample. Ignite the contents to carbonize the sample, heat strongly at about 5009C for 2 hours, and allow to cool in a desicator (silica gel): the amount of the residue is not more than 0.30z.
Pyrrolnitrin contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calcu-lated on the dried basis.
Pyrrolnitrin occurs as yellow to yellow-brown, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Pyrrolnitrin in ethanol (95) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pyrrolnitrin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Pyr-rolnitrin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Pyrrolnitrin RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
Related substances—Dissolve 0.10 g of Pyrrolnitrin in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 3 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 ‌mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of xylene, ethyl acetate and formic acid (18:2:1) to a distance of about 10 cm, and dry the plate at 809C for 30 minutes. Spray evenly diluted sulfuric acid (1 in 3) on the plate, and heat the plate at 1009C for 30 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution.‌ Loss on drying <2.41> Not more than 0.5z (1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure using light-resistant vessels. Weigh accurately an amount of Pyrrolnitrin and Pyrrolnitrin RS, equivalent to about 50 mg (potency) each, and dissolve separetely in diluted acetonitrile (3 in 5) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution, add diluted acetonitrile (3 in 5) to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of pyrrolnitrin to that of the internal standard. Amount [ mg (potency)] of pyrrolnitrin (C10H6Cl2N2O2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Pyrrolnitrin RS taken Internal standard solution—A solution of benzyl benzoate in diluted acetonitrile (3 in 5) (3 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (11:9). Flow rate: Adjust so that the retention time of pyrrolnitrin is about 9 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, pyrrolnitrin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of pyrrolnitrin to that of the internal standard is not more than 1.0z.
Quetiapine Fumarate contains not less than 98.
Quetiapine Fumarate occurs as a white pow-der. It is sparingly soluble in methanol, and slightly soluble in water and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Quetiapine Fumarate in a mixture of water and acetonitrile (1:1) (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Quetiapine Fumarate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Quetiapine Fumarate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Quetiapine Fumarate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 40 mg of Quetiapine Fumarate and 10 mg of fumaric acid for thin-layer chromatography in separate 10 mL of methanol, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of isopropyl ether, formic acid and water (90:7:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot having a larger Rf value among the spots obtained from the sample solution and the spot from the standard so-lution show the same Rf value.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Quetiapine Fumarate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances (i)—To 20 mg of Quetiapine Fumarate add 30 mL of the mobile phase, dissolve by sonicating, add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 5 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, and add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area by the automatic integra-tion method, and calculate the amount of each related sub-stance by the following equation: the amount is not more than 0.10z. For the area of the peaks, having a relative retention time of about 0.5 and about 0.9 to quetiapine, mul-tiply their correction factors, 0.6 and 0.9, respectively. Mobile phase: A mixture of methanol, diammonium hydrogen phosphate solution (33 in 12,500) and acetonitrile (70:21:9). Flow rate: Adjust so that the retention time of quetiapine is about 3.5 minutes. Time span of measurement: About 8 times as long as the retention time of quetiapine, beginning from about 1.2 times the retention time of quetiapine. Amount (z) of each related substance ＝ AT/AS × 1/2 System suitability— Test for required detectability: Pipet 5 mL of the standard AS: Peak area of quetiapine obtained with the standard solution AT: Each peak area other than quetiapine obtained with the sample solution Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.8 times as long as the retention time of quetiapine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of quetiapine obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 2.0z. —To 20 mg of Quetiapine Fumarate add 30 mL of a mixture of acetonitrile, water and the mobile phase (2:1:1), dissolve by sonicating, add the same mixture to make 50 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add the same mixture to make ex-actly 100 mL. Pipet 5 mL of this solution, add the same mix-ture to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amount of each related substance by the following equation: the amount is not more than 0.10z. For the area of the peak, having a rel-ative retention time of about 1.9 to quetiapine, multiply its correction factor, 0.8. Amount (z) of each related substance ＝ AT/AS × 1/2 AS: Peak area of quetiapine obtained from the standard solution AT: Each peak area other than quetiapine obtained from the sample solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. solution, and add a mixture of acetonitrile, water and the mobile phase (2:1:1) to make exactly 50 mL. Confirm that the peak area of quetiapine obtained with 50 mL of this solu-tion is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 2.0z. —The total amount of the related substances obtained in (i) and (ii) is not more than 0.5z. Water <2.48> Not more than 0.5z (Weigh accurately about 0.1 g of Quetiapine Fumarate, transfer to a centrifuge tube, add exactly 4 mL of methanol for water determination, shake vigorously for 1 minute, and centrifuge at 2000 round per minute for 5 minutes. Pipet 1 mL of the supernatant liq-uid and perform the test. Perform a blank determination in the same manner, and make any necessary correction. Coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 20 mg each of Quetiapine Fumarate and Quetiapine Fumarate RS (separately deter-mine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase to them, dissolve by sonicating, and add the mobile phase to make exactly 100 mL. Pipet 10 mL each of these solutions, add the mobile phase to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of quetiapine in each so-lution. Amount (mg) of quetiapine fumarate [(C21H25N3O2S)2.C4H4O4] ＝ MS × AT/AS MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.6 g of diammonium hydrogen phosphate in 1000 mL of water, and adjust to pH 6.5 with phosphoric acid. To 39 volumes of this solution add 54 ‌volumes of methanol and 7 volumes of acetonitrile.‌ Flow rate: Adjust so that the retention time of quetiapine is about 15 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 1.0z.
Quinapril Hydrochloride occurs as a white Amount (mg) of quetiapine (C21 of Quetiapine Fumarate Tablets H25N3O2S) in 1 tablet powder. It is very soluble in methanol, freely soluble in water and in ethanol (99.5), and soluble in acetic acid (100). ＝ MS × AT/AS × V/16 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, diammonium hydrogen phosphate solution (33 in 12,500) and acetonitrile (54:39:7). Flow rate: Adjust so that the retention time of quetiapine is about 15 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 1.0z.
(1) Determine the absorption spectrum of a solution of Quinapril Hydrochloride in methanol (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Quinapril Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Quinapril Hydrochloride (1 in 20) re-sponds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: ＋14.4 – ＋16.09(0.5 g calcu-lated on the anhydrous basis, methanol, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Quinapril Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Quinapril Hy-drochloride in 50 mL of a mixture of phosphate buffer solu-tion (pH 7.0) and acetonitrile for liquid chromatography (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chroma-tography (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peaks, having the relative retention time of about 0.5 and about 2.0 to quinapril, obtained from the sample solution are not larger than the peak area of quinapril from the standard solution, respectively, the area of peak other than quinapril and the peak mentioned above from the sample solution are not larger than 2/5 times the peak area of quinapril from the standard solution, and the total area of the peaks other than ‌quinapril from the sample solution is not larger than 3 times the peak area of quinapril from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: While keeping the temperature not below 259C, adjust to pH 2.0 of 0.2 mol/L potassium dihydrogen phosphate TS with perchloric acid. To 1000 mL of this solu-tion add 1000 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of quinapril is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of quinapril, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly 100 mL. Confirm that the peak area of quinapril obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quinapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quinapril is not more than 2.0z. Water <2.48> Not more than 1.0z (0.2 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Start to titrate within 3 minutes after dissolving Quinapril Hydrochloride. Weigh accurately about 0.5 g of Quinapril Hydrochloride, dissolve in 70 mL of acetic acid (100), add 4 mL of bismuth nitrate TS, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 47.50 mg of C25H30N2O5.HCl
Quinidine Sulfate Hydrate occurs as white crystals. It is odorless, and has a very bitter taste. It is freely soluble in ethanol (95) and in boiling water, sparingly soluble in water, and practically insoluble in diethyl ether. It, previously dried, is freely soluble in chlo-roform. It darkens gradually by light. D Optical rotation [a]20: ＋275 – ＋2879(after drying, 0.5 g, 0.1 mol/L hydrochloric acid VS, 25 mL, 100 mm).
(1) Dissolve 0.01 g of Quinidine Sulfate Hydrate in 10 mL of water and 2 to 3 drops of dilute sulfuric acid: a blue fluorescence is produced. To 5 mL of a solution of Quinidine Sulfate Hydrate (1 in 1000) add 1 to 2 drops of bromine TS, then add 1 mL of ammonia TS: a green color develops. To 5 mL of a solution of Quinidine Sulfate Hydrate (1 in 100) add 1 mL of silver nitrate TS, stir with a glass rod, and allow to stand for a short interval: a white precipitate is produced, and it dissolves on addition of nitric acid. Dissolve 0.4 g of Quinidine Sulfate Hydrate in 20 mL of water and 1 mL of dilute hydrochloric acid: the solution responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Quinidine Sulfate Hydrate in 100 mL of freshly boiled and cooled water: the pH of this solution is between 6.0 and 7.0.
(1) Chloroform-ethanol-insoluble substances—Warm 2.0 g of Quinidine Sulfate Hydrate with 15 mL of a mixture of chloroform and ethanol (99.5) (2:1) at about 509C for 10 minutes. After cooling, filter through a tared glass filter (G4) by gentle suction. Wash the residue with five 10-mL portions of a mixture of chloroform and ethanol (99.5) (2:1), and dry at 1059C for 1 hour: the mass of the residue is not more than 2.0 mg. Related substances—Dissolve 20 mg of Quinidine Sulfate Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dis-solve 25 mg of cinchonine in the mobile phase to make ex-actly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of the sample methanol, and add the mobile phase to make 50 mL. Pro-ceed with 50 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of quinidine, quinine, dihydroquinidine and dihy-droquinine in this order with a resolution between quinidine and quinine and that between quinine and dihydroquinidine being not less than 1.2, respectively. Detection sensitivity: Adjust the detection sensitivity so that the peak height of cinchonine obtained with 50 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About 2 times as long as the retention time of quinidine, beginning after the solvent peak. Readily carbonizable substances <1.15>—Take 0.20 g of Quinidine Sulfate Hydrate and perform the test: the solu-tion has no more color than Matching Fluid M. Loss on drying <2.41> Not more than 5.0z (1 g, 130 9C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Quinidine Sulfate Hydrate, previously dried, dissolve in 20 mL of acetic acid (100), and add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solu-tion changes from purple through blue to blue-green (indica-tor: 3 drops of crystal violet TS). Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.90 mg of (C20H24N2O2)2.H2SO4
Quinine Ethyl Carbonate occurs as white crys-tals. It is odorless, and tasteless at first but slowly develops a bitter taste. It is very soluble in methanol, freely soluble in ethanol (95) and in ethanol (99.5), soluble in diethyl ether, and practically insoluble in water. It dissolves in dilute hydrochloric acid.
(1) Determine the absorption spectrum of a solution of Quinine Ethyl Carbonate in methanol (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Qui-nine Ethyl Carbonate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －42.2 – －44.09(0.5 g calcu-lated on the anhydrous basis, methanol, 50 mL, 100 mm).
(1) Chloride—Dissolve 0.30 g of Quinine Ethyl Carbonate in 10 mL of dilute nitric acid and 20 mL of water. To 5 mL of the solution add 2 to 3 drops of silver nitrate TS: no color develops. Sulfate <1.14>—Dissolve 1.0 g of Quinine Ethyl Car- bonate in 5 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS, 5 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Quinine Ethyl Carbonate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 20 mg of Quinine Ethyl Carbonate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of quinine sulfate hydrate in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- adjust to pH 3.5 with diluted phosphoric acid (1 in 20). Flow rate: Adjust so that the retention time of the peak of quinine ethyl carbonate is about 20 minutes. Selection of column: Dissolve 5 mg each of Quinine Ethyl Carbonate and quinine sulfate hydrate in the mobile phase to make 50 mL. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of quinine, dihydroquinine, qui-nine ethyl carbonate and the main impurity of quinine ethyl carbonate in this order with the resolution between the peaks of quinine and dihydroquinine being not less than 2.7, and between the peaks of quinine and quinine ethyl carbonate being not less than 5. Detection sensitivity: Adjust the detection sensitivity so that the peak height of quinine obtained from 10 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About 2 times as long as the retention time of quinine ethyl carbonate. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Quinine Ethyl Car-bonate, dissolve in 60 mL of acetic acid (100), add 2 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 19.82 mg of C23H28N2O4
Quinine Hydrochloride Hydrate occurs as white crystals. It is odorless, and has a very bitter taste. It is very soluble in ethanol (99.5), freely soluble in acetic acid (100), in acetic anhydride and in ethanol (95), soluble in water, and practically insoluble in diethyl ether. It, previously dried, is freely soluble in chloroform. It gradually changes to brown by light.
(1) A solution of Quinine Hydrochloride Hydrate (1 in 50) shows no fluorescence. To 1 mL of the so-lution add 100 mL of water and 1 drop of dilute sulfuric acid: a blue fluorescence is produced. To 5 mL of a solution of Quinine Hydrochloride Hy-drate (1 in 1000) add 1 to 2 drops of bromine TS and 1 mL of ammonia TS: a green color develops. To 5 mL of a solution of Quinine Hydrochloride Hy-drate (1 in 50) add 1 mL of dilute nitric acid and 1 mL of sil-ver nitrate TS: a white precipitate is produced. Collect the precipitate, and add an excess of ammonia TS: it dissolves. D Optical rotation <2.49> [a]20: －245 – －2559(after drying, 0.5 g, 0.1 mol/L hydrochloric acid VS, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Quinine Hydrochloride Hy-drate in 100 mL of freshly boiled and cooled water: the pH of this solution is between 6.0 and 7.0.
(1) Sulfate <1.14>—Perform the test with 1.0 g of Quinine Hydrochloride Hydrate. Prepare the control solu-tion with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Barium—Dissolve 0.5 g of Quinine hydrochloride Hydrate in 10 mL of water by warming, and add 1 mL of di-lute sulfuric acid: no turbidity is produced. Chloroform-ethanol-insoluble substances—Warm 2.0 g of Quinine Hydrochloride Hydrate with 15 mL of a mixture of chloroform and ethanol (99.5) (2:1) at 509C for 10 minutes. After cooling, filter through a tared glass filter (G4) by gentle suction. Wash the residue with five 10-mL portions of a mixture of chloroform and ethanol (99.5) (2:1), dry at 1059C for 1 hour, and weigh: the mass of the residue so obtained is not more than 2.0 mg. Related substances—Dissolve 20 mg of Quinine Hy-drochloride Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of cinchonidine in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of the sample solution by the automatic integration method, and calculate the amount of dihydroquinine hydrochloride by the area per-centage method: it is not more than 10.0z. The total area of the peaks other than the main peak and the peaks mentioned above is not larger than the peak area of cinchonidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 25 cm in length, packed with octadecyl-silanized silica gel (10 mm in particle diameter). Column temperature: Room temperature. Mobile phase: A mixture of water, acetonitrile, methanesulfonic acid TS and a solution of diethylamine (1 in 10) (43:5:1:1). Flow rate: Adjust so that the retention time of quinine is about 10 minutes. Selection of column: Dissolve 10 mg each of Quinine Hy-drochloride Hydrate and quinidine sulfate hydrate in 5 mL of methanol, and add the mobile phase to make 50 mL. Pro-ceed with 50 mL of this solution under the above operating conditions. Use a column giving elution of quinidine, qui-nine, dihydroquinidine and dihydroquinine in this order with the resolution between quinidine and quinine, and that be-tween quinine and dihydroquinidine being not less than 1.2, respectively. Detection sensitivity: Adjust the detection sensitivity so that the peak height of cinchonidine from 50 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About 2 times as long as the retention time of quinine, beginning after the solvent peak. Readily carbonizable substances <1.15>—Perform the test with 0.25 g of Quinine Hydrochloride Hydrate. The so-lution has no more color than Matching Fluid M. Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Quinine Hydrochlo-ride Hydrate, previously dried, dissolve in 100 mL of a mix-ture of acetic anhydride and acetic acid (100) (7:3) by warm-ing, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 18.04 mg of C20H24N2O2.HCl
Quinine Sulfate Hydrate occurs as white, crys-tals or crystalline powder. It is odorless, and has a very bitter taste. It is freely soluble in acetic acid (100), slightly soluble in water, in ethanol (95), in ethanol (99.5) and in chloroform, and practically insoluble in diethyl ether. It gradually changes to brown by light.
(1) Determine the absorption spectrum of a solution of Quinine Sulfate Hydrate (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Qui-nine Sulfate Hydrate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. To 0.4 g of Quinine Sulfate Hydrate add 20 mL of water and 1 mL of dilute hydrochloric acid: the solution re-sponds to Qualitative Tests <1.09> for sulfate. D Optical rotation <2.49> [a]20: －235 – －2459(after drying, 0.5 g, 0.1 mol/L hydrochloric acid VS, 25 mL, 100 mm). pH <2.54> Shake 2.0 g of Quinine Sulfate Hydrate in 20 mL of freshly boiled and cooled water, and filter: the pH of this filtrate is between 5.5 and 7.0. actly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area from the sample solution by the automatic integration method, and calculate the amount of dihydroquinine sulfate by the area percentage method: it is not more than 5z. The total area of the peaks other than the main peak and the peaks mentioned above is not larger than the peak area of cinchonidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A column about 4 mm in inside diameter and about 25 cm in length, packed with octadecylsilanized silica gel (10 mm in particle diameter). Temperature: Room temperature. Mobile phase: A mixture of water, acetonitrile, methane sulfonic acid TS and a solution of diethylamine (1 in 10) (43:5:1:1). Flow rate: Adjust so that the retention time of quinine is about 10 minutes. Selection of column: Dissolve 0.01 g each of Quinine Sul-fate Hydrate and quinidine sulfate hydrate in 5 mL of methanol, and add the mobile phase to make 50 mL. Pro-ceed with 50 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of quinidine, quinine, dihydroquinidine and dihy-droquinine in this order with the resolution between quini-dine and quinine and that between quinine and dihy-droquinidine being not less than 1.2, respectively. Detection sensitivity: Adjust the detection sensitivity so that the peak height of cinchonidine obtained from 50 mL of the standard solution is between 5 mm and 10 mm. Time span of measurement: About 2 times as long as the retention time of quinine, beginning after the solvent peak. Loss on drying <2.41> 3.0z – 5.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Quinine Sulfate Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Chloroform-ethanol-insoluble substances—Warm 2.0 g of Quinine Sulfate Hydrate with 15 mL of a mixture of chloroform and ethanol (99.5) (2:1) at 509C for 10 minutes. After cooling, filter through a tared glass filter (G4) by gen-tle suction. Wash the residue with five 10-mL portions of a mixture of chloroform and ethanol (99.5) (2:1), dry at 1059C for 1 hour, and weigh: the mass of the residue is not more than 2.0 mg. Related substances—Dissolve 20 mg of Quinine Sul-fate Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dis-solve 25 mg of cinchonidine in the mobile phase to make ex-
Weigh accurately about 0.5 g of Quinine Sulfate Hy-drate, dissolve in 20 mL of acetic acid (100), add 80 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS until the color of the solution changes from pur-ple through blue to blue-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.90 mg of (C H N O ) .H SO 20 24 2 2 2 2 4 Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Quinine Sulfate Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Chloroform-ethanol-insoluble substances—Warm 2.0 g of Quinine Sulfate Hydrate with 15 mL of a mixture of chloroform and ethanol (99.5) (2:1) at 509C for 10 minutes. After cooling, filter through a tared glass filter (G4) by gen-tle suction. Wash the residue with five 10-mL portions of a mixture of chloroform and ethanol (99.5) (2:1), dry at 1059C for 1 hour, and weigh: the mass of the residue is not more than 2.0 mg. Related substances—Dissolve 20 mg of Quinine Sul-fate Hydrate in the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, dis-solve 25 mg of cinchonidine in the mobile phase to make ex-
Ranitidine Hydrochloride, when dried, contains not less than 97.
Ranitidine Hydrochloride occurs as a white to pale yellow, crystalline or fine granular powder. It is very soluble in water, freely soluble in methanol, and slightly soluble in ethanol (99.5). It is hygroscopic. It is gradually colored by light. Melting point: about 1409C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ranitidine Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ranitidine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ranitidine Hydrochloride as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Ranitidine Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Ranitidine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Ranitidine Hydrochloride in 100 mL of water is between 4.5 and 6.0.
(1) Clarity and color of solution—A solution of Ranitidine Hydrochloride (1 in 10) is clear and pale yellow to light yellow. Heavy metals <1.07>—Proceed with 2.0 g of Raniti- dine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ranitidine Hydrochloride according to Method 4, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 0.22 g of Ranitidine Hydrochloride in methanol to make ex-actly 10 mL, and use this solution as the sample solution. Pipet 0.5 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solu-tion (1). Pipet 6 mL, 4 mL, 2 mL and 1 mL of the standard solution (1), add to each methanol to make exactly 10 mL, and use these solutions as the standard solution (2), the standard solution (3), the standard solution (4) and the standard solution (5), respectively. Separately, dissolve 12.7 mg of ranitidinediamine in methanol to make exactly 10 mL, and use this solution as the standard solution (6). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solutions (1), (2), (3), (4) and (5) on a plate of silica gel for thin-layer chromatography. Separately, spot 10 mL of the sample solution on the plate, then spot 10 mL of the standard solution (6) on the spotted position of the sam-ple solution. Immediately develop the plate with a mixture of ethyl acetate, 2-propanol, ammonia solution (28) and water (25:15:5:1) to a distance of about 15 cm, and air-dry the plate. Allow the plate to stand in iodine vapor until the spot from the standard solution (5) appears: the spot obtained from the standard solution (6) is completely separated from the principal spot from the sample solution. The spot having Rf value of about 0.7 from the sample solution is not more intense than the spot from the standard solution (1), the spots other than the principal spot and the spot of Rf value of about 0.7 from the sample solution are not more intense than the spot from the standard solution (2), and the total amount of these related substances, calculated by compari-son with the spots from the standard solutions (1), (2), (3), (4) and (5), is not more than 1.0z. Loss on drying <2.41> Not more than 0.75z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). ‌Assay Weigh accurately about 20 mg of Ranitidine Hydro-chloride and Ranitidine Hydrochloride RS, previously dried, dissolve each in the mobile phase to make exactly 200 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ranitidine in each solution. Amount (mg) of ranitidine hydrochloride (C13H22N4O3S.HCl) ＝ MS × AT/AS MS: Amount (mg) of Ranitidine Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 322 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and diluted 0.5 mol/L ammonium acetate TS (1 in 5) (17:3). Flow rate: Adjust so that the retention time of ranitidine is about 5 minutes. System suitability— System performance: Dissolve 20 mg of Ranitidine Hydro-chloride and 5 mg of benzalphthalide in 200 mL of the mo-bile phase. When the procedure is run with 10 mL of this so-lution under the above operating conditions, benzalphthalide and ranitidine are eluted in this order with the resolution be-tween these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ranitidine is not more than 1.0z.
Rebamipide, when dried, contains not less than 99.
Rebamipide occurs as a white crystalline pow-der. It has a bitter taste. It is soluble in N,N-dimethylformamide, very slightly soluble in methanol and in ethanol (99.5), and practically in-soluble in water. A solution of Rebamipide in N,N-dimethylformamide (1 in 20) shows no optical rotation. Melting point: about 2919C (with decomposition).
(1) Determine the absorption spectrum of a solution of Rebamipide in methanol (7 in 1,000,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Reba-mipide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Rebamipide as directed under Flame Coloration Test <1.04> (2): a green color appears.
(1) Chloride <1.03>—Dissolve 0.5 g of Rebamipide in 40 mL of N,N-dimethylformamide, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.40 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of N,N-dimethylformamide, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.028z). Heavy metals <1.07>—Proceed with 2.0 g of Rebami- pide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Rebamipide m-chloro isomer—Dissolve 40 mg of Rebamipide in a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make 100 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make exactly 20 mL. Pipet 2 mL of this solution, add a mixture of water, 0.05 mol/L phosphate buffer solu-tion (pH 6.0) and methanol (7:7:6) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak of rebamipide m-chloro isomer, having the relative retention time of about 0.95 to re-bamipide obtained from the sample solution, is not larger than 3/8 times the area of the peak of rebamipide from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 300 mL of phosphate buffer solution (pH 6.2) add 750 mL of water. To 830 mL of this solution add 170 mL of acetonitrile. Flow rate: Adjust so that the retention time of rebamipide is about 20 minutes. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make ex-actly 25 mL. Confirm that the peak area of rebamipide ob-tained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. ‌System performance: To 1 mL of the sample solution add a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of rebamipide are not less than 11,000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rebamipide is not more than 2.0z. Related substances—Perform the test with exactly 10 mL each of the sample solution and standard solution ob-tained in (3) as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: each area of the peaks of rebamipide o-chloro isomer and debenzoylated isomer, having the relative reten-tion times of about 0.5 and about 0.7, respectively, to re-bamipide obtained from the sample solution, is not larger than 3/8 times the peak area of rebamipide from the stand-ard solution, the area of each peak other than rebamipide and the peak mentioned above from the sample solution is not larger than 1/4 times the peak area of rebamipide from the standard solution, and the total area of the peaks other than rebamipide from the sample solution is not larger than the peak area of rebamipide from the standard solution. For the peak area of rebamipide o-chloro isomer, multiply the correction factor, 1.4. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.44 g of sodium 1-decanesul-fonate in 1000 mL of water and to this solution add 1000 mL of methanol and 10 mL of phosphoric acid. Flow rate: Adjust so that the retention time of rebamipide is about 12 minutes. Time span of measurement: About 3 times as long as the retention time of rebamipide, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make ex-actly 50 mL. Confirm that the peak area of rebamipide ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 20 mg of 4-chlorobenzoate in methanol to make 50 mL. To 5 mL of this solution add 5 mL of the sample solution and a mixture of water, 0.05 mol/L phosphate buffer solution (pH 6.0) and methanol (7:7:6) to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, re-bamipide and 4-chlorobenzoate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rebamipide is not more than 2.0z. Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Rebamipide, previ-ously dried, dissolve in 60 mL of N,N-dimethylformamide, and titrate <2.50> with 0.1 mol/L potassium hydroxide VS until the color of the solution changes from pale yellow to colorless (indicator: 2 drops of phenol red TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L potassium hydroxide VS ＝ 37.08 mg of C19H15ClN2O4
Reserpine, when dried, contains not less than 96.
Reserpine occurs as white to light yellow, crys-tals or crystalline powder. It is freely soluble in acetic acid (100) and in chloroform, slightly soluble in acetonitrile, very slightly soluble in ethanol (95), and practically insoluble in water and in diethyl ether. It is affected by light.
(1) To 1 mg of Reserpine add 1 mL of vanillin-hydrochloric acid TS, and warm: a vivid red-purple color develops. Determine the absorption spectrum of a solution of Reserpine in acetonitrile (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Reserpine RS prepared in the same manner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. ‌Determine the infrared absorption spectrum of Reserpine, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of previously dried Reserpine RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －114 – －1279(after drying, 0.25 g, chloroform, 25 mL, 100 mm).
Related substances—Conduct this procedure with-out exposure to light, using light-resistant vessels. Dissolve 50 mg of Reserpine in 50 mL of acetonitrile, and use this so-lution as the sample solution. Pipet 3 mL of the sample solu-tion, add acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area from these solutions by the automatic integration method: the total area of the peaks other than reserpine ob-tained from the sample solution is not larger than the peak area of reserpine from the standard solution. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate (pH 3.0) and acetonitrile (13:7). Flow rate: Adjust so that the retention time of reserpine is about 20 minutes. Time span of measurement: About 2 times as long as the retention time of reserpine. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add acetonitorile to make exactly 50 mL. Confirm that the peak area of reserpine obtained with 10 mL of this solution is equivalent to 3 to 5z of that with 10 mL of the standard solution. System performance: Dissolve 0.01 g of Reserpine and 4 mg of butyl parahydroxybenzoate in 100 mL of acetonitrile. To 5 mL of this solution add acetonitrile to make 50 mL. When the procedure is run with 20 mL of this solution ac-cording to the operating conditions in the Assay, reserpine and butyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of reserpine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.2 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.2 g).
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 10 mg each of Reserpine and Reserpine RS, previously dried, and dissolve each in acetonitrile to make exactly 100 mL. Pipet 5 mL each of these solutions, add exactly 10 mL of the inter-nal standard solution, 5 mL of acetonitrile and water to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of reserpine to that of the internal standard. Amount (mg) of reserpine (C33H40N2O9) ＝ MS × QT/QS MS: Amount (mg) of Reserpine RS taken Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate (pH 3.0) and acetonitrile (11:9). Flow rate: Adjust so that the retention time of reserpine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, reserpine and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of reserpine to that of the internal standard is not more than 2.0z.
Residue on ignition <2.44> Not more than 0.2z (1 g).
Thiamine Chloride Hydrochloride Injection is a clear, colorless liquid. pH: 2.5 – 4.5
To a volume of Thiamine Chloride Hydro- ‌chloride Injection, equivalent to 0.05 g of Thiamine Chloride Hydrochloride, add water to make 25 mL. Proceed with 5 mL of this solution as directed in the Identification (1) under Thiamine Chloride Hydrochloride.‌ Bacterial endotoxins <4.01> Less than 6.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Rancidity—Thiamine Chloride Hydrochloride Pow-der has no unpleasant or rancid odor. It is tasteless.
Dilute with 0.001 mol/L hydrochloric acid TS if nec-essary, then measure exactly a volume of Thiamine Chloride Hydrochloride Injection, equivalent to about 20 mg of thia-mine chloride hydrochloride (C12H17ClN4OS.HCl), and add 20 mL of methanol and 0.001 mol/L hydrochloric acid TS to make exactly 100 mL. To 25 mL of this solution, exactly measured, add exactly 5 mL of the internal standard solu-tion, add 0.001 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Thiamine Chloride Hydro-chloride RS (separately determine the water <2.48> in the same manner as Thiamine Chloride Hydrochloride), and dis-solve in 0.001 mol/L hydrochloric acid TS to make exactly 50 mL. To 10 mL of this solution, exactly measured, add 20 mL of methanol and 0.001 mol/L hydrochloric acid TS to make exactly 100 mL. To 25 mL of this solution, exactly measured, add exactly 5 mL of the internal standard solu-tion, add 0.001 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Thiamine Chloride Hydrochloride. Amount (mg) of thiamine chloride hydrochloride (C12H17ClN4OS.HCl) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Thiamine Chloride Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of methyl benzoate in methanol (1 in 200).
Ribavirin, when dried, contains not less than 98.
Ribavirin occurs as a white crystalline powder. It is freely soluble in water and in N, N-dimethylfor-mamide, slightly soluble in methanol, and practically insolu- ble in ethanol (99.5). Melting point: 167 – 1719C Ribavirin shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Ribavirin (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ribavirin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of previ-ously dried Ribavirin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Ribavirin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: － 33.0 – － 37.09(after dry-ing, 0.1 g, water, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ribavirin according to Method 1, and perform the test. Pre-pare the control solution with 1.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ribavirin according to Method 5, and perform the test (not more than 2 ppm). Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of ‌the peak, having the relative retention time of about 0.85 to ribavirin, obtained from the sample solution, is not larger than 2/5 times the peak area of ribavirin from the standard solution, and the area of the peak other than ribavirin and other than the peak mentioned above from the sample solu-tion is not larger than 1/5 times the peak area of ribavirin from the standard solution. Furthermore, the total area of the peaks other than ribavirin and other than the peak men-tioned above from the sample solution is not larger than 2/5 259C. Mobile phase A: Dissolve 2.0 g of anhydrous sodium sul-fate in 300 mL of water, add 8 mL of phosphoric acid solu-tion (1 in 20) and water to make 2000 mL. Mobile phase B: A mixture of mobile phase A and aceto-nitrile for liquid chromatography (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. times the peak area of ribavirin from the standard solution, and the total area of the peaks other than ribavirin from the sample solution is not larger than the peak area of ribavirin Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) from the standard solution. For the area of the peaks, hav-ing the relative retention time of about 0.59 and about 0.85 to ribavirin, multiply their correction factors 0.6 and 1.7, re-spectively. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 35 minutes after the injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add water to make exactly 10 mL. Con-firm that the peak area of ribavirin obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: To 5 mL of the sample solution add 1 mL of sodium hydroxide TS, allow to stand for 30 minutes, and add 1 mL of 1 mol/L hydrochloric acid TS. To 1 mL of this solution add water to make 200 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, the resolution between the peak having the relative retention time of about 0.85 to ribavirin and the peak of ribavirin is not less than 4.0, and when the proce-dure is run with 5 mL of the standard solution under the above operating conditions, the symmetry factor of the peak of ribavirin is not more than 1,5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ribavirin is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Ribavirin and Ribavirin RS, both previously dried, equivalent to about 25 mg each, dissolve in water to make exactly 50 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ribavirin in each solution. Amount (mg) of ribavirin (C8H12N4O5) ＝ MS × AT/AS MS: Amount (mg) of Ribavirin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 0 – 15 100 0 15 – 25 100 → 0 0 → 100 25 – 35 0 100 Flow rate: 1.0 mL per minute. System suitability— System performance: To 5 mL of the standard solution add 1 mL of sodium hydroxide TS, allow to stand for 30 minutes, and add 1 mL of 1 mol/L hydrochloric acid TS. When the procedure is run with 5 mL of this solution under the above operating conditions, the resolution between the peak having the relative retention time of about 0.85 to ribavirin and the peak of ribavirin is not less than 4.0, and when the procedure is run with 5 mL of the standard solution under the above operating conditions, the symmetry factor of the peak of ribavirin is not more than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ribavirin is not more than 1.0z.
Ribostamycin Sulfate according to Method 1, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ribostamycin Sulfate according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.12 g of Ribostamycin Sulfate in water to make exactly 20 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g). Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) Medium under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Ribostamycin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 5 to 159C and use within 20 days. Take exactly a suitable amount of the standard stock solution be-fore use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration standard solution, respectively.
Ribostamycin Sulfate occurs as a white to yel-low-white powder. It is freely soluble in water, and practically insoluble in ethanol (95).
(1) Dissolve 20 mg of Ribostamycin Sulfate in 2 mL of phosphate buffer solution (pH 6.0), add 1 mL of ninhydrin TS, and boil: a blue-purple color develops. Dissolve 0.12 g each of Ribostamycin Sulfate and Ribostamycin Sulfate RS in 20 mL of water, and use these solutions as the sample solution and standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solu-tion of potassium dihydrogen phosphate (3 in 40) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS, and heat at 1009C for 10 minutes: the principal spots obtained from the sample solution and standard solution show a purple-brown color and the same Rf value. To 2 mL of a solution of Ribostamycin Sulfate (1 in 5) add 1 drop of barium chloride TS: a white turbidity is pro-duced. D Optical rotation <2.49> [a]20: ＋42 – ＋499(after drying, 0.25 g, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Ribostamycin Sulfate in 20 mL of water is between 6.0 and 8.0. ‌Purity (1) Clarity and color of solution—Dissolve 2.9 g of Ribostamycin Sulfate in 10 mL of water: the solution is clear. Determine the absorbance of this solution at 400 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: not more than 0.10. Heavy metals <1.07>—Proceed with 1.0 g of Ribostamycin Sulfate according to Method 1, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ribostamycin Sulfate according to Method 1, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.12 g of Ribostamycin Sulfate in water to make exactly 20 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solution of potassium dihydrogen phosphate (3 in 40) to a distance of about 10 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) Medium under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Ribostamycin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 5 to 159C and use within 20 days. Take exactly a suitable amount of the standard stock solution be-fore use, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Ribostamycin Sulfate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Rice Starch occurs as a white mass or pow-der. It is practically insoluble in water and in ethanol (99.5).◆
(1) Examined under a microscope <5.01> using a mixture of water and glycerin (1:1), Rice Starch presents polyhedral, simple grains 1 – 10 mm, mostly 4 – 6 mm, in size. These simple grains often gather in ellipsoidal, compound grains 50 – 100 mm in diameter. The granules have a poorly visible central hilum and there are no concen-tric striations. Between orthogonally orientated polarizing plates or prisms, the starch granules show a distinct black cross intersecting at the hilum. To 1 g of Rice Starch add 50 mL of water, boil for 1 minute, and allow to cool: a thin, cloudy mucilage is formed. To 1 mL of the mucilage obtained in (2) add 0.05 mL of diluted iodine TS (1 in 10): an orange-red to dark-blue color is produced which disappears on heating. pH <2.54> To 5.0 g of Rice Starch add 25 mL of freshly boiled and cooled water, and mix gently for 1 minute to achieve suspension. Allow to stand for 15 minutes: the pH of the solution is between 5.0 and 8.0.
(1) Iron—To 1.5 g of Rice Starch add 15 mL of 2 mol/L hydrochloric acid TS, mix, filter, and use the filtrate as the test solution. To 2.0 mL of Standard Iron Solution ‌add water to make 20 mL, and use this solution as the con-trol solution. Put 10 mL each of the test solution and the control solution in test tubes, add 2 mL of a solution of citric acid (1 in 5) and 0.1 mL of mercapto acetic acid, and mix. Add ammonia solution (28) to these solutions until the color of a litmus paper to change from red to blue, add water to make 20 mL, and mix. Transfer 10 mL each of these solu-tions into test tubes, allow to stand for 5 minutes, and com-pare the color of these solutions against a white background: the color of the test solution is not more intense than that of the control solution (not more than 10 ppm). Oxidizing substances—To 4.0 g of Rice Starch add 50 mL of water, shake for 5 minutes, and centrifuge. To 30 mL of the supernatant liquid add 1 mL of acetic acid (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow to stand for 25 to 30 minutes in the dark. Add 1 mL of starch TS, and titrate <2.50> with 0.002 mol/L sodium thiosulfate VS until the starch-iodine color disappears. Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Not more than 1.4 mL of 0.002 mol/L sodium thiosul-fate VS is required (not more than 20 ppm, calculated as hydrogen peroxide). Sulfur dioxide— Apparatus Use as shown in the figure. A: Three-necked round-bottom flask (500 mL) B: Cylindrical dropping funnel (100 mL) C: Condenser D: Test tube E: Tap Procedure Introduce 150 mL of water into the three-necked round-bottom flask, close the tap of the cylin-drical dropping funnel, and pass carbon dioxide through the whole system at a rate of 100 ± 5 mL per minute. Pass cool-ing water through the condenser, and place 10 mL of hydro-gen peroxide-sodium hydroxide TS in the test tube. After 15 minutes, remove the funnel without interrupting the stream of carbon dioxide, and introduce through the opening into the flask about 25 g of Rice Starch, accurately weighed, with the aid of 100 mL of water. Apply tap grease to the outside of the connection part of the funnel, and load the funnel. Close the tap of the funnel, pour 80 mL of 2 mol/L hydro-chloric acid TS into the funnel, open the tap to introduce the hydrochloric acid into the flask, and close the tap while several mL of the hydrochloric acid remains, in order to avoid losing sulfur dioxide. Place the flask in a water bath, and heat the mixture for 1 hour. Transfer the contents of the test tube with the aid of a little water to a wide-necked coni-cal flask. Heat on a water bath for 15 minutes and allow to cool. Add 0.1 mL of bromophenol blue TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 sec-onds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the amount of sulfur dioxide by applying the following formula: it is not more than 50 ppm. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 M: Amount (g) of Rice Starch taken V: Amount (mL) of 0.1 mol/L sodium hydroxide VS con-sumed (4) Foreign matter—Under a microscope <5.01>, Rice Starch does not contain starch granules of any other origin. It may contain a minute quantity, if any, of fragments of the tissue of the original plant.◆ Loss on drying <2.41> Not more than 15.0z (1 g, 1309C, 90 minutes). Residue on ignition <2.44> Not more than 0.6z (1 g).
Rifampicin is a derivative of a substance having an-tibacterial activity produced by the growth of Strep-tomyces mediterranei. It contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the dried basis.
Rifampicin occurs as orange-red to red-brown, crystals or crystalline powder. It is slightly soluble in water, in acetonitrile, in methanol and in ethanol (95).
(1) To 5 mL of a solution of Rifampicin in methanol (1 in 5000) add 0.05 mol/L phosphate buffer solu-tion (pH 7.0) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Rifampicin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Rifampicin as directed in the potassium bromide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Rifampicin RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Rifampicin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Rifampicin according to Method 3, and perform the test (not more than 2 ppm). Related substances—Perform the test immediately after preparing of the sample and standard solutions. Dis-solve 0.10 g of Rifampicin in 50 mL of acetonitrile, and use this solution as the sample stock solution. Pipet 5 mL of the sample stock solution, add citric acid-phosphate-acetonitrile TS to make exactly 50 mL, and use this solution as the sam-ple solution. Separately, pipet 1 mL of the sample stock so- ‌lution, and add acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add citric acid-phosphate-acetonitrile TS to make exactly 50 mL, and use this solution as the stand-ard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic inte-gration method: the area of the peak appeared at the relative retention time of about 0.7 to rifampicin obtained from the sample solution is not larger than 1.5 times the peak area of rifampicin from the standard solution, the area of the peak other than rifampicin and the peak mentioned above from the sample solution is not larger than the peak area of rifam-picin from the standard solution, and the total area of the peaks other than rifampicin and the peak mentioned above from the sample solution is not larger than 3.5 times the peak area of rifampicin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of rifampicin, beginning after the peak of the solvent. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 2 mL of the standard solution, and add citric acid-phosphate-acetoni-trile TS to make exactly 20 mL. Confirm that the peak area of rifampicin obtained with 50 mL of this solution is equiva-lent to 7 to 13z of that with 50 mL of the standard solution. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rifampicin is not more than 2.0z. Loss on drying <2.41> Not more than 2.0z (1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Rifampicin and Rifampicin RS, equivalent to about 40 mg (potency), and dissolve each in acetonitrile to make exactly 200 mL. Pipet 10 mL each of these solutions, add citric acid-phosphate-acetonitrile TS to make exactly 100 mL, and use these solu-tions as the sample solution and standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of rifampicin in each solution. Flow rate: Adjust so that the retention time of rifampicin is about 8 minutes. System suitability— System performance: To 5 mL of a solution of Rifampicin in acetonitrile (1 in 5000) add 1 mL of a solution of butyl parahydroxybenzoate in acetonitrile (1 in 5000) and citric acid-phosphate-acetonitrile TS to make 50 mL. When the procedure is run with 50 mL of this solution under the above operating conditions, butyl parahydroxybenzoate and rifam-picin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rifampicin is not more than 1.0z.
Rikkunshito Extract contains not less than 2.
Rikkunshito Extract is a light brown to brown powder or black-brown viscous extract. It has an odor and a sweet and bitter taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). (2) For preparation prescribed Atractylodes Rhizome—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylaminobenzalde-hyde TS for spraying, heated at 1059C for 5 minutes, and al-lowed to cool (Atractylodes Lancea Rhizome). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of hesperidin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, acetone, water and acetic acid (100) (10:6:3:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and allow to stand in an ammonia gas: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (Citrus Unshiu Peel). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 30 mL of the sample solution and 5 mL of JP XVIII Crude Drugs and Related Drugs / Rikkunshito Extract 2117 the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Ginsenoside Rb1—Weigh accurately about 2 g of the dry extract (or an amount of the viscous extract, equivalent to about 2 g of the dried substance), add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 15 mL of diluted methanol (3 in 5), repeat the same procedure. diameter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of acetonitrile, water and phos-phoric acid (400:100:1). Flow rate: 1.0 mL per minute (the retention time of gin-senoside Rb1 is about 16 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ginsenoside Rb1 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z. Hesperidin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted tetrahydrofuran (1 in 4), shake for 30 minutes, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 10 mg of hesperidin for assay, previously dried in a desiccator (silica gel) for more than 24 hours, dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add diluted tetra-hydrofuran (1 in 4) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of hesperidin in each solution. Combine the supernatant liquids, add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add Amount (mg) of hesperidin ＝ MS × AT/AS × 1/20 3 mL of sodium hydroxide TS, allow to stand for 30 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 20 mL. Apply exactly 5 mL of this solution to a column (about 10 mm in inside diameter and packed with 0.36 g of octadecylsilanized silica gel for pre-treatment (55 – 105 mm in particle size), washed just before use with methanol and then with diluted methanol (3 in 10)), and wash the column in sequence with 2 mL of diluted methanol (3 in 10), 1 mL of sodium carbonate TS and 10 mL of diluted methanol (3 in 10). Finally, elute with methanol to collect exactly 5 mL, and use this as the sample solution. Separately, weigh accurately about 10 mg of Gin-senoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solution. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of hesperidin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (82:18:1). Flow rate: 1.0 mL per minute (the retention time of hesperidin is about 15 minutes). System suitability— System performance: Dissolve 1 mg each of hesperidin for assay and naringin for thin-layer chromatography in diluted methanol (1 in 2) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, naringin and hesperidin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- MS: Amount (mg) of Ginsenoside Rb1 lated on the anhydrous basis Operating conditions— RS taken, calcu- ing conditions, the relative standard deviation of the peak area of hesperidin is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva- Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with carbamoyl groups bound silica gel for liquid chromatography (5 mm in particle lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 ‌2118‌ Rose Fruit / Crude Drugs and Related Drugs ‌mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Rilmazafone Hydrochloride Hydrate contains not less than 98.
Rilmazafone Hydrochloride Hydrate occurs as a white to pale yellow-white crystalline powder. It is very soluble in methanol, soluble in water, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Rilmazafone Hydrochloride Hydrate (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Rilmazafone Hydrochloride RS prepared in the same manner as the sam-ple solution: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Ril-mazafone Hydrochloride Hydrate as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Rilmazafone Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Rilmazafone Hydrochloride Hydrate (1 in 200) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Rilmazafone Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Rilmazafone Hydrochloride Hydrate in 50 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.87 to rilmazafone, obtained from the sample solution is not larger than the peak area of rilmazafone from the standard solu-tion, and the area of the peak other than rilmazafone and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of rilmazafone from the standard solution. Furthermore, the total area of the peaks other than rilmazafone from the sample solution is not larger than 2 times the peak area of rilmazafone from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: 0.02 mol/L phosphate buffer solution (pH 3.0). Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 3 3 – 20 75 → 70 25 → 30 20 – 30 70 → 50 30 → 50 30 – 45 Flow rate: 1.0 mL per minute. Time span of measurement: For 45 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of water and acetonitrile (1:1) to make exactly 20 mL. Confirm that the peak area of rilmaza-fone obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rilmazafone are not less than 20,000 and not more than 1.3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rilmazafone is not more than 2.0z. Water <2.48> 5.5 – 7.5z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 40 mg each of Rilmazafone Hydrochloride Hydrate and Rilmazafone Hydrochloride RS (separately determine the water <2.48> in the same manner as Rilmazafone Hydrochloride Hydrate), dissolve each in water to make exactly 200 mL. Pipet 10 mL each of these solu-tions, add exactly 20 mL of the internal standard solution to each solution, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of rilmazafone to that of the internal standard. ‌Amount (mg) of rilmazafone hydrochloride (C21H20Cl2N6O3.HCl) ＝ MS × QT/QS MS: Amount (mg) of Rilmazafone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and acetonitrile (1:1) (3 in 100,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.1 g of sodium 1-heptanesul-fonate in 1000 mL of water, and adjust to pH 3.0 with acetic acid (100). To 500 mL of this solution add 300 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of rilmaza-fone is about 5 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, rilmazafone and the internal standard are eluted in this order with the resolution between these peaks being not less than 13. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of rilmazafone to that of the internal stand-ard is not more than 1.0z.
Risperidone occurs as a white to pale yellow-white crystalline powder. It is sparingly soluble in methanol and in ethanol (99.5), very slightly soluble in 2-propanol, and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Risperidone in 2-propanol (1 in 40,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Risperidone, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Risperidone according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Risperidone in 10 mL of methanol, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than risperidone obtained from the sample solution is not larger than the peak area of risperidone from the standard solu-tion, and the total area of the peaks other than risperidone from the sample solution is not larger than 1.5 times the peak area of risperidone from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: A solution of ammonium acetate (1 in 0 – 2 70 30 2 – 17 70 → 30 30 → 70 17 – 22 30 70 Flow rate: 1.5 mL per minute. Time span of measurement: About 1.6 times as long as the retention time of risperidone. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add methanol to make exactly 20 mL. Confirm that the peak area of risperidone obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 10,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.16 g of Risperidone, dis-solve in 70 mL of a mixture of 2-butanone and acetic acid (100) (7:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.52 mg of C23H27FN4O2
Ritodrine Hydrochloride, when dried, contains not less than 98.
Ritodrine Hydrochloride occurs as a white crystalline powder. It is freely soluble in water, in methanol and in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. A solution of Ritodrine Hydrochloride (1 in 10) shows no optical rotation. It is gradually colored to light yellow by light. Melting point: about 1969C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ritodrine Hydrochloride in 0.01 mol/L hydro-chloric acid TS (1 in 20,000) as directed under Ultraviolet- Amount (mg) of risperidone (C ＝ MS × AT/AS × 1/25 23H27FN4O2) visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ritodrine Hydrochloride RS prepared in the same manner as MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (4:1) add 1.5 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 1.0z.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Ritodrine Hydrochloride in 10 mL of water is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Rito- drine Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Related substances—Dissolve 20 mg of Ritodrine Hy-drochloride in 20 mL of the mobile phase, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak of ritodrine threo-isomer, having the relative reten- ‌tion time of about 1.2 to ritodrine, obtained from the sample solution is not larger than 4/5 times the peak area of rito-drine from the standard solution, the area of the peak other than ritodrine and ritodrine threo-isomer from the sample solution is not larger than 3/10 times the peak area of rito-drine from the standard solution, and the total area of the peaks other than ritodrine and ritodrine threo-isomer from the sample solution is not larger than 4 times the peak area of ritodrine from the standard solution. Operating conditions— Column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Flow rate: Adjust so that the retention time of ritodrine is about 10 minutes. Time span of measurement: About 3 times as long as the retention time of ritodrine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 50 mL. Confirm that the peak area of ritodrine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: To 20 mg of Ritodrine Hydrochlo-ride add 50 mL of the mobile phase and 5.6 mL of sulfuric acid, and add the mobile phase to make 100 mL. Heat a portion of this solution at about 859C for about 2 hours, and allow to cool. Pipet 10 mL of this solution, and add exactly 10 mL of 2 mol/L sodium hydroxide TS. When the procedure is run with 10 mL of this solution under the above operating conditions, ritodrine and the threo-isomer are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ritodrine is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 30 mg each of Ritodrine Hy-drochloride and Ritodrine Hydrochloride RS, previously dried, and dissolve in methanol to make exactly 50 mL. Pipet 25 mL of these solutions, add exactly 5 mL of the in-ternal standard solution, then add water to make 50 mL, and use these solutions as the sample solution and standard solu-tion. Perform the test with 10 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ritodrine to that of the internal standard. Amount (mg) of ritodrine hydrochloride (C17H21NO3.HCl) ＝ MS × QT/QS MS: Amount (mg) of Ritodrine Hydrochloride RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.6 g of diammonium hydrogen phosphate and 1.1 g of sodium 1-heptanesulfonate in 700 mL of water, and add 300 mL of methanol. Adjust to pH 3.0 with phosphoric acid. Flow rate: Adjust so that the retention time of ritodrine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ritodrine and the internal standard are eluted in this order with the resolution between these peaks being not less then 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ritodrine to that of the internal standard is not more than 1.0z.
ロキサチジン酢酸エステル塩酸塩
Roxatidine Acetate Hydrochloride occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), and sparingly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Roxatidine Acetate Hydrochloride in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Roxati-dine Acetate Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Rox-atidine Acetate Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Roxatidine Acetate Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Roxatidine Acetate Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Roxatidine Acetate Hydrochlo-ride in 20 mL of water: the pH of this solution is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Roxatidine Acetate Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Roxati- dine Acetate Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Roxatidine ‌Acetate Hydrochloride in 10 mL of ethanol (99.5), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than roxatidine acetate obtained from sample solution is not larger than 1/5 times the peak area of roxati-dine acetate from the standard solution, and the total area of the peaks other than roxatidine acetate from the sample so-lution is not larger than 1/2 times the peak area of roxatidine acetate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with cyanopropylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of hexane, ethanol (99.5), triethylamine and acetic acid (100) (384:16:2:1). Flow rate: Adjust so that the retention time of roxatidine acetate is about 10 minutes. Time span of measurement: About 1.5 times as long as the retention time of roxatidine acetate, beginning after the sol-vent peak. System suitability— Test for required detectability: To 5 mL of the standard solution add ethanol (99.5) to make 10 mL, and use this so-lution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add ethanol (99.5) to make exactly 10 mL. Confirm that the peak area of roxatidine acetate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: Dissolve 50 mg of roxatidine acetate hydrochloride and 10 mg of benzoic acid in 25 mL of ethanol (99.5). When the procedure is run with 10 mL of this solution under the above operating conditions, benzoic acid and roxatidine acetate are eluted in this order with the reso-lution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of roxatidine acetate is not more than 1.0z. Loss on drying <2.41> Not more than 0.3z (1 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Roxatidine Acetate Hydrochloride, previously dried, dissolve in 5 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.49 mg of C19H28N2O4.HCl
Roxithromycin is a derivative of erythromycin. It contains not less than 970 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Roxithromycin occurs as a white crystalline powder. It is freely soluble in ethanol (95) and in acetone, soluble in methanol, and practically insoluble in water.
Determine the infrared absorption spectrum of Roxithromycin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Roxithromycin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －93 – －969(0.5 g calculated on the anhydrous basis, acetone, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Roxithromycin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve exactly 40 mg of Rox-ithromycin in the mobile phase A to make exactly 10 mL, and use this solution as the sample solution. Separately, dis-solve exactly 20 mg of Roxithromycin RS in the mobile phase A to make exactly 10 mL. Pipet 1 mL of this solution, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak having a relative retention time of about 1.05 to roxithromycin obtained from the sample solution is not larger than 2 times the peak area of roxithromycin from the standard solution. The area of the peak other than rox-ithromycin and the peak mentioned above from the sample solution is not larger than the peak area of roxithromycin ‌from the standard solution, and the total area of the peaks other than roxithromycin from the sample solution is not larger than 6 times the peak area of roxithromycin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: To 200 mL of a solution of ammonium dihydrogenphosphate (17 in 100) add 510 mL of water, and adjust to pH 5.3 with 2 mol/L sodium hydroxide TS. To this solution add 315 mL of acetonitrile for liquid chromato-graphy. Mobile phase B: A mixture of acetonitrile for liquid chro-matography and water (7:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: Amount [mg (potency)] of Roxithromycin RS taken Internal standard solution—A solution of isopropyl para-hydroxybenzoate in the mobile phase (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 49.1 g of ammonium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.3 with 2 mol/L sodium hydroxide TS. To 690 mL of this solu-tion add 310 mL of acetonitrile. Flow rate: Adjust so that the retention time of rox-ithromycin is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, roxithromycin and the internal standard are eluted in this order with the resolution between these peaks being Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) not less than 10. System repeatability: When the test is repeated 6 times 0 – 38 100 0 38 – 39 100 → 90 0 → 10 39 – 80 90 10 Flow rate: Adjust so that the retention time of rox-ithromycin is about 21 minutes. Time span of measurement: For 80 minutes after injection of the sample solution. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase A to make exactly 10 mL. Confirm that the peak area of roxithromycin obtained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of roxithromycin are not less than 9000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of roxithromycin is not more than 2.0z. Water <2.48> Not more than 3.0z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately an amount of Roxithromycin and Roxithromycin RS, equivalent to about 38 mg (potency), dis-solve them separately in a suitable amount of the mobile phase, then add exactly 1 mL of the internal standard, add the mobile phase to make 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of roxithromycin to that of the internal standard. Amount [mg (potency)] of roxithromycin (C41H76N2O15) ＝ MS × QT/QS × 1000 with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of roxithromycin to that of the internal stand-ard is not more than 1.0z.
Ryokeijutsukanto Extract contains not less than 1 mg and not more than 4 mg of (E )-cinnamic acid, and not less than 17 mg and not more than 51 mg of glycyrrhizic acid (C42H62O16: 822.
Ryokeijutsukanto Extract occurs as a brown powder or black-brown viscous extract. It has an odor, and a sweet first then bitter taste.
(1) To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solu-tion as the sample solution. Separately, dissolve 1 mg of (E )-cinnamic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of hexane, ethyl acetate, for-mic acid and water (60:40:4:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Cinna-mon Bark). For preparation prescribed Atractylodes Rhizome—To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the stand-ard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhi-zome). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of 1-butanol, and shake. Centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of dry extract (or an amount of the viscous extract, equivalent to 1.0 g of the dried substance) of Ryokeijutsukanto Extract as directed in the Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of dry extract (or an amount of the viscous extract, equiva-lent to 0.67 g of the dried substance) of Ryokeijutsukanto Extract according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 8.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g 1059C, 5 hours). Total ash <5.01> Not more than 8.0z, calculated on the dried basis.
(1) (E)-Cinnamic acid—Conduct this procedure using light-resistant vessels. Weigh accurately about 0.5 g of dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance) of Ryokeijutsukanto Extract, add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of (E)-cinnamic acid for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of (E)-cinnamic acid in each solu-tion. Amount (mg) of (E)-cinnamic acid ＝ MS × AT/AS × 1/20 MS: Amount (mg) of (E)-cinnamic acid for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave- JP XVIII Crude Drugs and Related Drugs / Safflower 2121 length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (750:250:1). Flow rate: 1.0 mL per minute (the retention time of (E)-cinnamic acid is about 12 minutes). ‌System suitability—‌ System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of (E)-cinnamic acid are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of (E)-cinnamic acid is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Saccharin occurs as colorless to white crystals or a white crystalline powder. It is sparingly soluble in ethanol (95), and slightly soluble in water. It dissolves in sodium hydroxide TS.
Determine the infrared absorption spectrum of Saccharin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 5.0 g of Saccharin in 25 mL of a solution of sodium acetate trihy-drate (1 in 5): the clarity of the solution is equivalent to that of water or a solution of sodium acetate trihydrate (1 in 5), or its degree of opalescence is not more than Reference sus-pension I, and it has the appearance of water in color or is not more intensely colored than a solution of sodium acetate trihydrate (1 in 5) or the following control solution. Control solution: Mix 3.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.4 mL of Copper (II) Sulfate CS, and add diluted dilute hydrochloric acid (1 in 10) to make 1000 mL. Heavy metals <1.07>—Proceed with 2.0 g of Saccha- rin according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Benzoate and salicylate—To 10 mL of a saturated so-lution of Saccharin in hot water add 3 drops of iron (III) chloride TS: no precipitate is formed, and no red-purple to purple color develops. o-Toluene sulfonamide—Dissolve 10 g of Saccharin in 70 mL of sodium hydroxide TS, and extract with three 30-mL portions of ethyl acetate. Combine all the ethyl ace-tate extracts, wash with 30 mL of a solution of sodium chlo-ride (1 in 4), dehydrate with 5 g of anhydrous sodium sul-fate, then evaporate the solvent. To the residue add exactly 5 mL of the internal standard solution to dissolve, and use this solution as the sample solution. Separately, dissolve 0.10 g of o-toluene sulfonamide in ethyl acetate to make exactly 100 mL. Pipet 1 mL of this solution, evaporate to dryness on a water bath, dissolve the residue in exactly 5 mL of the in-ternal standard solution, and use this solution as the stand-ard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chro-matography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of o-toluene sulfonamide to that of the internal standard: QT is not more than QS. Internal standard solution—A solution of caffeine in ethyl acetate (1 in 500). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with siliceous earth for gas chromatogra-phy coated 3z with diethylene glycol succinate polyester for gas chromatography (180 – 250 mm in particle diameter). Column temperature: A constant temperature of about 2009C. Temperature of injection port: A constant temperature of about 2259C. Temperature of detector: A constant temperature of about 2509C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of caffeine is about 6 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, the internal standard and o-toluene sulfonamide are ‌eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak height of o-toluene sulfonamide to that of the internal standard is not more than 2.0z. Readily carbonizable substances—Transfer 0.20 g of Saccharin to a Nessler tube, add 5 mL of sulfuric acid, mix to dissolve, and warm at 48 to 509C for 10 minutes: the solu-tion is not more intensely colored than Matching Fluid A, when compared both solutions against a white background by viewing transversely. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 50 mg of Saccharin, and dis-solve in a mixture of water and methanol (1:1) to make ex-actly 50 mL. Pipet 5 mL of this solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Saccharin RS (separately determine the loss on drying <2.41> in the same conditions as Saccha-rin), dissolve in a mixture of water and methanol (1:1) to make exactly 25 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of saccharin in each solu-tion. System suitability— System performance: Dissolve 25 mg of phthalic anhy-dride in a mixture of water and methanol (1:1) to make 25 mL. To 5 mL of this solution add 5 mL of the standard stock solution and a mixture of water and methanol (1:1) to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, phthalic an-hydride and saccharin are eluted in this order with the reso-lution between these peaks being not less than 1.5. When the procedure is run with 10 mL of the standard solution under the above operating conditions, the symmetry factor of the peak of saccharin is not more than 1.5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saccharin is not more than 0.73z.
Saibokuto Extract contains not less than 2 mg and not more than 8 mg of saikosaponin b2, not less than 90 mg and not more than 270 mg of baicalin (C21H18O11: 446.
Saibokuto Extract is a light brown powder or black-brown viscous extract, having a slightly odor and a slight sweet first, then a bitter taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1- JP XVIII Crude Drugs and Related Drugs / Saibokuto Extract 2123 butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and exa-mine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Scutellaria Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of magnolol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solution (Mag-nolia Bark). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginsenoside Rb1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of 0.1 mol/L hydrochloric acid TS, add 25 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of rosmarinic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (60:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solution (Perilla Herb). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than Saibokuto Extract / Crude Drugs and Related Drugs 9.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of sai-kosaponin b2 in each solution. Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. CS: Concentration (mg/mL) of saikosaponin b2 in saiko-saponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of saiko-saponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of baicalein for resolution check in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and bai-calein is not less than 1.5. System repeatability: When the test is repeated 6 times ‌JP XVIII Crude Drugs and Related Drugs / Saikokeishito Extract 2125 with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Method of preparation 1) 2) 3) 4) Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Bupleurum Root 5 g 5 g 5 g 5 g Pinellia Tuber 4 g 4 g 4 g 4 g Scutellaria Root 2 g 2 g 2 g 2 g Peony Root 2 g 2.5 g 2 g 2 g Jujube 2 g 2 g 2 g 2 g Ginseng 2 g 2 g 2 g 2 g Cinnamon Bark 2.5 g 2.5 g 2.5 g 2 g Glycyrrhiza 1.5 g 1.5 g 1.5 g 2 g Ginger 0.5 g 1 g 1 g 1 g Prepare a dry extract or viscous extract as directed under Extracts, according to the prescription 1) to 4), using the crude drugs shown above. Description Saikokeishito Extract is a yellow-brown pow-der or black-brown viscous extract, having a slightly odor and a slight sweet first, then a bitter and slightly pungent taste. Identification (1) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Saikokeishito Extract contains not less than 1.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). Lead—Take 5.0 g of the dry extract (or an amount of the viscous extract, equivalent to 5.0 g of the dried sub-stance) in a platinum, quartz or porcelain crucible, heat gently, and then incinerate by ignition at 450 to 5509C. After cooling, add a small amount of 2 mol/L nitric acid TS to the residue, filter if necessary, and wash the crucible several times with small portions of 2 mol/L nitric acid TS. Com-bine the washings and the filtrate, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sam-ple solution. Separately, to 2.5 mL of Standard Lead Solu-tion add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions: the absorbance of the sam-ple solution is not more than that of the standard solution (not more than 5 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: A lead hollow-cathode lamp. Wavelength: 283.3 nm. Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.5z (1 g, 1059C, 5 hours). JP XVIII Crude Drugs and Related Drugs / Saikokeishito Extract 2127 The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of sai-kosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 CS: Concentration (mg/mL) of saikosaponin b2 in saiko-saponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of sai-kosaponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as ‌2128 Saireito Extract / Crude Drugs and Related Drugs the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography and 1 mg of baicalein for resolu-tion check in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, two peaks other than glycyrrhizic acid are ob-served with the resolutions between the peak of glycyrrhizic acid and each of the two peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Saireito Extract contains not less than 2 mg and not more than 8 mg of saikosaponin b2, not less than 80 mg and not more than 240 mg of baicalin (C21H18O11: 446.
Saireito Extract occurs as a light yellow-brown powder. It has slightly a characteristic odor, and a sweet, then bitter taste.
(1) To 2.0 g of Saireito Extract add 10 mL of sodium hydroxide TS, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra- JP XVIII Crude Drugs and Related Drugs / Saireito Extract 2129 phy <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and examine under ul-traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). To 1.0 g of Saireito Extract add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pres-sure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 15 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). To 1.0 g of Saireito Extract add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pres-sure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Scutellaria Root). To 2.0 g of Saireito Extract add 10 mL of sodium hy-droxide TS, shake, then add 5 mL of 1-butanol, shake, cen-trifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginsenoside Rb1 RS or ginseno-side Rb1 for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Ginseng). To 2.0 g of Saireito Extract add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). To 2.0 g of Saireito Extract add 10 mL of sodium carbonate TS, shake, then add 10 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sam-ple solution. Separately, dissolve 1 mg of alisol A for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:10:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid-acetic acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Alisma Tuber). For preparation prescribed Atractylodes Rhizome—To 1.0 g of Saireito Extract add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of Saireito Extract add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solu-tion as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Exa-mine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylaminobenzaldehyde TS for spraying, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Saireito Extract / Crude Drugs and Related Drugs Lancea Rhizome). To 1.0 g of Saireito Extract add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pres-sure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of (E )-cinnamic acid for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of hexane, ethyl acetate, formic acid and water (60:40:4:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the dark purple spot from the standard solu-tion (Cinnamon Bark).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Saireito Extract as directed under Extract (4), and perform the test (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of Saireito Extract according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of Saireito Extract, add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of saikosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 Extract, add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Baica-lin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make ex-actly 100 mL. Pipet 5 mL of this solution, add diluted meth-anol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the CS: Concentration (mg/mL) of saikosaponin b2 saponin b2 standard TS for assay Operating conditions— in saiko- following i) or ii). Weigh accurately about 0.5 g of Saireito Extract, add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Flow rate: 1.0 mL per minute (the retention time of saiko-saponin b2 is about 12 minutes). System suitability— Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. (2) Baicalin—Weigh accurately about 0.1 g of Saireito MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. ‌JP XVIII Crude Drugs and Related Drugs / Salvia Miltiorrhiza Root 2131 Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography and 1 mg of baicalein for resolu-tion check in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, two peaks other than glycyrrhizic acid are ob-served with the resolutions between the peak of glycyrrhizic acid and each of the two peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of Saireito Extract, add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same man-ner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Com-bine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Salvia Miltiorrhiza Root Salviae Miltiorrhizae Radix
Anhydrous Sodium Sulfate occurs as white, crystals or powder. It is odorless and has a salty and slightly bitter taste. It is freely soluble in water, and practically insoluble in ethanol (99.5).
(1) A solution of Anhydrous Sodium Sul-fate (1 in 20) responds to Qualitative Tests <1.09> (1) for so-dium salt. (2) A solution of Anhydrous Sodium Sulfate (1 in 20) responds to Qualitative Tests <1.09> (1) for sulfate.
(1) Acidity or alkalinity—Dissolve 0.5 g of Anhy-drous Sodium Sulfate in 5 mL of freshly boiled and cooled water: the solution is clear and colorless, and neutral. Chloride <1.03>—Perform the test with 0.5 g of pre- viously dried Anhydrous Sodium Sulfate. Prepare the con-trol solution with 0.5 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 2.0 g of previ- ously dried Anhydrous Sodium Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of previously dried Anhydrous Sodium Sulfate according to Method 1, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (4 g, 1059C, 4 hours).
Weigh accurately about 0.4 g of previously dried Anhydrous Sodium Sulfate, dissolve in 200 mL of water, add 1 mL of hydrochloric acid, boil, and gradually add 8 mL of barium chloride TS. Heat the solution in a water bath for 1 hour. After cooling, filter through a filter paper for quan-titative analysis (No.5C), wash the residue on the filter paper with water until the washings do not give the turbidity with silver nitrate TS. After drying the residue together with the paper, ignite at 500 – 8009C to constant mass, and weigh the mass of the residue as the amount of barium sulfate (BaSO4: 233.39). Amount (mg) of sodium sulfate (Na2SO4) ＝ amount (mg) of barium sulfate (BaSO4) × 0.609
Sodium Sulfate Hydrate occurs as colorless or white, crystals or crystalline powder. It is odorless and has a cooling and salty taste. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is quickly efflorescent in air, soluble in its own water of crystallization at about 339C and lost the water at 1009C.
(1) A solution of Sodium Sulfate Hydrate (1 in 20) responds to Qualitative Tests <1.09> (1) for sodium salt. (2) A solution of Sodium Sulfate Hydrate (1 in 20) re-sponds to Qualitative Tests <1.09> (1) for sulfate.
(1) Acidity or alkalinity—Dissolve 0.5 g of So-dium Sulfate Hydrate in 5 mL of freshly boiled and cooled water: the solution is clear and colorless, and neutral. Chloride <1.03>—Perform the test with 0.5 g of pre- viously dried Sodium Sulfate Hydrate. Prepare the control solution with 0.5 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.036z). Heavy metals <1.07>—Proceed with 2.0 g of previ- ously dried Sodium Sulfate Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of previously dried Sodium Sulfate Hydrate according to Method 1, and perform the test (not more than 2 ppm). Loss on drying <2.41> 51.0 – 57.0z (4 g, 1059C, 4 hours).
Weigh accurately about 0.4 g of previously dried So-dium Sulfate Hydrate, dissolve in 200 mL of water, add 1 mL of hydrochloric acid, boil, and gradually add 8 mL of barium chloride TS. Heat the solution in a water bath for 1 hour. After cooling, filter through a filter paper for quan-titative analysis (No.5C), wash the residue on the filter paper with water until the washings do not give the turbidity with silver nitrate TS. After drying the residue together with the paper, ignite at 500 – 8009C to constant mass, and weigh the mass of the residue as the amount of barium sulfate (BaSO4: 233.39). Amount (mg) of sodium sulfate (Na2SO4) ＝ amount (mg) of barium sulfate (BaSO4) × 0.609
Salbutamol Sulfate, when dried, contains not less than 98.
Salbutamol Sulfate occurs as a white powder. It is freely soluble in water, slightly soluble in ethanol (95), and in acetic acid (100) and practically insoluble in diethyl ether. A solution of Salbutamol Sulfate (1 in 20) shows no opti-cal rotation.
(1) Determine the absorption spectrum of a solution of Salbutamol Sulfate in 0.1 mol/L hydrochloric acid TS (1 in 12,500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sal-butamol Sulfate, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A solution of Salbutamol Sulfate (1 in 20) responds to Qualitative Tests <1.09> for sulfate.
(1) Clarity and color of solution—Dissolve 1.0 g of Salbutamol Sulfate in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Sal- butamol Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 20 mg of Salbutamol Sulfate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 2-propanol, water and ammonia so-lution (28) (25:15:8:2) to a distance of about 15 cm, and air-dry the plate. Leave the plate in a well-closed vessel saturated with diethylamine vapor for 5 minutes, and spray evenly 4-nitrobenzenediazonium chloride TS: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution in color. Boron—Take 50 mg of Salbutamol Sulfate and 5.0 mL of the Standard Boron Solution, and transfer to a plati-num crucible. Add 5 mL of potassium carbonate-sodium carbonate TS, evaporate on a water bath to dryness, and dry at 1209C for 1 hour. Ignite the residue immediately. After cooling, add 0.5 mL of water and 3 mL of curcumin TS to the residue, warm gently in a water bath for 5 minutes. After cooling, add 3 mL of acetic acid-sulfuric acid TS, mix, and allow to stand for 30 minutes. Add ethanol (95) to make ex-actly 100 mL, and filter. Discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution and standard solution. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using ethanol (95) as the blank: the absorbance of the sample solution at 555 nm is not larger than that of the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, 1009C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.9 g of Salbutamol Sulfate, previously dried, and dissolve in 50 mL of acetic acid (100) by warming. After cooling, titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 57.67 mg of (C13H21NO3)2.H2SO4
The surface of Salicylic Acid Adhesive Plaster is whitish in color and adheres well to the skin.
Salicylic Acid Spirit is a clear, colorless liquid. Specific gravity d 20: about 0.86
The solution obtained in the Assay has a red-purple color. Determine the absorption spectrum of the solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 520 nm and 535 nm (salicylic acid). Alcohol number <1.01> Not less than 8.8 (Method 2).
Measure exactly 10 mL of Salicylic Acid Spirit, add 10 mL of ethanol (95) and water to make exactly 100 mL. Pipet 3 mL of this solution, and dilute with hydrochloric acid-potassium chloride buffer solution (pH 2.0) to make exactly 100 mL. Use this solution as the sample solution. Dissolve about 0.3 g of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours and accurately weighed, in 10 mL of ethanol (95) and water to make exactly 100 mL. Pipet 3 mL of this solution, dilute with hydrochlo-ric acid-potassium chloride buffer solution (pH 2.0) to make exactly 100 mL, and use this solution as the standard solu-tion. Pipet 10 mL each of the sample solution and standard solution, to each add exactly 5 mL of a solution of iron (III) nitrate enneahydrate (1 in 200), dilute with hydrochloric acid-potassium chloride buffer solution (pH 2.0) to exactly 25 mL. Determine the absorbances, AT and AS, of both solu-tions at 530 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a blank solution prepared in the same manner with water. Amount (mg) of salicylic acid (C H O ) Compound Salicylic Acid Spirit
Salicylic Acid, when dried, contains not less than 99.
Salicylic Acid occurs as white, crystals or crys-talline powder. It has a slightly acid, followed by an acrid taste. It is freely soluble in ethanol (95) and in acetone, and slightly soluble in water.
(1) A solution of Salicylic Acid (1 in 500) ‌responds to Qualitative Tests <1.09> (1) and (3) for salicylate. Determine the absorption spectrum of a solution of Salicylic Acid in ethanol (95) (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Sali-cylic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 5.0 g of Salicylic Acid in 90 mL of water by heating, cool, dilute with water to 100 mL, and filter. Discard the first 20 mL of the filtrate, take 30 mL of the subsequent filtrate, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.35 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.008z). Sulfate <1.14>—To 20 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test so-lution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Heavy metals <1.07>—Dissolve 2.0 g of Salicylic Acid in 25 mL of acetone, add 4 mL of sodium hydroxide TS, 2 mL of dilute acetic acid and water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 25 mL of acetone, 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Related substances—Dissolve 0.50 g of Salicylic Acid in the mobile phase to make exactly 100 mL, and use this so-lution as the sample solution. Separately, dissolve exactly 10 mg of phenol, exactly 25 mg of 4-hydroxyisophthalic acid and exactly 50 mg of parahydroxybenzoic acid in the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak areas of parahydroxybenzoic acid, 4-hydroxyisophthalic acid and phenol obtained from the sample solution are not larger than the area of each respective peak from the standard solu-tion, the area of the peak other than salicylic acid and the substances mentioned above is not larger than the peak area of 4-hydroxisophthalic acid from the standard solution, and the total area of peaks other than salicylic acid is not larger than 2 times the peak area of parahydroxybenzoic acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of water, methanol and acetic acid (100) (60:40:1). Flow rate: Adjust so that the retention time of salicylic acid is about 17 minutes. Time span of measurement: About 2 times as long as the retention time of salicylic acid, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak areas of parahydroxybenzoic acid, 4-hydroxyisophthalic acid and phenol obtained with 10 mL of this solution are equivalent to 14 to 26z of the area of each respective peak with 10 mL of the standard solution. System performance: Dissolve 10 mg of phenol, 25 mg of 4-hydroxyisophthalic acid and 50 mg of parahydroxybenzoic acid in 100 mL of the mobile phase. To 1 mL of this solution add the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, parahydroxybenzoic acid, 4-hydroxyisophthalic acid and phenol are eluted in this order with the resolution between the peaks of 4-hydroxyisophthalic acid and phenol being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of parahydroxybenzoic acid, 4-hydroxyisophthalic acid and phenol is not more than 2.0z, respectively. Loss on drying <2.41> Not more than 0.5z (2 g, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Salicylic Acid, pre-viously dried, dissolve in 25 mL of neutralized ethanol, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indica-tor: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 13.81 mg of C7H6O3
Santonin, when dried, contains not less than 98.
Santonin occurs as colorless crystals, or a white crystalline powder. It is freely soluble in chloroform, sparingly soluble in ethanol (95), and practically insoluble in water. It becomes yellow by light.
(1) Determine the absorption spectrum of a solution of Santonin in ethanol (95) (3 in 250,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of San-tonin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －170 – －1759(0.2 g, chlo-roform, 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Santonin according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Alkaloids—Boil 0.5 g of Santonin with 20 mL of di-luted sulfuric acid (1 in 100), cool, and filter. Dilute 10 mL of the filtrate with water to 30 mL, add 3 drops of iodine TS, and allow to stand for 3 hours: no turbidity is produced. Artemisin—Dissolve 1.0 g of powdered Santonin in 2 mL of chloroform by slight warming: the solution is clear and colorless, or any yellow color produced is not darker than Matching Fluid A. Phenols—Boil 0.20 g of Santonin with 10 mL of water, cool, and filter. To the filtrate add bromine TS until the color of the solution becomes yellow: no turbidity is pro-duced. Acid-coloring substances—Moisten 10 mg of Santo-nin with nitric acid: no color develops immediately. Moisten Santonin with sulfuric acid, previously cooled to 09C: no color is produced immediately. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.25 g of Santonin, previ- ously dried, dissolve in 10 mL of ethanol (95) by warming, add exactly 20 mL of 0.1 mol/L sodium hydroxide VS, and heat on a water bath under a reflux condenser for 5 minutes. Cool quickly, and titrate <2.50> the excess sodium hydroxide with 0.05 mol/L hydrochloric acid VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 24.63 mg of C15H18O3
Sarpogrelate Hydrochloride contains not less than 98.
Sarpogrelate Hydrochloride occurs as a white crystalline powder. It is slightly soluble in water and in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. A solution of Sarpogrelate Hydrochloride (1 in 100) shows no optical rotation. Sarpogrelate Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Sarpogrelate Hydrochloride in 0.01 mol/L hydrochloric acid TS (1 in 20,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Sarpogrelate Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sar-pogrelate Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Sarpogrelate Hydrochloride RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the Sarpogrelate Hydrochloride, or the Sar-pogrelate Hydrochloride and the Sarpogrelate Hydrochlo-ride RS separately with acetone by heating and suspending, filter and dry the crystals at 509C for 1 hour, and perform the test with the crystals. Dissolve 0.3 g of Sarpogrelate Hydrochloride in 6 mL of sodium hydroxide TS, shake well, allow to stand for 10 ‌minutes, and filter. To 1 mL of the filtrate add 1 mL of dilute nitric acid. This solution responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Sarpogrelate Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Sarpogrelate Hydrochloride according to Method 4, and perform the test (not more than 1 ppm). Related substances—Conduct this procedure within 3 hours after preparation of the sample solution. Dissolve 20 mg of Sarpogrelate Hydrochloride in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make ex-actly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of decomposed substance A, having the relative retention time about 0.82 to sarpogrelate, ob-tained from the sample solution is not larger than 1/5 times that of sarpogrelate from the standard solution, the area of the peak other than sarpogrelate and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of sarpogrelate from the standard solution, and the total area of the peaks other than sarpogrelate from the sample solution is not larger than 1/2 times the peak area of sarpogrelate from the standard solution. For the peak area of the decomposed substance A, multiply the correction factor, 0.78. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of sarpogrelate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 50 mL. Con-firm that the peak area of sarpogrelate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of Sarpogrelate Hy-drochloride in 20 mL of water, and use as the sarpogrelate hydrochloride stock solution. To 1 mL of the sarpogrelate hydrochloride stock solution add 2 mL of sodium hydroxide TS, shake thoroughly, allow to stand for 10 minutes, and add 3 mL of 1 mol/L hydrochloric acid TS. To this solution add 1 mL of the sarpogrelate hydrochloride stock solution, and add the mobile phase to make 50 mL. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, the decomposed substance A and sarpogre- Hydrochloride and Sarpogrelate Hydrochloride RS (separately determine the water <2.48> in the same manner as Sarpogrelate Hydrochloride), add to them exactly 2.5 mL of the internal standard solution, and dissolve them in the mo-bile phase to make 50 mL. To 5 mL each of these solutions add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of sarpogre-late to that of the internal standard. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × QT/QS MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (3 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and trifluoroacetic acid (1300:700:1). Flow rate: Adjust so that the retention time of sarpogre-late is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sarpogrelate and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of sarpogrelate to that of the internal standard is not more than 1.0z.
Oblong spike, 5 – 10 cm in length, 0.5 – 0.8 cm in diameter, purplish green-brown to green-brown in color. Spike, with calyx-tubes containing small labiate flower or often fruits; sometimes leaves under spike; leaf, linear or small lanceolate; stem, prismatic, purple-brown in color. Under a magnifying glass, it reveals short hairs. It has a characteristic aroma and slightly cool feeling on keeping in the mouth.
To 1 g of pulverized Schizonepeta Spike add 10 mL of ethyl acetate, shake for 15 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes. After cooling for more than 10 minutes under an adequate humidity, examine under ultraviolet light (main wavelength: 365 nm): two spots, one is a blue fluorescent spot with an Rf value of about 0.5 and the another is a yellow fluorescent spot with an Rf value of about 0.1, are observed. Total ash <5.05> Not more than 11.0z. Acid-insoluble ash <5.05> Not more than 3.0z. Extract content <5.05> Dilute ethanol-soluble extract: not less than 8.0z.
Scopolamine Butylbromide, when dried, contains not less than 98.
Scopolamine Butylbromide occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), soluble in ethanol (95), sparingly soluble in methanol, slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: about 1409C (with decomposition).
(1) To 1 mg of Scopolamine Butylbromide add 3 to 4 drops of fuming nitric acid, and evaporate on a water bath to dryness. After cooling, dissolve the residue in 1 mL of N,N-dimethylformamide, and add 6 drops of tet-raethylammonium hydroxide TS: a red-purple color devel-ops. Determine the absorption spectrum of a solution of Scopolamine Butylbromide (1 in 1000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Scopolamine Butylbromide, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Scopolamine Butylbromide (1 in 20) responds to Qualitative Tests <1.09> for bromide. D Optical rotation <2.49> [a]20: －18.0 – －20.09(after dry-ing, 1 g, water, 10 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Scopolamine Butylbromide in 10 mL of water: the pH of this solution is between 5.5 and 6.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Scopolamine Butylbromide in 10 mL of water: the solu-tion is clear, and has no more color than the following con-trol solution. Control solution: To 0.5 mL of Matching Fluid F add di-luted hydrochloric acid (1 in 40) to make 20 mL. Heavy metals <1.07>—Proceed with 2.0 g of Scopola- mine Butylbromide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Scopolamine Butylbromide in the mobile phase to make exactly 10 mL, and use this solution as the sample solution. Separately, dis- ‌solve 10 mg of scopolamine hydrobromide hydrate in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution (1). Pipet 5 mL of the standard solution (1), add the mobile phase to make ex-actly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution and standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the peak area of scopola-mine obtained from the sample solution is not larger than that from the standard solution (2), and each area of the peaks other than the peak appearing in the first elution and the peak of scopolamine and butylscopolamine from the sample solution are not larger than the peak area from the standard solution (1). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Scopolamine Hydrobromide Hydrate
スコポラミン臭化水素酸塩水和物
Scopolamine Hydrobromide Hydrate, when dried, contains not less than 98.
Scopolamine Hydrobromide Hydrate occurs as colorless or white crystals, or white granules or powder. It is odorless. It is freely soluble in water, sparingly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in diethyl ether.
(1) To 1 mg of Scopolamine Hydro-bromide Hydrate add 3 to 4 drops of fuming nitric acid, evaporate on a water bath to dryness, and cool. Dissolve the residue in 1 mL of N,N-dimethylformamide, and add 6 drops of tetraethylammonium hydroxide TS: a red-purple color is produced. (2) A solution of Scopolamine Hydrobromide Hydrate (1 in 20) responds to Qualitative Tests <1.09> for bromide. D Optical rotation <2.49> [a]20: －24.0 – －26.09(after dry-ing, 0.5 g, water, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Scopolamine Hydrobromide Hydrate in 10 mL of water: the solution is clear and colorless. Acidity—Dissolve 0.50 g of Scopolamine Hydro-bromide Hydrate in 15 mL of water, and add 0.50 mL of 0.02 mol/L sodium hydroxide VS and 1 drop of methyl red-methylene blue TS: a green color develops. Apoatropine—Dissolve 0.20 g of Scopolamine Hydrobromide Hydrate in 20 mL of water, add 0.60 mL of 0.002 mol/L potassium permanganate VS, and allow to stand for 5 minutes: the red color in the solution does not disappear. Related substances—Dissolve 0.15 g of Scopolamine Hydrobromide Hydrate in 3 mL of water, and use this solu-tion as the sample solution. To 1 mL of the sample solution add 2 to 3 drops of ammonia TS: no turbidity is produced. To 1 mL of the sample solution add 2 to 3 drops of potassium hydroxide TS: a transient white turbidity might be produced, and disappears clearly in a little while. Loss on drying <2.41> Not more than 13.0z [1.5 g, first dry in a desiccator (silica gel) for 24 hours, then dry at 1059C for 3 hours]. Residue on ignition <2.44> Not more than 0.1z (1 g). ‌Assay Weigh accurately about 0.5 g of Scopolamine Hydrobromide Hydrate, previously dried, in 10 mL of acetic acid (100) by warming. After cooling, add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction.‌ Each mL of 0.1 mol/L perchloric acid VS ＝ 38.43 mg of C17H21NO4.HBr
Weigh accurately about 0.8 g of Scopolamine Butyl-bromide, previously dried, dissolve in 40 mL of acetic acid (100) and 30 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.04 mg of C21H30BrNO4
Scopolia Extract and Tannic Acid Suppositories
Scopolia Extract and Tannic Acid Supposito-ries are light brown in color.
(1) To 2 Scopolia Extract and Tannic Acid Suppositories add 20 mL of diethyl ether, and dissolve the base of suppositories with shaking for 10 minutes. Shake thoroughly the mixture with 15 mL of water, separate the aqueous layer, and filter. To the filtrate add 10 mL of chlo-roform, shake well, and separate the chloroform layer. Take 5 mL of the chloroform solution, add 5 mL of ammonia TS, shake, and allow to stand: the ammonia layer shows a blue-green fluorescence. (2) To 1 mL of the aqueous layer obtained in (1) after ex-traction with diethyl ether, add 2 drops of iron (III) chloride TS: a blue-black color develops. Allow to stand: a blue-black precipitate is formed (tannic acid).
Scopolia Extract contains not less than 0.
Scopolia Extract is brown to dark brown in color. It has a characteristic odor, and a bitter taste. It dissolves in water with a slight turbidity.
(1) Dissolve 4 g of Scopolia Extract in 10 mL of water, add 8 mL of ammonia TS and 80 mL of diethyl ether, stopper tightly, shake for 1 hour, add 2.5 g of powdered tragacanth, shake vigorously, allow to stand for 5 minutes, and separate the diethyl ether layer into a porcelain dish. Evaporate the diethyl ether on a water bath, add 5 drops of fuming nitric acid, and evaporate on a water bath to dryness. After cooling, dissolve the residue in 1 mL of N,N-dimethylformamide, and add 5 to 6 drops of tetraethyl-ammonium hydroxide TS: a red-purple to purple color de-velops. (2) Mix 0.5 g of Scopolia Extract with 30 mL of ammo-nia TS in a flask, and transfer the mixture to a separator. Add 40 mL of ethyl acetate to the separator, and shake the mixture. After drain off the ethyl acetate layer, add 3 g of anhydrous sodium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent of the filtrate under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. Proceed as directed in Identification (2) under Scopolia Rhizome.
Heavy metals <1.07>—Prepare the test solution with 1.0 g of Scopolia Extract as directed in the Extracts (4), and perform the test (not more than 30 ppm).
Weigh accurately about 0.4 g of Scopolia Extract, place in a glass-stoppered centrifuge tube, add 15 mL of am-monia TS, and shake. Add 25 mL of diethyl ether, stopper tightly, shake for 15 minutes, centrifuge, and separate the diethyl ether layer. Repeat this procedure twice with the aqueous layer, using 25 mL each of diethyl ether. Combine all the extracts, and evaporate the solvent on a water bath. Dissolve the residue in 5 mL of the mobile phase, add exactly 3 mL of the internal standard solution, and add the mobile phase to make 25 mL. Proceed as directed under Scopolia Rhizome. Amount (mg) of hyoscyamine (C17H23NO3) ＝ MSA × QTA/QSA × 1/5 × 0.855 Amount (mg) of scopolamine (C17H21NO4) ＝ MSS × QTS/QSS × 1/25 × 0.789 MSA: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis MSS: Amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500).
Sevoflurane is a clear, colorless, and mobile liquid. It is miscible with ethanol (99.5). It is very slightly soluble in water. It is volatile and not inflammable. D Refractive index n20: 1.2745 – 1.2760 Boiling point: about 58.69C
Transfer about 1 mL of Sevoflurane to a gas cell having light path 10 cm in length, and determine the in-frared absorption spectrum as directed in the gas sampling method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Sevoflurane RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 20: 1.510 – 1.530
(1) Clarity and color of solution—Dissolve 1.0 g lated on the anhydrous basis. 4 3 7 of L-Serine in 10 mL of water: the solution is clear and color-less. Chloride <1.03>—Perform the test with 0.5 g of L-Serine. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.021z). Sulfate <1.14>—Perform the test with 0.6 g of L-Serine. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.028z). Ammonium <1.02>—Perform the test with 0.25 g of L-Serine. Prepare the control solution with 5.0 mL of Stand-ard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 2.0 g of L-Serine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of L-Serine according to Method 1, and perform the test ac-cording to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of L-Serine in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex- Description Sevoflurane is a clear, colorless, and mobile liquid. It is miscible with ethanol (99.5). It is very slightly soluble in water. It is volatile and not inflammable. D Refractive index n20: 1.2745 – 1.2760 Boiling point: about 58.69C Identification Transfer about 1 mL of Sevoflurane to a gas cell having light path 10 cm in length, and determine the in-frared absorption spectrum as directed in the gas sampling method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Sevoflurane RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Specific gravity <2.56> d 20: 1.510 – 1.530 Purity (1) Acidity or alkalinity—To 50 mL of Sevoflu-rane with 50 mL of freshly boiled and cooled water vigor-ously for 3 minutes. Separate the water layer and use this solution as the sample solution. To 20 mL of the sample solution add 1 drop of bromocresol purple TS and 0.10 mL of 0.01 mol/L sodium hydroxide VS: a red-purple color develops. To 20 mL of the sample solution add 1 drop of bromocresol purple TS and 0.6 mL of 0.01 mol/L hydro-chloric acid VS: a yellow color is produced. Soluble fluoride—To 6 g of Sevoflurane add 12 mL of diluted 0.01 mol/L sodium hydroxide TS (1 in 20), and shake for 10 minutes. Transfer 4.0 mL of diluted 0.01 mol/L sodium hydroxide solution (1 in 20) layer into a Nessler tube. Add 30 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1), add water to make 50 mL, allow to stand for 60 minutes, and use this solution as the sample so-lution. Separately, transfer 0.2 mL of the fluorine standard solution and 4.0 mL of diluted 0.01 mol/L sodium hydrox-ide TS (1 in 20) into a Nessler tube, and add 30 mL of a mix-ture of alizarin complexone TS, acetic acid-potassium ace-tate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1). Proceed in the same manner as directed for the preparation of the sample solution, and use this solution as the standard solution. Determine the absorbances of the sample solution and standard solution at 600 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared with 4.0 mL of diluted 0.01 mol/L so-dium hydroxide TS (1 in 20) in the same manner, as the blank: the absorbance of the sample solution is not more than that of the standard solution (not more than 1 ppm). Fluorine standard solution: Dissolve exactly 2.21 g of so-dium fluoride in water to make exactly 1000 mL. Pipet 10 mL of this solution and add water to make exactly 1000 mL. symmetry factor of the peak of sevoflurane are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 2 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of Sevoflurane is not more than 5.0z. (4) Residue on evaporation—Evaporate 10 mL of Sevoflurane, exactly measured, on a water bath to dryness, and dry at 1059C for 2 hours: the mass of the residue is not more than 1.0 mg. Water <2.48> Not more than 0.2 w/vz (5 mL, volumetric titration, direct titration).
Pipet 5 mL each of Sevoflurane and Sevoflurane RS (separately determine the water <2.48> in the same manner as Sevoflurane), to each add exactly 5 mL of dimethoxy-methane as an internal standard, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of sevoflurane to that of the internal standard. Amount (mg) of sevoflurane (C4H3F7O) ＝ VS × QT/QS × 1000 × 1.521 VS: Amount (mL) of Sevoflurane RS taken, calculated on the anhydrous basis 1.521: Specific gravity of Sevoflurane (d 20) Each mL of this solution contains 0.01 mg of fluorine (F). Related substances—Perform the test with 2 mL of Sevoflurane as directed under Gas Chromatography <2.02> according to the following conditions. Determine each peak area by the automatic integration method and calculate the amount of them by the area percentage method: the amount of the peak of hexafluoroisopropyl methyl ether, having the relative retention time of about 0.84 to sevoflurane, is not more than 0.005z, the amount of each peak other than the peaks of sevoflurane and hexafluoroisopropyl methyl ether is not more than 0.0025z, and the total amount of the peaks other than the peaks of sevoflurane and hexafluoroisopropyl methyl ether is not more than 0.005z. Operating conditions— Detector, column, injection port temperature, detector temperature, carrier gas and split ratio: Proceed as directed in the operating conditions in the Assay. Column temperature: Inject at a constant temperature of about 409C, maintain the temperature for 10 minutes, raise at a rate of 109C per minute to 2009C, and maintain at a constant temperature of about 2009C. Flow rate: Adjust so that the retention time of sevoflurane is about 7 minutes. Time span of measurement: About 6 times as long as the retention time of sevoflurane. System suitability— Test for required detectability: To 20 mL of Sevoflurane add o-xylene to make 20 mL. To 1 mL of this solution add o-xylene to make 20 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for sys-tem suitability test and add o-xylene to make exactly 10 mL. Confirm that the peak area of sevoflurane obtained with 2 mL of this solution is equivalent to 7 to 13z of the peak area of sevoflurane with 2 mL of the solution for system suita-bility test. System performance: When the procedure is run with 2 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica column 0.32 mm in inside diameter and 30 m in length, coated inside with cyanopropyl methyl-phenyl silicone for gas chromatography in 1.8 mm thickness. Column temperature: 409C. Injection port temperature: A constant temperature of about 2009C. Detector temperature: A constant temperature of about 2259C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of Sevoflu-rane is about 3 minutes. Split ratio: 1:20. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, sevoflurane and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of sevoflurane to that of the internal standard is not more than 1.0 z.
Beef Tallow occurs as a white, uniform mass. It has a characteristic odor and a mild taste. It is freely soluble in diethyl ether and in petroleum ether, very slightly soluble in ethanol (95), and practically insoluble in water. It is breakable at a low temperature, but softens above 309C. Melting point: 42 – 509C Acid value <1.13> Not more than 2.0. Saponification value <1.13> 193 – 200 Iodine value <1.13> 33 – 50 (When the sample is insoluble in 20 mL of cyclohexane, dissolve it by shaking a glass-stoppered flask in warm water. Then, if insoluble, increase the volume of solvent.)
(1) Moisture and coloration—Beef Tallow (5.0 g), melted by heating on a water bath, forms a clear liquid, from which no water separates. In a 10-mm thick layer of the liquid, it is colorless or slightly yellow. Alkalinity—To 2.0 g of Beef Tallow add 10 mL of water, melt by heating on a water bath, and shake vigor-ously. After cooling, add 1 drop of phenolphthalein TS to the separated aqueous layer: no color develops. Chloride—To 1.5 g of Beef Tallow add 30 mL of ethanol (95), heat for 10 minutes under a reflux condenser, and filter after cooling. To 20 mL of the filtrate add 5 drops of a solution of silver nitrate in ethanol (95) (1 in 50): the turbidity of the mixture does not exceed that of the following control solution. Control solution: To 1.0 mL of 0.01 mol/L hydrochloric acid VS add ethanol (95) to make 20 mL, then add 5 drops of an ethanolic solution of silver nitrate (1 in 50).
Shakuyakukanzoto Extract contains not less than 50 mg and not more than 150 mg of paeoniflorin parenchyma; in the interior, endosperm and cotyledon; epidermal cells contain orbicular crystals of calcium oxalate (C23H 28O 11: 480.
Shakuyakukanzoto Extract occurs as a light brown powder or brown viscous extract. It has slightly an odor, and a sweet taste.
Grind a suitable amount of Sesame. To 1.0 g of the ground add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 1 mg of sesamin for thin-layer chromatography in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (10:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard so-lution. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) according to Method 3, and perform the test (not more than 2 ppm). Loss on drying <2.41> The dry extract: Not more than 8.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 9.0z, calculated on the dried basis.
(1) Paeoniflorin—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equiva-lent to 0.2 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. (2) Glycyrrhizic acid—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.2 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Shimbuto Extract contains not less than 26 mg and raphy.
Shimbuto Extract occurs as light yellow-brown to brown powder. It has a characteristic odor and a hot and bitter taste.
(1) To 2.0 g of Shimbuto Extract, add 10 mL of water, shake, then add 5 mL of 1-butanol, shake, cen-trifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog- To 1.0 g of Shimbuto Extract, add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly diluted sulfu-ric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution (Atrac-tylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of Shimbuto Extract, add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solu-tion as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Exa-mine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylaminobenzaldehyde TS for spraying, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhizome). To 1.0 g of Shimbuto Extract, add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pres-sure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). To 3.0 g of Shimbuto Extract, add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes, centrifuge, and take the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of aceto- Shimbuto Extract / Crude Drugs and Related Drugs nitrile to the residue, and use this solution as the sample so-lution. Separately, dissolve 1 mg of benzoylmesaconine hy-drochloride for thin-layer chromatography in 10 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample so-lution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Processed Aconite Root or Pow-dered Processed Aconite Root).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Shimbuto Extract as directed in the Extracts (4), and perform the test (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 0.67 g of Shimbuto Extract according to Method 3, and perform the test (not more than 3 ppm). Aconitum diester alkaloids (aconitine, jesaconitine, hypaconitine and mesaconitine)—Weigh accurately 1.0 g of Shimbuto Extract, add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS and shake for mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between these peaks is not less than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5z. Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z.
(1) Paeoniflorin—Weigh accurately about 0.5 g of Shimbuto Extract, add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C H O ) 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the ＝ MS × AT/AS × 1/2 23 28 11 same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centri-fuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, pro-ceed in the same manner, and repeat the procedure twice. Combine all the extracts, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue with exactly 10 mL of a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1). Centrifuge this solution, and use the supernatant liquid as the sample solution. Sepa-rately, pipet 1 mL of aconitum diester alkaloids standard so-lution for purity, add a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make ex-actly 10 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions: the heights of the peaks corresponding to aconitine, jesaconi-tine, hypaconitine and mesaconitine from the sample solu-tion are not higher than the respective heights corresponding to aconitine, jesaconitine, hypaconitine and mesaconitine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of mesaconitine is about 31 minutes). System suitability— System performance: When the procedure is run with 20 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. [6]-gingerol—Weigh accurately about 0.5 g of Shim-buto Extract, add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of [6]-gingerol for assay, dissolve in diluted methanol to make exactly 100 mL. Pipet 5 mL of this solution, add meth-anol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of [6]-gingerol in each solution. ‌JP XVIII Crude Drugs and Related Drugs / Shosaikoto Extract 2147 Amount (mg) of [6]-gingerol ＝ MS × AT/AS × 1/20 MS: Amount (mg) of [6]-gingerol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (620:380:1). Flow rate: 1.0 mL per minute (the retention time of [6]-gingerol is about 15 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-gingerol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-gingerol is not more than 1.5z. Total alkaloids—Weigh accurately about 1 g of Shimbuto Extract, add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. Centrifuge this solution, remove the diethyl ether layer, then add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centri-fuge, and take the diethyl ether layer. To the aqueous layer, add 1.0 mL of ammonia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the ex-tracts, and evaporate the solvent under low pressure (in vacuo). Dissolve the residue with a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1) to make exactly 10 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Perform the test with exactly 20 mL each of the sample solution and the aconitum monoester alkaloids standard solution TS for assay as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas of benzoylmesaconine, benzoylhypaconine and 14-anisoylaconine, ATM and ASM, ATH and ASH, as well as ATA and ASA, in each solution, respectively. Amount (mg) of benzoylmesaconine hydrochloride ＝ CSM × ATM/ASM × 10 Amount (mg) of benzoylhypaconine hydrochloride ＝ CSH × ATH/ASH × 10 Amount (mg) of 14-anisoylaconine hydrochloride ＝ CSA × ATA/ASA × 10 CSM: Concentration (mg/mL) of benzoylmesaconine hy-drochloride for assay in aconitum monoester alkaloids standard solution TS for assay CSH: Concentration (mg/mL) of benzoylhypaconine hy-drochloride for assay in aconitum monoester alkaloids standard solution TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for benzoylmesaconine and benzoylhypaco-nine; 254 nm for 14-anisoylaconine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: 1.0 mL per minute (the retention time of ben-zoylmesaconine is about 15 minutes). System suitability— System performance: When the procedure is run with 20 mL of the aconitum monoester alkaloids standard solution TS for assay under the above operating conditions, the num-ber of theoretical plates and the symmetry factor of the peak of benzoylmesaconine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the aconitum monoester alkaloids standard solution TS for assay under the above operating conditions, the relative standard deviation of the peak areas of ben-zoylmesaconine, benzoylhypaconine and 14-anisoylaconine is not more than 1.5z.
Shosaikoto Extract contains not less than 2 mg and not more than 8 mg of saikosaponin b2, not less than 80 mg and not more than 240 mg of baicalin (C21H18O11: 446.
Shosaikoto Extract occurs as a light brown to grayish brown powder or black-grayish brown viscous ex-tract. It has a slight odor, and a sweet first then slightly pun-gent and bitter taste.
(1) Shake 2.0 g of the dry extract (or 6.0 g CSA : Concentration (mg/mL) of 14-anisoylaconine hydro-chloride for assay in aconitum monoester alkaloids standard solution TS for assay of the viscous extract) with 10 mL of sodium hydroxide TS, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu- Shosaikoto Extract / Crude Drugs and Related Drugs tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and exa-mine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 15 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, air-dry the plate. Spray evenly iron (III) chlo- (5) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to about 1.0 g of dried substance) as di-rected under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separaterly, use saikosaponin b2 standard TS for assay as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of sai-kosaponin b2 in each solution. ride-methanol TS on the plate: one of the several spots ob-tained from the sample solution has the same color tone and Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 Rf value with the yellow-brown spot from the standard solu-tion (Scutellaria Root). (4) Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, then add 5 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Ginseno-side Rb1 RS or ginsenoside Rb1 for thin-layer chromatogra-phy in 1 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solu-tion (Ginseng). CS: Concentration (mg/mL) of saikosaponin b2 in saiko-saponin b2 standard TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of sai-kosaponin b2 is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 JP XVIII Crude Drugs and Related Drugs / Shosaikoto Extract 2149 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry ex-tract (or an amount of the viscous extract, equivalent to about 0.1 g of dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute (the retention time of baica-lin is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of baicalein for resolution check in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and bai-calein is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of ethyl acetate and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the ethyl acetate layer, add 20 mL of ethyl acetate, proceed in the same manner as described above, and remove the ethyl acetate layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be- ‌2150 Shoseiryuto Extract / Crude Drugs and Related Drugs tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Shoseiryuto Extract contains not less than 8 mg and not more than 24 mg of the total alkaloids (ephedrine and pseudoephedrine), not less than 26 mg and not more than 78 mg of paeoniflorin (C23H28O11: 480.
Shoseiryuto Extract occurs as a light brown to brown powder or black-brown viscous extract. It has a char-acteristic odor and a acid first then pungent taste.
(1) Shake 1.0 g of dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol and shake, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solu-tion as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid(100) (4:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot is observed at an Rf value of about 0.5 (Ephedra Herb). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Peony Root). For preparation prescribed Processed Ginger—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of [6]-shogaol for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 1 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of cyclohexane and ethyl ace-tate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Processed Ginger). For preparation prescribed Ginger—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and spray water: one of the several spots obtained from the sam-ple solution has the same color tone and Rf value with the blue-green to grayish green spot from the standard solution (Ginger). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza). Perform the test according to the following i) or ii) (Cinnamon Bark). Put 10 g of the dry extract (or 30 g of the viscous ex-tract) in a 300-mL hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect the apparatus for essential oil determination, and heat under a reflux condenser. The grad-uated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to JP XVIII Crude Drugs and Related Drugs / Shoseiryuto Extract 2151 the graduated tube. After heating under reflux for 1 hour, separate the hexane layer, and use the layer as the sample so-lution. Separately, dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution and 2 mL the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,4-dinitrophenylhydrazine TS on the plate: one of the several spots obtained from the sample so-lution has the same color tone and Rf value with the yellow-orange spot from the standard solution. Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, then add 5 mL of hexane, shake, centrifuge, and use the hexane layer as the sample so-lution. Separately, dissolve 1 mg of (E )-2-methoxycinnamal-dehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 20 mL of the sample solution and 2 mL the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution. Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of asarinin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown spot from the standard solution (Asiasarum Root). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of sodium hydroxide TS, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solu-tion as the sample solution. Separately, dissolve 1 mg of schisandrin for thin-layer chromatography in 1 mL of meth-anol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple spot from the standard solution (Schisandra Fruit).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous extract, equivalent to 1.0 g of dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). Cadmium—Take 5.0 g of the dry extract (or an amount of the viscous extract, equivalent to 5.0 g of the dried substance) in a platinum, quartz or porcelain crucible, heat weakly, then incinerate by ignition at 4509C. After cooling, add a small amount of 2 mol/L nitric acid TS to the residue, filter if necessary, wash the crucible several times with small portions of 2 mol/L nitric acid TS, combine the filtrate and washings, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sample solution. Separately, to 5.0 mL of Standard Cadmium Solution add 2 mol/L nitric acid TS to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23>: the absor-bance of the sample solution is not more than that of the standard solution (not more than 1 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: Cadmium hollow-cathode lamp. Wavelength: 228.8 nm. Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 12.0z, calculated on the dried basis.
(1) Total alkaloids (ephedrine and pseudoephe-drine)—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of dried substance), add 20 mL of diethyl ether, shake, then add 3.0 mL of 0.1 mol/L hydrochloric acid TS, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ethler layer. To the aqueous layer add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. In addition, repeat twice in the same manner for the aqueous layer using 1.0 mL of am-monia TS and 20 mL of diethyl ether. Combine all the ex-tracts, evaporate the solvent under low pressure (in vacuo), dissolve the residue in diluted methanol (1 in 2) to make ex-actly 50 mL. Centrifuge this solution, and use the superna-tant liquid as the sample solution. Separately, weigh accu-rately about 10 mg of ephedrine hydrochloride for assay of crude drugs, previously dried at 1059C for 3 hours, and dis-solve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine the peak areas, ATE and ATP, of ephe-drine and pseudoephedrine obtained from the sample solu-tion, and the peak area, AS, of ephedrine from the standard solution. Shoseiryuto Extract / Crude Drugs and Related Drugs Amount (mg) of total alkaloids (ephedrine and pseudoephedrine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drugs taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, and add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: 1.0 mL per minute (the retention time of ephe-drine is about 27 minutes). System suitability— System performance: Dissolve 1 mg each of ephedrine hy-drochloride for assay of crude drugs and pseudoephedrine under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Glycyrrhizic acid—Perform the test according to the following i) or ii). Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS, (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. hydrochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephe- Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) drine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 1.5z. (2) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoniflorin RS, (separately deter-mined the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. Dissolve 1 mg of (E )-cinnamaldehyde for thin-layer chromatography in 50 mL of methanol. To 2 mL of this solution add 2 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of glycyrrhizic acid and (E )-cinnamaldehyde is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. After centrifugation, re-move the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as described above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the su- ‌JP XVIII Crude Drugs and Related Drugs / Smilax Rhizome 2153‌‌ pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and take the su-pernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solu-tion. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Silodosin occurs as a white to pale yellow-white powder. It is freely soluble in methanol and in ethanol (99.5), and very slightly soluble in water. It gradually becomes yellow-white on exposure to light. D Optical rotation [a]20: －13 – －179(0.2 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm). Melting point: 105 – 1099C Silodosin shows crystal polymorphism.
(1) Prepare the test solution with 10 mg of Silodosin as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Silodosin in methanol (1 in 20,000) as directed under Ultravi-olet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Silodosin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Silodosin as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Silodosin RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the Reference Standard according to the method otherwise specified, filter and dry the crystals, and perform the test using the crystals.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Silodosin in a platinum crucible according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Silodosin in 100 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the peak of related substance A, having the relative retention time of about 1.3 to silodo-sin, obtained from the sample solution is not larger than 3/20 times the peak area of silodosin from the standard solu-tion, the peak areas of related substance B and related sub-stance C, having the relative retention time of about 1.6 and about 2.0, respectively, is not larger than 1/16 times the peak area of silodosin from the standard solution, and the area of the peak other than silodosin and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of silodosin from the standard solution. In addi-tion, the total area of the peaks other than silodosin from the sample solution is not larger than 7/20 times the peak area of silodosin from the standard solution. For the areas of the peaks of related substance A, related substance B, and related substance C, multiply the correction factor 0.6, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 3.9 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water, and adjust to pH 3.4 with diluted phosphoric acid (1 in 10). Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 15 – 35 75 → 50 25 → 50 35 – 45 Flow rate: Adjust so that the retention time of silodosin is about 13 minutes. Time span of measurement: About 3 times as long as the retention time of silodosin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add methanol to make exactly 20 mL. Confirm that the peak area of silodosin obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: Thinly spread out an amount of Silodosin in a petri dish, exposure to a 4000 lx light for not less than 24 hours using a D65 fluorescent lamp, and dissolve 4 mg of this sample in 8 mL of methanol. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, the resolution between the peaks of silodo-sin and related substance A is not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.5z. Enantiomer—Conduct this procedure using light-resistant vessels. Dissolve 0.1 g of Silodosin in 10 mL of ethanol (99.5), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (99.5) to make exactly 200 mL. Pipet 3 mL of this solution, add ethanol (99.5) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak of the enantiomer, having the relative retention time of about 0.8 to silodosin, obtained from the sample solution is not larger than the peak area of silodosin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel coated with cellulose tris(4-methylbenzoate) for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of hexane, diethylamine and ethanol (99.5) (93:10:7). Flow rate: Adjust so that the retention time of silodosin is about 29 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of silodosin are not less than 1000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of silodosin is not more than 5z. Water <2.48> Not more than 0.1z, using a water vaporiza-tion device (heating temperature: 1509C; heating time: 2 minutes) (1.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Conduct this procedure using light-resistant vessels. Weigh accurately about 50 mg each of Silodosin and Silodo-sin RS (separately determine the water <2.48> in the same manner as Silodosin), dissolve each in methanol to make ex-actly 100 mL. Pipet 5 mL of both solutions, add exactly 5 mL of the internal standard solution to them, add methanol to make 25 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of silodosin to that of the internal standard. Amount (mg) of silodosin (C25H32F3N3O4) ＝ MS × QT/QS MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in methanol (1 in 8000). Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.9 g of sodium dihydrogen phos-phate dihydrate in 1000 mL of water, and adjust to pH 3.4 with diluted phosphoric acid (1 in 10). To 730 mL of this so-lution add 270 mL of acetonitrile. Flow rate: Adjust so that the retention time of silodosin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, silodosin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of silodosin to that of the internal standard is not more than 1.0z.
Silver Nitrate, when dried, contains not less than 99.
Silver Nitrate occurs as lustrous, colorless or white crystals. It is very soluble in water, soluble in ethanol (95), and practically insoluble in diethyl ether. It gradually turns grayish black by light.
A solution of Silver Nitrate (1 in 50) re-sponds to Qualitative Tests <1.09> for silver salt and for nitrate.
(1) Clarity and color of solution, and acidity or alkalinity—Dissolve 1.0 g of Silver Nitrate in 10 mL of freshly boiled and cooled water: the solution is clear and colorless. It is neutral. (2) Bismuth, copper and lead—To 5 mL of a solution of Silver Nitrate (1 in 10) add 3 mL of ammonia TS: the solu-tion is clear and colorless. Loss on drying <2.41> Not more than 0.20z (2 g, silica gel, light resistant, 4 hours).
Weigh accurately about 0.7 g of Silver Nitrate, pre-viously powdered and dried, dissolve in 50 mL of water, add 2 mL of nitric acid, and titrate <2.50> with 0.1 mol/L ammo-nium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 16.99 mg of AgNO3
Silver Protein is a compound of silver and proteins. It contains not less than 7.
Silver Protein occurs as a light yellow-brown to brown powder. It is odorless. It (1 g) dissolves slowly in 2 mL of water. It is practically insoluble in ethanol (95), in diethyl ether and in chloroform. The pH of a solution of 1.0 g of Silver Protein in 10 mL of water is between 7.0 and 8.5. It is slightly hygroscopic. It is affected by light.
(1) To 10 mL of a solution of Silver Pro-tein (1 in 100) add 2 mL of dilute hydrochloric acid, shake frequently for 5 minutes, and filter. To the filtrate add 5 mL of a solution of sodium hydroxide (1 in 10), and add 2 mL of diluted copper (II) sulfate TS (2 in 25): a purple color devel-ops. To 5 mL of a solution of Silver Protein (1 in 100) add dropwise iron (III) chloride TS: the color of the solution fades and a precipitate is gradually formed. Incinerate 0.2 g of Silver Protein by strong heating, dissolve the residue in 1 mL of nitric acid by warming, and add 10 mL of water: this solution responds to Qualitative Tests <1.09> (1) for silver salt.
Silver salt—Dissolve 0.10 g of Silver Protein in 10 mL of water, and filter. To the filtrate add 1 mL of potas- sium chromate TS: no turbidity is produced.
Transfer about 1 g of Silver Protein, accurately weighed, to a 100-mL decomposition flask, add 10 mL of sulfuric acid, cover the flask with a funnel, and boil for 5 minutes. Cool, add dropwise 3 mL of nitric acid with cau-tion, and heat for 30 minutes without boiling. Cool, add 1 mL of nitric acid, boil, and, if necessary, repeat this opera-tion until the solution becomes colorless. After cooling, transfer the solution to a 250-mL conical flask with 100 mL of water, and titrate <2.50> with 0.1 mol/L ammonium thiocyanate VS (indicator: 3 mL of ammonium iron (III) sul-fate TS). Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 10.79 mg of Ag
Simvastatin occurs as a white crystalline pow-der. It is freely soluble in acetonitrile, in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Simvastatin in acetonitrile (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24> and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Simvastatin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sim-vastatin as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Simvastatin RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋285 – ＋3009(50 mg calcu-lated on the dried basis, acetonitrile, 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1 g of Simvastatin in 10 mL of methanol: the solution is clear. Per-form the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 440 nm is not more than 0.10. Heavy metals <1.07>—To 1.0 g of Simvastatin add 2 mL of sulfuric acid, and heat gently to carbonize. After cooling, add 2 mL of nitric acid and 1 mL of sulfuric acid, heat gently until the white fumes no more evolve, and heat to incinerate at 500 to 6009C. If the incineration is not accom-plished, add 0.5 mL of nitric acid, heat in the same manner as above, and ignite at 500 to 6009C to incinerate com-pletely. After cooling, add 2 mL of hydrochloric acid, proceed with this solution according to Method 2, and per-form the test. Prepare the control solution by using the same quantities of the same reagents as directed for the prepara-tion of the test solution, and add 2.0 mL of Standard Lead solution and water to make 50 mL (not more than 20 ppm). Related substances—Dissolve 30 mg of Simvastatin in 20 mL of a mixture of acetonitrile and 0.01 mol/L potas-sium dihydrogen phosphate TS (pH 4.0) (3:2), and use this ‌solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area from the sample solution by the auto- 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of simvastatin in each solution. matic integration method, and calculate the amount of them by the area percentage method: the amounts of the peaks, Amount (mg) of simvastatin (C 25H 38O5) ＝ MS × AT/AS having the relative retention times of about 0.45, about 0.80, about 2.42, and about 3.80 to simvastatin are not more than 0.2z, respectively; the amount of the peak, having a relative retention time of about 2.38 is not more than 0.3z; the amount of the peak, having a relative retention time of about 0.60 is not more than 0.4z; and the amount of each peak other than simvastatin and the peaks mentioned above is not more than 0.1z. Furthermore, the total amount of the peaks other than simvastatin and the peak with relative retention time of about 0.60 is not more than 1.0z. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase A: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (1:1). Mobile phase B: A solution of phosphoric acid in aceto-nitrile for liquid chromatography (1 in 1000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: Amount (mg) of Simvastatin RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 33 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (1:1). Flow rate: Adjust so that the retention time of simvastatin is about 3 minutes. System suitability— System performance: Dissolve 3 mg of lovastatin in 2 mL of the standard solution. When the procedure is run with 5 mL of this solution under the above operating conditions, lovastatin and simvastatin are eluted in this order with the Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times 0 – 4.5 100 0 4.5 – 4.6 100 → 95 0 → 5 4.6 – 8.0 95 → 25 5 → 75 8.0 – 11.5 25 75 Flow rate: 3.0 mL per minute. Time span of measurement: About 5 times as long as the retention time of simvastatin. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 0.5 mL of the sample solution, add a mixture of acetonitrile and 0.01 mol/L po-tassium dihydrogen phosphate TS (pH 4.0) (3:2), to make 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suita-bility test, add a mixture of acetonitrile and 0.01 mol/L potassium dihydrogen phosphate TS (pH 4.0) (3:2), to make exactly 10 mL. Confirm that the peak area of simvastatin obtained with 5 mL of this solution is equivalent to 16 to 24z of that with 5 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above conditions, the relative standard deviation of the peak area of simvastatin is not more than 1.0z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 30 mg each of Simvastatin and Simvastatin RS (previously determine the loss on drying <2.41> under the same conditions as Simvastatin), dissolve each in a mixture of acetonitrile and 0.01 mol/L potassium dihydrogen phosphate TS (pH 4.0) (3:2), to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of simvastatin is not more than 1.0z.
Containers—Tight containers. Storage—Under nitrogen atmosphere.
Sitagliptin Phosphate Hydrate contains not less than 98.
Sitagliptin Phosphate Hydrate occurs as a white powder. It is soluble in water, sparingly soluble in methanol, very slightly soluble in acetonitrile and in ethanol (99.5). Identiˆcation (1) Determine the absorption spectrum of a solution of Sitagliptin Phosphate Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Sitagliptin Phosphate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sitagliptin Phosphate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Sitagliptin Phosphate RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Al-ternatively, perform the test by the potassium bromide disk method or the ATR method, and compare the spectrum with make exactly 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of sitagliptin is not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sitagliptin is not more than 2.0z. Enantiomer—Dissolve 80 mg of Sitagliptin Phos-phate Hydrate in a mixture of methanol and water (9:1) to make 10 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine the total peak area, AT, of sitagliptin and related substance A (enantiomer), having the relative retention time of about 0.9 to sitagliptin and the peak area, AS, of related substance A (enantiomer), and cal-culate the amount of the enantiomer by the following equa-tion: not more than 0.5z. Amount (z) of enantiomer ＝ AS/AT × 100 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel coated with amylose tris-(3,5-dimethylphenylcarbamate) coated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. ‌Mobile phase: A mixture of ethanol (99.5), heptane, water and diethylamine (600:400:1:1). Flow rate: 0.8 mL per minute. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, and dissolve in a mixture of methanol and water (9:1) to make exactly 100 mL. Pipet 1 mL of this solution, add a mixture of methanol and water (9:1) to make exactly 10 mL. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the SN ratio of the peak of sitagliptin is not less than 10. System performance: Dissolve 8 mg of Sitagliptin Phos-phate for System Suitability RS in 1 mL of a mixture of methanol and water (9:1). When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of related substance A (enan-tiomer) and sitagliptin is not less than 1.5. Water <2.48> 3.3 – 3.7z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g, plati-num crucible).
(1) Heavy metals—Being speciˆed separately when the drug is granted approval based on the Law. (2) Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make exactly 1000 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area by the automatic integra-tion method: the area of the peak other than sitagliptin ob-tained from the sample solution is not larger than the peak area of sitagliptin from the standard solution, and the total area of the peaks other than sitagliptin from the sample solu-tion is not larger than 5 times the peak area of sitagliptin from the standard solution. For this calculation the peak area not larger than 1/2 times the peak area of sitagliptin from the standard solution is excluded. Operating conditions— Detector, column, column temperature, mobile phase and ‰ow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5.5 times as long as the retention time of sitagliptin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of diluted phosphoric acid (1 in 16 15 6 5 3 4 2 1000) and acetonitrile for liquid chromatography (19:1) to (3R)-3-Amino-1-[3-(tri‰uoromethyl)-5,6-dihydro[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl]- 4-(2,4,5-tri‰uorophenyl)butan-1-one monophosphate monohydrate [654671-77-9] Sitagliptin Phosphate Hydrate contains not less than 98.0z and not more than 102.0z of sitagliptin phos-phate (C16H15F6N5O.H3PO4: 505.31), calculated on the anhydrous basis. Description Sitagliptin Phosphate Hydrate occurs as a white powder. It is soluble in water, sparingly soluble in methanol, very slightly soluble in acetonitrile and in ethanol (99.5). Identiˆcation (1) Determine the absorption spectrum of a solution of Sitagliptin Phosphate Hydrate (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Sitagliptin Phosphate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sitagliptin Phosphate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Sitagliptin Phosphate RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Al-ternatively, perform the test by the potassium bromide disk method or the ATR method, and compare the spectrum with make exactly 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of sitagliptin is not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sitagliptin is not more than 2.0z. Enantiomer—Dissolve 80 mg of Sitagliptin Phos-phate Hydrate in a mixture of methanol and water (9:1) to make 10 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine the total peak area, AT, of sitagliptin and related substance A (enantiomer), having the relative retention time of about 0.9 to sitagliptin and the peak area, AS, of related substance A (enantiomer), and cal-culate the amount of the enantiomer by the following equa-tion: not more than 0.5z. Amount (z) of enantiomer ＝ AS/AT × 100 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with silica gel coated with amylose tris-(3,5-dimethylphenylcarbamate) coated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. ‌Mobile phase: A mixture of ethanol (99.5), heptane, water and diethylamine (600:400:1:1). Flow rate: 0.8 mL per minute. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, and dissolve in a mixture of methanol and water (9:1) to make exactly 100 mL. Pipet 1 mL of this solution, add a mixture of methanol and water (9:1) to make exactly 10 mL. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the SN ratio of the peak of sitagliptin is not less than 10. System performance: Dissolve 8 mg of Sitagliptin Phos-phate for System Suitability RS in 1 mL of a mixture of methanol and water (9:1). When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peaks of related substance A (enan-tiomer) and sitagliptin is not less than 1.5. Water <2.48> 3.3 – 3.7z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g, plati-num crucible).
Weigh accurately about 20 mg each of Sitagliptin Phosphate Hydrate and Sitagliptin Phosphate RS (separately determine the water <2.48> in the same manner as Sitagliptin Phosphate Hydrate), dissolve each in a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chro-matography (19:1) to make exactly 200 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of sitagliptin in each solution. Amount (mg) of sitagliptin phosphate (C16H15F6N5O.H3PO4) tion becomes clear. Use the supernatant liquid as the solu-tion for system suitability test. When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the resolution between sitagliptin and the peak having the relative retention time of about 1.2 to sitagliptin is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sitagliptin is not more than 1.0z.
ソルビタンセスキオレイン酸エステル
Sorbitan Sesquioleate is a pale yellow to light yellow-brown, viscous oily liquid. It has a faint, characteris-tic odor and a slightly bitter taste. It is freely soluble in diethyl ether, slightly soluble in ethanol (95), and very slightly soluble in methanol. It is dispersed as fine oily drops in water.
(1) To 0.5 g of Sorbitan Sesquioleate add 5 mL of ethanol (95) and 5 mL of dilute sulfuric acid, and heat on a water bath for 30 minutes. Cool, shake with 5 mL of petroleum ether, and allow to stand, and separate the upper layer and the lower layer. Shake 2 mL of the lower layer with 2 mL of freshly prepared catechol solution (1 in 10), then with 5 mL of sulfuric acid: a red to red-brown color devel-ops. (2) Heat the upper layer obtained in (1) on a water bath, and evaporate petroleum ether. To the residue add 2 mL of diluted nitric acid (1 in 2), and then add 0.5 g of potassium nitrite between 309C and 359C with stirring: the solution develops an opalescence, and, when cooled, crystals are formed. Specific gravity <1.13> d 25: 0.960 – 1.020 Saponification value <1.13> 150 – 168
(1) Acidity—To 2.0 g of Sorbitan Sesquioleate add 50 mL of neutralized ethanol, and heat on a water bath nearly to boiling with stirring once or twice. Cool, add 4.3 mL of 0.1 mol/L sodium hydroxide VS and 5 drops of phe-nolphthalein TS: a red color develops. Heavy metals <1.07>—Proceed with 1.0 g of Sorbitan Sesquioleate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sorbitan Sesquioleate according to Method 2, and per- ‌form the test (not more than 2 ppm).‌ Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration, stir for 30 minutes). Residue on ignition <2.44> Not more than 1.0z (1 g).
スペクチノマイシン塩酸塩水和物
Spectinomycin Hydrochloride Hydrate is the hydro-chloride of a substance having antibacterial activity produced by the growth of Streptomyces spectabilis. It contains not less than 763 mg (potency) and not more than 831 mg (potency) per mg, calculated on the anhydrous basis.
Spectinomycin Hydrochloride Hydrate occurs as a white to light yellow-white crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (95).
(1) To 5 mL of a solution of Spectinomy-cin Hydrochloride Hydrate (1 in 100) add gently anthrone TS: a blue to blue-green color is produced at the zone of con-tact. Determine the infrared absorption spectra of Spec-tinomycin Hydrochloride Hydrate and Spectinomycin Hy-drochloride RS as directed in the paste method under Infra-red Spectrophotometry <2.25>, and compare these spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. To 3 mL of a solution of Spectinomycin Hydrochlo-ride Hydrate (1 in 150) add 1 drop of silver nitrate TS: a white turbidity is produced. D Optical rotation <2.49> [a]20: ＋15 – ＋219(2.1 g calculated on the anhydrous basis, water, 25 mL, 200 mm). pH <2.54> Dissolve 0.10 g of Spectinomycin Hydrochloride Hydrate in 10 mL of water: the pH of the solution is between 4.0 and 5.6.
Related substances—Dissolve 0.20 g of Spectinomy-cin Hydrochloride Hydrate in 5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-propanol, water, pyridine ‌and acetic acid (100) (10:8:1:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly alkaline 1.6z potassium periodate-0.2z potassium permanganate TS: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Water <2.48> Not less than 16.0z and not more than 20.0z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 1.0z (1 g).
Weigh accurately an amount of both Spectinomycin Hydrochloride Hydrate and Spectinomycin Hydrochloride RS, equivalent to about 20 mg (potency), add exactly 10 mL of the internal standard solution to them, add 1 mL of 1,1,1,3,3,3-hexamethyldisilazan, allow to stand at room tem-perature for 1 hour, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the test with 1 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions, and calculate the ratios, QT and QS, of the peak area of spectinomycin to that of the in-ternal standard. Amount [mg (potency)] of spectinomycin (C14H24N2O7) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Spectinomycin Hydrochlo-ride RS taken Internal standard solution—A solution of triphenylantimony in N,N-dimethylformamide (1 in 500). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 60 cm in length, packed with 150 to 180 mm siliceous earth for gas chromatography coated in 5z with 5z phenyl-methyl silicone polymer for gas chromatography. Column temperature: A constant temperature of about 1909C. Injection port temperature: A constant temperature of about 2159C. Detector temperature: A constant temperature of about 2209C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of spec-tinomycin is about 10 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, the internal standard and spectinomycin are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of spectinomycin to that of the internal standard is not more than 1.5z.
Spiramycin Acetate occurs as a white to light yellow-white powder. It is very soluble in acetonitrile and in methanol, freely soluble in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Spiramycin Acetate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Spiramycin Acetate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Content ratio of the active principle Dissolve 25 mg of Spiramycin Acetate in 25 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the areas, AII, AIII, AIV, AV, AVI and AVII, of the peaks of spiramycin II acetate, spiramycin III acetate, spiramycin IV acetate, spiramycin V acetate, spiramycin VI acetate and spiramycin VII acetate, respectively, by the auto-matic integration method, and calculate the ratios of the amounts of AII, AIV and the total of AIII and AV to the total amount of all these peaks: the amount of AII is 30 – 45z, AIV is 30 – 45z, and the total of AIII and AV is not more than 25z. The relative retention times of spiramycin III ace-tate, spiramycin IV acetate, spiramycin V acetate, spiramy-cin VI acetate and spiramycin VII acetate to spiramycin II acetate are about 1.3, about 1.7, about 2.3, about 0.85 and about 1.4, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, 0.02 mol/L po-tassium dihydrogen phosphate TS and a solution of dipotas-sium hydrogen phosphate (87 in 25,000) (26:7:7). Flow rate: Adjust so that the retention time of spiramycin II acetate is about 10 minutes. System suitability— System performance: Dissolve 25 mg of Spiramycin II Acetate RS in the mobile phase to make 100 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of spiramycin II acetate are not less than 14,500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of spiramycin II acetate is not more than 2.0z.
Heavy metals <1.07>—Proceed with 1.0 g of Spiramycin Acetate according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 3.0z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.5z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Spiramycin II Acetate RS, equivalent to about 50 mg (po-tency), dissolve in 20 mL of methanol, add 0.1 mol/L phos-phate buffer solution for antibiotics (pH 8.0) to make ex-actly 50 mL, and use this solution as the standard stock solu-tion. Keep the standard stock solution at not exceeding 59C, and use within 3 days. Take exactly a suitable amount of the ‌standard stock solution before use, add 0.1 mol/L phos-phate buffer solution for antibiotics (pH 8.0) to make solu-tions so that each mL contains 80 mg (potency) and 20 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solu-tion, respectively.‌ Sample solutions—Weigh accurately an amount of Spiramycin Acetate, equivalent to about 50 mg (potency), dissolve in 20 mL of methanol, and add 0.1 mol/L phos-phate buffer solution for antibiotics (pH 8.0) to make ex-actly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) to make solutions so that each mL contains 80 mg (po-tency) and 20 mg (potency), and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
Stearic Acid is a mixture consisting mainly of stearic overall diameter at right angles to the rod. The inner tube with its jacket is supported centrally in a 1-L beaker contain-ing a suitable cooling liquid to within 20 mm of the top. A thermometer is supported in the cooling bath. Place in the inner tube sufficient quantity of the liquid or previously melted substance to be examined, to cover the thermometer bulb and determine the approximate congeal-ing point by cooling rapidly. Place the inner tube in a bath about 59C above the approximate congealing point until all but the last traces of crystals are melted. Fill the beaker with water or a saturated solution of sodium chloride, at a tem-perature about 59C lower than the expected congealing point, insert the inner tube into the outer tube, ensuring that some seed crystals are present, and stir thoroughly until solidification takes place. Note the highest temperature ob-served during solidification. The apparatus directed under Congealing Point Determi- nation <2.42> is also can be used. Transfer the melted sample into sample container B up to the marked line C. Adjust the immersion line H of thermometer F to the same level of the meniscus of the sample, and then determine the approximate congealing point by cooling rapidly. Place the sample con-tainer B in a bath at a temperature about 59C above the ap-proximate congealing point until all but the last traces of crystals are melted. Fill bath D with water or a saturated so-lution of sodium chloride, at a temperature about 59C lower than the expected congealing point, and set the sample con-tainer B in A. Ensuring that some seed crystals are present, stir thoroughly until solidification takes place. Note the highest temperature observed during solidification.◆ The congealing point of stearic acid 50 is 53 – 599C, of stearic acid 70 is 57 – 649C, and of stearic acid 95 is 64 – acid (C 18H36 O2: 284.48) and palmitic acid (C 16H32 O2: 699C. 256.42) obtained from fats or oils of vegetable or animal origin. It occurs as three types, stearic acid 50, stearic acid 70 and stearic acid 95, composed with different fatty acid composition. Each type contains respectively the amount of stearic acid and the sum of stearic acid and palmitic acid as shown in the following table.
Stearic acid occurs as white, unctuous masses, crystalline masses or powder. It has a faint, fatty odor. It is soluble in ethanol (99.5), and practically insoluble in water.◆ Congealing point The apparatus consists of a test tube about 25 mm in diameter and 150 mm long placed inside a test tube about 40 mm in diameter and 160 mm long. The in-ner tube is closed by a stopper which carries a thermometer about 175 mm long and graduated in 0.29C fixed so that the upper end of◆ the bulb is about 15 mm above the bot- tom of the tube. The stopper has a hole allowing the passage of the stem of a stirrer made from a glass rod or other suita-ble material formed at one end into a loop of about 18 mm adding the 0.1 mol/L sodium thiosulfate VS dropwise until the color is discharged. Perform a blank determination in the same manner. When the iodine value is calculated by the following equation, that of stearic acid 50 and 70 is not more than 4.0, and of stearic acid 95 is not more than 1.5. Iodine value ＝ (a － b) × 1.269/M M: Amount (g) of Stearic Acid taken a: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con-sumed in the blank determination b: Volume (mL) of 0.1 mol/L sodium thiosulfate VS con-sumed in the test
(1) Acidity—Melt 5.0 g, shake for 2 minutes with 10 mL of hot carbon dioxide-free water, cool slowly and filter. To the filtrate add 0.05 mL of methyl orange TS: no red color develops. (2) Heavy metals <1.07>—Proceed with 1.0 g of Stearic Acid according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ ‌Residue on ignition <2.44> Not more than 0.1z (1 g).◆ not more than 3.0z. Furthermore, the relative standard de-
Place 0.100 g of Stearic Acid in a ◆small◆ conical flask fitted with a reflux condenser. Add 5.0 mL of boron trifluoride-methanol TS, ◆shake, and◆ boil under reflux for about 10 minutes ◆to dissolve.◆ Add 4 mL of heptane through the condenser, and boil again under reflux for 10 minutes. Allow to cool, add 20 mL of a saturated solution of sodium chloride, shake and allow the layers to separate. Remove 2 mL of the separated heptane layer, and dry it over about 0.2 g of anhydrous sodium sulphate, ◆previously washed with heptane.◆ Take 1.0 mL of the dried heptane layer in a 10-mL volumetric flask, add heptane to make up to 10 mL, and use this solution as the sample solution. Per- viation of the ratio of the peak area of methyl palmitate to the peak area of methyl stearate obtained from the 6-time repetition is not more than 1.0z.
Stearyl Alcohol occurs as a white unctuous matter. It has a faint, characteristic odor. It is tasteless. It is freely soluble in ethanol (95), in ethanol (99.5) and in diethyl ether, and practically insoluble in water.
(1) Clarity of solution—Dissolve 3.0 g of Stearyl Alcohol in 25 mL of ethanol (99.5) by warming: the solution is clear. (2) Alkalinity—To the solution obtained in (1) add 2 drops of phenolphthalein TS: no red color develops. Residue on ignition <2.44> Not more than 0.05z (2 g).
Sterile Purified Water in Containers is a clear and colorless liquid, having no odor.
Potassium permanganate-reducing substances—To 100 mL of Sterile Purified Water in Containers add 10 mL of dilute sulfuric acid, boil, then add 0.10 mL of 0.02 mol/L potassium permanganate VS, and boil again for 10 minutes: the red color of the solution does not disappear. Conductivity <2.51> When the test is performed according to the following method, the conductivity (259C) is not more than 25 mS・cm－1 for containers with a nominal volume of 10 mL or less, and not more than 5 mS・cm－1 for containers with a nominal volume greater than 10 mL. Transfer a suitable amount of Sterile Purified Water in Containers to a beaker, and stir the water specimen. Adjust the temperature to 25 ± 19C, and begin agitating the water specimen vigorously, while observing its conductivity period-ically. When the change in conductivity becomes not greater than 0.1 mS・cm－1 per 5 minutes, adopt the observed value as the conductivity of the water specimen. Sterility <4.06> It meets the requirements.
Containers—Hermetic containers. Plastic containers for aqueous injections can be used in place of hermetic containers.
To 0.4 g of 0.1z Reserpine Powder add 20 mL of acetonitrile, shake for 30 minutes, and centrifuge. De-termine the absorption spectrum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 265 nm and 269 nm, and between 294 nm and 298 nm. Dissolution Being specified separately when the drug is granted approval based on the Law.
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately a quantity of 0.1z Reserpine Powder, equivalent to about 0.5 mg of reserpine (C33H40N2O9), disperse in 12 mL of water, add ex-actly 10 mL of the internal standard solution and 10 mL of acetonitrile, and dissolve by warming at 509C for 15 minutes, then add water to make 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Reserpine RS, previously dried at 609C in vacuum for 3 hours, dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 10 mL of the internal standard solution, 5 mL of acetonitrile and water to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Reserpine. Amount (mg) of reserpine (C33H40N2O9) ＝ MS × QT/QS × 1/20 MS: Amount (mg) of Reserpine RS taken Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 50,000).
Streptomycin Sulfate occurs as a white to light yellow-white powder. It is freely soluble in water, and very slightly soluble in ethanol (95).
(1) Dissolve 50 mg of Streptomycin Sulfate in 5 mL of water, add 1 mL of ninhydrin TS and 0.5 mL of pyridine, and heat for 10 minutes: a purple color is devel-oped. Dissolve 10 mg each of Streptomycin Sulfate and Streptomycin Sulfate RS in 10 mL of water, and use these solutions as the sample solution and standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solu-tion of potassium dihydrogen phosphate (7 in 100) to a dis-tance of about 12 cm, and air-dry the plate. Spray evenly a mixture of a solution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heat the plate at about 1509C for about 5 minutes: the principal spots obtained from the sample solu-tion and the standard solution show the same in color tone and Rf value. A solution of Streptomycin Sulfate (1 in 5) responds to Qualitative Tests <1.09> for sulfate. D Optical rotation <2.49> [a]20: －79 – －889(0.5 g calculated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving wash the inside of the condenser with a suitable amount of a mixture of methanol and sulfuric acid (97:3), and add a mix-ture of methanol and sulfuric acid (97:3) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of methanol and sulfuric acid (97:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatog-raphy, develop with a mixture of toluene, methanol and ace-tic acid (100) (2:1:1) to a distance of 13 to 15 cm, and air-dry the plate. Spray evenly a mixture of a solution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heat the plate at 1109C for 5 minutes: the spot obtained from the sample so-lution corresponding to the spot from the standard solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
(1) Clarity and color of solution—Dissolve 1.0 g of Streptomycin Sulfate in 5 mL of water: the solution is clear, and its absorbance at 400 nm determined as directed under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.17. Heavy metals <1.07>—Proceed with 2.0 g of Strep- tomycin Sulfate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Streptomycin Sulfate according to Method 3 and perform the test (not more than 1 ppm). Related substances—Dissolve exactly 0.20 g of Strep-tomycin Sulfate in a mixture of methanol and sulfuric acid (97:3) to make 5 mL, and heat under a reflux condenser for 1 hour. After cooling, wash the inside of the condenser with a suitable amount of a mixture of methanol and sulfuric acid (97:3), add a mixture of methanol and sulfuric acid (97:3) to make exactly 20 mL, and use this solution as the sample so-lution. Separately, dissolve exactly 36 mg of D-mannose in a (C21H39N7O 12)2.3H2SO4: 1457.38 mixture of methanol and sulfuric acid (97:3) to make 5 mL, and heat under a reflux condenser for 1 hour. After cooling, 2-Deoxy-2-methylamino-a-L-glucopyranosyl-(1→2)- 5-deoxy-3-C-formyl-a-L-lyxofuranosyl-(1→4)-N,N?-diamidino-D-streptamine sesquisulfate [3810-74-0] Streptomycin Sulfate is the sulfate of an amino-glycoside substance having antibacterial activity pro-duced by the growth of Streptomyces griseus. It contains not less than 740 mg (potency) and not more than 820 mg (potency) per mg, calculated on the dried basis. The potency of Streptomycin Sulfate is expressed as mass (potency) of streptomycin (C21H39N7O12: 581.57). Description Streptomycin Sulfate occurs as a white to light yellow-white powder. It is freely soluble in water, and very slightly soluble in ethanol (95). Identification (1) Dissolve 50 mg of Streptomycin Sulfate in 5 mL of water, add 1 mL of ninhydrin TS and 0.5 mL of pyridine, and heat for 10 minutes: a purple color is devel-oped. Dissolve 10 mg each of Streptomycin Sulfate and Streptomycin Sulfate RS in 10 mL of water, and use these solutions as the sample solution and standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a solu-tion of potassium dihydrogen phosphate (7 in 100) to a dis-tance of about 12 cm, and air-dry the plate. Spray evenly a mixture of a solution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heat the plate at about 1509C for about 5 minutes: the principal spots obtained from the sample solu-tion and the standard solution show the same in color tone and Rf value. A solution of Streptomycin Sulfate (1 in 5) responds to Qualitative Tests <1.09> for sulfate. D Optical rotation <2.49> [a]20: －79 – －889(0.5 g calculated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving wash the inside of the condenser with a suitable amount of a mixture of methanol and sulfuric acid (97:3), and add a mix-ture of methanol and sulfuric acid (97:3) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of methanol and sulfuric acid (97:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatog-raphy, develop with a mixture of toluene, methanol and ace-tic acid (100) (2:1:1) to a distance of 13 to 15 cm, and air-dry the plate. Spray evenly a mixture of a solution of 1,3-dihydroxynaphthalene in ethanol (95) (1 in 500) and diluted sulfuric acid (1 in 5) (1:1) on the plate, and heat the plate at 1109C for 5 minutes: the spot obtained from the sample so-lution corresponding to the spot from the standard solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 5.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Residue on ignition <2.44> Not more than 1.0z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer, having pH 7.8 – 8.0 after sterilization. Standard solutions—Weigh accurately an amount of Streptomycin Sulfate RS, previously dried, equivalent to about 20 mg (potency), dissolve in diluted phosphate buffer solution (pH 6.0) (1 in 2) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution between 59C and 159C, and use within 30 days. Take exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solu-tion (pH 8.0) to make solutions so that each mL contains 8 mg (potency) and 2 mg (potency), and use these solutions as the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Streptomycin Sulfate, equivalent to about 20 mg (potency), ‌dissolve in water to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 8 mg (potency) and 2 mg (potency), and use these so-lutions as the high concentration sample solution and the low concentration sample solution, respectively.‌
Sucralfate Hydrate contains not less than 17.
Sucralfate Hydrate occurs as a white powder. It is odorless and tasteless. It is practically insoluble in water, in hot water, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid and in sulfuric acid-sodium hydroxide TS.
(1) To 0.05 g of Sucralfate Hydrate in a small test tube add 0.05 g of fresh pieces of sodium, and melt by careful heating. Immerse the test tube immediately in 100 mL of water, break the test tube, shake well, and filter. To 5 mL of the filtrate add 1 drop of sodium pentacyanonitro-sylferrate (III) TS: a red-purple color develops. Dissolve 40 mg of Sucralfate Hydrate in 2 mL of di-lute sulfuric acid, and add gently 2 mL of anthrone TS to make 2 layers: a blue color develops at the zone of contact, and gradually changes to blue-green. Dissolve 0.5 g of Sucralfate Hydrate in 10 mL of di-lute hydrochloric acid: the solution responds to Qualitative Tests <1.09> for aluminum.
(1) Clarity and color of solution—Dissolve 1.0 g of Sucralfate Hydrate in 10 mL of dilute sulfuric acid: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of Sucralfate Hy- drate in 30 mL of dilute nitric acid, and heat gently to boil-ing. After cooling, add water to make 100 mL, and to 10 mL of this solution add 3 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.70 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.50z). Heavy metals <1.07>—Dissolve 1.0 g of Sucralfate Hydrate in 20 mL of a solution of sodium chloride (1 in 5) and 1 mL of dilute hydrochloric acid, and to this solution add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution as follows: evaporate 1 mL of di-lute hydrochloric acid on a water bath to dryness, and add 20 mL of a solution of sodium chloride (1 in 5), 2 mL of di-lute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Dissolve 1.0 g of Sucralfate Hydrate in 5 mL of dilute hydrochloric acid, use this solution as the test solution, and perform the test (not more than 2 ppm). Free aluminum—To 3.0 g of Sucralfate Hydrate add 50 mL of water, heat in a water bath for 5 minutes, cool, and filter. Wash the residue with four 5-mL portions of water, combine the filtrate with the washings, add 2 mL of dilute hydrochloric acid, and heat in a water bath for 30 minutes. After cooling, neutralize the solution with sodium hydroxide TS, add water to make exactly 100 mL, and use this solution as the sample solution. Pipet 50 mL of the sample solution, add exactly 25 mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS and 20 mL of acetic acid-ammonium acetate buffer solution (pH 4.5) and boil for 5 minutes. After cooling, add 50 mL of ethanol (95), and titrate <2.50> the excess disodium dihydrogen ethylene-diamine tetraacetate with 0.05 mol/L zinc acetate VS until the color of the solution changes from green-purple through purple to red (indicator: 3 mL of dithizone TS). Perform a blank determination in the same manner (not more than 0.2 z). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.349 mg of Al Related substances—Proceed with 50 mL of the sam-ple solution obtained in the Assay (2) Sucrose octasulfate ester as directed in the Assay (2) Sucrose octasulfate ester, and perform the test as directed under Liquid Chromatogra-phy <2.01>. Determine the peak area of sucrose octasulfate ester obtained from the sample solution and that of a related substance with the relative retention time about 0.7 to su-crose octasulfate ester by the automatic integration method, and calculate the ratio of the peak area of the related sub-stance to that of sucrose octasulfate ester: it is not more than 0.1. Detection sensitivity: Adjust so that the peak height of su-crose octasulfate ester from 50 mL of the standard solution obtained in the Assay (2) Sucrose octasulfate ester composes 60 to 100z of the full scale. Loss on drying <2.41> Not more than 14.0z (1 g, 1059C, 3 hours). Acid-consuming capacity Weigh accurately about 0.25 g of Sucralfate Hydrate, previously dried, place in a 200-mL glass-stoppered conical flask, add exactly 100 mL of 0.1 mol/L hydrochloric acid VS, stopper the flask tightly, and shake at 37 ± 29C for exactly 1 hour (150 shakings per minute, amplitude: 20 mm). After cooling in water for 5 minutes, pipet 10 mL of the supernatant liquid, and titrate <2.50> the excess acid with 0.1 mol/L sodium hydroxide VS until the pH becomes 3.5. Perform a blank determination in the same manner. The amount of 0.1 mol/L hydrochloric acid VS consumed per g of Sucralfate Hydrate is not less than 130 mL.
(1) Aluminum—Weigh accurately about 1 g of Sucralfate Hydrate, dissolve in 10 mL of dilute hydrochloric acid by warming on a water bath, cool, and add water to make exactly 250 mL. Pipet 25 mL of this solution, add ex-actly 25 mL of 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS and 20 mL of acetic acid-ammoni-um acetate buffer solution (pH 4.5) and boil for 5 minutes. After cooling, add 50 mL of ethanol (95), and titrate <2.50> the excess disodium dihydrogen ethylenediamine tetraacetate with 0.05 mol/L zinc acetate VS until the color of the solu-tion changes from green-purple through purple to red (indi- cator: 3 mL of dithizone TS). Perform a blank determina-tion in the same manner. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.349 mg of Al (2) Sucrose octasulfate ester—Weigh accurately about 0.55 g of Sucralfate Hydrate, add exactly 10 mL of sulfuric acid-sodium hydroxide TS, shake vigorously, and dissolve by sonicating at below 309C for 5 minutes. To this solution add 0.1 mol/L sodium hydroxide VS to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.25 g of Potassium Sucrose Octasul-fate RS, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Prepare rapidly the sample solution and the standard solution, and perform the test immediately. Pipet 50 mL each of the sample solution and standard solution, and perform the test as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of sucrose octasulfate ester in each solution. Amount (mg) of sucrose octasulfate ester (C12H22O35S8) ＝ MS × AT/AS × 0.763 MS: Amount (mg) of Potassium Sucrose Octasulfate RS taken, calculated on the anhydrous basis Operating conditions— Detector: A differential refractometer. Column: A stainless steel column about 4 mm in inside di-ameter and about 30 cm in length, packed with amino-propylsilanized silica gel for liquid chromatography (about 8 mm in particle diameter). Column temperature: Room temperature. Mobile phase: Dissolve a suitable amount (26 to 132 g) of ammonium sulfate in 1000 mL of water, and adjust with phosphoric acid to pH 3.5. Allow a solution of Potassium Sucrose Octasulfate RS in dilute hydrochloric acid (1 in 100) to stand at 609C for 10 minutes, cool, and perform the test immediately. Adjust the amount of ammonium sulfate in the mobile phase so that the peak of a related substance with the relative retention time about 0.7 to sucrose octasulfate ester almost returns to the base line, and the peak of sucrose oc-tasulfate ester elutes most rapidly. Flow rate: Adjust so that the retention time of sucrose oc-tasulfate ester is between 6 and 11 minutes. Selection of column: Allow a solution of Potassium Su-crose Octasulfate RS in dilute hydrochloric acid (1 in 100) to stand at 609C for 10 minutes, cool, and proceed immediately with 50 mL of this solution under the above operating condi-tions. Use a column with a resolution being not less than 1.5 between sucrose octasulfate ester and a related substance with the relative retention time about 0.7 to sucrose octasul-fate ester. System repeatability: Repeat the test 6 times with the standard solution under the above operating conditions: the relative standard deviation of the peak area of sucrose oc-tasulfate ester is not more than 2.0z.
Sucrose is a white crystalline powder, or lus-trous colorless or white crystals. It is very soluble in water, and practically insoluble in ethanol (99.5).◆
Determine the infrared absorption spec-trum of Sucrose as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ D Optical rotation <2.49> [a]20: ＋66.3 – ＋67.09(26 g, water, 100 mL, ◆100 mm◆).
(1) Clarity and color of solution—Dissolve 100 g of White Soft Sugar in 100 mL of water, take 50 mL of this solution in a Nessler tube, and view transversely the Nessler tube against a white background: the solution is colorless or only slightly yellow and has no blue color. Fill the solution in the Nessler tube, stopper, and allow to stand for 2 days: no precipitate is produced. Chloride <1.03>—To 10.0 g of White Soft Sugar add water to make 100 mL, and use this solution as the sample solution. To 20 mL of the sample solution add 6 mL of di-lute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.005z). Sulfate <1.14>—To 40 mL of the sample solution ob- tained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.006z). Calcium—To 10 mL of the sample solution obtained in (2) add 1 mL of ammonium oxalate TS: this solution shows immediately no change. Heavy metals <1.07>—Proceed with 5.0 g of White Soft Sugar according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of White Soft Sugar according to Method 1, and perform the test (not more than 2 ppm). Invert sugar—Dissolve 5.0 g of White Soft Sugar in water to make 100 mL, filter if necessary, and use this solu-tion as the sample solution. Separately place 100 mL of alka-line copper (II) sulfate TS in a 300-mL beaker, cover the b-D-Fructofuranosyl a-D-glucopyranoside [57–50–1] This monograph is harmonized with the European Phar-macopoeia and the U.S.Pharmacopeia. The parts of the text that are not harmonized are marked with symbols (◆ ◆). Information on the harmonization with the European Pharmacopoeia and the U.S. Pharmacopeia is available on the website of the Pharmaceuticals and Medical Devices Agency. Sucrose contains no additives. For Sucrose used for preparation of the parenteral infusions, the label states the purpose. Description Sucrose is a white crystalline powder, or lus-trous colorless or white crystals. It is very soluble in water, and practically insoluble in ethanol (99.5).◆ Identification Determine the infrared absorption spec-trum of Sucrose as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ D Optical rotation <2.49> [a]20: ＋66.3 – ＋67.09(26 g, water, 100 mL, ◆100 mm◆). Purity ◆(1) Color value—Dissolve 50.0 g of Sucrose in 50.0 mL of water, filter through a membrane filter with 0.45 mm in pore size, degas, and use this solution as the sample solution. Measure the absorbance of the sample solution at 420 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a cell of at least 4 cm (a cell length of ‌10 cm or more is preferred), and calculate the color value by the following equation: not more than 45. Color value ＝ A × 1000/b/c A: Absorbance measured at 420 nm b: Path length (cm) D c: Concentration (g/mL) of Sucrose in the sample solu-tion, calculated from the refractive index (n20) obtained as directed under Refractive Index Determination <2.45>. Use the following table and interpolate the value, if necessary. n20 c (g/mL) 1.4138 0.570 1.4159 0.585 1.4179 0.600 1.4200 0.615 1.4221 0.630 1.4243 0.645 1.4264 0.661 D System suitability— System repeatability: When the test is repeated 2 times with the sample solution, the difference between 2 results is not larger than 3.◆ Clarity of solution—Dissolve 50.0 g of Sucrose in water to make 100 mL, and use this solution as the sample solution: the sample solution is clear, and its clarity is not different from water, or its opalescence is not more than that of reference suspension 1. Sulfite Enzyme reaction: Sulfite is oxidized by sulfite oxidase to sulfuric acid and hydrogen peroxide which in turn is reduced by nicotinamide adenine dinucleotide peroxidase in the presence of nicotinamide adenine dinucleotide reduced form (NADH). The amount of NADH oxidized is propor-tional to the amount of sulfite. Calculate the amount of oxi-dized NADH from the degree of reduction of the absorbance at 340 nm. A suitable kit may be used. Procedure: Dissolve 4.0 g of Sucrose in freshly pre-pared distilled water to make exactly 10 mL, and use this so-lution as the sample solution. Separately, dissolve 4.0 g of Sucrose in freshly prepared distilled water, add exactly 0.5 mL of Standard Sulfite Solution, then add freshly prepared distilled water to make exactly 10 mL, and use this solution as the standard solution. Use freshly prepared distilled water as a blank. Separately, introduce 2.0 mL each of the sample solution, the standard solution and the blank in 10-mm cells, add 1.00 mL of b-nicotinamide adenine dinucleotide reduced form TS and 10 mL of NADH peroxidase TS, stir with a plastic stirring rod, and allow to stand at 20 – 259C for 5 minutes. Measure the absorbance of these solutions at 340 nm, AT1, AS1 and AB1, as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Then, to these solutions add 50 mL each of sulfite oxidase TS, stir, allow to stand at 20 – 259C for 30 minutes, then measure the absorbance of these solutions in the same manner as above, AT2, AS2 and AB2: the result of (AT1 – AT2) – (AB1 – AB2) is not larger than half the result of (AS1 – AS2) – (AB1 – AB2) (not more than 10 ppm expressed as SO2). Reducing sugars—Transfer 5 mL of the sample solu-tion obtained in (2) to a test-tube about 150 mm long and about 16 mm in diameter, add 5 mL of water, 1.0 mL of 1 mol/L sodium hydroxide VS and 1.0 mL of methylene blue TS, mix, and heat in a water bath. After exactly 2 minutes, take the tube out of the bath, and examine the solution immediately: the blue color does not disappear completely. Ignore any blue color at the air and solution interface. Conductivity <2.51> Dissolve 31.3 g of Sucrose in freshly prepared distilled water to make 100 mL, and use this solu-tion as the sample solution. Measure the conductivity of the sample solution (k1 (mS･cm－1)) while gently stirring with a magnetic stirrer at 20 ± 0.19C. Measure the conductivity of the water used for preparing the sample solution (k2 (mS･cm－1)) in the same manner as above. The measured con-ductivity must be stable within 1z in the rate of change per 30 seconds. Calculate the corrected conductivity of the sam-ple solution (kC) by the following expression: kC is not more than 35 mS･cm－1. kC (mS･cm－1) ＝ k1 － 0.35k2 Loss on drying <2.41> Not more than 0.1z (2 g, 1059C, 3 hours). Dextrins For Sucrose used to prepare parenteral infusions, to 2 mL of the sample solution obtained in the Purity (2) add 8 mL of water, 0.05 mL of 2 mol/L hydrochloric acid and 0.05 mL of iodine TS: the solution remains yellow. Bacterial endotoxins <4.01> Less than 0.25 EU/mg, for Sucrose used to prepare parenteral infusions.
Sulfadiazine Silver, when dried, contains not less than 99.
Sulfadiazine Silver occurs as a white to pale yellow crystalline powder. It is odorless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in ammonia TS. It is gradually colored by light. Melting point: about 2759C (with decomposition).
Determine the infrared absorption spectrum of Sulfadiazine Silver, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Sulfadiazine Silver RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Nitrate—To 250 mL of water add 1.0 g of Sul-fadiazine Silver, shake well for 50 minutes, filter, and use this filtrate as the sample solution. Separately, weigh accu-rately 0.25 g of potassium nitrate, and dissolve in water to make exactly 2000 mL. Pipet 5 mL of this solution and add water to make exactly 200 mL, and use this solution as the standard solution. Pipet 2.0 mL each of the sample solution ‌and standard solution, and add 5 mL of a solution of disodium chromotropate dihydrate in sulfuric acid (1 in 10,000) and sulfuric acid to make exactly 10 mL. Determine the absorbances, AT and AS, of the sample solution and standard solution at 408 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared with exactly 2.0 mL of water in the same manner, as the blank: AT is not larger than AS (not more than 0.05z). (2) Related substances—Dissolve 50 mg of Sulfadiazine Silver in 5 mL of a mixture of ethanol (95) and ammonia so-lution (28) (3:2), and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add a mixture of ethanol (95) and ammonia solution (28) (3:2) to make exactly 20 mL. Pipet 2 mL of this solution, add a mixture of ethanol (95) and ammonia solution (28) (3:2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia solu-tion (28) (10:5:2) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot and spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 809C, 4 hours). Residue on ignition <2.44> 41 – 45z (1 g). Silver content Weigh accurately about 50 mg of Sulfadia-zine Silver, previously dried, dissolve in 2 mL of nitric acid, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the sample solution. Measure accurately a suitable quantity of Standard Silver Solution for Atomic Absorption Spectrophotometry, dilute with water to make solutions con-taining 1.0 to 2.0 mg of silver (Ag:107.87) per mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions, and calculate the silver con-tent of the sample solution from the calibration curve ob-tained from the absorbances of the standard solutions: it contains not less than 28.7z and not more than 30.8z of silver. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A silver hollow cathode lamp. Wavelength: 328.1 nm.
Weigh accurately about 0.1 g each of Sulfadiazine Silver and Sulfadiazine Silver RS, each previously dried, and add ammonia TS to make exactly 100 mL, respectively. Pipet 1 mL each of these solutions, add water to make ex-actly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 255 nm, as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using a solution, prepared with ex-actly 1 mL of ammonia TS and a sufficient water to make exactly 100 mL, as the blank.
Sulfisoxazole occurs as white, crystals or crys-talline powder. It is odorless, and has a slightly bitter taste. It is freely soluble in pyridine and in n-butylamine, soluble in methanol, sparingly soluble in ethanol (95), slightly solu-ble in acetic acid (100), and very slightly soluble in water and in diethyl ether. It dissolves in dilute hydrochloric acid, in sodium hydrox-ide TS and in ammonia TS. It is gradually colored by light.
(1) Dissolve 0.01 g of Sulfisoxazole in 1 mL of dilute hydrochloric acid and 4 mL of water: the solu-tion responds to Qualitative Tests <1.09> for primary aro-matic amines. Dissolve 0.02 g of Sulfisoxazole in 5 mL of water and 1 mL of n-butylamine, add 2 to 3 drops of copper (II) sulfate TS, and shake well. Add 5 mL of chloroform, shake, and al-low to stand: a blue-green color develops in the chloroform layer. Dissolve 0.01 g of Sulfisoxazole in 1 mL of pyridine, add 2 drops of copper (II) sulfate TS, and shake. Add 3 mL of water and 5 mL of chloroform, shake, and allow to stand: a light yellow-brown color develops in the chloroform layer. To 0.5 g of Sulfisoxazole add 2 mL of acetic acid (100), dissolve by heating under a reflux condenser, add 1 mL of acetic anhydride, and boil for 10 minutes. Add 10 mL of water, cool, and alkalize with about 7 mL of a solution of sodium hydroxide (3 in 10). Filter, if necessary, immediately acidify by adding acetic acid (100) dropwise, collect the pro-duced precipitate, recrystallize from methanol, and dry at 1059C for 1 hour: the crystals melt <2.60> between 2089C and 2109C.
(1) Clarity and color of solution—Dissolve 1.0 g of Sulfisoxazole in 5 mL of sodium hydroxide TS and 20 mL of water: the solution is clear and colorless to pale yellow. Acidity—To 1.0 g of Sulfisoxazole add 50 mL of water, warm at 709C for 5 minutes, allow to stand in an ice bath for 1 hour, and filter. To 25 mL of the filtrate add 2 drops of methyl red TS and 0.20 mL of 0.1 mol/L sodium hydroxide VS: a yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of Sulfisox- azole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.5z (2 g, 1059C, ‌hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Sulfisoxazole, previ-ously dried, dissolve in 50 mL of methanol by warming, cool and titrate <2.50> with 0.2 mol/L sodium hydroxide VS (in-dicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner using a mixture of 50 mL of methanol and 18 mL of water, and make any necessary correction. Each mL of 0.2 mol/L sodium hydroxide VS ＝ 53.46 mg of C11H13N3O3S
Sulfamethizole, when dried, contains not less than 99.
Sulfamethizole occurs as white to yellowish white, crystals or crystalline powder. It is odorless. It is slightly soluble in ethanol (95), and in acetic acid (100) and practical insoluble in water and in diethyl ether. It dissolves in dilute hydrochloric acid and in sodium hy-droxide TS. It is gradually colored by light.
Determine the infrared absorption spectrum of Sulfamethizole, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Clarity and color of solution—Dissolve 0.5 g of Sulfamethizole in 3 mL of sodium hydroxide TS and 20 mL of water: the solution is clear and colorless. Acidity—To 1.0 g of Sulfamethizole add 50 mL of water, warm at 709C for 5 minutes, allow to stand for 1 hour in an ice bath, and filter. To 25 mL of the filtrate add 2 drops of methyl red TS and 0.60 mL of 0.1 mol/L sodium hydroxide VS: a yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of Sul- famethizole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sulfamethizole according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Sulfamethi-zole in 10 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetone to make exactly 50 mL, then pipet 5 mL of this solution, add acetone to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer Amount (mg) of sulfadiazine silver (C ＝ MS × AT/AS 10H9AgN4O2S) chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (20:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light MS: Amount (mg) of Sulfadiazine Silver RS taken (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more ‌intense than the spot from the standard solution.‌ Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Sulfamethizole, previously dried, dissolve in 5 mL of hydrochloric acid and 50 mL of water, add 10 mL of a solution of potassium bro-mide (3 in 10), cool below 159C, and titrate <2.50> with 0.1 mol/L sodium nitrite VS according to the potentiometric titration method or the amperometric titration method. Each mL of 0.1 mol/L sodium nitrite VS ＝ 27.03 mg of C9H10N4O2S2
Sulfamethoxazole, when dried, contains not less than 99.
Sulfamethoxazole occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in N,N-dimethylformamide, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, and very slightly soluble in water. It dissolves in sodium hydroxide TS. It is gradually colored by light.
Determine the infrared absorption spectrum of Sulfamethoxazole, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Sulfamethoxazole in 5 mL of sodium hydroxide TS, and add 20 mL of water: the solution is clear and colorless. Acidity—To 1.0 g of Sulfamethoxazole add 50 mL of water, heat at 709C for 5 minutes, allow to stand in ice water for 1 hour, and filter. To 25 mL of the filtrate add 2 drops of methyl red TS and 0.60 mL of 0.1 mol/L sodium hydroxide VS: a yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of Sul- famethoxazole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sulfamethoxazole according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Sulfamethox-azole in 10 mL of a solution of ammonia solution (28) in methanol (1 in 50), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add a solution of ammonia solution (28) in methanol (1 in 50) to make exactly 10 mL. Pipet 1 mL of this solution, add a solution of ammo-nia solution (28) in methanol (1 in 50) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetonitrile and di-luted ammonia solution (28) (7 in 100) (10:8:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Sulfamethoxazole, previously dried, dissolve in 30 mL of N,N-dimethyl-formamide, add 10 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until a light blue color is pro-duced (indicator: 0.5 mL of thymolphthalein TS). Sepa-rately, perform a blank determination in the same manner with a mixture of 30 mL of N,N-dimethylformamide and 26 mL of water, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 25.33 mg of C10H11N3O3S
Sulfamonomethoxine Hydrate occurs as white to pale yellow, crystals, granules or crystalline powder. It is odorless. It is soluble in acetone, slightly soluble in ethanol (95), very slightly soluble in diethyl ether, and practically insolu-ble in water. It dissolves in dilute hydrochloric acid and in sodium hy-droxide TS. ‌It is gradually colored by light.
Determine the infrared absorption spectrum of Sulfamonomethoxine Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Sulfamonomethoxine Hydrate in 5 mL of sodium hydrox-ide TS and 20 mL of water: the solution is clear and colorless to pale yellow. Dissolve 0.5 g of Sulfamonomethoxine Hy-drate in 5 mL of sodium hydroxide TS, and heat: no tur-bidity is produced. After cooling, add 5 mL of acetone: the solution is clear. Heavy metals <1.07>—Proceed with 1.0 g of Sul- famonomethoxine Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sulfamonomethoxine Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.02 g of Sul-famonomethoxine Hydrate in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol and ammonia solution (28) (4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not larger and not more intense than the spot from the standard solution. Loss on drying <2.41> 4.5 – 6.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.10z (1 g).
Weigh accurately about 0.5 g of Sulfamono-methoxine Hydrate, previously dried, dissolve in 5 mL of hy-drochloric acid and 50 mL of water, add 10 mL of a solution of potassium bromide (3 in 10), cool below 159C, and titrate <2.50> with 0.1 mol/L sodium nitrite VS (potentiometric titration or amperometric titration). Each mL of 0.1 mol/L sodium nitrite VS ＝ 28.03 mg of C11H12N4O3S
Salazosulfapyridine occurs as a yellow to yel-low-brown fine powder. It is odorless and tasteless. It is sparingly soluble in pyridine, slightly soluble in ethanol (95), practically insoluble in water, in chloroform and in diethyl ether. It dissolves in sodium hydroxide TS. Melting point: 240 – 2499C (with decomposition).
(1) Dissolve 0.1 g of Salazosulfapyridine in 20 mL of dilute sodium hydroxide TS: a red-brown color de-velops. This color gradually fades upon gradual addition of 0.5 g of sodium hydrosulfite with shaking. Use this solution in the following tests (2) to (4). To 1 mL of the solution obtained in (1) add 40 mL of water, neutralize with 0.1 mol/L hydrochloric acid TS, and add water to make 50 mL. To 5 mL of this solution add 2 to 3 drops of dilute iron (III) chloride TS: a red color develops and changes to purple, then fades when dilute hydrochloric acid is added dropwise. The solution obtained in (1) responds to Qualitative Tests <1.09> for primary aromatic amines. To 1 mL of the solution obtained in (1) add 1 mL of pyridine and 2 drops of copper (II) sulfate TS, and shake. Add 3 mL of water and 5 mL of chloroform, shake, and al-low to stand: a green color develops in the chloroform layer. Determine the absorption spectrum of a solution of Salazosulfapyridine in dilute sodium hydroxide TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths.
(1) Chloride <1.03>—Dissolve 2.0 g of Salazosul-fapyridine in 12 mL of sodium hydroxide TS and 36 mL of water, add 2 mL of nitric acid, shake, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of mL of hydrochloric acid, shake, and filter. To 25 mL of the filtrate add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Salazosulfapyridine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Take 1.0 g of Salazosulfapyridine in a decomposition flask, add 20 mL of nitric acid, and heat gently until it becomes fluid. After cooling, add 5 mL of sul-furic acid, and heat until white fumes are evolved. Add, if necessary, 5 mL of nitric acid after cooling, and heat again. Repeat this operation until the solution becomes colorless to slightly yellow. After cooling, add 15 mL of a saturated solu-tion of ammonium oxalate monohydrate, and heat until white fumes are evolved again. After cooling, add water to make 25 mL. Perform the test with 5 mL of this solution as the test solution: the color of the test solution is not deeper than that of the following color standard. Color standard: Proceed in the same manner without Salazosulfapyridine, transfer 5 mL of the obtained solution to a generator bottle, add exactly 2 mL of Standard Arsenic Solution, and proceed in the same manner as the test solu-tion with this solution (not more than 10 ppm). Related substances—Dissolve 0.20 g of Salazosul-fapyridine in 20 mL of pyridine, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add pyridine to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with diluted methanol (9 in 10) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Salicylic acid—To 0.10 g of Salazosulfapyridine add 15 mL of diethyl ether, and shake vigorously. Add 5 mL of dilute hydrochloric acid, shake vigorously for 3 minutes, col-lect the diethyl ether layer, and filter. To the water layer add 15 mL of diethyl ether, shake vigorously for 3 minutes, col-lect the diethyl ether layer, filter, and combine the filtrates. Wash the residue on the filter paper with a small quantity of diethyl ether, and combine the washings and the filtrate. Evaporate the diethyl ether with the aid of air-stream at room temperature. To the residue add dilute ammonium iron (III) sulfate TS, shake, and filter, if necessary. Wash the residue on the filter paper with a small quantity of dilute am-monium iron (III) sulfate TS, combine the washings and the filtrate, add dilute ammonium iron (III) sulfate TS to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours, dissolve in dilute ammonium iron (III) sulfate TS to make exactly 400 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 535 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: salicylic acid content is not more than 0.5z. Content (z) of salicylic acid (C H O ) 0.01 mol/L hydrochloric acid VS (not more than 0.014z). (2) Sulfate <1.14>—Dissolve 2.0 g of Salazosulfapyridine ＝ MS × AT/AS × 1/20 7 6 3 in 12 mL of sodium hydroxide TS and 36 mL of water, add 2 MS: Amount (mg) of salicylic acid for assay taken ‌Loss on drying <2.41> Not more than 2.0z (1 g, 1059C, 4 hours).‌ Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 20 mg of Salazosulfapyri-dine, previously dried, and perform the test as directed in the procedure of determination for sulfur under Oxygen Flask Combustion Method <1.06>, using 10 mL of diluted hydro-gen peroxide (30) (1 in 40) as an absorbing liquid. Each mL of 0.005 mol/L barium perchlorate VS ＝ 1.992 mg of C18H14N4O5S
Sulfur and Camphor Lotion is a light yellow suspension. A part of the components separates out on standing.
(1) To 5 mL of well shaken Sulfur and Camphor Lotion add 25 mL of water, and centrifuge [use this supernatant liquid for test (3)]. To 0.02 g of the precipi-tate add 2 mL of pyridine and 0.2 mL of sodium hydrogen carbonate TS, and boil: a blue color develops (sulfur). To 10 mL of well shaken Sulfur and Camphor Lotion add 5 mL of diethyl ether, and mix. Separate the diethyl ether layer, and filter through a pledget of cotton. Wash the cotton with a small portion of diethyl ether, combine the washings with the filtrate, and distil cautiously on a water bath to remove the diethyl ether. Dissolve the residue in 1 mL of methanol, add 1 mL of 2,4-dinitrophenylhydrazine TS, and heat for about 2 minutes on a water bath. Cool, dilute with water to make about 5 mL, and allow to stand. Filter the produced precipitate through a glass filter (G4), and wash the residue on the filter with water until the last washing is colorless. Dissolve the residue in 10 mL of ethanol (95), add 5 mL of sodium hydroxide TS, and allow to stand for 2 minutes: a red color develops (d-camphor or dl-cam-phor). The supernatant liquid obtained in (1) responds to Qualitative Tests <1.09> (2) and (3) for calcium salt.
Sulfur, when dried, contains not less than 99.
Sulfur occurs as a light yellow to yellow pow-der. It is odorless and tasteless. It is freely soluble in carbon disulfide, and practically in-soluble in water, in ethanol (95) and in diethyl ether.
(1) Ignite Sulfur: it burns with a blue flame and gives a pungent odor of sulfur dioxide. Dissolve 5 mg of Sulfur in 5 mL of sodium hydroxide TS by heating in a water bath, cool, and add 1 drop of sodi- um pentacyanonitrosylferrate (III) TS: a blue-purple color develops. Boil 1 mg of Sulfur with 2 mL of pyridine and 0.2 mL of sodium hydrogen carbonate TS: a blue color devel-ops.
(1) Clarity of solution—Dissolve 1.0 g of Sulfur in a mixture of 20 mL of a solution of sodium hydroxide (1 in 6) and 2 mL of ethanol (95) by boiling: the solution is clear. Dissolve 2.0 g of Sulfur in 10 mL of carbon disulfide: the so-lution is almost clear or slightly opalescent. Acidity or alkalinity—Shake 2.0 g of Sulfur with 50 mL of freshly boiled and cooled water, and add 2 drops of phenolphthalein TS: no red color develops. Further add 1.0 mL of 0.1 mol/L sodium hydroxide VS: a red color devel-ops. Arsenic <1.11>—Prepare the test solution with 0.20 g of Sulfur according to Method 3, and perform the test (not more than 10 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, not more than 0.67 kPa, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.4 g of Sulfur, previously dried, dissolve in 20 mL of potassium hydroxide-ethanol TS and 10 mL of water by boiling, cool, and add water to make exactly 100 mL. Transfer exactly 25 mL of the solution to a 400-mL beaker, add 50 mL of hydrogen peroxide TS, and heat on a water bath for 1 hour. Acidify the solution with dilute hydrochloric acid, add 200 mL of water, heat to boil, add hot barium chloride TS dropwise until no more precipi-tate is formed, and heat on a water bath for 1 hour. Collect the precipitate, and wash with water until the last washing shows no opalescence with silver nitrate TS. Dry the precipi-tate, heat strongly to constant mass, and weigh as barium sulfate (BaSO4: 233.39). Perform a blank determination in the same manner, and make any necessary correction. Amount (mg) of sulfur (S) ＝ amount (mg) of barium sulfate (BaSO4) × 0.13739
Sulindac, when dried, contains not less than 99.
Sulindac occurs as a yellow crystalline powder. It is sparingly soluble in methanol and in ethanol (99.5), and practically insoluble in water. A solution of Sulindac in methanol (1 in 100) shows no optical rotation. Melting point: about 1849C (with decomposition).
(1) Dissolve 15 mg of Sulindac in 1000 mL of a solution of hydrochloric acid in methanol (1 in 120). De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24> and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Sulin-dac as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Sulindac according to Method 2 and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sulindac according to Method 3 and perform the test (not more than 2 ppm). Related substances—Dissolve 0.25 g of Sulindac in 10 mL of methanol and use this solution as the sample solution. Pipet 1 mL of the sample solution and add methanol to make exactly 100 mL. Pipet 5 mL, 4 mL and 2 mL of this solution, to each add methanol to make exactly 10 mL, and ‌use these solutions as the standard solutions (1), (2) and (3). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 4 mL each of the sample solution, standard solution (1), (2) and (3) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (97:3) to a dis-tance of about 17 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion (1), and the total intensity of spots other than the princi-pal spot from the sample solution is not more than 1.0z cal-culated on the basis of intensities of the spots from the standard solution (1), (2) and (3).‌ Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum not exceeding 0.7 kPa, 1009C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.3 g of Sulindac, previ-ously dried, dissolve in 50 mL of methanol and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titra-tion). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Purity (1) Clarity and color of solution—Dissolve 2.0 g of Sulpiride in 7 mL of dilute acetic acid, and add water to make 20 mL: the solution is clear. Perform the test with the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank: the absorbance at a wavelength of 450 nm does not exceed 0.020. Heavy metals <1.07>—Proceed with 2.0 g of Sulpiride as directed under Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Sulpiride in 10 mL of methanol, and use this solution as the sample solu-tion. Dilute 1 mL of the sample solution, accurately meas-ured, with methanol to make exactly 100 mL. Dilute 2 mL of this solution, accurately measured, with methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: ＝ 35.64 mg of C 20H 17FO3S 254 nm): the number of the spots other than the principal spot obtained from the sample solution is not more than 2,
Sulpiride, when dried, contains not less than 98.
Sulpiride is a white crystalline powder. It is freely soluble in acetic acid (100) and in dilute acetic acid, sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It is soluble in 0.05 mol/L sulfuric acid TS. A solution of Sulpiride in methanol (1 in 100) shows no optical rotation. Melting point: about 1789C (with decomposition).
(1) Dissolve 0.1 g of Sulpiride in 0.05 mol/L sulfuric acid TS to make 100 mL. Dilute 5 mL of the solution with water to make 100 mL. Determine the absorp-tion spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Sulpi-ride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spec-
Dissolve about 0.4 g of Sulpiride, previously dried and accurately weighed, in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS) until the color of the solution changes from violet through blue to bluish green. Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 34.14 mg of C H N O S 15 23 3 4 Description Sulpiride is a white crystalline powder. It is freely soluble in acetic acid (100) and in dilute acetic acid, sparingly soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It is soluble in 0.05 mol/L sulfuric acid TS. A solution of Sulpiride in methanol (1 in 100) shows no optical rotation. Melting point: about 1789C (with decomposition). Identification (1) Dissolve 0.1 g of Sulpiride in 0.05 mol/L sulfuric acid TS to make 100 mL. Dilute 5 mL of the solution with water to make 100 mL. Determine the absorp-tion spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Sulpi-ride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spec-
Sulpyrine Hydrate occurs as white to light yel-low, crystals or crystalline powder. It is odorless, and has a bitter taste. It is very soluble in water, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. It is colored by light.
(1) Add 2 drops of dilute sulfuric acid and 1 mL of chlorinated lime TS to 3 mL of a solution of Sulpy-rine Hydrate (1 in 15): a deep blue color develops at first, but the color immediately turns red, then gradually changes to yellow. Boil 5 mL of a solution of Sulpyrine Hydrate (1 in 25) with 3 mL of dilute hydrochloric acid: the odor of sulfur dioxide is perceptible at first, and on further boiling, the odor of formaldehyde is perceptible. A solution of Sulpyrine Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity of solution, and acidity or alkalinity—Dissolve 1.0 g of Sulpyrine Hydrate in 10 mL of water: the solution is clear and neutral. Sulfate <1.14>—Dissolve 0.20 g of Sulpyrine Hydrate in 0.05 mol/L hydrochloric acid VS to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS and 0.05 mol/L hydrochloric acid VS to make 50 mL (not more than 0.120z). Heavy metals <1.07>—Proceed with 1.0 g of Sulpy- rine Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Merbuline—Transfer 0.10 g of Sulpyrine Hydrate with 2 mL of water and 1 mL of dilute sulfuric acid into a flask, cover with a funnel, and boil gently for 15 minutes. Cool, add 2 mL of a solution of sodium acetate trihydrate (1 in 2) and water to make 5 mL, shake this solution with 5 mL of benzaldehyde-saturated solution, and allow to stand for 5 minutes: the solution is clear. Chloroform-soluble substances—Mix, by frequent shaking, 1.0 g of Sulpyrine Hydrate and 10 mL of chlo-roform for 30 minutes. Collect the precipitate, wash with two 5-mL portions of chloroform, combine the washings with the filtrate, and evaporate on a water bath to dryness. Dry the residue at 1059C for 4 hours: the mass of the residue is not more than 5.0 mg. Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, Each mL of 0.05 mol/L iodine VS ＝ 16.67 mg of C13H16N3NaO4S
Weigh accurately about 0.25 g of Sulpyrine Hy-drate, dissolve in 100 mL of diluted hydrochloric acid (1 in 20), previously cooled below 109C. Titrate <2.50> immedi-ately with 0.05 mol/L iodine VS while keeping the tempera-ture between 59C and 109C, until the color of the solution remains blue upon shaking vigorously for 1 minute after the addition of 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). C13H16N3NaO4S.H2O: 351.35 Monosodium [(1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazol- 4-yl)(methyl)amino]methanesulfonate monohydrate [5907-38-0] Sulpyrine Hydrate contains not less than 98.5z of sulpyrine (C13H16N3NaO4S: 333.34), calculated on the dried basis. Description Sulpyrine Hydrate occurs as white to light yel-low, crystals or crystalline powder. It is odorless, and has a bitter taste. It is very soluble in water, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. It is colored by light. Identification (1) Add 2 drops of dilute sulfuric acid and 1 mL of chlorinated lime TS to 3 mL of a solution of Sulpy-rine Hydrate (1 in 15): a deep blue color develops at first, but the color immediately turns red, then gradually changes to yellow. Boil 5 mL of a solution of Sulpyrine Hydrate (1 in 25) with 3 mL of dilute hydrochloric acid: the odor of sulfur dioxide is perceptible at first, and on further boiling, the odor of formaldehyde is perceptible. A solution of Sulpyrine Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. Purity (1) Clarity of solution, and acidity or alkalinity—Dissolve 1.0 g of Sulpyrine Hydrate in 10 mL of water: the solution is clear and neutral. Sulfate <1.14>—Dissolve 0.20 g of Sulpyrine Hydrate in 0.05 mol/L hydrochloric acid VS to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS and 0.05 mol/L hydrochloric acid VS to make 50 mL (not more than 0.120z). Heavy metals <1.07>—Proceed with 1.0 g of Sulpy- rine Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Merbuline—Transfer 0.10 g of Sulpyrine Hydrate with 2 mL of water and 1 mL of dilute sulfuric acid into a flask, cover with a funnel, and boil gently for 15 minutes. Cool, add 2 mL of a solution of sodium acetate trihydrate (1 in 2) and water to make 5 mL, shake this solution with 5 mL of benzaldehyde-saturated solution, and allow to stand for 5 minutes: the solution is clear. Chloroform-soluble substances—Mix, by frequent shaking, 1.0 g of Sulpyrine Hydrate and 10 mL of chlo-roform for 30 minutes. Collect the precipitate, wash with two 5-mL portions of chloroform, combine the washings with the filtrate, and evaporate on a water bath to dryness. Dry the residue at 1059C for 4 hours: the mass of the residue is not more than 5.0 mg. Loss on drying <2.41> Not more than 6.0z (1 g, 1059C, Each mL of 0.05 mol/L iodine VS ＝ 16.67 mg of C13H16N3NaO4S
スルタミシリントシル酸塩水和物
Sultamicillin Tosilate Hydrate contains not less than 698 mg (potency) and not more than 800 mg (potency) per mg, calculated on the anhydrous and residual sol-vent-free basis.
Sultamicillin Tosilate Hydrate occurs as a white to yellowish white crystalline powder. It is freely soluble in acetonitrile, in methanol and in ethanol (99.5), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Sultamicillin Tosilate Hydrate in methanol (1 in 1000) as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Sultamicillin Tosi-late RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Sul-tamicillin Tosilate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Sultamicillin Tosilate RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋173 – ＋1879(0.5 g calcu-lated on the anhydrous bases, a mixture of water and aceto- nitrile (3:2), 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Sultamicillin Tosilate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL Standard Lead Solution (not more than 20 ppm). Ampicillin—Perform the procedure rapidly. Weigh accurately about 20 mg of Sultamicillin Tosilate Hydrate, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Ampicillin RS, equivalent to about 20 mg (potency), dissolve in the mobile phase to make exactly 100 mL. Pipet 6 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak area of ampicillin by the au-tomatic integration method: the peak area obtained from the sample solution is not larger than that from the standard so-lution. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 3.12 g of sodium dihydrogen phos-phate dihydrate in about 750 mL of water, adjust to pH 3.0 with diluted phosphoric acid (1 in 10), and add water to make 1000 mL. To 80 mL of acetonitrile for liquid chroma-tography add this solution to make 1000 mL. Flow rate: Adjust so that the retention time of ampicillin is about 14 minutes. System suitability— System performance: Dissolve 12 mg of Ampicillin RS, 4 mg of Sulbactam RS and 4 mg of p-toluenesulfonic acid monohydrate in 1000 mL of the mobile phase. When the procedure is run with 25 mL of this solution under the above operating conditions, sulbactam, p-toluenesulfonic acid and ampicillin are eluted in this order, and the resolutions be-tween these peaks are not less than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ampicillin is not more than 2.0z. Sulbactam—Perform the procedure rapidly. Weigh accurately about 20 mg of Sultamicillin Tosilate Hydrate, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Sulbactam RS, equivalent to about 20 mg (potency), dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak area of sulbactam of each solution by the automatic integration method: the peak area obtained from the sample solution is not larger than that from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (2). System suitability— Proceed as directed in the system suitability in the Purity (2). Penicilloic acids—Weigh accurately about 25 mg of Sultamicillin Tosilate Hydrate, dissolve in 1 mL of aceto-nitrile, and add 25 mL of 0.02 mol/L phosphate buffer solu- ‌tion (pH 3.0) in a 100-mL glass-stoppered flask. Add exactly 5 mL of 0.005 mol/L iodine VS, and allow to stand the stop- that of the internal standard in each solution. Amount [mg (potency)] of sultamicillin (C H N O S ) pered flask for 5 minutes. Titrate <2.50> with 0.005 mol/L sodium thiosulfate VS (indicator: 1.0 mL of starch TS). Per- ＝ MS × QT/QS × 1000 25 30 4 9 2 form a blank determination in the same manner, and make any necessary correction. Calculate the amount of penicilloic acid (C25H34N4O11S2: 630.69) by using the following equa-tion: it is not more than 3.0z. Each mL of 0.005 mol/L sodium thiosulfate VS ＝ 0.2585 mg of C25H34N4O11S2 Residual solvent <2.46>—Weigh accurately about 0.1 g of Sultamicillin Tosilate Hydrate, dissolve in 2 mL of methanol, add water to make exactly 20 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 1 g of ethyl acetate, and mix with water to make ex-actly 200 mL. Pipet 2 mL of this solution, add 10 mL of methanol, then add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions, and determine the peak areas, AT and AS, of ethyl acetate in each solution. Calculate the amount of ethyl acetate by the following equation: not more than 2.0z. Amount (z) of ethyl acetate ＝ MS/MT × AT/AS × 1/5 MS: Amount (mg) of ethyl acetate taken MT: Amount (mg) of the Sultamicillin Tosilate Hydrate taken Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A column 3 mm in inside diameter and 1 m in length, packed with porous styrene-divinylbenzene copoly-mer for gas chromatography (average pore diameter: 0.0085 mm, 300 – 400 m2/g) (150 to 180 mm in particle diameter). Column temperature: A constant temperature of about 1559C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethyl ace-tate is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of ethyl acetate are not less than 500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ethyl acetate is not more than 5z. Water <2.48> 4.0 – 6.0z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Perform the procedure rapidly. Weigh accurately an amount of Sultamicillin Tosilate Hydrate and Sultamicillin Tosilate RS, equivalent to about 50 mg (potency), dissolve each in the mobile phase to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of sultamicillin to MS: Amount [mg (potency)] of Sultamicillin Tosilate RS taken Internal standard solution—A solution of isopropyl-4-aminobenzoate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 3.12 g of sodium dihydrogen-phosphate in about 750 mL of water, adjust to pH 3.0 with diluted phosphoric acid (1 in 10), and add water to make 1000 mL. To 400 mL of acetonitrile for liquid chromatogra-phy add this solution to make 1000 mL. Flow rate: Adjust so that the retention time of sultamicil-lin is about 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, p-toluenesulfonic acid, sultamicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sultamicillin is not more than 2.0z.
Sultiame occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste. It is very soluble in N,N-dimethylformamide, freely solu-ble in n-butylamine, slightly soluble in methanol and in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether. It dissolves in sodium hydroxide TS.
(1) Dissolve 0.02 g of Sultiame in 5 mL of water and 1 mL of n-butylamine, add 2 to 3 drops of copper (II) sulfate TS, and shake well. To this solution add 5 mL of chloroform, shake, and allow to stand: a green color devel-ops in the chloroform layer. Mix 0.1 g of Sultiame with 0.5 g of sodium carbonate decahydrate, and melt carefully: the gas evolved changes moistened red litmus paper to blue. After cooling, crush the fused substance with a glass rod, stir with 10 mL of water, and filter. To 4 mL of the filtrate add 2 drops of hydrogen peroxide (30), 5 mL of diluted hydrochloric acid (1 in 5) and 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Sultiame according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sultiame according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Sultiame in methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of sulfanilamide in methanol to make exactly 100 mL. Pipet 10 mL of this so-lution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia solution (28) (30:8:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Chloride <1.03>—Dissolve 1.0 g of Sultiame in 20 mL of sodium hydroxide TS by warming, cool, and add 2 mL of acetic acid (100) and water to make 100 mL. After shaking, filter, and discard the first 10 mL of the filtrate. To 40 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 8 mL of sodium hydroxide TS, 0.8 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.022z). (2) Sulfate <1.14>—Dissolve 1.0 g of Sultiame in 20 mL of sodium hydroxide TS by warming, cool, and add 8 mL of dilute hydrochloric acid and water to make 100 mL. After shaking, filter, and discard the first 10 mL of the filtrate. To 40 mL of the subsequent filtrate add 1 mL of dilute hydro-chloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 8 mL of sodium hydroxide TS, 4.2 mL of dilute hy-drochloric acid and water to make 50 mL (not more than
Weigh accurately about 0.8 g of Sultiame, previ-ously dried, dissolve in 70 mL of N,N-dimethylformamide, and titrate <2.50> with 0.2 mol/L tetramethylammonium hy-droxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.2 mol/L tetramethylammonium hydroxide VS 2 to 3 drops of barium chloride TS: a white precipitate is formed. ＝ 58.07 mg of C10H14 N2O4S2 Determine the absorption spectrum of a solution of Sultiame in methanol (1 in 100,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Melting point <2.60> 185 – 1889C Purity (1) Chloride <1.03>—Dissolve 1.0 g of Sultiame in 20 mL of sodium hydroxide TS by warming, cool, and add 2 mL of acetic acid (100) and water to make 100 mL. After shaking, filter, and discard the first 10 mL of the filtrate. To 40 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 8 mL of sodium hydroxide TS, 0.8 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.022z). (2) Sulfate <1.14>—Dissolve 1.0 g of Sultiame in 20 mL of sodium hydroxide TS by warming, cool, and add 8 mL of dilute hydrochloric acid and water to make 100 mL. After shaking, filter, and discard the first 10 mL of the filtrate. To 40 mL of the subsequent filtrate add 1 mL of dilute hydro-chloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.40 mL of 0.005 mol/L sulfuric acid VS add 8 mL of sodium hydroxide TS, 4.2 mL of dilute hy-drochloric acid and water to make 50 mL (not more than
Suxamethonium Chloride Hydrate contains not less than 98.
Suxamethonium Chloride Hydrate occurs as a white crystalline powder. It is freely soluble in water, in methanol and in acetic acid (100), slightly soluble in ethanol (95), very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether.
(1) Determine the infrared absorption spec- ‌trum of Suxamethonium Chloride Hydrate as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. (2) A solution of Suxamethonium Chloride Hydrate (1 in 20) responds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution of 0.1 g of Suxamethoni-um Chloride Hydrate in 10 mL of water is between 4.0 and 5.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Suxamethonium Chloride Hydrate in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve 0.25 g of Suxamethoni-um Chloride Hydrate in 5 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of a solution of ammonium acetate (1 in 100), acetone, 1-butanol and formic acid (20:20:20:1) to a distance of about 10 cm, and dry the plate at 1059C for 15 minutes. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate, and allow to stand for 15 minutes: the spots other than the principal spot from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 8.0 – 10.0z (0.4 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Suxamethonium Chloride Hydrate, dissolve in 80 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Each mL of 0.1 mol/L perchloric acid VS the sample solution. Separately, dissolve 0.05 g of sux-amethonium chloride for thin-later chromatography in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of a solution of ammonium acetate (1 in 100), acetone, 1-butanol and formic acid (20:20:20:1) to a distance of about 10 cm, and dry the plate at 1059C for 15 minutes. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate: the spots obtained from the sample solution and standard solution are blue-purple in color and have simi-lar Rf value. pH <2.54> 3.0 – 5.0 Purity Hydrolysis products—Perform the preliminary neutralization with 0.1 mol/L sodium hydroxide VS in the Assay: not more than 0.7 mL of 0.1 mol/L sodium hydrox-ide VS is required for each 200 mg of Suxamethonium Chlo-ride (C14H30Cl2N2O4) taken. Bacterial endotoxins <4.01> Less than 2.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Transfer to a separator an accurately measured volume of Suxamethonium Chloride Injection, equivalent to about 0.2 g of suxamethonium chloride (C14H30Cl2N2O4), add 30 mL of freshly boiled and cooled water, and wash the solution with five 20-mL portions of diethyl ether. Combine the diethyl ether washings, and extract the combined diethyl ether layer with two 10-mL portions of freshly boiled and cooled water. Wash the combined water extracts with two 10-mL portions of diethyl ether. Combine the solution and ＝ 18.07 mg of C14H30 Cl2N2O4 the water extracts, add 2 drops of bromothymol blue TS, and neutralize with 0.1 mol/L sodium hydroxide VS. Add
Tacalcitol Hydrate contains not less than 97.
Tacalcitol Hydrate occurs as white, crystals or crystalline powder. It is very soluble in methanol and in ethanol (99.5), and practically insoluble in water. It decomposes on exposure to light. Melting point: about 1009C. Place Tacalcitol Hydrate in a capillary tube and immediately flame-seal, put the tube in a bath heated at a temperature of about 109C below the predicted melting point, then start the determination by ris-ing the temperature at the rate of 19C per minute.
(1) Determine the absorption spectrum of a solution of Tacalcitol Hydrate in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Tacalcitol RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tacalcitol Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Tacalcitol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋58 – ＋639(25 mg calcu-lated on the anhydrous basis, ethanol (99.5), 5 mL, 100 mm).
(1) 1a,24(S)-Dihydroxycolecalciferol — Conduct this procedure avoiding contact to the air as possible and using light-resistant vessels. Dissolve 1 mg of Tacalcitol Hydrate in 20 mL of methanol, and use this solution as the sample solution. Perform the test with 30 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak area of tacalcitol, Aa, and the area of a peak, having the rel-ative retention time of about 1.1 to tacalcitol, Ab, by the au-tomatic integration method: Ab/(Aa ＋ Ab) is not more than 0.02. ＝ 18.07 mg of C14H30 Cl2N2O4 Operating conditions— Detector: An ultraviolet absorption photometer (wave-
Conduct this procedure avoiding contact to the air as possible and using light-resistant vessels. Weigh accurately about 1 mg each of Tacalcitol Hydrate and Tacalcitol RS (separately determine the water <2.48> in the same manner as Tacalcitol Hydrate), and dissolve each in methanol to make exactly 50 mL. Pipet 5 mL each of these solutions, add methanol to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 40 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak area, AT and AS, of tacalcitol in each so-lution. Amount (mg) of tacalcitol (C27H44O3) ＝ MS × AT/AS MS: Amount (mg) of Tacalcitol RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile and water (3:1). Flow rate: Adjust so that the retention time of tacalcitol is about 10 minutes. System suitability— System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of tacalcitol are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tacalcitol is not more than 1.0z.
Containers—Hermetic containers. length: 265 nm).
Tacalcitol Lotion contains not less than 90.
Perform the test with 30 mL each of the sam-ple solution and standard solution, both are obtained in the Assay, as directed under Liquid Chromatography <2.01> ac-cording to the following conditions: the retention time of the principal peaks in the chromatograms obtained from the sample solution and standard solution is the same, and both adsorption spectra of these peaks exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: A photodiode array detector (wavelength: 265 nm; spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay.
Weigh accurately an amount of Tacalcitol Lotion, equivalent to about 2 mg of tacalcitol (C27H44O3), add exactly 4 mL of methanol and exactly 1 mL of the internal standard solution, and shake. Add 5 mL of hexane, shake thoroughly for 30 minutes, centrifuge at 49C, filter the lower layer through a membrane filter with a pore size not exceeding 0.2 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 1 mg of Tacalcitol RS (separately de-termine the water <2.48> in the same manner as Tacalcitol Hydrate), and dissolve in methanol to make exactly 20 mL. Pipet 1 mL of this solution, and add methanol to make ex-actly 100 mL. Pipet 4 mL of this solution, add exactly 1 mL of the internal standard solution, shake, add 5 mL of hexane and shake well for 30 minutes, then centrifuge at 49C, filter the lower layer through a membrane filter with a pore size ‌not exceeding 0.2 mm, and use the filtrate as the standard so-lution. Perform the test with 30 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tacalcitol to that of the internal standard. Amount (mg) of tacalcitol (C27H44O3) ＝ MS × QT/QS × 2 MS: Amount (mg) of Tacalcitol RS taken, calculated on the anhydrous basis Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (3 in 2,500,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of acetonitrile for liquid chroma-tography and diluted 0.25 mol/L acetic acid TS (1 in 10) (13:7). Flow rate: Adjust so that the retention time of tacalcitol is about 18 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the internal standard and tacalcitol are eluted in this order with the resolution between these peaks being not less than 14. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tacalcitol to that of the internal standard is not more than 2.0z.
Tacrolimus Hydrate contains not less than 98.
Tacrolimus Hydrate occurs as white, crystals or crystalline powder. It is very soluble in methanol and in ethanol (99.5), freely soluble in N,N-dimethylformamide and in ethanol (95), and practically insoluble in water.
(1) Dissolve 5 mg of Tacrolimus Hydrate in 1 mL of ethanol (95), add 1 mL of 1,3-dinitrobenzene TS and 1 mL of sodium hydroxide TS, and shake: a red-purple color develops. (2) Determine the infrared absorption spectrum of Tacrolimus Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum or the spectrum of Tacrolimus RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]25: －112 – －1179(0.2 g calcu-lated on the anhydrous basis, N,N-dimethylformamide, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Tacrolimus Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> 1.9 – 2.5z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer Being specified separately when the drug is granted approval based on the Law.
Weigh accurately about 25 mg each of Tacrolimus Hydrate and Tacrolimus RS (separately determine the water <2.48> in the same manner as Tacrolimus Hydrate) and dis- ‌solve each in 15 mL of ethanol (99.5), to each add exactly 10 mL of the internal standard solution, add 25 mL of water, allow to stand for 6 hours, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tacrolimus to that of the internal standard. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Tacrolimus Capsules add exactly 3V/5 mL of the internal standard solution, then add ethanol (99.5) to make V mL so that each mL contains about 0.1 mg of tacrolimus (C44H69NO12), and sonicate for 10 minutes with occasional shaking. Centrifuge this solution, take 2 mL of the supernatant liquid, add 2 mL of water, allow to stand 6 Amount (mg) of tacrolimus (C 44H69NO12 ) ＝ M S × QT/QS hours, and use this solution as the sample solution. Sepa-rately, weigh accurately about 25 mg of Tacrolimus RS MS: Amount (mg) of Tacrolimus RS taken, calculated on the anhydrous basis Internal standard solution—A solution of heptyl parahy-droxybenzoate in ethanol (99.5) (3 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, 2-propanol for liquid chromatography and tetrahydrofuran for liquid chromatog-raphy (5:2:2). Flow rate: Adjust so that the retention time of tacrolimus is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, tacrolimus and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tacrolimus to that of internal standard is not more than 1.0z.
Talampicillin Hydrochloride is the hydrochloride of ampicillin phthalidyl ester. It contains not less than 600 mg (potency) and not more than 700 mg (potency) per mg, calculated on the anhydrous basis.
Talampicillin Hydrochloride occurs as a white to light yellow-white powder. It is very soluble in methanol, and freely soluble in water and in ethanol (99.5).
(1) To 1 mL of a solution of Talampicillin Hydrochloride (1 in 30) add 1 mL of sodium hydroxide TS, mix, allow to stand for 5 minutes, and add 2 mL of dilute sulfuric acid and 2 to 3 drops of 2,4-dinitrophenylhydrazine TS: an orange-yellow precipitate is formed. Determine the infrared absorption spectrum of Talampicillin Hydrochloride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Talampicillin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. To 10 mL of a solution of Talampicillin Hydrochlo-ride (1 in 300) add 1 mL of dilute nitric acid, and add silver nitrate TS: a white precipitate is formed. D Optical rotation <2.49> [a]20: ＋151 – ＋1719(0.2 g calcu-lated on the anhydrous basis, ethanol (99.5), 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Talampicillin Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Talampicillin Hydrochloride according to Method 4, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Talampicillin Hydrochloride in ethanol (99.5) to make exactly 10 mL, and use this solution as the sample solution. Pipet 1 mL, 2 mL and 3 mL of the sample solution, add ethanol (99.5) to each to make exactly 100 mL, and use these solutions as the standard solution (1), the standard solution (2) and the standard solution (3), respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of tetrahydrofuran, ethyl acetate, water and ethanol (95) (4:4:2:1) to a distance of about 13 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (99.5) (1 in 500) on the plate, and heat the plate at 1109C for 5 minutes: the spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution (3), and the total of the amount of each spot other than the principal spot from the sample solution, which is calculated by the comparison with the spots from the standard solutions (1), (2) and (3), is not more than 5z. 2-Formylbenzoic acid—Dissolve 50 mg of Talam-picillin Hydrochloride in ethanol (99.5) to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of 2-formylbenzoic acid in ethanol (99.5) to make exactly 100 mL. Pipet 5 mL of this solution, add ethanol (99.5) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of chloroform and acetic acid (100) (4:1) to a distance of about 13 cm, and air-dry the plate. Spray evenly a solution of 2,4-dinitrophenylhydrazine in diluted sulfuric acid (6 in 25) (1 in 500): the spot of 2-formylbenzoic acid obtained from the sample solution is not more intense than that from the standard solution. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Talampicillin Hy-drochloride and Talampicillin Hydrochloride RS, equivalent to about 20 mg (potency), dissolve in water to make exactly 20 mL each, and use these solutions as the sample solution and the standard solution. The standard solution should be prepared before use. Pipet 2 mL each of the sample solution and standard solution in separate 100-mL glass-stoppered flasks, add 2.0 mL of sodium hydroxide TS, and allow them to stand for exactly 15 minutes. Add 2.0 mL of diluted hy-drochloric acid (1 in 10) and exactly 10 mL of 0.005 mol/L iodine VS, allow them to stand for exactly 15 minutes, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS until the color of the solution is disappeared. If necessary, add 0.2 to 0.5 mL of starch TS. Separately, pipet 2 mL each of the sample solution and the standard solution in separate 100-mL glass-stoppered flasks, add exactly 10 mL of 0.005 mol/L iodine VS, titrate <2.50> with 0.01 mol/L sodium thiosulfate VS until the color of the solution is disappeared, and make any necessary correction. For this titration, add 0.2 to 0.5 mL of starch TS, if necessary. Calculate the ‌amount (mL) of 0.005 mol/L iodine VS, VT and VS, con-sumed by the sample solution and the standard solution, re-spectively. the original mass, and filter. Centrifuge, if necessary, until the filtrate becomes clear. Evaporate 20 mL of the filtrate to dryness, and dry the residue at 1059C for 1 hour: the mass of Amount [ mg (potency)] of ampicillin (C H N O S) the residue is not more than 4.0 mg.◆ ＝ MS × VT/VS × 1000 16 19 3 4 Iron—Weigh accurately about 10 g of Talc, add 50 mL of 0.5 mol/L hydrochloric acid TS gently while stirring, MS: Amount [mg (potency)] of Talampicillin Hydrochlo-ride RS taken
Taltirelin Hydrate contains not less than 98.
Taltirelin Hydrate occurs as white, crystals or crystalline powder. It is freely soluble in water, in ethanol (99.5) and in acetic acid (100), and soluble in methanol. It dissolves in 1 mol/L hydrochloric acid TS. It shows crystal polymorphism.
(1) Dissolve 30 mg of Taltirelin Hydrate in 10 mL of water. To 0.5 mL of this solution add 2 mL of a solution of 4-nitrobenzenediazonium fluoroborate (1 in 2000) and 3 mL of boric acid-potassium chloride-sodium hy-droxide buffer solution (pH 9.0): a red color is produced. (2) Determine the infrared absorption spectrum of Tal-tirelin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －22.5 – －24.59(1 g calcu-lated on the anhydrous basis, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Taltirelin Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 10 mg of Taltirelin Hy-drate in 20 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 20 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calcu-late their amounts by the area percentage method: the amount of the peak other than taltirelin is not more than 0.1 z, and the total amount of the peaks other than taltirelin is not more than 0.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos- ‌phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of taltirelin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of taltirelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of taltirelin is not more than 2.0z. Water <2.48> 14.0 – 15.5z (0.2 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Taltirelin Hydrate, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of solution changes from violet through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.54 mg of C17H23N7O5
Tamoxifen Citrate, when dried, contains not less than 99.
Tamoxifen Citrate occurs as a white crystalline powder. It is freely soluble in acetic acid (100), sparingly soluble in methanol, and slightly soluble in water and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Tamoxifen Citrate in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tamoxifen Citrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Tamoxifen Citrate (1 in 100) responds to Qualitative Tests <1.09> (1) for citrate.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Tamoxifen Citrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Conduct this procedure rapidly, using light-resistant vessels. Dissolve 15 mg of Tamoxifen Citrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than tamoxifen obtained from the sample solution is not larger than 3/10 times the peak area of tamoxifen from the standard solution, and the total area of the peaks other than tamoxifen from the sample solution is not larger than 4/5 times the peak area of tamoxifen from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.8 g of N,N-dimethyl-n-octyla-mine in 1000 mL of water. Separately, dissolve 0.9 g of so-dium dihydrogen phosphate dihydrate in 1000 mL of water. Mix these solutions, and adjust to pH 3.0 with phosphoric acid. To 600 mL of this solution add 400 mL of acetonitrile. Flow rate: Adjust so that the retention time of tamoxifen is about 21 minutes. Time span of measurement: About 2.5 times as long as the retention time of tamoxifen, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of tamoxifen obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of tamoxifen are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tamoxifen is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Tamoxifen Citrate, previously dried, dissolve in 150 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentio-metric titration). Perform the blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 56.36 mg of C26H29NO.C6H8O7
Tamsulosin Hydrochloride occurs as white crystals. It is freely soluble in formic acid, sparingly soluble in water, slightly soluble in acetic acid (100), and very slightly soluble in ethanol (99.5). Melting point: about 2309C (with decomposition).
(1) Determine the absorption spectrum of a solution of Tamsulosin Hydrochloride (3 in 160,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tam-sulosin Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of an ice cooled solution of Tamsulosin Hy-drochloride (3 in 400) add 3 mL of dilute nitric acid, shake well, allow to stand at room temperature for 30 minutes, and filter: the filtrate responds to Qualitative Tests <1.09> for chloride. D Optical rotation <2.49> [a]20: －17.5 – －20.59(after dry-ing, 0.15 g, water, warming, after cooling, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tamsulosin Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances— Dissolve 50 mg of Tamsulosin Hydrochloride in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 2.5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than tamsulosin obtained from the sample solution is not larger than 1/2 times the peak area of tamsulosin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.4 mL of perchloric acid and 1.5 g of sodium hydroxide in 950 mL of water, adjust the pH to 2.0 with sodium hydroxide TS, and add water to make 1000 mL. To 700 mL of this solution add 300 mL of aceto-nitrile for liquid chromatography. Flow rate: Adjust so that the retention time of tamsulosin is about 6 minutes. Time span of measurement: Until tamsulosin is eluted, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of tamsulosin obtained with 10 mL of this solution is equivalent to 1.4 to 2.6z of that with 10 mL of the standard solution. System performance: Dissolve 5 mg of Tamsulosin Hydro-chloride and 10 mg of propyl parahydroxybenzoate in 20 mL of the mobile phase. To 2 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, tam-sulosin and propyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tamsulosin is not more than 4.0z. Perform the test with exactly 10 mL each of the sam-ple solution and standard solution which are obtained in above (i) as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than tamsulosin obtained from the sample so-lution is not larger than 1/2 times the peak area of tamsulo-sin from the standard solution. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Purity (2) (i). Mobile phase: Dissolve 4.4 mL of perchloric acid and 1.5 g of sodium hydroxide in 950 mL of water, adjust the pH to 2.0 with sodium hydroxide TS, and add water to make 1000 mL. To this solution add 1000 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of tamsulosin is about 2.5 minutes. Time span of measurement: About 5 times as long as the retention time of tamsulosin, beginning after the peak of tamsulosin. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) (i). Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase used in the Purity (2) (i) to make exactly 50 mL. Confirm that the peak area of tamsulosin obtained with 10 mL of this solution is equivalent to 1.4 to 2.6z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tamsulosin is not more than 4.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, ‌hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Tamsulosin Hydro-chloride, previously dried, dissolve in 5 mL of formic acid, add 75 mL of a mixture of acetic acid (100) and acetic anhy-dride (3:2), and immediately titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 44.50 mg of C20H28N2O5S.HCl
Tannic Acid occurs as a yellow-white to light brown amorphous powder, glistening leaflets, or spongy masses. It is odorless or has a faint, characteristic odor, and has a strongly astringent taste. It is very soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether.
(1) To 5 mL of a solution of Tannic Acid (1 in 400) add 2 drops of iron (III) chloride TS: a blue-black color develops. Allow the solution to stand: a blue-black precipitate is produced. (2) To 5 mL of a solution of Tannic Acid (1 in 20) add 1 drop each of albumin TS, gelatin TS, or 1 mL of starch TS: a precipitate is produced in each solution.
(1) Gum, dextrin and sucrose—Dissolve 3.0 g of Tannic Acid in 15 mL of boiling water: the solution is clear or slightly turbid. Cool, and filter the solution. To 5 mL of the filtrate add 5 mL of ethanol (95): no turbidity is pro-duced. Add further 3 mL of diethyl ether to this solution: no turbidity is produced. (2) Resinous substances—To 5 mL of the filtrate ob-tained in (1) add 10 mL of water: no turbidity is produced. Loss on drying <2.41> Not more than 12.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 1.0z (0.5 g).
Tartaric Acid, when dried, contains not less than 99.
Tartaric Acid occurs as colorless crystals or a white crystalline powder. It is odorless, and has a strong acid taste. It is very soluble in water, freely soluble in ethanol (95), and slightly soluble in diethyl ether. A solution of Tartaric Acid (1 in 10) is dextrorotatory.
(1) Ignite Tartaric Acid gradually: it decomposes and an odor of burning sugar is perceptible. (2) A solution of Tartaric Acid (1 in 10) changes blue litmus paper to red, and responds to Qualitative Tests <1.09> for tartrate.
(1) Sulfate <1.14>—Perform the test with 0.5 g of Tartaric Acid. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Oxalate—Dissolve 1.0 g of Tartaric Acid in 10 mL of water, and add 2 mL of calcium chloride TS: no turbidity is produced. Heavy metals <1.07>—Proceed with 2.0 g of Tartaric Acid according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Calcium—Neutralize a solution of 1.0 g of Tartaric Acid in 10 mL of water with ammonia TS, and add 1 mL of ammonium oxalate TS: no turbidity is produced. Arsenic <1.11>—Prepare the test solution with 2.0 g of Tartaric Acid according to Method 1, and perform the test (not more than 1 ppm). Loss on drying <2.41> Not more than 0.5z (3 g, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.05z (1 g).
Weigh accurately about 1.5 g of Tartaric Acid, pre-viously dried, dissolve in 40 mL of water, and titrate <2.50> with 1 mol/L sodium hydroxide VS (indicator: 2 drops of phenolphthalein TS). Each mL of 1 mol/L sodium hydroxide VS ＝ 75.05 mg of C4H6O6
Taurine, when dried, contains not less than 99.
Taurine occurs as colorless or white crystals, or a white crystalline powder. It is soluble in water, and practically insoluble in ethanol (99.5). The pH of a solution prepared by dissolving 1.0 g of Tau-rine in 20 mL of freshly boiled and cooled water is between 4.1 and 5.6.
Determine the infrared absorption spectrum of Taurine as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Taurine in 20 mL of water is clear and colorless. Chloride <1.03>—Perform the test with 1.0 g of Tau- rine. Prepare the control solution with 0.30 mL of 0.01 ‌mol/L hydrochloric acid VS (not more than 0.011z). Sulfate <1.14>—Perform the test with 2.0 g of Tau-rine. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Ammonium <1.02>—Perform the test with 0.25 g of Taurine. Prepare the control solution with 5.0 mL of Stand-ard Ammonium Solution (not more than 0.02z). Heavy metals <1.07>—Proceed with 2.0 g of Taurine according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Iron <1.10>—Prepare the test solution with 2.0 g of Taurine according to Method 1, and perform the test accord-ing to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 10 ppm). Related substances—Dissolve 1.0 g of Taurine in 50 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of water, ethanol (99.5), 1-butanol and acetic acid (100) (150:150:100:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin-butanol TS on the plate, and heat the plate at 1059C for 5 minutes: the spot other than the principle spot obtained from the sample solu-tion is not more than one spot, and it is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.20z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Taurine, previously dried, dissolve in 50 mL of water, add 5 mL of formalde-hyde solution, and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 12.52 mg of C2H7NO3S
Tazobactam contains not less than 980 mg (potency) and not more than 1020 mg (potency) per 1 mg, calcu-lated on the anhydrous basis.
Tazobactam occurs as a white to pale yellow-white crystalline powder. It is freely soluble in dimethylsulfoxide and in N,N-dimethylformamide, and slightly soluble in water, in metha-nol and in ethanol (99.5). It dissolves in a solution of sodium hydrogen carbonate (3 in 100).
(1) Determine the infrared absorption spec-trum of Tazobactam as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Tazobactam RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Tazobac-tam in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 35) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethyl-silane for nuclear magnetic resonance spectroscopy as an in-ternal reference compound: it exhibits a singlet signal A at around d 1.3 ppm, and doublet signals, B and C, at around d 7.8 ppm and at around d 8.1 ppm. The ratio of the inte-grated intensity of each signal, A:B:C, is about 3:1:1. D Optical rotation <2.49> [a]20: ＋162 – ＋1679(1 g calculated on the anhydrous basis, N,N-dimethylformamide, 100 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Tazobactam in 10 mL of sodium hydrogen carbonate (3 in 100): the solution is clear. Perform the test with the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 420 nm is not more than 0.14. Heavy metals <1.07>—Proceed with 1.0 g of Tazobactam according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—This operation must be per-formed quickly. Dissolve 50 mg of Tazobactam in 20 mL of the mobile phase, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 1 mL of the standard solution (1), add the mobile phase to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with exactly 50 mL each of the sample solution, the standard solutions (1) and (2) as directed under Liquid Chromatogra- ‌phy <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integra-tion method: the area of the peak, having the relative reten-tion time of about 0.17 to tazobactam, obtained from the sample solution is not larger than 4/5 times the peak area of tazobactam from the standard solution (1), the area of the peak other than tazobactam and the peak having the relative retention time of about 0.17 from the sample solution is not larger than the peak area of tazobactam from the standard solution (2), and the total area of the peaks other than tazobactam and the peak having the relative retention time of about 0.17 from the sample solution is not larger than 2 times the peak area of tazobactam from the standard solu-tion (2). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of tazobactam. System suitability— Test for required detectability: Pipet 1 mL of the standard solution (1), and add the mobile phase to make exactly 20 mL. Confirm that the peak area of tazobactam obtained with 50 mL of this solution is equivalent to 3 to 7z of that of tazobactam with 50 mL of the standard (1). System performance: When the procedure is run with 50 mL of the standard solution (1) under the above operating conditions, the number of theoretical plates and the symme-try factor of the peak of tazobactam are not less than 2000 and 0.8 – 1.2, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviations of the peak area of tazobactam is not more than 1.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (3:1) instead of methanol for water determination). Residue on ignition <2.44> Not more than 0.1z (1 g). Bacterial endotoxins <4.01> Less than 0.04 EU/mg (po-tency).
Weigh accurately an amount of Tazobactam and Tazobactam RS, equivalent to about 50 mg (potency), dis-solve each in exactly 10 mL of the internal standard solution, add water to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under the Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak areas of tazobactam to that of the internal standard. Amount [mg (potency)] of tazobactam (C10H12N4O5S) ＝ MS × (QT/QS) × 1000 MS: Amount [mg (potency)] of Tazobactam RS taken Internal standard solution—A solution of phenylalanine (1 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.32 g of diammonium hydrogen phosphate in 750 mL of water, adjust the pH to 2.5 with phosphoric acid, add water to make 1000 mL, and add 25 mL of acetonitrile. Flow rate: Adjust so that the retention time of tazobactam is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and tazobactam are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of tazobactam to that of the internal stand-ard is not more than 1.0z.
Teceleukin (Genetical Recombination) is a recom-binant human interleukin-2, and is a protein consisting of 134 amino acid residues with methionine at the N-terminus. It is a solution. It contains potency between 7.
Teceleukin (Genetical Recombination) occurs as a clear and colorless liquid.
(1) Pipet an appropriate amount of Teceleukin (Genetical Recombination), add an accurate amount of potency measuring medium for teceleukin so that each mL contains about 200 units, and use this solution as the sample stock solution. Dilute reference anti-interleukin-2 antibody for teceleukin with potency measuring medium for teceleukin to a concentration of approximately 200 neutral units per mL and use this solution as the interleukin-2 neu-tral antibody solution. Accurately add an equivalent volume of the interleukin-2 neutral antibody solution to the sample stock solution, shake, and then leave for 1 hour in a 379C incubator in air containing 5z carbon dioxide. This solution is used as the sample solution. Prepare a standard solution by accurately adding an equivalent volume of potency meas- uring medium for teceleukin to the sample stock solution, mixing, and then processing in the same way. Process the sample and standard solutions according to the assay method, determine their respective dilution coefficients, DN and DT, and then calculate the neutralization rate, which should be at least 90z, using the following formula. solution is used as the amino acid standard solution. Sepa-rately, weigh accurately about 17 mg of L-cysteic acid and dissolve in sodium citrate solution for dilution to make ex-actly 50 mL. Pipet 1 mL of this solution and add sodium citrate solution for dilution to make exactly 100 mL. This so-lution is used as the cysteic acid standard solution. Neutralization rate (z) ＝ (DT － DN)/DT × 100 (ii) Amino acid analysis—Accurately measure 0.25 mL each of the sample solution (1), the sample solution (2), the If the mean values of the absorbance of the maximum up-take control solution and absorbance of the minimum up-take control solution do not fit the standard curve, the neutralization coefficient is to be determined within the following range. amino acid standard solution, and the cysteic acid standard solution, perform the test by Liquid Chromatography <2.01> under the following conditions, and confirm the peaks of amino acids appeared on the chromatogram obtained from the sample solution (1). Also, measure the peak area of each Neutralization coefficient (z) ＞ (DT － 2)/DT × 100 amino acid in the sample solution (1) and the amino acid standard solution, and taking the molar number of alanine (2) When hydrolyzed according to modified Method 2 and Method 4 as directed in 1. Hydrolysis of Protein and Peptide, and performed the test according to Method 1 in 2. Methodologies of Amino Acid Analysis under Amino Acid Analysis of Proteins <2.04>, the molar ratios of the respec-tive amino acids are as follows: aspartic acid is 11.4 to 12.6, glutamic acid is 17.1 to 18.9, proline is 4.5 to 5.5, glycine is 1.8 to 2.2, cysteine 2.7 to 3.3, methionine is 4.5 to 5.5, leu- cine is 20.9 to 23.1, tyrosine is 2.7 to 3.3, phenylalanine is 5.4 to 6.6, lysine is 10.5 to 11.6, histidine is 2.7 to 3.3, tryp- tophan is 0.7 to 1.2, and arginine is 3.6 to 4.4. Furthermore, the peaks of the constituent 18 amino acids are observed in the chromatogram obtained from the sample solution (1). Hydrolysis—Place a volume of Teceleukin (Genetical Recombination) corresponding to approximately 50 mg of protein in 2 test tubes for hydrolysis, evaporate to dryness under vacuum, and use one as the sample (1). To the other, add 50 mL of a mixture of formic acid and hydrogen peroxide (30) (9:1) that has been left at room temperature for 1 hour, cool for 4 hours in ice, add 0.5 mL of water, and then evaporate to dryness under vacuum to give the sample (2). To 1.3 mL of methanesulfonic acid add 3.7 mL of water, mix well, and dissolve 10 mg of 3-(2-aminoethyl)in-dole, to make a 4 mol/L methanesulfonic acid solution. Dissolve 39.2 g of trisodium citrate dihydrate, 33 mL of hydrochloric acid, 40 mL of thiodiglycol, and 4 mL of lauromacrogol solution (1 in 4) in 700 mL of water, adjust the pH to 2.2, add water to make 1000 mL, add 100 mL of capric acid, and mix to make a sodium citrate solution for dilution. Add 50 mL of freshly prepared 4 mol/L methane-sulfonic acid to the sample (1) and sample (2), cool to －709C, and then deaerate under vacuum. Heat to 1159C ± 29C for 24 hours after sealing these test tubes under reduced pressure. After cooling, unseal, add 50 mL of 4 mol/L sodi-um hydroxide TS followed by 0.4 mL of sodium citrate solu-tion for dilution to make the sample solution (1) and sample solution (2). Separately, weigh accurately 0.25 mmol amounts of L-aspartic acid, L-threonine, L-serine, L-glutamic in the sample solution (1) as 5.0, determine the concentra-tions of aspartic acid, glutamic acid, proline, glycine, methionine, leucine, tyrosine, phenylalanine, lysine, histi-dine, tryptophan, and arginine and then calculate the molar ratio for each amino acid. Also, measure the cysteic acid peak areas of the sample solution (2) and the cysteic acid standard solution, determine the concentration of the cys-teine, and calculate the molar ratio of cysteine taking the molar number of alanine in the sample solution (2) as 5.0. Operating conditions— Detector: Visible absorption photometer [wavelengths: 440 nm (proline) and 570 nm (amino acids other than pro-line)]. Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with a strongly acidic ion ex-change resin for liquid chromatography consisting of poly-styrene to which sulfonate group binds (5 mm in particle diameter). Column temperature: A constant temperature of about 509C when the sample is injected. After a certain time, in-crease the temperature to a constant temperature of about 629C. Reaction temperature: A constant temperature of about 989C. Time for color formation: Approximately 2 minutes. Mobile phase: After preparing mobile phases A, B, and C according to the following table, add 0.1 mL of capric acid to each. Mobile phase Mobile phase Mobile phase A B C Citric acid 18.70 g monohydrate 10.50 g 7.10 g Trisodium citrate 7.74 g 14.71 g 26.67 g dihydrate Sodium chloride 7.07 g 2.92 g 54.35 g Ethanol (99.5) 60 mL — — Benzyl alcohol — — 10 mL Thiodiglycol 5 mL 5 mL — acid, L-proline, glycine, L-alanine, L-valine, L-methionine, Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL L-isoleucine, L-leucine, L-tyrosine, L-phenylalanine, L-lysine hydrochloride, ammonium chloride, L-histidine hydrochlo- Water Appropriate amount Appropriate amount Appropriate amount ride monohydrate, and L-arginine hydrochloride as well as 0.125 mmol of L-cysteine, and then dissolve in 0.1 mol/L hy-drochloric acid TS to make exactly 100 mL. This solution is used as the amino acid standard stock solution. Pipet 1 mL of the amino acid standard stock solution, and add sodium citrate solution for dilution to make exactly 25 mL. This so-lution is used as solution A. Weigh accurately about 20 mg of L-tryptophan and dissolve in water to make exactly 1000 mL. This solution is used as solution B. Pipet 10 mL of both solution A and solution B, combine together, and add sodi-um citrate solution for dilution to make exactly 50 mL. This pH 3.2 4.3 4.7 Total volume 1000 mL 1000 mL 1000 mL Changing mobile phases and column temperature: When operating under the above conditions using 0.25 mL of the amino acid standard solution, the amino acids will elute in the following order; aspartic acid, threonine, serine, glutam-ic acid, proline, glycine, alanine, cystine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, ammo-nia, histidine, tryptophan, and arginine. Switchover to the mobile phases A, B, and C, in sequence so that the resolu-tion between the peaks of cystine and valine is 2.0 or more and that between ammonia and histidine is 1.5 or more. Also, increase the temperature after a constant length of time so that the resolution between the peaks of glutamic acid and proline is at least 2.0. Reaction reagents: Dissolve 408 g of lithium acetate dihy-drate in water, and add 100 mL of acetic acid (100) and water to make 1000 mL. To this solution add 1200 mL of dimethylsulfoxide and 800 mL of 2-methoxyethanol. This solution is used as solution (I). Separately, mix together 600 mL of dimethylsulfoxide and 400 mL of 2-methoxyethanol and then add 80 g of ninhydrin and 0.15 g of sodium boro-hydride. This solution is used as solution (II). After gassing 3000 mL of the solution (I) for 20 minutes with nitrogen, rapidly add 1000 mL of the solution (II) and then mix by gassing for 10 minutes with nitrogen. Mobile phase flow rate: About 0.275 mL per minute. Reaction reagent flow rate: About 0.3 mL per minute. System suitability— System performance: When the procedure is run with 0.25 mL of the amino acid standard solution under the above operating conditions, the resolution between the peaks of threonine and serine is at least 1.5. Molecular mass Dissolve 0.242 g of 2-amino-2-hydrox-ymethyl-1,3-propanediol, 5.0 g of sodium lauryl sulfate, and 74 mg of disodium dihydrogen ethylenediamine tetraacetate dihydrate in 60 mL of water. After adjusting the pH to 8.0 with 1 mol/L hydrochloric acid TS, add water to make 100 mL. This solution is used as the molecular mass determina-tion buffer solution. Pipet 20 mL of Teceleukin (Genetical Recombination), add exactly 20 mL of the molecular mass determination buffer solution and exactly 2 mL of 2-mercap-toethanol, and then heat at 90 to 1009C for 5 minutes on a water bath without allowing any water evaporation from the mixture. After cooling, add exactly 1 mL of bromophenol blue solution (1 in 2000) and then shake. This solution is used as the sample solution. Separately, pipet 5 mL of mo-lecular mass marker for teceleukin, and add exactly 50 mL of water, exactly 55 mL of the molecular mass determination buffer solution, and exactly 5 mL of 2-mercaptoethanol, and then heat at 90 to 1009C for 5 minutes on a water bath with-out allowing any water evaporation from the mixture. After cooling, add exactly 1 mL of bromophenol blue solution (1 in 2000), and shake well. This solution is used as the molecular mass standard solution. When conducting a test using SDS-polyacrylamide gel electrophoresis with 1 mL each of the sample solution and the molecular mass standard solution, the molecular mass of the main band is between 14,000 and 16,000. Operating conditions— Equipment: Horizontal electrophoresis vessel with a cool-ing unit, a device that accumulates load voltage over time, and a direct current power source device that controls the amperage, voltage, wattage. Spotting of solutions: Solutions are spotted on concentrat-ing gel of polyacrylamide gel sheets. Electrophoresis conditions Polyacrylamide gel sheet: Polyester sheet to which a polyacrylamide gel (width, about 43 mm, length, about 50 mm, and thickness, about 0.5 mm) is closely adhered. The polyacrylamide gel consists of a concentrating gel with a gel support concentration of 7.5z and a 3z degree of crosslinking and a separating gel with corresponding values of 20z and 2z. The gel contains tris-acetate buffer (pH 6.5). Buffer solution for electrode: Prepared by dissolving 35.83 g of tricine, 24.23 g of 2-amino-2-hydroxymethyl-1,3-propanediol, and 5.5 g of sodium lauryl sulfate in water to make 1000 mL. Cooling temperature of gel support plate: 159C. Running conditions Pre-electrophoresis and electrophoresis: The voltage, amperage, and wattage should not exceed 250 V, 10 mA, and 3 W, respectively. The amperage and wattage should be proportional to the number of polyacrylamide sheets. Immediately after adding sample: The voltage, am-perage, and wattage should not exceed 250 V, 1 mA, and 3 W, respectively. The amperage and wattage should be pro-portional to the number of polyacrylamide sheets. Electrophoresis time Before adding sample: Until value of load voltage inte-grated to time reaches 60 V･h. Immediately after adding sample: Until value of load voltage integrated to time reaches 1 V･h. Main electrophoresis: Until value of load voltage inte-grated to time reaches 140 V･h. Fixation and staining Dissolve 25 g of anhydrous sodium carbonate and 0.8 mL of formaldehyde solution in water to make 1000 mL. This solution is used as the developing solution. After immersing the polyacrylamide gel sheet in a mixture of ethanol (99.5), water and acetic acid (100) (5:4:1) for 2 minutes, immerse for 2 minutes in a mixture of water, ethanol (99.5) and acetic acid (100) (17:2:1). Change the mixture, immerse for another 4 minutes, immerse in water for 2 minutes to rinse the polyacrylamide gel sheet, and change the water to immerse for 2 minutes. This proce-dure is carried out with warming to 509C. Next, while warming at 409C, immerse for 10 to 15 minutes in diluted silver nitrate TS (1 in 7), warm to 309C, and gently rinse the polyacrylamide gel sheet with water. While warming at 309C, immerse the polyacrylamide gel sheet in freshly pre-pared developing solution. After obtaining adequate color formation, immerse the polyacrylamide gel sheet in di-luted acetic acid (100) (1 in 20) to terminate the color for-mation. Estimation of molecular mass Plot graphs for each band obtained from the molecular mass standard solution, distance from the border of the concentrating gel and separating gel, and the logarithm of the molecular mass of proteins in each band. Calculate the molecular mass by reading the corresponding position of the major band obtained from the sample solution on the graph. Isoelectric point The isoelectric point determined from the electrophoresis position is 7.4 to 7.9 when 3 mL of Teceleu-kin (Genetical Recombination) and 8 mL of isoelectric mar-ker for teceleukin are tested by the polyacrylamide gel isoe-lectric method. Operating conditions— Equipment: Horizontal electrophoretic vessel with a cool-ing unit and direct current power source that can perform constant wattage control. Preparation of polyacrylamide gel: Dissolve 1.62 g of acrylamide and 50 mg of N,N?-methylenebisacrylamide in water to make 25 mL. Accurately measure 7.5 mL of this so-lution, 2 mL of a 10 mL solution prepared by adding water to 5 g of glycerin, and 0.64 mL of a pH 3 to pH 10 ampho-teric electrolyte solution, and degas under reduced pressure while stirring thoroughly. Next, accurately measure 74 mL of freshly prepared ammonium peroxodisulfate solution (1 in 50), 3 mL of N,N,N?,N?-tetramethylethylenediamine, and 50 mL of freshly prepared riboflavin sodium phosphate solution (1 in 1000), stir well, immediately pour on a gel preparation plate (10 cm wide, 11 cm long, and 0.8 mm thick), and then expose to a fluorescent light source for 60 minutes to gelate. Spotting Add Teceleukin (Genetical Recombination) or isoelec-tric marker for teceleukin 30 minutes after starting elec-trophoresis to wells in gel plates to which plastic tape (3.5 mm wide, 3.5 mm long, 0.4 mm thick) has been applied in advance and that have undergone gelation. Electrophoresis conditions Cathode solution: Sodium hydroxide TS. Anode solution: DL-Aspartic acid solution (133 in 25,000). Cooling temperature of gel support plate: 2 ± 19C. Running conditions: After starting the electrophoresis, a constant wattage of 10 W for 20 minutes and 20 W there-after. However, the voltage should be 3000 V or less. Running time: 120 to 140 minutes while blowing Nitrogen into the electrophoresis vessel. Fixation and washing Dissolve 28.75 g of trichloroacetic acid and 8.65 g of 5-sulfosalicylic acid dihydrate in 75 mL of methanol and 175 mL of water. Immerse the gel in this solution for 60 minutes to fix the protein to the gel. After fixation, im-merse for 10 minutes in a mixture of water, ethanol (99.5) and acetic acid (100) (67:25:8). Staining and decolorization Dissolve 0.11 g of Coomassie brilliant blue G-250 in 25 mL of ethanol (99.5), and add 8 mL of acetic acid (100) and water to make 100 mL. This solution is used as the staining solution. Immerse the gel for 10 minutes while warming at 609C in freshly filtered staining solution. After staining, decolorize by immersing in a mixture of water, ethanol (99.5) and acetic acid (100) (67:25:8). Determination of isoelectric point Plot the protein isoelectric points and the distance from the cathode of each band obtained from the isoelectric markers for teceleukin. Calculate the isoelectric point from the corresponding position of the major bands ob-tained from the sample solution. pH <2.54> 2.7 – 3.5
(1) Desmethionyl form—To Teceleukin (Genetical Recombination) add water so that each mL contains about 0.17 mg of protein, and use this solution as the sample solu-tion. Perform the test with 1.2 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak area, A2, of teceleukin and the peak area of the desmethionyl form hav-ing the relative retention time of about 0.8 to teceleukin, A1, by the automatic integration method. The amount of the desmethionyl form is not more than 1.0z when determined using the following formula. of water, adjust the pH to 9.0 with 1 mol/L hydrochloric acid TS, and then add water to make 500 mL. Mobile phase B: Add 300 mL of water to 2.6 mL of a pH 6 to 9 amphoteric electrolyte solution and 0.5 mL of a pH 8 to 10.5 amphoteric electrolyte solution, adjust to pH 7 with diluted hydrochloric acid (9 in 100), and then add water to make 400 mL. Switching mobile phases and sample injection: Inject the sample solution while running the mobile phase A. Repeat-edly inject 10 times a sample solution volume of 0.11 mL fol-lowed by a single injection of 100 mL. After injecting the entire volume and running mobile phase A for 60 minutes, switch to mobile phase B. After measuring the sample solu-tion and after running 1 mol/L sodium chloride TS for 10 minutes for postreatment and cleaning of the columns, inject 100 mL of sodium hydroxide TS while running the mobile phase A and then 55 minutes later start injection of the next sample solution. Flow rate: Adjust the flow of the mobile phase B so that the retention time for teceleukin is 45 to 65 minutes. Measure the retention time from the point at which the mobile phase is switched to the mobile phase B. System suitability— System performance: Dissolve in water a mixture of two kinds of equine heart-derived myoglobin whose isoelectric points are 6.76 and 7.16 to make a concentration of approxi-mately 0.5 mg/mL. Mix together 50 mL of this solution, 50 mL of Teceleukin (Genetical Recombination), and 1.47 mL of water. When the procedure is run with 1.2 mL of this so-lution under the above operating conditions, myoglobin and teceleukin are eluted in this order, and their respective peaks are completely separated. Dimer—Prepare a sample solution by adding 20 mL of 0.2z sodium laurylsulfate TS to 20 mL of Teceleukin (Genetical Recombination). Perform the test as directed under Liquid Chromatography <2.01> with 20 mL of the sample solution according to the following conditions. Determine the teceleukin peak area, A2, and the peak area, A1, of the dimer with the relative retention time of 0.8 to 0.9 in relation to teceleukin, by the automatic integration method. The amount of the dimer is not more than 1.0z by the following formula. Amount (z) of dimer ＝ A1/(A1 ＋ A2) × 100 Operating conditions— Detector: Ultraviolet absorption photometer (wavelength: 220 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with glycol etherifized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 0.1 mol/L sodium phosphate buffer (pH 7.0) to make 1000 mL. Amount (z) of desmethionyl form ＝ A1/(A1 Operating conditions— ＋ A2) × 100 Flow rate: Adjust so that the retention time of teceleukin is 30 – 40 minutes. System suitability— Detector: Ultraviolet absorption photometer (wavelength: 280 nm). Columns: Two stainless steel columns with inside diame-ters of 7.5 mm and lengths of 7.5 cm connected in sequence and packed with 10 mm synthetic polymer bound to diethylaminoethyl base for liquid chromatography. Column temperature: A constant temperature of about 259C. Mobile phase A: Mix 0.658 g of diethanolamine in 400 mL System performance: Add 20 mL of 0.2z sodium lauryl sulfate TS to 20 mL of a solution consisting of 5 mg of carbonic anhydrase and 5 mg of a-lactoalbumin dissolved in 100 mL of water. When the procedure is run with 20 mL of this solution under the above operating conditions, carbonic anhydrase and a-lactoalbumin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: To exactly 1 mL of the sample solu-tion add the mobile phase to make exactly 20 mL. To exactly 1 mL of this solution add the mobile phase to make exactly 10 mL. When the test is repeated 3 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the teceleukin peak area is not more than 7z. Tetracycline hydrochloride—Serially subculture through 2 passages at 35 to 379C the test bacteria Kocuria rhizophila ATCC9341 in a slant culture of test bacteria in-oculation media for teceleukin and then dilute this 100-fold by adding sterilized purified water. This solution is used as the test bacteria solution. Store the test bacteria solution at 59C or less and use the solution within 5 days. Dilute the test bacteria solution serially by adding sterilized purified water, add an appropriate amount to 100 mL of normal agar medi-um for teceleukin, conduct a preliminary test, and determine the amount of tetracycline hydrochloride that shows an inhi-bition zone corresponding to standard solution containing 0.5 mg (potency) of tetracycline hydrochloride (C22H24N2O8. HCl) in 1 mL. Add this amount to 100 mL of normal agar culated by the area percent method, the total amount of peaks other than the teceleukin and solvent peaks is not more than 1.0z. Operating conditions— Detector: Ultraviolet absorption photometer (wavelength: 220 nm). Column: A stainless steel column with an inside diameter of 4.6 mm and 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: A solution of trifluoroacetic acid in a mixture of water and acetonitrile (19:1) (1 in 1000). Mobile phase B: A solution of trifluoroacetic acid in acetonitrile (7 in 10,000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as follows. medium for teceleukin dissolved and then cooled to 45 to 509C and mix. Pipet 25 mL of this solution into square Petri dishes (135 × 95 mm) and spread horizontally to solidify. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Prepare plates for testing by making an appropriate number of wells in this agar medium. The volume of the test bacteria solution to which 100 mL of normal agar medium for teceleukin has been added is 0.25 to 1.0 mL. Pipet an appropriate amount of Tetracycline Hydrochloride RS and dilute accurately with water to make a clear solution with a concentration of 1 mg (potency) of tetracycline hydrochlo-ride (C22H24N2O8.HCl) per mL. Pipet an appropriate amount of this solution and dilute precisely with water to make standard solutions with concentrations of 4, 2, 1 and 0.5 mg (potency)/mL. Separately, dilute Teceleukin (Geneti-cal Recombination) with diluted acetic acid (100) (3 in 1000), if necessary, or alternatively concentrate under reduced pres-sure, to make a sample solution with a protein concentration of 0.8 to 1.2 mg/mL. Pipet 25 mL of the sample solution and each standard solution, and add each to the wells in the same test plate. Repeat the same procedure for at least 3 more test plates. Leave the test plates at room temperature for 30 to 60 minutes and then incubate at 35 to 379C for 16 to 18 hours. Measure the inhibitions zones to a diameter of 0.25 mm. Determine the mean among the test plates for each of the solutions. Prepare a standard curve by plotting a graph with the con-centration of each standard solution in logarithmic scale on the horizontal axis and the diameter of the inhibition zone on the vertical axis. Match the diameter of the inhibition zone of teceleukin from the standard curve and determine A, the concentration of tetracycline hydrochloride. When the amount of tetracycline hydrochloride per mg of protein is determined by the following formula, the amount is not more than 0.7 mg. However, if an inhibition zone is not seen, or is seen but the diameter is smaller than 0.5 mg/mL on the standard curve, A is taken as being 0.5 mg/mL or less. Amount [mg (potency)] of tetracycline hydrochloride (C22H24N2O8.HCl) per mg of protein ＝ A/P P: The protein concentration (mg/mL) of the sample solu-tion. Other related proteins—Perform the test with 5 mL of Teceleukin (Genetical Recombination) as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and measure the area of each peak by the automatic integration method. When the amounts of the peak are cal- 0 – 12 60 → 50 40 → 50 12 – 25 50 50 25 – 45 50 → 0 50 → 100 45 – 50 0 100 Flow rate: 1.0 mL per minute. Time span of measurement: About 1.2 times as long as the retention time of teceleukin. System suitability— System performance: Add 3.8 mL of water and 16.6 mL of polysorbate 80 solution (1 in 100) to 83.6 mL of Teceleukin (Genetical Recombination) and let stand for at least 1 hour. When the procedure is run with 5 mL of this solution under the above operating conditions, there is complete separation between the teceleukin peak and the peak with the relative retention time of about 0.98 to the teceleukin. Host cell proteins—Being specified separately when the drug is granted approval based on the Law. Host cell DNA—Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 5EU per mg of pro-tein. Acetic acid Pipet 0.25 mL of Teceleukin (Genetical Recom-bination) and add exactly 0.25 mL of the internal standard solution to make the sample solution. Separately, pipet 3 mL of acetic acid (100) and add water to make exactly 100 mL. Pipet 10 mL of this solution and add water to make exactly 100 mL. Pipet 2 mL of this solution and add exactly 2 mL of the internal standard solution to make the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions. Calculate the ratios of the peak area of acetic acid to that of the internal standard, QT and QS, and the amount of acetic acid (C2H4 O2) in 1 mL of Teceleukin (Genetical Recombination) calcu-lated by the following formula is between 2.85 mg and 3.15 mg. Amount (mg) of acetic acid (C2H4O2) in 1 mL of Teceleukin (Genetical Recombination) ＝ QT/QS × 1.5 × 1.049 × 2 1.5: Concentration (mL/mL) of acetic acid (100) in the standard solution ‌1.049: Density (mg/mL) of acetic acid (100) at 259C 2: Dilution factor Internal standard solution—Diluted propionic acid (1 in 500). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column with an inside diameter of 1.2 mm and 40 m in length, whose inside is covered with chemically-bound polyethylene glycol for gas chromato-graphy 1.0 mm in thickness. Column temperature: A constant temperature of about 1109C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of acetic acid is about 8 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, acetic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 1 mL of standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of acetic acid to that of the internal standard is not more than 5z. Specific activity Pipet an appropriate amount of Teceleu-kin (Genetical Recombination), and add water accurately so that each mL contains about 0.1 mg. This solution is used as the sample solution. Separately, pipet about 25 mg of human serum albumin for assay, dissolve in water to make exactly 50 mL. Pipet an appropriate amount of this solution, and accurately dilute with water to make standard solutions with concentrations of 0.05, 0.10, and 0.15 mg per mL. Pipet 1 mL each of the sample solution, the standard solutions, and water, add 2.5 mL of alkaline copper solution, mix, leave for at least 10 minutes to dissolve, add exactly 2.5 mL of water and 0.5 mL of diluted Folin TS (1 in 2), immediately shake vigorously, and then leave at 379C for 30 minutes. Perform the test with these solutions, with water as a control, as directed under Ultraviolet-visible Spectrophotometry <2.24>, and measure the absorbance at 750 nm. With the concentra-tion of the standard solution as the x-axis and the absor-bance as the y-axis, perform linear regression using their re-spective reciprocals, and calculate the protein content. Calculate the ratio of the potency determined by Assay and the protein content.
Pipet an appropriate amount of Teceleukin (Geneti-cal Recombination) and, depending on the cell sensitivity, di-lute precisely by adding potency measuring medium for teceleukin to a constant concentration of 10 to 50 units/mL (estimated value). This solution is used as the sample solu-tion. Separately, dissolve Interleukin-2 RS in 1 mL of steri-lized purified water, and, depending on the cell sensitivity, dilute precisely by adding potency measuring medium for teceleukin to a constant concentration of 10 to 50 units/mL. This solution is used as the standard solution. Add exactly 50 mL of potency measuring medium for teceleukin to all but 8 wells in a 96-well microplate. Add exactly 50 mL of the sam-ple solution and standard solution to 2 wells each containing potency measuring medium for teceleukin. From these 4 wells, remove exactly 50 mL and add to 4 other wells contain-ing potency measuring medium for teceleukin. From these 4 wells, remove exactly 50 mL and add to 4 other wells contain-ing potency measuring medium for teceleukin and repeat this procedure to prepare 2 wells that contain each of 1/2, 1/4, 1/8, 1/16, 1/32, 1/64, 1/128, and 1/256 dilutions of the sample solution and standard solution. Add 50 mL of the standard solution to each of the 8 empty wells to make maxi-mum uptake controls. The 8 wells containing only potency measuring medium for teceleukin serve as the minimum up-take controls. After adding exactly 50 mL of cell suspension solution for teceleukin to each well in the microplate, leave for 15 to 17 hours in an incubator at 379C filled with air con-taining 5z carbon dioxide. After adding exactly 25 mL of MTT TS to each of the wells in the microplate, leave for 4 hours in an incubator at 379C filled with air containing 5z carbon dioxide. Transfer the culture medium in all of the wells to empty wells into another microplate. To each of the empty wells from which the culture medium was removed, add 100 mL of hydrochloric acid-2-propanol TS, and then shake the plates horizontally for 5 minutes to elute the pig-ment. After returning the transferred culture medium to each original well, perform the test with the solution in each well, determine the difference in absorption at wavelengths of 560 nm and 690 nm, and calculate the mean values of the identical respective solutions in the two wells (dilution solu-tions of the sample solution and standard solutions) as well as the 8 wells containing the maximum or minimum uptake controls. Prepare standard curves by plotting the values ob-tained from each dilution solution of the sample solution, with the dilution coefficient of the sample solution on the microplate in logarithmic scale on the horizontal axis and the absorbance on the vertical axis. Determine the mean absor-bance values of the maximum and minimum uptake con-trols, find the values on the standard curve, and then calcu-late the dilution coefficient, DT. Perform the same plot for the dilution solution of the standard solution, calculate the dilution coefficient, DS, and then calculate the potency in 1 mL by the following formula. Teceleukin potency (unit) in 1 mL of Teceleukin (Genetical Recombination) ＝ S × DT/DS × d S: Concentration of standard solution (unit/mL) d: Dilution coefficient when sample solution prepared
Containers—Tight containers Storage—Store at －709C or lower.
Tegafur, when dried, contains not less than 98.
Tegafur occurs as a white crystalline powder. It is soluble in methanol, and sparingly soluble in water and in ethanol (95). It dissolves in dilute sodium hydroxide TS. A solution of Tegafur in methanol (1 in 50) shows no opti-cal rotation. It shows crystal polymorphism.
(1) Prepare the test solution with 0.01 g of Tegafur as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L so-dium hydroxide TS and 20 mL of water as an absorbing liq-uid: the test solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Tegafur in 0.01 mol/L sodium hydroxide TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tegafur, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample with a mixture of methanol and acetone (1:1), filter and dry the crystals, and perform the test with the crystals. pH <2.54> Dissolve 0.5 g of Tegafur in 50 mL of water: the pH of this solution is between 4.2 and 5.2.
(1) Clarity and color of solution—Dissolve 0.2 g of Tegafur in 10 mL of dilute sodium hydroxide TS: the so-lution is clear and colorless. Chloride <1.03>—Dissolve 0.8 g of Tegafur in 40 mL of water by warming, cool, filter if necessary, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the con-trol solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Heavy metals <1.07>—Dissolve 1.0 g of Tegafur in 40 mL of water by warming, cool, filter if necessary, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). ‌Arsenic <1.11>—Prepare the test solution in a plati-num crucible with 1.0 g of Tegafur according to Method 4, incinerating by ignition between 7509C and 8509C, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Tegafur in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of chloroform and ethanol (95) (5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g, plati-num crucible).
Weigh accurately about 0.15 g of Tegafur, previ-ously dried, place in an iodine bottle, dissolve in 75 mL of water, and add exactly 25 mL of 1/60 mol/L potassium bro-mate VS. Add rapidly 1.0 g of potassium bromide and 12 mL of hydrochloric acid, stopper the bottle tightly at once, and allow to stand for 30 minutes with occasional shaking. To this solution add 1.6 g of potassium iodide, shake gently, allow to stand for exactly 5 minutes, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indi-cator: 2 mL of starch TS). Perform a blank determination in the same manner. Each mL of 1/60 mol/L potassium bromate VS ＝ 10.01 mg of C8H9FN2O3
Teicoplanin A2-1 C88H95Cl2N9O33: 1877.64 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2- Acetylamino-2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-56-[2-(4Z )-dec-4-enoylamino-2-deoxy-b- D-glucopyranosyloxy]-6,11,40,44-tetrahydroxy-42-(a-D- mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo-2,3,16,17,18,19,35,36,37,38,48,49,50,50a- tetradecahydro-1H,15H,34H-20,23:30,33-dietheno-3,18:35,48-bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H- [1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]-benzoxadiazacyclotetracosine-38-carboxylic acid [91032-34-7] Teicoplanin A2-2 C88H97Cl2N9O33: 1879.66 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2- Acetylamino-2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-56-[2-deoxy-2-(8-methylnonanoylamino)-b-D-glucopyranosyloxy]-6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo-2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]-benzoxadiazacyclotetracosine-38-carboxylic acid [91032-26-7] Teicoplanin A2-3 C88H97Cl2N9O33: 1879.66 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2-Acetylamino- deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-56-(2-decanoylamino-2-deoxy-b-D-glucopyranosyloxy)-6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo- 2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]-benzoxadiazacyclotetracosine-38-carboxylic acid [91032-36-9] Teicoplanin A2-4 C89H99Cl2N9O33: 1893.68 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2- Acetylamino-2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-56-[2-deoxy-2-(8-methyldecanoylamino)-b-D-glucopyranosyloxy]-6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo-2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]-benzoxadiazacyclotetracosine-38-carboxylic acid [91032-37-0] Teicoplanin A2-5 C89H99Cl2N9O33: 1893.68 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2- Acetylamino-2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-56-[2-deoxy-2-(9-methyldecanoylamino)-b-D-glucopyranosyloxy]-6,11,40,44-tetrahydroxy-42-(a-D-mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo-2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]- It contains not less than 900 mg (potency) and not more than 1120 mg (potency) per 1 mg, calculated on the anhydrous, sodium chloride-free and residual solvent-free basis.
Teicoplanin occurs as a white to light yellowish white powder. It is freely soluble in water, sparingly soluble in N,N-dimethylformamide, and practically insoluble in acetonitrile, in methanol, in ethanol (95), in acetone, in acetic acid (100) and in diethyl ether.
(1) To 1 mL of a solution of Teicoplanin (1 in 100) add 2 mL of ninhydrin TS, and warm for 5 minutes: a blue-purple color develops. To 1 mL of a solution of Teicoplanin (3 in 100) add slowly 2 mL of anthrone TS, and shake gently: a dark brown color develops. Determine the infrared absorption spectra of Tei-coplanin and Teicoplanin RS as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the spectrum of Tei- coplanin RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. pH <2.54> Dissolve 0.5 g of Teicoplanin in 10 mL of water: the pH of the solution is between 6.3 and 7.7. Content ratio of the active principle Dissolve about 20 mg of Teicoplanin in water to make 10 mL, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the sum of peak areas of teicoplanin A2 group, Sa, the sum of peak areas of teicoplanin A3 group, Sb, and the sum of peak areas of other contents, Sc from the sample solution by the automatic integration method. Calculate the content ratio of them by the formula given below: teicoplanin A2 group, teicoplanin A3 group, and the other are not less than 80.0z, not more than 15.0z and not more than 5.0z, re-spectively. The elution order of each content and the relative reten-tion time of each content to teicoplanin A2-2 are shown in the following table. benzoxadiazacyclotetracosine-38-carboxylic acid [91032-38-1] Name of content Elution order Relative retention time Teicoplanin A3-1 C72H68Cl2N8O28: 1564.25 (3S,15R,18R,34R,35S,38S,48R,50aR)-34-(2- Acetylamino-2-deoxy-b-D-glucopyranosyloxy)-15-amino-22,31-dichloro-6,11,40,44,56-pentahydroxy-42-(a-D-mannopyranosyloxy)-2,16,36,50,51,59-hexaoxo-2,3,16,17,18,19,35,36,37,38,48,49,50,50a-tetradecahydro- 1H,15H,34H-20,23:30,33-dietheno-3,18:35,48- bis(iminomethano)-4,8:10,14:25,28:43,47-tetrametheno-28H-[1,14,6,22]dioxadiazacyclooctacosino[4,5-m][10,2,16]-benzoxadiazacyclotetracosine-38-carboxylic acid [93616-27-4] [61036-62-2, Teicoplanin] Teicoplanin is a mixture of glycopeptide substances having antibacterial activity produced by the growth of Actinoplanes teichomyceticus. teicoplanin A3 group ≦0.42 teicoplanin A3-1 1 0.29 teicoplanin A2 group 0.42＜,≦1.25 teicoplanin A2-1 2 0.91 teicoplanin A2-2 3 1.00 teicoplanin A2-3 4 1.04 teicoplanin A2-4 5 1.17 teicoplanin A2-5 6 1.20 others 1.25＜ Content ratio (z) of teicoplanin A2 group ＝ Sa/(Sa ＋ 0.83Sb ＋ Sc) × 100 Content ratio (z) of teicoplanin A3 group ＝ 0.83Sb/(Sa ＋ 0.83Sb ＋ Sc) × 100 Content ratio (z) of others ＝ Sc/(Sa ＋ 0.83Sb ＋ Sc) × 100 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 7.80 g of sodium dihydrogen phosphate dihydrate in 1650 mL of water, add 300 mL of acetonitrile, adjust pH to 6.0 with sodium hydroxide TS, and add water to make 2000 mL. Mobile phase B: Dissolve 7.80 g of sodium dihydrogen phosphate dihydrate in 550 mL of water, add 1400 mL of acetonitrile, adjust the pH to 6.0 with sodium hydroxide TS, and add water to make 2000 mL. Flowing of mobile phase: Flow mobile phase A for 10 minutes before injection. After injection, control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 32 100 → 70 0 → 30 32 – 40 70 → 50 30 → 50 40 – 42 50 → 100 50 → 0 Flow rate: About 1.8 mL per minute. Time span of measurement: About 1.7 times as long as the retention time of teicoplanin A2-2, beginning after the solvent peak. System suitability— Test for required detectability: Confirm that peak height of teicoplanin A2-2 obtained with the sample solution is equivalent to 90z of the full scale. System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, the symmetry factor of the peak of teicoplanin A3-1 is not more than 2.2. System repeatability: When the test is repeated 3 times with 20 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of teicoplanin A2-2 is not more than 2.0z.
(1) Clarity and color of solution—Dissolve 0.8 g of Teicoplanin in 10 mL of water: the solution is clear. Per-form the test with this solution according to Method 1 under Methods for Color Matching <2.65>: the color is not more colored than Matching Fluids BY3 and B4. Sodium chloride—Weigh accurately about 0.5 g of Teicoplanin, dissolve in 50 mL of water, titrate <2.50> with 0.1 mol/L silver nitrate VS (indicator: 1 mL of potassium chromate TS), and calculate an amount of sodium chloride: not more than 5.0z. Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl Heavy metals <1.07>—Place 2.0 g of Teicoplanin in a quartz or porcelain crucible, cover loosely with a lid, and heat gently to carbonize. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid, heat cautiously until white fumes are no longer evolved, and incinerate by ignition be-tween 5009C and 6009C. If a carbonized substance remains, add 2 mL of nitric acid and 5 drops of sulfuric acid, heat in the same manner as above, and incinerate by ignition be-tween 5009C and 6009C. Cool, then proceed according to Method 2, and perform the test. The control solution is pre-pared as follows: Evaporate a mixture of 4 mL of nitric acid, 10 drops of sulfuric acid and 2 mL of hydrochloric acid on a water bath, further evaporate to dryness on a sand bath, and moisten the residue with 3 drops of hydrochloric acid. Then proceed in the same manner as the test solution, and add 1.0 mL of Standard Lead Solution and water to make 50 mL (not more than 5 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Teicoplanin according to Method 3, and perform the test (not more than 2 ppm). Residual solvents <2.46>—Weigh accurately about g of Teicoplanin, dissolve in N,N-dimethylformamide to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 1 g each of methanol and acetone, and add N,N-dimethylformamide to make exactly 100 mL. Pipet 1 mL of this solution, add N,N-dimethylformamide to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 4 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions. Determine the peak area of methanol, A1, and the peak area of acetone, A2, obtained from the sample solution, and the peak area of methanol, AS1, and the peak area of acetone, AS2, from the standard solution by the automatic integration method, and calculate the amounts of methanol and acetone by the following formula: not more than 0.5z and not more than 1.0z, respectively. Amount (z) of methanol ＝ MS1 × A1/AS1 × 0.001 × 1/MT × 100 Amount (z) of acetone ＝ MS2 × A2/AS2 × 0.001 × 1/MT × 100 MS1: Amount (g) of methanol taken MS2: Amount (g) of acetone taken MT: Amount (g) of Teicoplanin taken Operating conditions— Detector: A Hydrogen flame-ionization detector. Column: A glass column 2 mm in inside diameter and 3 m in length, packed with graphite carbon for gas chromatogra-phy, 150 to 180 mm in particle diameter, coated with 0.1z of polyethylene glycol esterified. Column temperature: Inject the sample at a constant tem-perature of about 709C, maintain the temperature for 4 minutes, then program to raise the temperature to 2109C at the rate of 89C per minute. Detector temperature: A constant temperature of about 2409C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention times of methanol and acetone are about 2 minutes and 5 minutes, respectively. System suitability— Test for required detectability: Confirm that the peak height of acetone obtained from 4 mL of the standard solu-tion is equivalent to about the full scale. System performance: When the procedure is run with 4 mL of the standard solution under the above operating condi-tions, methanol and acetone are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 3 times with 4 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas of acetone is not more than 3z. Water <2.48> Not more than 15.0z (0.2 g, volumetric titration, direct titration). ‌Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Teicoplanin RS equivalent to about 50 mg (potency), dis-solve in phosphate buffer solution (pH 6.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at not exceeding 59C and use within 14 days. Take exactly a suitable amount of this solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 160 mg (potency) and 40 mg (potency), and use these solutions as the high con-centration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Teicoplanin equivalent to about 50 mg (potency), dissolve in phosphate buffer solution (pH 6.0) to make exactly 50 mL. Take exactly a suitable amount of this solution, add phos-phate buffer solution (pH 6.0) to make solutions so that each mL contains 160 mg (potency) and 40 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Containers—Tight containers. Storage—Light-resistant, and at a temperature between
Telmisartan, when dried, contains not less than 99.
Telmisartan occurs as a white to pale yellow crystalline powder. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) It is freely soluble in formic acid, slightly soluble in metha-nol, very slightly soluble in ethanol (99.5), and practically in-soluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Telmisartan in methanol (7 in 1,000,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tel-misartan as directed in the potassium bromide disk method 0 – 25 70 → 20 30 → 80 Flow rate: 1.0 mL per minute. Time span of measurement: About 2 times as long as the retention time of telmisartan, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add methanol to make exactly 100 mL. Confirm that the peak area of telmisartan obtained with 2 mL of this solution is equivalent to 3.5 to 6.5z of that with 2 mL of the standard solution. ‌System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of telmisartan are not less than 45,000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of telmisartan is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Telmisartan according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 10 ppm). (2) Related substances—To 25 mg of Telmisartan add 5 mL of methanol and 0.1 mL of sodium hydroxide TS, and dissolve by sonicating. To this solution add methanol to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the peak, having the relative retention time of about 1.7 to telmisartan, obtained from the sample solution is not larger than 1/5 times the peak area of telmisartan from the standard solution, the area of the peak other than telmisartan and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of telmisartan from the standard solution, and the total area of the peaks other than telmisartan from the sample so-lution is not larger than the peak area of telmisartan from the standard solution. For the area of the peak, having the relative retention time of about 0.7, multiply its correction factor 1.2. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2.0 g of potassium dihydrogen phosphate and 3.4 g of sodium 1-pentanesulfonate in 1000 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (1 in 10). Mobile phase B: A mixture of acetonitrile and methanol (4:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Description Telmisartan occurs as a white to pale yellow crystalline powder. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) It is freely soluble in formic acid, slightly soluble in metha-nol, very slightly soluble in ethanol (99.5), and practically in-soluble in water. It shows crystal polymorphism. Identification (1) Determine the absorption spectrum of a solution of Telmisartan in methanol (7 in 1,000,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tel-misartan as directed in the potassium bromide disk method 0 – 25 70 → 20 30 → 80 Flow rate: 1.0 mL per minute. Time span of measurement: About 2 times as long as the retention time of telmisartan, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add methanol to make exactly 100 mL. Confirm that the peak area of telmisartan obtained with 2 mL of this solution is equivalent to 3.5 to 6.5z of that with 2 mL of the standard solution. ‌System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of telmisartan are not less than 45,000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of telmisartan is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.19 g of Telmisartan, pre-viously dried, dissolve in 5 mL of formic acid, add 75 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 25.73 mg of C33H30N4O2
Temocapril Hydrochloride contains not less than 99.
Temocapril Hydrochloride occurs as a white crystalline powder. It is freely soluble in ethanol (99.5), and vey slightly solu-ble in water.
(1) Determine the absorption spectrum of a solution of Temocapril Hydrochloride in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Temocapril Hydrochloride as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Temocapril Hydrochloride in ethanol (99.5) (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride. Optical rotation <2.49> [a]20: ＋60 – ＋649(0.2 g calculated ＝ MS × AT/AS 7 8 3 4 2 D on the anhydrous basis, ethanol (99.5), 20 mL, 100 mm). MS: Amount (mg) of Hydrochlorothiazide RS taken Dissolving solution: Dissolve 2 g of ammonium dihydro- gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Temocapril Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Temocapril Hydrochloride in 100 mL of diluted acetonitrile (1 in 2), and ‌use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted acetonitrile (1 in 2) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: Temocapril Hydrochloride Tablets
Teprenone occurs as a colorless to pale yellow clear oily liquid, with slight, characteristic odor. It is miscible with ethanol (99.5), with ethyl acetate and with hexane. It is practically insoluble in water. It is oxidized by air, and gradually turns yellow.
(1) To 2 mL of a solution of Teprenone in ethanol (99.5) (1 in 100) add 1 mL of a solution of phos-phomolybdic acid n-hydrate in acetic acid (100) (1 in 100), heat in a water bath for 5 minutes, and continue heating with addition of 5 to 6 drops of sulfuric acid: blue to bluish green color develops. To 2 mL of a solution of Teprenone in ethanol (99.5) (1 in 100) add 2 mL of 2,4-dinitrophenylhydrazine TS, and shake: a yellow to orange-yellow precipitate is formed. Determine the infrared absorption spectrum of Teprenone as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Teprenone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.485 – 1.491 Specific gravity <2.56> d 20: 0.882 – 0.890
(1) Clarity and color of solution—To 1.0 mL of Teprenone add 9 mL of ethanol (99.5) and shake: the solu-tion is clear, and its absorbance at 400 nm determined as di-rected under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.02. Heavy metals <1.07>—Proceed with 1.0 g of Tepre- none according to Method 2 and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 30 mg of Teprenone in 6 mL of hexane, and use this solution as the sample solution. Perform the test with 3 mL of the sample solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions. Determine each peak area from the sample solution by the automatic integration method and calculate the amounts of them by the area percentage method: the peak area of the di-cis isomer of teprenone, having the rela-tive retention time of about 0.8 to the all-trans isomer of teprenone, is not more than 0.5z, and each area of the peaks other than the mono-cis and all-trans isomers of the teprenone and the other than mentioned above is not more than 0.2z. Furthermore, the total area of the peaks other than the mono-cis, all-trans and di-cis isomers of teprenone is not more than 1.0z. Operating conditions— Detector, column, column temperature, carrier gas and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time for the all-trans isomer of teprenone, begin-ning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add hexane to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the so-lution for system suitability test, and add hexane to make ex-actly 10 mL. Confirm that the sum of the peak areas of the mono-cis and all-trans isomers of teprenone obtained with 3 mL of this solution is 7 to 13z of the peak areas of the mono-cis and all-trans isomers of teprenone with 3 mL of the solution for system suitability test. System performance: When the procedure is run with 3 mL of the solution for system suitability test under the above op-erating conditions, the mono-cis and all-trans isomers of teprenone are eluted in this order with the resolution be-tween these peaks being not less than 1.1. System repeatability: When the test is repeated 6 times with 3 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the sum of the peak areas of the mono-cis and all-trans isomers of teprenone is not more than 3.0z. Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 30 mg of Teprenone in 6 mL of hexane, and use this solution as the sample solution. Per-form the test with 3 mL of the sample solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions. Determine the areas of two adjacent peaks, Aa and Ab, having retention times of about 18 minutes, where Aa is the peak area of the mono-cis isomer, having the shorter retention time, and Ab is the peak area of the all-trans isomer, having the longer retention time: Aa/Ab is 0.60 to 0.70. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Purity (3).
Weigh accurately the mass of not less than 20 Temocapril Hydrochloride Tablets, and powder. Weigh ac- C23 H38O: 330.55 curately a portion of the powder, equivalent to about 10 mg of temocapril hydrochloride (C23H28N2O5S2.HCl), add ex-actly 20 mL of the internal standard solution, and sonicate for 10 minutes. Furthermore, shake for 10 minutes, and cen-trifuge. To 2 mL of the supernatant liquid add diluted aceto-nitrile (1 in 2) to make 20 mL, and use this solution as the (5E,9E,13E )-6,10,14,18-Tetramethylnonadeca- 5,9,13,17-tetraen-2-one (5Z,9E,13E )-6,10,14,18-Tetramethylnonadeca-5,9,13,17-tetraen-2-one [6809-52-5] Teprenone contains not less than 97.0z and not sample solution. Separately, weigh accurately about 50 mg of temocapril hydrochloride for assay (separately determine more than 101.0z of teprenone (C 23H38 O). the water <2.48> in the same manner as Temocapril Hydro-chloride), and dissolve in diluted acetonitrile (1 in 2) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 10 mL of the internal standard solution, then add diluted aceto-nitrile (1 in 2) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq- Teprenone is comprised of mono-cis and all-trans isomers, with their ratio being about 2:3. Description Teprenone occurs as a colorless to pale yellow clear oily liquid, with slight, characteristic odor. It is miscible with ethanol (99.5), with ethyl acetate and with hexane. It is practically insoluble in water. It is oxidized by air, and gradually turns yellow. Identification (1) To 2 mL of a solution of Teprenone in ethanol (99.5) (1 in 100) add 1 mL of a solution of phos-phomolybdic acid n-hydrate in acetic acid (100) (1 in 100), heat in a water bath for 5 minutes, and continue heating with addition of 5 to 6 drops of sulfuric acid: blue to bluish green color develops. To 2 mL of a solution of Teprenone in ethanol (99.5) (1 in 100) add 2 mL of 2,4-dinitrophenylhydrazine TS, and shake: a yellow to orange-yellow precipitate is formed. Determine the infrared absorption spectrum of Teprenone as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Teprenone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.485 – 1.491 Specific gravity <2.56> d 20: 0.882 – 0.890 Purity (1) Clarity and color of solution—To 1.0 mL of Teprenone add 9 mL of ethanol (99.5) and shake: the solu-tion is clear, and its absorbance at 400 nm determined as di-rected under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.02. Heavy metals <1.07>—Proceed with 1.0 g of Tepre- none according to Method 2 and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 30 mg of Teprenone in 6 mL of hexane, and use this solution as the sample solution. Perform the test with 3 mL of the sample solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions. Determine each peak area from the sample solution by the automatic integration method and calculate the amounts of them by the area percentage method: the peak area of the di-cis isomer of teprenone, having the rela-tive retention time of about 0.8 to the all-trans isomer of teprenone, is not more than 0.5z, and each area of the peaks other than the mono-cis and all-trans isomers of the teprenone and the other than mentioned above is not more than 0.2z. Furthermore, the total area of the peaks other than the mono-cis, all-trans and di-cis isomers of teprenone is not more than 1.0z. Operating conditions— Detector, column, column temperature, carrier gas and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time for the all-trans isomer of teprenone, begin-ning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add hexane to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the so-lution for system suitability test, and add hexane to make ex-actly 10 mL. Confirm that the sum of the peak areas of the mono-cis and all-trans isomers of teprenone obtained with 3 mL of this solution is 7 to 13z of the peak areas of the mono-cis and all-trans isomers of teprenone with 3 mL of the solution for system suitability test. System performance: When the procedure is run with 3 mL of the solution for system suitability test under the above op-erating conditions, the mono-cis and all-trans isomers of teprenone are eluted in this order with the resolution be-tween these peaks being not less than 1.1. System repeatability: When the test is repeated 6 times with 3 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the sum of the peak areas of the mono-cis and all-trans isomers of teprenone is not more than 3.0z. Residue on ignition <2.44> Not more than 0.1z (1 g). Isomer ratio Dissolve 30 mg of Teprenone in 6 mL of hexane, and use this solution as the sample solution. Per-form the test with 3 mL of the sample solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions. Determine the areas of two adjacent peaks, Aa and Ab, having retention times of about 18 minutes, where Aa is the peak area of the mono-cis isomer, having the shorter retention time, and Ab is the peak area of the all-trans isomer, having the longer retention time: Aa/Ab is 0.60 to 0.70. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Purity (3). Assay Weigh accurately about 50 mg each of Teprenone and Teprenone RS, dissolve each in exactly 5 mL of the in-ternal standard solution, add ethyl acetate to make 50 mL, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with 3 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of teprenone (sum of the peak areas of mono-cis and all-trans isomers) to that of the internal standard. Amount (mg) of teprenone (C23H38O) ＝ MS × QT/QS MS: Amount (mg) of Teprenone RS taken Internal standard solution—A solution of di-n-butyl phtha-late in ethyl acetate (1 in 200). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 2 m in length, packed with 149 to 177 mm silica-gel for gas chro-matography coated in 5z with polyethylene glycol 2-nitroterephthalate for gas chromatography. Column temperature: A constant temperature of about 2359C. Carrier gas: Nitrogen or helium. Flow rate: Adjust so that the retention time of the peak of all-trans isomer of teprenone, having the larger retention time among the adjacent two main peaks appearing at a retention time of about 18 minutes, is about 19 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, the internal standard and the mono-cis and all-trans isomers of teprenone are eluted in this order with the resolu-tion between the mono-cis and all-trans isomers being not less than 1.1. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the sum of the peak areas of the mono-cis and all-trans isomers of teprenone to that of the internal standard is not more than 1.0z.
Containers—Tight Containers. Storage—Under Nitrogen atmosphere at 2 to 89C.
Terbinafine Hydrochloride occurs as a white to pale yellow-white crystalline powder. It is freely soluble in methanol, in ethanol (99.5) and in acetic acid (100), and slightly soluble in water. The pH of a solution of 1.0 g of Terbinafine Hydrochlo- ride in 1000 mL of water is 3.5 to 4.5. Melting point: about 2059C (with decomposition).
(1) Determine the absorption spectrum of a solution of Terbinafine Hydrochloride in methanol (1 in 40,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Ter-binafine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Terbinafine Hydrochloride in ethanol (99.5) (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Terbinafine Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Terbinafine Hydro-chloride in 100 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add a mixture of water and acetonitrile (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of water and acetonitrile (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of a dimer, having the relative retention time of about 1.7 to terbinafine obtained from the sample solution is not larger than 1/2 times the peak area of terbinafine from the standard solution, the area of the peaks other than terbinafine and the dimer from the sample solution is not larger than the peak area of terbina-fine from the standard solution, and the total area of the peaks other than terbinafine is not larger than 3 times the peak area of terbinafine from the standard solution. Flow rate: Adjust so that the retention time of terbinafine is about 15 minutes. Time span of measurement: About 2 times as long as the retention time of terbinafine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water and acetonitrile (1:1) to make exactly 20 mL. Confirm that the peak area of terbina-fine obtained with 20 mL of this solution is equivalent to 18 to 32z of that with 20 mL of the standard solution. System performance: Dissolve 20 mg of Terbinafine Hy-drochloride in 20 mL of a mixture of water and acetonitrile (1:1), and irradiate under a short-wave lamp (main wave-length: 254 nm) for 1 hour. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution between the peak of cis-terbinafine, having the relative retention time of about 0.94 to terbinafine, and the peak of terbinafine is not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of terbinafine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.26 g of Terbinafine Hy-drochloride, previously dried, dissolve in 5 mL of acetic acid (100), add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 32.79 mg of C21H25N.HCl
Terbutaline Sulfate contains not less than 98.
Terbutaline Sulfate is white to brownish white, crystals or crystalline powder. It is odorless or has a faint odor of acetic acid. It is freely soluble in water, and practically insoluble in acetonitrile, in ethanol (95), in acetic acid (100), in chlo-roform, and in diethyl ether. It is gradually colored by light and by air. Melting point: about 2559C (with decomposition).
(1) Dissolve 1 mg of Terbutaline Sulfate in 1 mL of water, and add 5 mL of Tris buffer solution (pH 9.5), 0.5 mL of 4-aminoantipyrine solution (1 in 50) and 2 drops of potassium hexacyanoferrate (III) solution (2 in 25): a reddish purple color is produced. ‌Determine the absorption spectrum of a solution of Terbutaline Sulfate in 0.01 mol/L hydrochloric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. This maximum can be biphasic. A solution of Terbutaline Sulfate (1 in 50) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 0.10 g of Terbutaline Sulfate in 10 mL of water: the pH of this solution is between 4.0 and 4.8.
(1) Clarity and color of solution—Dissolve 0.10 g of Terbutaline Sulfate in 10 mL of water: the solution is clear and colorless or pale yellow. Chloride <1.03>—Perform the test with 2.0 g of Terbutaline Sulfate. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.004z). Acetic acid—Dissolve 0.50 g of Terbutaline Sulfate in a solution of phosphoric acid (59 in 1000) to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 1.50 g of acetic acid (100) in a solution of phos-phoric acid (59 in 1000) to make exactly 100 mL. Dilute 2 mL of this solution, accurately measured, with a solution of phosphoric acid (59 in 1000) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> ac-cording to the following conditions. Determine the peak areas, AT and AS, of acetic acid in each solution: AT is not larger than AS. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with 10z of macrogol 6000 on 180- to 250-mm terephthalic acid for gas chromatography. Column temperature: A constant temperature at about 1209C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of acetic acid is about 5 minutes. System suitability— System performance: Mix 0.05 g each of acetic acid (100) and propionic acid in 100 mL of diluted phosphoric acid (59 in 1000). When the procedure is run with 2 mL of this solu-tion under the above conditions, acetic acid and propionic acid are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak areas of acetic acid is not more than 3.0z. 3,5-Dihydroxy-v-tert-butylaminoacetophenone sul-fate—Dissolve 0.50 g of Terbutaline Sulfate in 0.01 mol/L hydrochloric acid TS to make exactly 25 mL, and perform the test as directed under Ultraviolet-visible Spectropho-tometry <2.24>: the absorbance at a wavelength of 330 nm does not exceed 0.47. Heavy metals <1.07>—Proceed with 2.0 g of Terbuta-line Sulfate as directed under Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Terbutaline Sulfate according to method 3, and perform the test (not more than 2 ppm). Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Terbutaline Sulfate, dissolve in 50 mL of a mixture of acetonitrile and acetic acid (100) (1:1) by stirring and warming. Allow to cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration, substituting a saturated solution of potas-sium chloride in methanol for the internal fluid). Each mL of 0.1 mol/L perchloric acid VS ＝ 54.87 mg of (C12H19NO3)2.H2SO4
テストステロンエナント酸エステル
Testosterone Enanthate, when dried, contains not less than 95.
Testosterone Enanthate occurs as white to pale yellow, crystals or crystalline powder, or a pale yellow-brown viscous liquid. It is odorless or has a slight, character-istic odor. It is freely soluble in ethanol (99.5), and practically insolu-ble in water. Melting point: about 369C
Heat 25 mg of Testosterone Enanthate with 2 mL of a solution of potassium hydroxide in methanol (1 in 100) under a reflux condenser on a water bath for 1 hour, cool, and add 10 mL of water. Collect the produced precipi-tate by suction, wash with water until the last washing is neu-tral, and dry the precipitate in a desiccator (in vacuum, phos-phorus (V) oxide) for 4 hours: the precipitate melts <2.60> between 1519C and 1579C. D Optical rotation <2.49> [a]25: ＋76 – ＋869(after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm).
Acidity—Dissolve 0.5 g of Testosterone Enanthate in 10 mL of ethanol (95) which has previously been rendered neutral to bromothymol blue TS, and add 2 drops of bromo-thymol blue TS and 0.50 mL of 0.01 mol/L sodium hydrox-ide VS: the color of the solution is light blue. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately about 0.1 g of Testosterone Enan-thate, previously dried, and dissolve in ethanol (95) to make exactly 100 mL. Measure exactly 10 mL of this solution, and ‌dilute with ethanol (95) to make exactly 100 mL. Measure ex-actly 10 mL of this solution, and dilute with ethanol (95) to make exactly 100 mL. Perform the test as directed under Ultraviolet-visible Spectrophotometry <2.24> with this solu-tion. Determine the absorbance A of this solution at the wavelength of maximum absorption at about 241 nm.‌ Amount (mg) of testosterone enanthate (C26H40O3) ＝ A/426 × 100,000
テストステロンプロピオン酸エステル
Testosterone Propionate, when dried, contains not less than 97.
Testosterone Propionate occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (95), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Testosterone Propionate in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Testosterone Propionate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. (2) Determine the infrared absorption spectrum of Testosterone Propionate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Testosterone Propionate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋83 – ＋909(after drying, 0.1 g, ethanol (95), 10 mL, 100 mm).
Related substances—Dissolve 40 mg of Testosterone Propionate in 2 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, phosphorus (V) oxide, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g).
Weigh accurately each about 10 mg of Testosterone Propionate and Testosterone Propionate RS, previously dried, and dissolve in methanol to make exactly 100 mL. To ‌exactly 5 mL of these solutions add exactly 5 mL of the inter-nal standard solution and methanol to make 20 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of testosterone propionate to that of the internal standard. Amount (mg) of testosterone propionate (C22H32O3) ＝ MS × QT/QS MS: Amount (mg) of Testosterone Propionate RS taken Internal standard solution—A solution of progesterone in methanol (9 in 100,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 241 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile and water (7:3). Flow rate: Adjust so that the retention time of testoster- one propionate is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and testosterone propionate are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of testosterone propionate to that of the internal standard is not more than 1.0z.
Tetracaine Hydrochloride, when dried, contains not less than 98.
Tetracaine Hydrochloride occurs as white, crystals or crystalline powder. It is odorless, and has a slightly bitter taste followed by a sense of numbness on the tongue. It is very soluble in formic acid, freely soluble in water, soluble in ethanol (95), sparingly soluble in ethanol (99.5), slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. A solution of Tetracaine Hydrochloride (1 in 10) is neu-tral. Melting point: about 1489C.
Heavy metals <1.07>—Proceed with 1.0 g of Tetracaine Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Tetracaine Hydro-chloride, previously dried, dissolve in 2 mL of formic acid, add 80 mL of acetic anhydride, allow to stand at 309C on a water bath for 15 minutes, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.08 mg of C15H24N2O2.HCl
Tetracycline Hydrochloride is the hydrochloride of a tetracycline substance having antibacterial activity produced by the growth of Streptomyces aureofaciens. It contains not less than 950 mg (potency) and not more than 1010 mg (potency) per mg, calculated on the dried basis.
Tetracycline Hydrochloride occurs as a yellow to pale brownish yellow crystalline powder. It is freely soluble in water, and sparingly soluble in ethanol (95).
(1) Dissolve 0.5 g of Tetracaine Hydro- chloride in 50 mL of water, add 5 mL of ammonia TS, shake, and allow to stand in a cold place. Collect the precipi-tate, wash with water until the washings become neutral, and dry in a desiccator (silica gel) for 24 hours: it melts <2.60> be-tween 429C and 449C. Dissolve 0.1 g of Tetracaine Hydrochloride in 8 mL of water, and add 3 mL of ammonium thiocyanate TS: a crystalline precipitate is produced. Collect the precipitate, recrystallize from water, and dry at 809C for 2 hours: it melts <2.60> between 1309C and 1329C. Determine the absorption spectrum of a solution of Tetracaine Hydrochloride in ethanol (99.5) (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Tetracaine Hydrochloride (1 in 10) re-sponds to Qualitative Tests <1.09> for chloride. Description Tetracycline Hydrochloride occurs as a yellow to pale brownish yellow crystalline powder. It is freely soluble in water, and sparingly soluble in ethanol (95). Identification (1) Determine the absorption spectrum of a solution of Tetracycline Hydrochloride (1 in 62,500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Tetracycline Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tetracycline Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Tetracycline Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. ‌A solution of Tetracycline Hydrochloride (1 in 100) responds to Qualitative Tests <1.09> (2) for chloride.‌ pH <2.54> Dissolve 1.0 g of Tetracycline Hydrochloride in 100 mL of water: the pH of the solution is between 1.8 and 2.8.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tetracycline Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 25 mg of Tetracycline Hydrochloride in 50 mL of 0.01 mol/L hydrochloric acid TS, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak other than tetracy-cline obtained from the sample solution is not larger than the peak area of tetracycline from the standard solution, and the total area of the peaks other than tetracycline from the sam-ple solution is not larger than 3 times the peak area of tetracycline from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 7 times as long as the retention time of tetracycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 3 mL of the standard solution, add 0.1 mol/L hydrochloric acid TS to make ex-actly 100 mL, and confirm that the peak area of tetracycline obtained with 20 mL of this solution is equivalent to 1 to 5z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tetracycline is not more than 1.0z. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1.0 g).
Weigh accurately an amount of Tetracycline Hydro-chloride and Tetracycline Hydrochloride RS, equivalent to about 25 mg (potency), and dissolve each in 0.1 mol/L hy-drochloric acid TS to make exactly 50 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with exactly 20 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of tetracycline in each solution. Amount [ mg (potency)] of tetracycline hydrochloride (C22H24N2O8.HCl) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Tetracycline Hydrochlo-ride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with stylene-divinylbenzene copolymer for liquid chromatography (0.01 mm in pore di-ameter). Column temperature: A constant temperature of about 609C. Mobile phase: Dissolve 3.5 g of dipotassium hydrogen-phosphate, 2.0 g of tetrabutylammonium hydrogensulfate and 0.4 g of disodium dihydrogen ethylenediamine tetraace-tate dihydrate in 300 mL of water, adjust to pH 9.0 with so-dium hydroxide TS, add 90.0 g of t-butyl alcohol, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of tetracycline is about 5 minutes. System suitability— System performance: Dissolve 0.05 g of Tetracycline Hy-drochloride RS in water to make 25 mL. Heat 5 mL of this solution on a water bath for 60 minutes, then add water to make 25 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the retention time of 4-epitetracycline is about 3 minutes, and 4-epitetracycline and tetracycline are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When, the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of tetracycline is not more than 1.0z.
Theophylline, when dried, contains not less than 99.
(1) Determine the absorption spectrum of a solution of Theophylline in 0.1 mol/L hydrochloric acid TS (1 in 200,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of The-ophylline, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Acidity—To 0.5 g of Theophylline add 75 mL of water, 2.0 mL of 0.01 mol/L sodium hydroxide VS and 1 drop of methyl red TS: a yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of The- ophylline according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Theophylline according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Theophylline in 3 mL of N,N-dimethylformamide, add 10 mL of metha-nol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of acetone, chloroform, methanol, 1-butanol and ammonia solution (28) (3:3:2:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Theophylline, pre-viously dried, and dissolve in 100 mL of water, add exactly 20 mL of 0.1 mol/L silver nitrate VS, shake the mixture, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 18.02 mg of C7H8N4O2
Thiamazole, when dried, contains not less than 98.
Thiamazole occurs as white to pale yellow-white, crystals or crystalline powder. It has a faint, charac-teristic odor, and has a bitter taste. It is freely soluble in water and in ethanol (95), and slightly soluble in diethyl ether. The pH of a solution of 1.0 g of Thiamazole in 50 mL of water is between 5.0 and 7.0.
(1) Dissolve 5 mg of Thiamazole in 1 mL of water, shake with 1 mL of sodium hydroxide TS, and add 3 drops of sodium pentacyanonitrosylferrate (III) TS: a yel-low color develops, and it gradually changes to yellow-green to green. To this solution add 1 mL of acetic acid (31): it changes to blue. To 2 mL of a solution of Thiamazole (1 in 200) add 1 mL of sodium carbonate TS and 1 mL of diluted Folin's TS (1 in 5): a deep blue color develops.
(1) Selenium—Proceed with 0.10 g of Thiamazole as directed under Oxygen Flask Combustion Method <1.06>, using 25 mL of diluted nitric acid (1 in 30) as the absorbing liquid, and prepare the test solution. Apply a small amount of water to the upper part of apparatus A, pull out C care-fully, and transfer the test solution to a beaker. Wash C, B and the inner side of A with 25 mL of water, and combine the washings with the test solution. Boil gently for 10 minutes, cool to room temperature, add water to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh exactly 40 mg of selenium, dissolve in 100 mL of diluted nitric acid (1 in 2), heat to dissolve on a water bath if necessary, and add water to make exactly 1000 mL. Pipet 5 mL of this solution, and add water to make exactly 200 mL. To 2 mL of this solution, exactly measured, add di-luted nitric acid (1 in 60) to make exactly 50 mL, and use this solution as the standard solution. Pipet 40 mL each of the sample solution and standard solution into separate beakers, and adjust each solution with ammonia solution (28) to a pH of 1.8 to 2.2. To each solution add 0.2 g of hydroxylammo-nium chloride, shake gently to dissolve. To these solutions add 5 mL of a solution prepared by dissolving 0.10 g of 2,3-diaminonaphthalene and 0.5 g of hydroxylammonium chlo-ride in 0.1 mol/L hydrochloric acid TS to make 100 mL, shake, and allow to stand for 100 minutes. Transfer these so-lutions to corresponding separators, rinse the beakers with 10 mL of water, combine the rinsings in the respective sepa-rators, shake well with 5.0 mL of cyclohexane for 2 minutes, and extract. Centrifuge the cyclohexane extracts to remove any water remaining in these solutions. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared with 40 mL of diluted nitric acid (1 in 60) in the same manner as the blank. ‌The absorbance of of the sample solution at the wavelength of maximum absorbance at about 378 nm does not exceed the absorbance of the standard solution.‌ Heavy metals <1.07>—Proceed with 1.0 g of Thiama- zole according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Thiamazole according to Method 1, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Thiamazole, previ-ously dried, dissolve in 75 mL of water, add 15 mL of 0.1 mol/L sodium hydroxide VS from a burette, and add 30 mL of 0.1 mol/L silver nitrate VS with stirring. Add 1 mL of bromothymol blue TS, and titrate <2.50> with 0.1 mol/L so-dium hydroxide VS, until a persistent blue-green color is produced. Determine the total volume of 0.1 mol/L sodium hydroxide VS consumed. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 11.42 mg of C4H6N2S
Thianthol consists of dimethylthianthrene and ditol-uene disulfide. It contains not less than 23.
Thianthol is a yellowish, viscous liquid. It has a faint, agreeable odor. It is freely soluble in diethyl ether, slightly soluble in ethanol (95), and practically insoluble in water. It, when cold, may separate crystals, which melt on warm-ing. Specific gravity d 20: 1.19 – 1.23
To 0.1 g of Thianthol add cautiously 5 mL of sulfuric acid: a blue-purple color develops. Add 5 to 6 drops of nitric acid to the solution: the color of the solution changes to yellow-red with evolution of gas.
(1) Acidity or alkalinity—Shake 10 g of Thianthol with 20 mL of water, allow to stand, and separate the water layer. The solution is neutral. (2) Sulfate—To 10 mL of the water layer obtained in (1) add 2 to 3 drops of barium chloride TS: no opalescence is produced. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 10 mg of Thianthol, and proceed as directed in the sulfur determination of Oxygen Flask Combustion Method <1.06>, using a mixture of 5 mL of diluted sodium hydroxide TS (1 in 10) and 1.0 mL of hydrogen peroxide TS as an absorbing liquid.
Thioridazine Hydrochloride, when dried, contains not less than 99.
Thioridazine Hydrochloride occurs as a white to pale yellow crystalline powder. It is odorless, and has a bitter taste. It is freely soluble in water, in methanol, in ethanol (95) and in acetic acid (100), sparingly soluble in acetic anhy-dride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Thioridazine Hydrochlo-ride in 100 mL of water is between 4.2 and 5.2. It is gradually colored by light.
(1) Dissolve 0.01 g of Thioridazine Hydro-chloride in 2 mL of sulfuric acid: a deep blue color develops. Dissolve 0.01 g of Thioridazine Hydrochloride in 2 mL of water, and add 1 drop of cerium (IV) tetraammonium sulfate TS: a blue color develops, and the color disappears on the addition of excess of the reagent. Determine the infrared absorption spectrum of Thioridazine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Thioridazine Hydrochloride (1 in 100) add 2 mL of ammonia TS, and heat on a water bath for 5 minutes. After cooling, filter, and acidify the fil-trate with dilute nitric acid: the solution responds to Qualita-tive Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Thioridazine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Thioridazine Hydrochloride, according to Method 3, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure without exposure to light. Dissolve 0.10 g of Thioridazine Hydro- chloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, 2-propanol and ammonia solution (28) (74:25:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Thioridazine Hy-drochloride, previously dried, dissolve in 80 mL of a mixture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.70 mg of C21H26N2S2.HCl
Thrombin is prepared from prothrombin obtained from blood of man or bull, through interaction with added thromboplastin in the presence of calcium ions, sterilized and lyophilized. It contains not less than 80z and not more than 150z of the labeled Units of thrombin.
Thrombin is a white to light yellow, amor-phous substance. Thrombin (500 Units) dissolves in 1.0 mL of isotonic sodi-um chloride solution clearly or with slight turbidity within 1 minute. Loss on drying <2.41> Not more than 3z (50 mg, in vacu-um, phosphorus (V) oxide, 4 hours). Sterility <4.06> It meets the requirement.
(i) Fibrinogen solution—Weigh accurately about 30 mg of fibrinogen, and dissolve in 3 mL of isotonic sodium chloride solution. Allow the solution to clot sufficiently with frequent shaking after the addition of about 3 Units of thrombin. Wash the precipitated clot thoroughly until the washings yield no turbidity on addition of silver nitrate TS, weigh the clot after drying at 1059C for 3 hours, and calcu-late the percentage of the clot in the fibrinogen. Dissolve the fibrinogen in isotonic sodium chloride solution so that the clot should be 0.20z, adjust the pH of the solution between 7.0 and 7.4 by addition of 0.05 mol/L dibasic sodium phos-phate TS (or if necessary, use 0.5 mol/L disodium hydrogen-phosphate TS), and dilute with isotonic sodium chloride so-lution to make a 0.10z solution. (ii) Procedure—Dissolve Thrombin RS in isotonic sodi-um chloride solution, and prepare four kinds of standard so-lutions which contain 4.0, 5.0, 6.2, and 7.5 Units in 1 mL. Transfer accurately 0.10 mL each of the standard solutions maintained at a given degree ± 19C between 209C and 309C to a small test tube, 10 mm in inside diameter, 100 mm in length, blow out 0.90 mL of the fibrinogen solution at the same temperature into the test tube from a pipet, start a stop watch simultaneously, shake the tube constantly, and deter-mine the time for the first appearance of clot. Calculate the average values of five determinations for the four kinds of standard solutions, respectively. If the deviation between the maximum and the minimum values of five determinations is more than 10z of the average value, reject the whole run, and try the experiment again. The concentration of the standard solution may be changed appropriately within the range between 14 and 60 seconds of the clotting time. The determination proceeds at the same temperature described above. Next, weigh accurately the whole contents of a single container of Thrombin, dissolve it in isotonic sodium chlo-ride solution to provide a solution which is presumed to con-tain about 5 Units in each mL, treat 0.10 mL of the solution with the same reagents in the same manner five times, deter-mine the clotting times, and calculate the average value. Plot the average values of the clotting times of the four kinds of the standard solutions on a logarithmic graph, using Units as the abscissa and clotting times as the ordinate, and draw a calibration line which best fits the four plotted points. Using this line, read the Units U from the average value of the clot-ting times of the sample solution. Units of 1 container of Thrombin ＝ U × 10 × V V: The number of mL of the volume in which the contents of 1 container of Thrombin has been dissolved Calculate the units for 1 mg of the contents.
Containers—Hermetic containers. Storage—Not exceeding 109C.
Thymol occurs as colorless crystals or white crystalline masses. It has an aromatic odor, and has a burn-ing taste. It is very soluble in acetic acid (100), freely soluble in ethanol (95) and in diethyl ether, and slightly soluble in water. It sinks in water, but when warmed, it melts and rises to the surface of water.
(1) To 1 mL of a solution of Thymol in acetic acid (100) (1 in 300) add 6 drops of sulfuric acid and 1 drop of nitric acid: a blue-green color develops by reflected light and a red-purple color develops by transmitted light. Dissolve 1 g of Thymol in 5 mL of a solution of sodi-um hydroxide (1 in 10) by heating in a water bath, and con-tinue heating for several minutes: a light yellow-red color slowly develops. Allow this solution to stand at room tem-perature: the color changes to dark yellow-brown. Shake this solution with 2 to 3 drops of chloroform: a purple color gradually develops. Triturate Thymol with an equal mass of camphor or menthol: the mixture liquefies.
(1) Non-volatile residue—Volatilize 2.0 g of Thymol by heating on a water bath, and dry the residue at 1059C for 2 hours: the mass is not more than 1.0 mg. (2) Other phenols—Shake vigorously 1.0 g of Thymol with 20 mL of warm water for 1 minute, and filter. To 5 mL of the filtrate add 1 drop of iron (III) chloride TS: a green color may develop, but no blue to purple color develops.
Weigh accurately about 0.5 g of Thymol, dissolve in 10 mL of sodium hydroxide TS, and add water to make ex-actly 100 mL. Measure exactly 10 mL of the solution into an iodine flask, add 50 mL of water and 20 mL of dilute sulfu-ric acid, and cool in ice water for 30 minutes. Add exactly 20 mL of 0.05 mol/L bromine VS, stopper tightly immediately, allow to stand for 30 minutes in ice water with occasional shaking in a dark place, add 14 mL of potassium iodide TS and 5 mL of chloroform, stopper tightly, shake vigorously, and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Stopper tight-ly, shake vigorously near the end point, and continue the titration until the blue color in the chloroform layer disap-pears. Perform a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 3.756 mg of C10H14O
Tiapride Hydrochloride, when dried, contains not less than 99.
Tiapride Hydrochloride occurs as a white to pale yellow-white, crystal or crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), soluble in methanol, slightly soluble in ethanol (99.5) and very slightly soluble in acetic anhydride. It dissolves in 0.1 mol/L hydrochloric acid TS.
(1) Determine the absorption spectrum of a solution of Tiapride Hydrochloride in 0.1 mol/L hydrochlo-ric acid TS (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tiapride Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>: both spectra exhibit similar intensities of absorption at the same wave numbers. from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Tiapride Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 36.49 mg of C15H24N2O4S.HCl
Tiaramide Hydrochloride occurs as a white crystalline powder. It is odorless. It is freely soluble in water, slightly soluble in ethanol (95) and in acetic acid (100), and practically insoluble in acetic anhydride and in diethyl ether. The pH of a solution of 1.0 g of Tiaramide Hydrochloride in 20 mL of water is between 3.0 and 4.5. Melting point: about 2659C (with decomposition).
(1) Dissolve 5 mg of Tiaramide Hydrochlo-ride in 5 mL of 0.1 mol/L hydrochloric acid TS, and add 3 drops of Dragendorff's TS: an orange precipitate is formed. Determine the infrared absorption spectrum of Tiaramide Hydrochloride, previously dried, as directed in the potassium chloride disk method under the Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Tiaramide Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 0.5 g of Tiaramide Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Tiaramide Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Tiaramide Hydrochloride according to Method 1, and perform the test. In the procedure, add 20 mL of diluted hy-drochloric acid (1 in 2) (not more than 2 ppm). Related substances—Dissolve 0.20 g of Tiaramide Hydrochloride in 10 mL of diluted ethanol (7 in 10), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, and add diluted ethanol (7 in 10) to make ex-actly 100 mL. Pipet 2 mL of this solution, add diluted ethanol (7 in 10) to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. After air-drying, immediately devel-op the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 10 cm, air-dry the plate, and then dry at 1009C for 30 minutes. After cooling, examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot and the spot of the starting point obtained from the sample solution are not more intense than the spot from the standard solution. Allow the plate to stand in iodine vapor for 30 minutes: the spots other than the principal spot and the spot of the start-ing point from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Tiaramide Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from red through purple to blue-purple (indicator: 3 drops of neutral red TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.23 mg of C15H18ClN3O3S.HCl ‌Containers and storage Containers—Well-closed contain-ers. Tiaramide Hydrochloride Tablets
Ticlopidine Hydrochloride occurs as a white to pale yellow-white crystalline powder. It is freely soluble in acetic acid (100), soluble in water and in methanol, sparingly soluble in ethanol (95), and practi-cally insoluble in diethyl ether.
(1) Determine the infrared absorption spec-trum of Ticlopidine Hydrochloride as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. (2) A solution of Ticlopidine Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ticlopidine Hydrochloride according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of drochloride in 19.0 mL of water, add 1.0 mL of 4 mol/L so-dium hydroxide TS, shake well, centrifuge, and filter the su-pernatant liquid. To 5.0 mL of the filtrate add 5.0 mL of acetylacetone TS, mix, and warm at 409C for 40 minutes: the solution has no more color than the following control so-lution. Control solution: Weigh exactly 0.54 g of formaldehyde solution, and add water to make exactly 1000 mL. To exactly 10 mL of this solution add water to make exactly 1000 mL. Prepare before use. To 8.0 mL of this solution add water to make 20.0 mL, and filter. To 5.0 mL of the filtrate add 5.0 mL of acetylacetone TS, and proceed in the same manner. Water <2.48> Not more than 1.0z (0.3 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Ticlopidine Hydro-chloride, dissolve in 20 mL of acetic acid (100), add 40 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.03 mg of C14H14ClNS.HCl
Timepidium Bromide Hydrate contains not less than 98.
Timepidium Bromide Hydrate occurs as white, crystals or crystalline powder. It is very soluble in methanol and in acetic acid (100), freely soluble in ethanol (99.5), sparingly soluble in water and in acetic anhydride, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Timepidium Bromide Hydrate in 100 mL of freshly boiled and cooled water is between 5.3 and 6.3. A solution of Timepidium Bromide Hydrate in methanol (1 in 20) shows no optical rotation.
(1) To 1 mL of a solution of Timepidium Bromide Hydrate (1 in 100) add 1 mL of ninhydrin-sulfuric acid TS: a red purple color develops. ‌Determine the absorption spectrum of a solution of Timepidium Bromide Hydrate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Timepidium Bromide Hydrate as directed in the potassium bromide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at Timolol Maleate
Timolol Maleate, when dried, contains not less than 98.
Timolol Maleate occurs as a white to pale yel-low-white crystalline powder. It is freely soluble in acetic acid (100), and soluble in water and in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. Melting point: about 1979C (with decomposition).
(1) Determine the absorption spectrum of a solution of Timolol Maleate in 0.1 mol/L hydrochloric acid TS (3 in 100,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Timolol Maleate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 5 mL of a solution of Timolol Maleate (1 in 500) add 1 drop of potassium permanganate TS: the red color of the TS disappears immediately. D Optical rotation <2.49> [a]20: －5.7 – －6.29(after drying, 1.25 g, 1 mol/L hydrochloric acid TS, 25 mL, 100 mm). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Timolol Maleate in 20 mL of water is between 3.8 and 4.3.
(1) Clarity and color of solution—Dissolve 0.10 g of Timepidium Bromide Hydrate in 10 mL of water: the so-lution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Timepidium Bromide Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Timepidium Bromide Hydrate in 10 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol, water, acetic acid (100) and ethyl acetate (5:4:1:1:1) to a distance of about 13 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the princi-pal spot obtained from the sample solution are not more intense than the spot from the standard solution. Water <2.48> 3.5 – 5.0z (0.4 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Timepidium Bromide Hydrate, dissolve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (2:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 40.04 mg of C17H22BrNOS2
Bitter Tincture is a yellow-brown liquid. It has a characteristic aroma and a bitter taste. Specific gravity d 20: about 0.90
(1) To 1 mL of Bitter Tincture add 5 mL of methanol, then add 0.1 g of magnesium in ribbon form and 1 mL of hydrochloric acid, and allow to stand: the solution is red-purple in color. (2) Use Bitter Tincture as the sample solution. Separate-ly, to 0.5 g of pulverized Bitter Orange Peel add 10 mL of methanol, shake for 5 minutes, ˆlter, and use the ˆltrate as the standard solution (1). Proceed with 0.5 g each of pulver-ized Swertia Herb and Japanese Zanthoxylum Peel in the same manner, and use the solutions so obtained as the stan-dard solution (2) and the standard solution (3). Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution and 5 mL each of standard solutions (1), (2) and (3) on a plate of silica gel with ‰uorescent indicator for thin-layer chro-matography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): one of the sever-al spots obtained from the sample solution has the same Rf value with the clear spot at an Rf value of about 0.7 from the standard solution (3). Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: two of the several spots from the sam-ple solution show the same color tone and Rf value with the clear spot at an Rf value of about 0.4 from the standard so-lution (1), and the clear spot at an Rf value of about 0.35 from the standard solution (2). Alcohol number <1.01> Not less than 6.9 (Method 2).
Tinidazole, when dried, contains not less than 98.
Tinidazole occurs as a light yellow crystalline powder. It is soluble in acetic anhydride and in acetone, sparingly soluble in methanol, slightly soluble in ethanol (99.5), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Tinidazole in methanol (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tinidazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Sulfate <1.14>—To 2.0 g of Tinidazole add 100 mL of water, boil for 5 minutes, cool, add water to make 100 mL, and filter. Take 25 mL of the filtrate, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Use this solution as the test solution, and perform the test. Pre-pare the control solution with 0.45 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.043z). Heavy metals <1.07>—Proceed with 1.0 g of Tinida- zole according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Tinidazole according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 50 mg of Tinidazole in 2 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate and diethylamine (19:1) to a distance of about 10 cm, air-dry the plate, heat the plate at 1009C for 5 minute, and cool. Exa-mine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, ‌2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.35 g of Tinidazole, previ-ously dried, dissolve in 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.73 mg of C8H13N3O4S
Tipepidine Hibenzate, when dried, contains not less than 98.
Tipepidine Hibenzate occurs as a white to light yellow crystalline powder. It is odorless and tasteless. It is freely soluble in acetic acid (100), slightly soluble in methanol and in ethanol (95), very slightly soluble in water, and practically insoluble in diethyl ether.
(1) Dissolve 0.01 g of Tipepidine Hibenzate in 5 mL of sulfuric acid: an orange-red color develops. Dissolve 0.3 g of Tipepidine Hibenzate in 10 mL of sodium hydroxide TS and 5 mL of water, and extract with two 20-mL portions of chloroform. Wash the chloroform extracts with 10 mL of water, and filter the chloroform layer. Evaporate the filtrate on a water bath to dryness, and dissolve the residue in 0.5 mL of 1 mol/L hydrochloric acid TS and 5 mL of water. To 2 mL of this solution add 5 mL of Reinecke salt TS: a light red precipitate is formed. Determine the absorption spectrum of a solution of Tipepidine Hibenzate in ethanol (99.5) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum; both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tipepidine Hibenzate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Tipepidine Hibenzate in 10 mL of acetic acid (100): the solution is clear. Perform the test with this solution as di- rected under Ultraviolet-visible Spectrophotometry <2.24>: its absorbance at 400 nm is not more than 0.16. Heavy metals <1.07>—Proceed with 2.0 g of Tipepi-dine Hibenzate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Tipepidine Hibenzate according to Method 3, and per-form the test (not more than 2 ppm). Related substances—(i) Dissolve 10 mg of Tipepi-dine Hibenzate in 20 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the total area of peaks other than hibenzic acid and tipepidine obtained from the sample solution is not larger than the peak area of the tipepidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of a solution of ammonium ace-tate (1 in 100) and tetrahydrofuran (32:13). Flow rate: Adjust so that the retention time of tipepidine is about 12 minutes. Time span of measurement: As long as the retention time of tipepidine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of tipepidine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg of Tipepidine Hiben-zate and 3 mg of propyl parahydroxybenzoate in 100 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, hibenzic acid, tipepidine and propyl parahydroxybenzoate are eluted in this order with the resolution between the peaks of tipepi-dine and propyl parahydroxybenzoate being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tipepidine is not more than 1.5z. (ii) Dissolve 10 mg of Tipepidine Hibenzate in 20 mL of the mobile phase, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the total area of peaks other than hibenzic acid and tipepidine obtained from the sample solution is not larger than 1/2 times the peak area of the tipepidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and a solution of ammonium acetate (1 in 500) (13:7). Flow rate: Adjust so that the retention time of tipepidine is about 10 minutes. Time span of measurement: Two times as long as the retention time of tipepidine, beginning after the peak of tipepidine. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of tipepidine obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 12 mg of Tipepidine Hiben-zate and 4 mg of xanthene in 50 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, hibenzic acid, tipepidine and xanthene are eluted in this order with the resolution between the peaks of tipepidine and xanthene being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tipepidine is not more than 3.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 609C, in vacuum, phosphorus (V) oxide, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 1 g of Tipepidine Hiben-zate, previously dried, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each ml of 0.1 mol/L perchloric acid VS ＝ 51.77 mg of C H NS .C H O mL of Reinecke salt TS: a light red precipitate is formed. (2) To a quantity of powdered Tipepidine Hibenzate Tablets, equivalent to 11 mg of Tipepidine Hibenzate, add 30 mL of ethanol (99.5), and warm for 10 minutes with occa-sional shaking. After cooling, add ethanol (99.5) to make 50 mL, and ˆlter. To 1 mL of the ˆltrate add ethanol (99.5) to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 280 nm and 286 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Tipepidine Hibenzate Tablets add 5 mL of diluted acetic acid (100) (1 in 2) and 15 mL of methanol per 11 mg of tipepidine hibenzate (C15H17NS2.C14H10O4), and warm for 15 minutes with occasional shaking. After cooling, add diluted methanol (1 in 2) to make exactly V mL so that each mL contains about 0.44 mg of tipepidine hibenzate (C15H17NS2.C14H10O4), and filter. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of the internal standard solution, then add diluted methanol (1 in 2) to make 25 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of tipepidine hibenzate (C15H17NS2.C14H10O4) ＝ MS × QT/QS × V/50 MS: Amount (mg) of tipepidine hibenzate for assay taken Internal standard solution—A solution of dibucaine hydro-chloride in methanol (1 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Tipepidine Hibenzate Tablets is not less than 80z. Start the test with 1 tablet of Tipepidine Hibenzate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, filter, discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately 15 17 2 14 10 4 about 0.11 g of tipepidine hibenzate for assay, previously
Titanium Oxide, when dried, contains not less than 98.
Titanium Oxide occurs as a white powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (99.5) and in diethyl ether. It dissolves in hot sulfuric acid and in hydrofluoric acid, and does not dissolve in hydrochloric acid, in nitric acid and in dilute sulfuric acid. sample stock solution. Take 25 mL of the sample stock solu-tion to a separator, add 10 mL of a solution of ammonium sulfate (2 in 5) and 5 drops of thymol blue TS, neutralize with ammonia TS, and add 2.5 mL of ammonia TS. To this solution add exactly 20 mL of a solution of dithizone in n-butyl acetate (1 in 500), shake for 10 minutes, and use this n-butyl acetate solution as the sample solution. Separately, place 6.0 mL of Standard Lead Solution in a platinum cruci-ble, proceed as directed in the sample solution, and use this solution as the standard solution. Determine the absorbances of the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions: the absorbance of the sam-ple solution is smaller than that of the standard solution (not more than 60 ppm). Gas: Combustible gas—Acetylene gas or hydrogen gas. Supporting gas—Air. Lamp: Lead hollow-cathode lamp. Wavelength: 283.3 nm. Arsenic <1.11>—Perform the test with 20 mL of the sample stock solution obtained in (1) as the test solution: the color is not deeper than the following color standard. Color standard: Proceed in the same manner without Titanium Oxide, transfer 20 mL of the obtained solution to a generator bottle, add 2.0 mL of Standard Arsenic Solution, and proceed in the same manner as the test with the test solu-tion (not more than 10 ppm). Water-soluble substances—Shake thoroughly 4.0 g of Titanium Oxide with 50 mL of water, and allow to stand overnight. Shake thoroughly with 2 mL of ammonium chlo-ride TS, add further 2 mL of ammonium chloride TS if nec-essary, and allow titanium oxide to settle. Add water to make 200 mL, shake thoroughly, and filter through double filter paper. Discard the first 10 mL of the filtrate, evaporate 100 mL of the clear filtrate on a water bath, and heat strongly at 8009C to constant mass: the mass of the residue is not more than 5.0 mg. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately about 0.2 g of Titanium Oxide, previously dried, transfer to a crucible, and add 3 g of potas-sium disulfate. Cover, and heat gently at first, gradually raise the temperature, and then heat the fused contents for 30 minutes. Continue heating for 30 minutes at a higher tem-perature to make the fused mixture a deep yellow-red, almost clear liquid. Cool, transfer the contents of the cruci-ble to a 250-mL beaker, wash the crucible with a mixture of ‌75 mL of water and 2.5 mL of sulfuric acid into the beaker, and heat on a water bath until the solution becomes almost clear. Dissolve 2 g of L-tartaric acid in the solution, add 2 to 3 drops of bromothymol blue TS, neutralize with ammonia TS, and acidify with 1 to 2 mL of diluted sulfuric acid (1 in 2). Pass hydrogen sulfide sufficiently through the solution, add 30 mL of ammonia TS, again saturate the solution with hydrogen sulfide, allow to stand for 10 minutes, and filter. Wash the precipitate on the filter paper with ten 25-mL por-tions of a mixture of ammonium L-tartrate solution (1 in 100) and ammonium sulfide TS (9:1). When the precipitate is filtered and washed, prevent iron (II) sulfide from oxidation by filling the solution on the filter paper. Combine the fil-trate and the washings, add 40 mL of diluted sulfuric acid (1 in 2), and boil to expel hydrogen sulfide. Cool, and dilute with water to make 400 mL. Add gradually 40 mL of cupfer-ron TS to the solution with stirring, and allow to stand. After sedimentation of a yellow precipitate, add again cup-ferron TS until a white precipitate is produced. Filter by slight suction using quantitative filter paper, wash with twenty portions of diluted hydrochloric acid (1 in 10), and remove water by stronger suction at the last washing. Dry the precipitate together with the filter paper at 709C, trans-fer to a tared crucible, and heat very gently at first, and raise the temperature gradually after smoke stops evolving. Heat strongly between 9009C and 9509C to constant mass, cool, and weigh as titanium oxide (TiO2).
Tizanidine Hydrochloride occurs as a white to light yellow-white crystalline powder. It is soluble in water, slightly soluble in ethanol (99.5), and practically insoluble in acetic anhydride and in acetic acid (100). Melting point: about 2909C (with decomposition).
(1) Determine the absorption spectrum of a solution of Tizanidine Hydrochloride in diluted 1 mol/L ammonia TS (1 in 10) (1 in 125,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tizanidine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: Flow rate: Adjust so that the retention time of tizanidine is about 7 minute. Time span of measurement: About 4 times as long as the retention time of tizanidine, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mixture of water and acetonitrile (17:3) to make exactly 10 mL. Confirm that the peak area of tizanidine obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: Dissolve 2 mg each of Tizanidine Hydrochloride and p-toluenesulfonic acid monohydrate in 100 mL of the mixture of water and acetonitrile (17:3). When the procedure is run with 10 mL of this solution under the above operating conditions, p-toluenesulfonic acid and tizanidine are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times ‌with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tizanidine is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tizanidine Hydrochloride according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 60 mg of Tizanidine Hydrochloride in 10 mL of a mixture of water and aceto-nitrile (17:3), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and acetonitrile (17:3) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than tizanidine obtained from the sample solution is not larger than 1/5 times the peak area of tizanidine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm for about 3 minutes after sample injection and 318 nm subsequently). Column: A stainless steel column 4.6 mm in inside diame-ter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water and formic acid (200:1), adjusted to pH 8.5 with ammonia water (28). Mobile phase B: A mixture of acetonitrile and the mobile phase A (4:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) C9H8ClN5S.HCl: 290.17 0 – 10 81 → 68 19 → 32 5-Chloro-N-(4,5-dihydro-1H-imidazol-2-yl)- 10 – 13 2,1,3-benzothiadiazole-4-amine monohydrochloride 13 – 26 68 → 10 32 → 90 [64461-82-1] 26 – 28 Tizanidine Hydrochloride, when dried, contains not less than 99.0z and not more than 101.0z of tizanidine hydrochloride (C9H8ClN5S.HCl). Description Tizanidine Hydrochloride occurs as a white to light yellow-white crystalline powder. It is soluble in water, slightly soluble in ethanol (99.5), and practically insoluble in acetic anhydride and in acetic acid (100). Melting point: about 2909C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Tizanidine Hydrochloride in diluted 1 mol/L ammonia TS (1 in 10) (1 in 125,000) as directed under Ultra-violet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tizanidine Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: Flow rate: Adjust so that the retention time of tizanidine is about 7 minute. Time span of measurement: About 4 times as long as the retention time of tizanidine, beginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mixture of water and acetonitrile (17:3) to make exactly 10 mL. Confirm that the peak area of tizanidine obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: Dissolve 2 mg each of Tizanidine Hydrochloride and p-toluenesulfonic acid monohydrate in 100 mL of the mixture of water and acetonitrile (17:3). When the procedure is run with 10 mL of this solution under the above operating conditions, p-toluenesulfonic acid and tizanidine are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times ‌with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tizanidine is not more than 2.0z. Loss on drying <2.41> Not more than 0.2z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Tizanidine Hydro-chloride, previously dried, dissolve in 60 mL of a mixture of acetic anhydride and acetic acid (100) (7:3) with the aid of warming. After cooling, titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.02 mg of C9H8ClN5S.HCl
Tobramycin is an aminoglycoside substance having antibacterial activity produced by the growth of Strep-tomyces tenebrarius. It contains not less than 900 mg (potency) and not more than 1060 mg (potency) per mg, calculated on the anhydrous basis.
Tobramycin occurs as a white to pale yellow-white powder. It is very soluble in water, freely soluble in formamide, slightly soluble in methanol, and very slightly soluble in ethanol (95). It is hygroscopic.
(1) Determine the 1H spectrum of a solu-tion of Tobramycin in heavy water for nuclear magnetic resonance spectroscopy (1 in 125) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropanesulfonate for nuclear magnetic reso-nance spectroscopy as an internal reference compound: it exhibits a doublet signal A at around d 5.1 ppm, a multiplet signal B between d 2.6 ppm and d 4.0 ppm, and a multiplet signal C between d 1.0 ppm and d 2.1 ppm. The ratio of the integrated intensity of these signals, A:B:C, is about 1:8:2. (2) Dissolve 10 mg each of Tobramycin and Tobramycin RS in 1 mL of water, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 4 mL of the sample solution and standard solu- tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia TS, 1-butanol and methanol (5:5:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 1009C for 5 minutes: the Rf values of the principal spots obtained from the sample solution and the standard solution are the same. D Optical rotation <2.49> [a]20: ＋138 – ＋1489(1 g calculated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.10 g of Tobramycin in 10 mL of water is between 9.5 and 11.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Tobramycin in 10 mL of water: the solution is clear, and its absorbance at 400 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.05. Heavy metals <1.07>—Proceed with 1.0 g of Tobramycin according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Related substances—Dissolve 80 mg of Tobramycin in 10 mL of diluted ammonia solution (28) (1 in 250), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add diluted ammonia solution (28) (1 in 250) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ammonia solution (28), ethanol (95) and 2-butanone (1:1:1) to a distance of about 10 cm, air-dry the plate, then further dry at 1109C for 10 minutes. Immedi-ately spray evenly a mixture of water and sodium hypochlorite TS (4:1) on the plate, air-dry the plate, then spray potassium iodide-starch TS on the plate: the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Water <2.48> Not more than 11.0z (0.1 g, volumetric titration, direct titration). Use a mixture of formamide for water determination and methanol for water determination (3:1) instead of methanol for water determination. Residue on ignition <2.44> Not more than 1.0z (0.5 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Tobramycin RS, equivalent to about 25 mg (potency), dis-solve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL, and use this solution as the standard stock solution. Keep the standard stock solution between 59C and 159C, and use within 30 days. Take exactly a suita-ble amount of the standard stock solution before use, add ‌0.1 mol/L phosphate buffer solution (pH 8.0) to make solu-tions so that each mL contains 8 mg (potency) and 2 mg (po-tency), and use these solutions as the high concentration standard solution and the low concentration standard solu-tion, respectively.‌ Sample solutions—Weigh accurately an amount of Tobramycin, equivalent to about 25 mg (potency), and dis-solve in 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 25 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 8 mg (potency) and 2 mg (potency), and use these solutions as the high con-centration sample solution and the low concentration sample solution, respectively.
Todralazine Hydrochloride Hydrate contains not less than 98.
Todralazine Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It has a slight, charac-teristic odor, and has a bitter taste. It is very soluble in formic acid, freely soluble in metha-nol, soluble in water, sparingly soluble in ethanol (95), and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Todralazine Hydrochlo-ride Hydrate in 200 mL of water is between 3.0 and 4.0.
(1) To 2 mL of a solution of Todralazine Hydrochloride Hydrate (1 in 200) add 5 mL of silver nitrate-ammonia TS: the solution becomes turbid, and a black pre-cipitate is formed. Determine the absorption spectrum of a solution of Todralazine Hydrochloride Hydrate in 0.1 mol/L hydro-chloric acid TS (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Todralazine Hydrochloride Hydrate as directed in the potas-sium chloride disk method under the Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Todralazine Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> (1) for chloride.
(1) Clarity and color of solution—Dissolve 0.30 g of Todralazine Hydrochloride Hydrate in 10 mL of water: the solution is clear and colorless to pale yellow. Sulfate <1.14>—Proceed the test with 2.0 g of Todralazine Hydrochloride Hydrate. Prepare the control so-lution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.012z). Heavy metals <1.07>—Proceed with 1.0 g of Todrala- zine Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Todralazine Hydrochloride Hydrate according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Todralazine Hydrochloride Hydrate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the total area of the peaks other than todralazine obtained from the sample solution is not larger than the peak area of todrala-zine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.10 g of sodium 1-heptane sul-fonate in 1000 mL of diluted methanol (2 in 5). Adjust the pH of the solution to between 3.0 and 3.5 with acetic acid (100). Flow rate: Adjust so that the retention time of todralazine is about 8 minutes. Time span of measurement: About twice as long as the retention time of todralazine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 25 mL. Confirm that the peak area of todralazine obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: Dissolve 5 mg each of Todralazine Hydrochloride Hydrate and potassium hydrogen phthalate in 100 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating condi-tions, phthalic acid and todralazine are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of todralazine is not more than 2.0z. Water <2.48> 6.0 – 7.5z (0.5 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Todralazine Hydro-chloride Hydrate, dissolve in 5 mL of formic acid, add 70 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L ‌perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction.‌ Each mL of 0.1 mol/L perchloric acid VS ＝ 26.87 mg of C11H12N4O2.HCl
Tofisopam, when dried, contains not less than 98.
Tofisopam occurs as a pale yellow-white crys-talline powder. It is freely soluble in acetic acid (100), soluble in acetone, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. A solution of Tofisopam in ethanol (95) (1 in 100) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Tofisopam in ethanol (95) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tofisopam, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tofisopam according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Tofisopam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.05 g of Tofisopam in 10 mL of acetone, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add acetone to make exactly 25 mL, pipet 1 mL of this solution, add ace-tone to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetone, methanol and formic acid (24:12:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Tofisopam, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.25 mg of C22H26N2O4
Tokakujokito Extract contains not less than 38 mg and not more than 152 mg of amygdalin, not less than 1 mg and not more than 4 mg of (E )-cinnamic acid, the standard solution on a plate of silica gel for thin-layer chromatography.
Tokakujokito Extract occurs as a green-yellow-brown to dark brown powder. It has characteristic odor and, salty, slightly astringent, and then slightly sweet taste.
(1) To 1.0 g of Tokakujokito Extract add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solu-tion. Separately, dissolve 2 mg of amygdalin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-propanol, ethyl acetate and water (4:4:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the green-brown spot from the standard solution (Peach Kernel). Perform the test according to the following (i) or (ii) (Cinnamon Bark). Put 10 g of Tokakujokito Extract in a 300-mL of hard-glass flask, add 100 mL of water and 1 mL of silicone resin, connect the apparatus for essential oil determination, and heat to boil under a reflux condenser. The graduated tube of the apparatus is to be previously filled with water to the standard line, and 2 mL of hexane is added to the grad-uated tube. After heating under reflux for 1 hour, separate the hexane layer, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of (E)-cinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 40 mL of the sample solution and 2 mL of dissolve 1 mg of (E)-2-methoxycinnamaldehyde for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 40 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the bluish white fluorescent spot from the standard solution. To 1.0 g of Tokakujokito Extract add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, centri-fuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg of rhein for thin-layer chromatog-raphy in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the orange fluorescent spot from the standard solution (Rhubarb). To 1.0 g of Tokakujokito Extract add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centri-fuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chroma-tography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Tokakujokito Extract as directed in Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of Tokakujokito Extract according to Method 3, and per-form the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 8.0z (1 g, 1059C, 5 hours). Total ash <5.01> Not less than 20.0z and more than 40.0z.
(1) Amygdalin—Weigh accurately about 0.5 g of JP XVIII Crude Drugs and Related Drugs / Tokakujokito Extract 2161 Tokakujokito Extract, add exactly 50 mL of diluted metha-nol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, elute through a column prepared previously with 2 g of polyamide for column chromatography using water to make exactly 20 mL of effluent , and use this solution as the sample solution. Separately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desiccator (silica gel) for 24 hours or more, and dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (800:200:1). Flow rate: 1.0 mL per minute (the retention time of (E)-cinnamic acid is about 22 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry Amount (mg) of amygdalin ＝ MS × AT/AS × 4 factor of the peak of (E)-cinnamic acid are not less than 5000 and not more than 1.5, respectively. MS: Amount (mg) of amygdalin for assay taken Operation conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). Systemic suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amygdalin is not more than 1.5z. (E)-Cinnamic acid—Conduct this procedure using light-resistant vessels. Weigh accurately about 0.5 g of Tokakujokito Extract, add 20 mL of diethyl ether and 10 mL of water, shake for 10 minutes, centrifuge, and separate the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, proceed in the same manner as above, and repeat this procedure two more times. Combine all the ex-tracts, evaporate the solvent under low pressure (in vacuo), dissolve the residue in diluted methanol (1 in 2) to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of (E)-cinnamic acid for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of (E)-cinnamic acid in each solution. Amount (mg) of (E)-cinnamic acid ＝ MS × AT/AS × 1/20 MS: Amount (mg) of (E)-cinnamic acid for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave- System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of (E)-cinnamic acid is not more than 1.5z. Sennoside A—Weigh accurately about 0.5 g of Tokakujokito Extract, add 20 mL of ethyl acetate and 10 mL of water, shake for 10 minutes, centrifuge, remove the ethyl acetate layer, then add 20 mL of ethyl acetate, proceed in the same manner as above, and remove the ethyl acetate layer. To the aqueous layer obtained add 10 mL of metha-nol, shake for 30 minutes, centrifuge, and separate the su-pernatant liquid. To the residue add 20 mL of diluted metha-nol (1 in 2), shake for 5 minutes, centrifuge, and separate the supernatant liquid. Combine all the supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 5 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of sennoside A in each solution. Amount (mg) of sennoside A (C42H38O20) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (840:160:1). Flow rate: 1.0 mL per minute (the retention time of senno-side A is about 20 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sennoside A are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌2162 Tokishakuyakusan Extract / Crude Drugs and Related Drugs area of sennoside A is not more than 1.5z. Rhein—Weigh accurately about 0.5 g of Tokakujoki-to Extract, add 80 mL of water, shake, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add 20 mL of iron (III) chloride TS, heat under a reflux condenser for 30 minutes, add 3 mL of hydrochloric acid, and heat in addition under a reflux condenser for 30 minutes. After cooling, extract three times with 25 mL each of diethyl ether, combine all the extracts, evaporate the solvent under low pressure (in vacuo), dissolve the residue to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 5 mg of rhein for assay, and dissolve in acetone to make exactly 100 mL. Pipet 10 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of rhein in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— Amount (mg) of rhein ＝ MS × AT/AS × 4/5 System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute MS: Amount (mg) of rhein for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (650:350:1). Flow rate: 1.0 mL per minute (the retention time of rhein is about 17 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rhein are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Tokishakuyakusan Extract contains not less than 0.
Tokishakuyakusan Extract is a light brown to JP XVIII Crude Drugs and Related Drugs / Tokishakuyakusan Extract 2163 brown powder or black-brown viscous extract. It has a char-acteristic odor, and a slight sweet taste at first and a bitter taste later. Identiˆcation (1) Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 15 mL of water and 5 mL of 0.1 mol/L hydrochloric acid TS, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solu-tion (Japanese Angelica Root; Cnidium Rhizome). Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the purple spot from the standard solution (Peony Root). For preparation prescribed Atractylodes Rhizome—Shake 1.0 g of the dry extract (or 3.0 g of the viscous extract) with 10 mL of water, add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Sepa-rately, dissolve 1 mg of atractylenolide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the stand-ard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 10 mL of water, add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhi-zome). Shake 2.0 g of the dry extract (or 6.0 g of the viscous extract) with 20 mL of water and 2 mL of ammonia solution (28), add 20 mL of a mixture of hexane and ethyl acetate (20:1), shake, and centrifuge. Separate the upper layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of methanol to the residue, and use this solution as the sample solution. Separately, dissolve 1 mg of alisol A for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl for-mate, water and formic acid (30:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzalde-hyde-sulfuric acid-acetic acid TS on the plate, heat the plate at 1059C for 5 minutes, allow to cool, and examine under ul-traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Alisma Tuber).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 9.5z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not more than 10.0z, calculated on the dried basis.
(1) (E)-Ferulic acid—Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, weigh accurately about 10 mg of (E)-ferulic acid for assay, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Pipet 2 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of (E)-ferulic acid in each solution. under low pressure (in vacuo), add 0.5 mL of hexane to the residue, and use this solution as the sample solution. Per- Amount (mg) of (E)-ferulic acid ＝ MS × AT/AS × 1/50 form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam- MS: Amount (mg) of (E)-ferulic acid for assay taken Tokishakuyakusan Extract / Crude Drugs and Related Drugs Operating conditions— Detector: An ultraviolet absorption photometer (wavelength: 320 nm). Column: A stainless steel column 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate in 1000 mL of water, and add 2 mL of phosphoric acid. To 850 mL of this solution add 150 mL of acetonitrile. Flow rate: 1.0 mL per minute [the retention time of (E)- ferulic acid is about 10 minutes]. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of (E)-ferulic acid are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of (E)-ferulic acid is not more than 1.5z. (2) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Paeoniflorin RS (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of atractylenolide III for assay, pre-viously dried in a desiccator (silica gel) for more than 24 hours, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of atrac-tylenolide III in each solution. Amount (mg) of atractylenolide III ＝ MS × AT/AS × 1/40 MS: Amount (mg) of atractylenolide III for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (550:450:1). Flow rate: 1.0 mL per minute (the retention time of atrac-tylenolide III is about 10 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of atractylenolide III are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of atractylenolide III is not more than 1.5z. (4) Atractylodin—Conduct this procedure without ex-posure to light, using light-resistant vessels. Weigh accu-rately about 0.5 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.5 g of the dried substance), add exactly 50 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Perform the test with exactly 10 mL each of the sample solu-tion and atractylodin TS for assay as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of atrac-tylodin in each solution. Mobile phase: A mixture of water, acetonitrile and phos- phoric acid (850:150:1). Amount (mg) of atractylodin ＝ CS × AT/AS × 50 Flow rate: 1.0 mL per minute (the retention time of paeoniflorin is about 9 minutes). System suitability— System performance: Dissolve 1 mg of albiflorin in 10 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. (3) Atractylenolide III—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva- CS: Concentration (mg/mL) of atractylodin in atrac-tylodin TS for assay Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 330 mL of a mixture of water and phos-phoric acid (55:1) add 670 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of atrac-tylodin is about 13 minutes). ‌JP XVIII Crude Drugs and Related Drugs / Tribulus Fruit 2165‌‌ System suitability— System performance: When the procedure is run with 10 mL of atractylodin TS for assay under the above operating conditions, the number of theoretical plates and the symme-try factor of the peak of atractylodin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of atractylodin TS for assay under the above op-erating conditions, the relative standard deviation of the peak area of atractylodin is not more than 1.5z.
Tolbutamide, when dried, contains not less than 99.
Tolbutamide occurs as white, crystals or crys-talline powder. It is odorless or has a slight, characteristic odor. It is tasteless. It is soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water.
(1) Boil 0.2 g of Tolbutamide with 8 mL of diluted sulfuric acid (1 in 3) under a reflux condenser for 30 minutes. Cool the solution in ice water, collect the precipi-tated crystals, recrystallize from water, and dry at 1059C for 3 hours: the crystals melt <2.60> between 1359C and 1399C. (2) Render the filtrate obtained in (1) alkaline with about 20 mL of a solution of sodium hydroxide (1 in 5), and heat: an ammonia-like odor is perceptible.
(1) Acidity—Warm 3.0 g of Tolbutamide with 150 mL of water at 709C for 5 minutes, allow to stand for 1 hour in ice water, and filter. To 25 mL of the filtrate add 2 drops of methyl red TS and 0.20 mL of 0.1 mol/L sodium hydrox-ide VS: a yellow color develops. Chloride <1.03>—To 40 mL of the filtrate obtained in add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.25 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.011z). Sulfate <1.14>—To 40 mL of the filtrate obtained in add 1 mL of dilute hydrochloric acid and water to make ‌50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.021z).‌ Heavy metals <1.07>—Proceed with 2.0 g of Tol- butamide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Dissolution rate (z) with respect to the labeled amount of tolbutamide (C12H18N2O3S) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Tolbutamide RS taken C: Labeled amount (mg) of tolbutamide (C12H18N2O3S) in 1 tablet
Weigh accurately and powder not less than 20 Tol- butamide Tablets. Weigh accurately a portion of the pow-der, equivalent to about 0.5 g of tolbutamide (C H N O S), 12 18 2 3 Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Tolbutamide, previ-ously dried, and dissolve in 30 mL of neutralized ethanol. Add 20 mL of water, and titrate <2.50> with 0.1 mol/L sodi-um hydroxide VS (indicator: 3 drops of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 27.04 mg of C12H18N2O3S
Tolnaftate, when dried, contains not less than Tablets, equivalent to 0.
Tolnaftate occurs as a white powder. It is odorless. It is freely soluble in chloroform, sparingly soluble in diethyl ether, slightly soluble in methanol and in ethanol (95), and practically insoluble in water.
Shake a quantity of powdered Tolbutamide O-Naphthalen-2-yl N-methyl-N-(3- methylphenyl)thiocarbamate [2398-96-1] Tolnaftate, when dried, contains not less than Tablets, equivalent to 0.5 g of Tolbutamide, with 50 mL of chloroform, filter, and evaporate the filtrate to dryness. Pro- 98.0z of tolnaftate (C 19H 17NOS). ceed with the residue as directed in the Identification under Tolbutamide. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of phosphate buffer solution (pH 7.4) as the dissolu-tion medium, the dissolution rate in 30 minutes of Tol-butamide Tablets is not less than 80z. Start the test with 1 tablet of Tolbutamide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 10 mg of tolbutamide (C12H18N2O3S), and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of Tolbutamide RS, previously dried at 1059C for 3 hours, dissolve in 10 mL of methanol, and add the dissolution medium to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the control, and determine the absorbances, AT and AS, at 226 nm. Description Tolnaftate occurs as a white powder. It is odorless. It is freely soluble in chloroform, sparingly soluble in diethyl ether, slightly soluble in methanol and in ethanol (95), and practically insoluble in water. Identification (1) To 0.2 g of Tolnaftate add 20 mL of potassium hydroxide-ethanol TS and 5 mL of water, and heat under a reflux condenser for 3 hours. After cooling, to 10 mL of this solution add 2 mL of acetic acid (100), and shake with 1 mL of lead (II) acetate TS: a black precipitate is formed. Determine the absorption spectrum of a solution of Tolnaftate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Tolnaftate RS prepared in the same manner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tol-naftate, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Tolnaftate RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Carbonize 1.0 g of Tol-naftate by gentle heating. After cooling, add 5 mL of nitric ‌acid and 1 mL of sulfuric acid, and heat until white fumes are evolved. After cooling, add 2 mL of nitric acid, and heat until white fumes are evolved. After cooling, add 2 mL of nitric acid and 0.5 mL of perchloric acid, and heat gradually until white fumes are evolved. Repeat this procedure twice, and heat until white fumes are no longer evolved. Incinerate the residue by igniting between 5009C and 6009C for 1 hour. Proceed according to Method 2, and perform the test with 50 mL of the test solution so obtained. Prepare the control so-lution as follows: to 11 mL of nitric acid add 1 mL of sulfu-ric acid, 1 mL of perchloric acid and 2 mL of hydrochloric acid, proceed in the same manner as the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). Related substances—Dissolve 0.50 g of Tolnaftate in 10 mL of chloroform, and use this solution as the sample so-lution. Pipet 2 mL of the sample solution, and add chlo-roform to make exactly 100 mL. Pipet 5 mL of this solution, add chloroform to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with toluene to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 5 minutes, and examine under ultraviolet light (wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Loss on drying <2.41> Not more than 0.5z (1 g, in vacuum at a pressure not exceeding 0.67 kPa, 659C, 3 hours). Residue on ignition <2.44> Weigh accurately about 2 g of Tolnaftate, and carbonize by gradual heating. Moisten the substance with 1 mL of sulfuric acid, heat gradually until white fumes are no longer evolved, and ignite between 4509C and 5509C for about 2 hours to constant mass: the residue is not more than 0.1z.
Weigh accurately about 50 mg of Tolnaftate and Tolnaftate RS, previously dried, dissolve each in 200 mL of methanol by warming in a water bath, cool, and add metha-nol to make exactly 250 mL. Pipet 5 mL each of the solu-tions, to each add methanol to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absorbances, AT and AS, of the sample solution and standard solution at 257 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Tolnaftate Solution
Tolperisone Hydrochloride, when dried, contains not less than 98.
Tolperisone Hydrochloride occurs as a white crystalline powder. It has a slight, characteristic odor. It is very soluble in acetic acid (100), freely soluble in water and in ethanol (95), soluble in acetic anhydride, slightly soluble in acetone, and practically insoluble in diethyl ether. The pH of a solution of 1.0 g of Tolperisone Hydrochlo-ride in 20 mL of water is between 4.5 and 5.5. It is hygroscopic. Melting point: 167 – 1749C
(1) Dissolve 0.2 g of Tolperisone Hydro-chloride in 2 mL of ethanol (95), add 2 mL of 1,3-dinitrobenzene TS and 2 mL of sodium hydroxide TS, and heat: a red color develops. To 5 mL of a solution of Tolperisone Hydrochloride (1 in 20) add 2 to 3 drops of iodine TS: a red-brown precipi-tate is produced. Dissolve 0.5 g of Tolperisone Hydrochloride in 5 mL of water, add 2 mL of ammonia TS, and filter. Acidify 5 mL of the filtrate with dilute nitric acid: the solution responds to Qualitative Tests <1.09> for chloride. 1 cm Absorbance <2.24> E1z (257 nm): 555 – 585 (after drying, 5 mg, ethanol (95), 500 mL). and shake vigorously for 30 minutes. Immediately after al-lowing to stand, separate the isooctane-carbon disulfide mix-ture layer, and dehydrate with anhydrous sodium sulfate. Perform the test with these solutions as directed under Ultra-violet-visible Spectrophotometry <2.24>: the absorbance of the sample solution at 438 nm is not more than that of the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Tolperisone Hydro-chloride, previously dried, dissolve in 70 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.18 mg C16H23NO.HCl
トスフロキサシントシル酸塩水和物
Tosufloxacin Tosilate Hydrate contains not less than 98.
Tosufloxacin Tosilate Hydrate occurs as a white to pale yellowish white crystalline powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in methanol, and practically insoluble in water and in ethanol (99.5). A solution of Tosufloxacin Tosilate Hydrate in methanol (1 in 100) shows no optical rotation. Melting point: about 2549C (with decomposition).
(1) Tosufloxacin Tosilate Hydrate shows a light bluish-white fluorescence under ultraviolet light (main wavelength 254 nm). Proceed 10 mg of Tosufloxacin Tosilate Hydrate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Tosufloxacin Tosilate Hydrate in a mixture of methanol and sodium hydroxide TS (49:1) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Tosufloxacin Tosilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tosufloxacin Tosilate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Tosufloxacin Tosilate RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers.
(1) Clarity and color of solution—Dissolve 1.0 g of Tolperisone Hydrochloride in 10 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Perform the test with 4.0 g of Tol- perisone Hydrochloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.005z). Heavy metals <1.07>—Proceed with 1.0 g of Tolperi- sone Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Piperidine hydrochloride—Dissolve 0.20 g of Tol-perisone Hydrochloride in water to make exactly 10 mL, and use this solution as the sample solution. Separately, dissolve 20 mg of piperidine hydrochloride in water to make exactly 1000 mL, and use this solution as the standard solution. Transfer 5.0 mL each of the sample solution and standard solution to different separators, add 0.1 mL each of a solu-tion of copper (II) sulfate pentahydrate (1 in 20), then add 0.1 mL each of ammonia solution (28) and exactly 10 mL each of a mixture of isooctane and carbon disulfide (3:1), lated on the anhydrous basis. Description Tosufloxacin Tosilate Hydrate occurs as a white to pale yellowish white crystalline powder. It is freely soluble in N,N-dimethylformamide, sparingly soluble in methanol, and practically insoluble in water and in ethanol (99.5). A solution of Tosufloxacin Tosilate Hydrate in methanol (1 in 100) shows no optical rotation. Melting point: about 2549C (with decomposition). Identification (1) Tosufloxacin Tosilate Hydrate shows a light bluish-white fluorescence under ultraviolet light (main wavelength 254 nm). Proceed 10 mg of Tosufloxacin Tosilate Hydrate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution responds to Qualitative Tests <1.09> (2) for fluoride. Determine the absorption spectrum of a solution of Tosufloxacin Tosilate Hydrate in a mixture of methanol and sodium hydroxide TS (49:1) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Tosufloxacin Tosilate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tosufloxacin Tosilate Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Tosufloxacin Tosilate RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. Purity (1) Chloride <1.03>—Dissolve 1.0 g of Tosufloxa-cin Tosilate Hydrate in 40 mL of N,N-dimethylformamide, and add 6 mL of dilute nitric acid and N,N-dimethylfor-mamide to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS, 6 mL of dilute nitric acid and N,N-dimethylformamide to make 50 mL (not more than 0.007z). Heavy metals <1.07>—Proceed with 1.0 g of Tosufloxacin Tosilate Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Tosufloxacin Tosilate Hydrate according to Method 4, and perform the test under the condition of the ignition tem-perature being between 7509C and 8509C, and add 10 mL of diluted hydrochloric acid to residue (not more than 2 ppm). Related substances—Dissolve 10 mg of Tosufloxacin Tosilate Hydrate in 12 mL of mobile phase B, add water to make 25 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add mobile phase A to make exactly 100 mL. Pipet 2 mL of this solution, add mobile phase A to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of each peak other than tosylic acid and tosufloxacin obtained from the sample solu-tion is not larger than 3/4 times the peak area of tosufloxa-cin from the standard solution, and the total area of the peaks other than tosylic acid and tosufloxacin from the sam-ple solution is not larger than 2.5 times the peak area of tosufloxacin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: To 300 to 500 mL of water add slowly 100 mL of methanesulfonic acid under ice-cooling, add slowly 100 mL of triethylamine under ice-cooling too, and add water to make 1000 mL. To 10 mL of this solution add 143 mL of water, 40 mL of acetonitrile and 7 mL of 1 mol/L dipotassium hydrogen phosphate TS for buffer solution. Mobile phase B: To 300 to 500 mL of water add slowly 100 mL of methanesulfonic acid under ice-cooling, add slowly 100 mL of triethylamine under ice-cooling too, and add water to make 1000 mL. To 10 mL of this solution add 100 mL of acetonitrile, 83 mL of water and 7 mL of 1 mol/L dipotassium hydrogen phosphate TS for buffer solution. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 1 1 – 16 100 → 0 0 → 100 16 – 35 Flow rate: 0.5 mL per minute. Time span of measurement: About 5 times as long as the retention time of tosufloxacin. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add mobile phase A to make exactly 20 mL. Confirm that the peak area of tosufloxacin obtained with 20 mL of this solution is equivalent to 18 to 32z of the peak area of tosufloxacin with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of tosufloxacin are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tosufloxacin is not more than 2.0z. Water <2.48> 2.5 – 3.5z (30 mg, coulometric titration).
Weigh accurately about 30 mg each of Tosufloxacin Tosilate Hydrate and Tosufloxacin Tosilate RS (separately determine the water <2.48> in the same manner as Tosufloxa-cin Tosilate Hydrate), and dissolve each in methanol to make exactly 100 mL. Pipet 20 mL each of these solutions, to each add exactly 4 mL of the internal standard solution and methanol to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tosufloxacin to that of the internal standard. Amount (mg) of tosufloxacin tosilate (C19H15F3N4O3.C7H8O3S) ＝ MS × QT/QS MS: Amount (mg) of Tosufloxacin Tosilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To a mixture of 0.02 mol/L phosphate buffer solution (pH 3.5) and a solution of dibutylamine in methanol (1 in 2500) (3:1) add diluted phosphoric acid (1 in 10) to adjust the pH to 3.5. Flow rate: Adjust so that the retention time of tosufloxa-cin is about 20 minutes. System suitability— System performance: When the procedure is run with 10 ‌mL of the standard solution under the above operating con-ditions, the internal standard and tosufloxacin are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tosufloxacin to that of the internal standard is not more than 1.0z.
Powdered Tragacanth occurs as a white to yellowish white powder. It is odorless, tasteless and mucilaginous. Under a microscope <5.01>, it, immersed in olive oil or liquid paraffin, reveals numerous angular fragments with a small amount of the circular or irregular lamellae or of starch grains. Starch grains are spherical to elliptical, mostly simple and occasionally 2- to 4-compound grains, simple grain, 3 – 25 mm in diameter. The fragments are swollen and altered with water.
(1) To 1 g of Powdered Tragacanth add 50 mL of water: a nearly uniform, slightly turbid mucilage is formed. (2) To Powdered Tragacanth add dilute iodine TS, and examine the mixture microscopically <5.01>: a few blue-colored starch grains are observable.
Karaya gum—Boil 1 g of Powdered Tragacanth with 20 mL of water until a mucilage is formed, add 5 mL of hydrochloric acid, and again boil the mixture for 5 minutes: no light red to red color develops. Total ash <5.01> Not more than 4.0z.
Tragacanth occurs as curved, flattened or lamellate fragments, 0.5 – 3 mm in thickness. It is white to light yellow in color, translucent, and horny in texture. It is easily broken, and swells in water. Odorless; tasteless and mucilaginous.
(1) To 1 g of pulverized Tragacanth add 50 mL of water: a nearly uniform, slightly turbid mucilage is formed. (2) To pulverized Tragacanth add dilute iodine TS, and examine the mixture microscopically <5.01>: a few blue-colored starch grains are observable.
Karaya gum—Boil 1 g of Tragacanth with 20 mL of water until a mucilage is formed, add 5 mL of hydrochloric acid, and again boil the mixture for 5 minutes: no light red to red color develops. Total ash <5.01> Not more than 4.0z.
Tramadol Hydrochloride occurs as a white crystalline powder. It is very soluble in water, and freely soluble in methanol, in ethanol (95) and in acetic acid (100). A solution of Tramadol Hydrochloride (1 in 20) shows no optical rotation. Melting point: 180 – 1849C Tramadol Hydrochloride shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Tramadol Hydrochloride in ethanol (95) (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Tramadol Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Tramadol Hydrochloride (1 in 100) re-sponds to Qualitative Tests <1.09> (2) for chloride.
(1) Acidity or alkalinity—Dissolve 1.0 g of Tramadol Hydrochloride in water to make 20 mL. To 10 mL of this solution add 0.2 mL of methyl red TS for acidity or alkalinity test and 0.2 mL of 0.01 mol/L hydrochloric acid VS: a red color develops. To this solution add 0.01 mol/L sodium hydroxide VS until the color of the solution changes from red to yellow: the consumed volume is not more than 0.4 mL. Heavy metals <1.07>—Proceed with 1.0 g of Tramadol Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—(i) Dissolve 0.10 g of Tramadol Hydrochloride in 2 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add methanol to make exactly 500 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Allow the plate to stand in am-monia vapor for 20 minutes, develop with a mixture of tolu-ene, isopropanol and ammonia solution (28) (80:19:1) to a distance of about 15 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 1 hour, and examine under ultraviolet light (main wavelength: 254 nm): the spot at the Rf value of about 0.5 obtained from the sample solution is not more intense than the spot from the standard solution. (ii) Dissolve 0.15 g of Tramadol Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak having the relative retention time of about 0.9 to tramadol obtained from the sample solution is not larger than 1/5 times the peak area of tramadol from the standard solution, the area of the peak other than tramadol and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of tramadol from the standard solution, and the total area of the peaks other than tramadol from the sample solution is not larger than 2/5 times the peak area of tramadol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted trifluoroacetic acid (1 in 500) and acetonitrile (141:59). Flow rate: Adjust so that the retention time of tramadol is about 5 minutes. Time span of measurement: About 4 times as long as the retention time of tramadol, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add the mobile phase to make exactly 20 mL. Con-firm that the peak area of tramadol obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of tramadol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tramadol is not more than 2.0z. Water <2.48> Not more than 0.5z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.18 g of Tramadol Hydro-chloride, dissolve in 25 mL of acetic acid(100), add 10 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. ‌Each mL of 0.1 mol/L perchloric acid VS ＝ 29.98 mg of C16H25NO2.HCl
Tranexamic Acid, when dried, contains not less than 98.
Tranexamic Acid occurs as white, crystals or crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).
Determine the infrared absorption spectrum of Tranexamic Acid as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Tranexamic Acid RS: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Tranexamic Acid in 20 mL of water is between 7.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Tranexamic Acid in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 1.0 g of Tranexamic Acid. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals—Dissolve 2.0 g of Tranexamic Acid in water to make 20 mL, and use this solution as the sample stock solution. To 12 mL of the sample stock solution add 2 mL of hydrochloric acid-ammonium acetate buffer solution (pH 3.5), mix, add 1.2 mL of thioacetamide TS, mix imme-diately, and use this solution as the sample solution. Sepa-rately, proceed in the same manner as above with a mixture of 1 mL of Standard Lead Solution, 2 mL of the sample stock solution and 9 mL of water, and use the solution so obtained as the standard solution. Separately, proceed in the same manner with a mixture of 10 mL of water and 2 mL of the sample stock solution, and use the solution so obtained as the control solution. Conform that the color of the stand-ard solution is slightly darker than that of the control solu-tion. Compare the sample solution and the standard solution 2 minutes after they are prepared: the color of the sample so-lution is not more intense than that of the standard solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution by dissolv- ing 1.0 g of Tranexamic Acid in 10 mL of water, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Tranexamic Acid in 20 mL of water, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area multiplied by correc-tion factor 1.2 of the peak, having the relative retention time of about 1.5 to tranexamic acid obtained from sample solu-tion, is not larger than 2/5 times the peak area of tranexamic acid from the standard solution, and the area of the peak, having the relative retention time of about 2.1, is not larger than 1/5 times the peak area of tranexamic acid from the standard solution. The area of each peak other than tranex-amic acid and the peaks mentioned above is not larger than 1/5 times the peak area of tranexamic acid from the stand-ard solution. For the area of the peaks, having the relative retention time of about 1.1 and about 1.3, multiply their cor-rection factors 0.005 and 0.006, respectively. The total area of the peaks other than tranexamic acid from the sample so-lution is not larger than the peak area of tranexamic acid from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of tranexamic acid, beginning after the sol-vent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 5 mL of the standard solution add water to make exactly 25 mL. Con-firm that the peak area of tranexamic acid obtained with 20 mL of this solution is equivalent to 14 to 26z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 7z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Tranexamic Acid and Tranexamic Acid RS, previously dried, dissolve in water to make exactly 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of tranexamic acid in each solution. Amount (mg) of tranexamic acid (C8H15NO2) ＝ MS × AT/AS MS: Amount (mg) of Tranexamic Acid RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 11.0 g of anhydrous sodium dihy- ‌drogen phosphate in 500 mL of water, and add 5 mL of triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid or diluted phosphoric acid (1 in 10), add water to make 600 mL, and add 400 mL of methanol. Flow rate: Adjust so that the retention time of tranexamic acid is about 20 minutes. System suitability— System performance: To 5 mL of the standard solution add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in 10,000) and water to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, tranexamic acid and 4-(aminomethyl)benzoic acid are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 0.6z.
Tranilast, when dried, contains not less than 99.
Tranilast occurs as light yellow, crystals or crystalline powder. It is freely soluble in N,N-dimethylformamide, slightly soluble in acetonitrile, in methanol and in ethanol (99.5), and practically insoluble in water. It gradually becomes light yellow-brown on exposure to light. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Tranilast in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tranilast, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Tranilast according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Conduct this procedure without exposure to light, using light-resistant vessels. Dissolve 50 mg of Tranilast in 50 mL of acetonitrile, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, and add acetonitrile to make exactly 50 mL. Pipet 1 mL of this solution, add acetonitrile to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than tranilast obtained from the sample solution is not larger than the peak area of tranilast from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of tranilast is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of tranilast, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of tranilast are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of tranilast is not more than 3.0z. Chloroform—Weigh accurately about 1 g of Tranilast, dissolve in exactly 5 mL of a solution, prepared by adding N,N-dimethylformamide to exactly 1 mL of the in-ternal standard solution to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 3 g of chloroform, and add N,N-dimethylfor-mamide to make exactly 100 mL. Pipet 1 mL of this solu-tion, add exactly 1 mL of the internal standard solution, add N,N-dimethylformamide to make 100 mL, and use this solu-tion as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and calculate the ratios, QT and QS, of the peak area of chloroform to that of the internal standard: the amount of chloroform is not more than 0.006z. Amount (z) of chloroform ＝ MS/MT × QT/QS × 1/20 MS: Amount (g) of chloroform taken MT: Amount (g) of Tranilast taken Internal standard solution—A solution of trichloroethylene in N,N-dimethylformamide (1 in 50). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with porous styrene-divinylbenzene copolymer for gas chromatography (0.3 – 0.4 mm in mean pore size, not exceeding 50 m2/g) (150 – 180 mm in particle diameter). Column temperature: A constant temperature of about 1609C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of chloroform is about 2 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, chloroform and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating ‌conditions, the relative standard deviation of the ratio of the peak area of chloroform to that of the internal standard is not more than 1.0z. Loss on drying <2.41> Not more than 0.3z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.4 g of Tranilast, previ-ously dried, dissolve in 25 mL of N,N-dimethylformamide, add 25 mL of water, and titrate <2.50> with 0.1 mol/L so-dium hydroxide VS, until a 30-seconds persistent light-red color is obtained (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 32.73 mg of C18H17NO5
Trapidil, when dried, contains not less than 98.
Trapidil occurs as a white to pale yellow-white crystalline powder. ‌It is very soluble in water and in methanol, freely soluble in ethanol (95), in acetic anhydride and in acetic acid (100), and sparingly soluble in diethyl ether. The pH of a solution of 1.0 g of Trapidil in 100 mL of water is between 6.5 and 7.5.
(1) To 5 mL of a solution of Trapidil (1 in 50) add 3 drops of Dragendorff's TS: an orange color devel-ops. (2) Determine the absorption spectrum of a solution of Trapidil (1 in 125,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wavelengths.
Trehalose Hydrate contains not less than 98.
Trehalose Hydrate occurs as white crystals or crystalline powder. It is freely soluble in water, and slightly soluble in metha-nol and in ethanol (99.5).
(1) To 1 mL of a solution of Trehalose Hy-drate (2 in 5) add 5 – 6 drops of a solution of 1-naphthol in ethanol (95) (1 in 20), shake thoroughly, and add gently 2 mL of sulfuric acid: a purple color appears at the zone of contact. Mix 2 mL of a solution of Trehalose Hydrate (1 in 25) with 1 mL of dilute hydrochloric acid, and allow stand-ing for 20 minutes at room temperature. Then add 4 mL of sodium hydroxide TS and 2 mL of a solution of glycine (1 in 25), and heat in a water bath for 10 minutes: no brown color appears. Determine the infrared absorption spectrum of Tre-halose Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Trehalose RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋197 – ＋2019(10 g calculated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> The pH of a solution of 1 g of Trehalose Hy-drate in 10 mL of water is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve 2.5 g of Trapidil in 10 mL of water: the solution is clear and color-less to pale yellow. Chloride <1.03>—Perform the test with 0.5 g of Trapidil. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Ammonium—Place 0.05 g of Trapidil in a glass-stoppered conical flask, thoroughly moisten with 10 drops of sodium hydroxide TS, and stopper the flask. Allow it to stand at 379C for 15 minutes: the gas evolved does not change moistened red litmus paper to blue. Heavy metals <1.07>—Dissolve 1.0 g of Trapidil in 40 mL of water, and add 1.5 mL of dilute hydrochloric acid, 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 1.0 mL of Standard Lead So-lution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Trapidil according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Trapidil in 4 mL of methanol, and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 1 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, ethanol (95) and ace-tic acid (100) (85:13:2) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 60 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Trapidil, previously dried, dissolve in 20 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.53 mg of C10H15N5 C12H22O11.2H2O: 378.33 a-D-Glucopyranosyl a-D-glucopyranoside dihydrate [6138-23-4] Trehalose Hydrate contains not less than 98.0z and not more than 101.0z of trehalose (C12H22O11: 342.30), calculated on the anhydrous basis. Description Trehalose Hydrate occurs as white crystals or crystalline powder. It is freely soluble in water, and slightly soluble in metha-nol and in ethanol (99.5). Identification (1) To 1 mL of a solution of Trehalose Hy-drate (2 in 5) add 5 – 6 drops of a solution of 1-naphthol in ethanol (95) (1 in 20), shake thoroughly, and add gently 2 mL of sulfuric acid: a purple color appears at the zone of contact. Mix 2 mL of a solution of Trehalose Hydrate (1 in 25) with 1 mL of dilute hydrochloric acid, and allow stand-ing for 20 minutes at room temperature. Then add 4 mL of sodium hydroxide TS and 2 mL of a solution of glycine (1 in 25), and heat in a water bath for 10 minutes: no brown color appears. Determine the infrared absorption spectrum of Tre-halose Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Trehalose RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋197 – ＋2019(10 g calculated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> The pH of a solution of 1 g of Trehalose Hy-drate in 10 mL of water is between 4.5 and 6.5. Purity (1) Chloride <1.03>—Perform the test with 2.0 g of Trehalose Hydrate. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Sulfate <1.14>—Perform the test with 2.0 g of Treha- lose Hydrate. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Heavy metals <1.07>—Proceed with 5.0 g of Treha-lose Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Related substances—Dissolve 0.5 g of Trehalose Hy-drate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard ‌solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine each peak area by the automatic integra-tion method: the total area of the peaks which are eluted be-fore the peak of trehalose and the total area of the peaks which are eluted after the peak of trehalose obtained from the sample solution are both not larger than 1/2 times the peak area of trehalose from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions under the Assay. Time span of measurement: About 2 times as long as the retention time of trehalose. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add water to make exactly 10 mL. Con-firm that the peak area of trehalose obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System repeatability: To exactly 5 mL of the standard so-lution add water to make exactly 10 mL. When the test is repeated 6 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of trehalose is not more than 1.0z. Dextrin, soluble starch, and sulfite—Dissolve 1.0 g of Trehalose Hydrate in 10 mL of water and add 1 drop of iodine TS: a yellow color appears, which is changed to blue on addition of 1 drop of starch TS. Nitrogen—Perform the test with accurately weighed Trehalose Hydrate of about 5 g as directed under Nitrogen Determination <1.08>, using 30 mL of sulfuric acid for the degradation and adding 45 mL of sodium hydroxide solution (2 in 5): the amount of nitrogen (N: 14.01) is not more than 0.005z. Water <2.48> Not less than 9.0z and not more than 11.0z (0.1 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (2 g). Assay Weigh accurately about 0.2 g each of Trehalose Hy-drate and Trehalose RS (separately determine the water <2.48> in the same manner as Trehalose Hydrate), dissolve each in 6 mL of water, add exactly 2 mL each of the internal standard solution, add water to make them 20 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of trehalose to that of the internal standard. Amount (mg) of trehalose (C12H22O11) ＝ MS × QT/QS MS: Amount (mg) of Trehalose RS taken, calculated on the anhydrous basis Internal standard solution—A solution of glycerin (1 in 10). Operating conditions— Detector: A differential refractometer. Column: A stainless steel column 8 mm in inside diameter and 30 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography consist of styrene-divinylbenzene copolymer carrying sulfonic acid groups (6 mm in particle diameter). Column temperature: A constant temperature of about 809C. Mobile phase: Water. Flow rate: Adjust so that the retention time of trehalose is about 15 minutes. System suitability— System performance: Dissolve 0.1 g each of maltotriose and glucose in 10 mL of the standard solution, add 1 mL of the internal standard solution, and add water to make 20 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, maltotriose, treha-lose, glucose and the internal standard are eluted in this order, and the resolution between the peaks of maltotriose and trehalose is not less than 1.5, the resolution between the peaks of trehalose and glucose is not less than 4, and the resolution between the peaks of glucose and the internal standard is not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of trehalose to that of the internal standard is not more than 1.0z.
Trepibutone, when dried, contains not less than 98.
Trepibutone occurs as white to yellowish white, crystals or crystalline powder. It is odorless, and is tasteless or has a slight, characteristic aftertaste. It is soluble in acetone, sparingly soluble in ethanol (95), slightly soluble in diethyl ether, and practically insoluble in water. It dissolves in sodium hydroxide TS.
(1) Determine the absorption spectrum of a solution of Trepibutone in diluted dilute sodium hydroxide TS (1 in 10) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. (2) Determine the 1H spectrum of a solution of Trepibu-tone in deuterated chloroform for the nuclear magnetic reso-nance spectroscopy (1 in 10), using tetramethylsilane for the nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits a sharp multiplet signal A at around d 1.5 ppm, a triplet signal B at around d 2.7 ppm, a triplet signal C at around d 3.3 ppm, a multiplet signal D at around d 4.2 ppm, a sharp singlet signal E at around d 6.4 ppm, a sharp singlet signal F at around d 7.4 ppm, and a singlet signal G at around d 10.5 ppm. The ratio of integrated intensity of each signal, A:B:C:D:E:F:G, is about 9:2:2:6:1:1:1. ‌
(1) Chloride <1.03>—Dissolve 0.5 g of Trepibu-tone in 30 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Heavy metals <1.07>—Proceed with 1.0 g of Trepibu- tone according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Trepibutone in 10 mL of acetone, and use this solution as the sample so-lution. Pipet 2 mL of the sample solution, add acetone to make exactly 100 mL. To exactly 10 mL of this solution add acetone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of isopro-pylether, acetone, water and formic acid (100:30:3:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solu-tion are not more intense than the spot from the standard so-lution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Trepibutone, previ-ously dried, dissolve in 50 mL of ethanol (95), add 50 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (indicator: 5 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L sodium hydroxide VS It is freely soluble in N,N-dimethylformamide, slightly soluble in methanol and in ethanol (95), and practically in-soluble in water. Melting point: about 2649C (with decomposition). It shows crystal polymorphism. Identification (1) Dissolve 1 mg of Triamcinolone in 6 mL of ethanol (95), add 5 mL of 2,6-di-tert-butylcresol TS and 5 mL of sodium hydroxide TS, and heat on a water bath for 30 minutes under a reflux condenser: a red-purple color develops. Add 5 mL of water and 1 mL of Fehling's TS to 0.01 g of Triamcinolone, and heat: a red precipitate is pro-duced. Proceed with 0.01 g of Triamcinolone as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution re-sponds to Qualitative Tests <1.09> for fluoride. Determine the infrared absorption spectrum of Triamcinolone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Triamcinolone RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve 0.1 g each of Triamcinolone and Triam-cinolone RS in 7 mL of a mixture of 2-propanol and water (2:1), respectively, by warming. Allow the solutions to cool in ice to effect crystals, filter, then wash the formed crystals with two 10-mL portions of water, and repeat the test on the dried crystals. D Optical rotation <2.49> [a]20: ＋65 – ＋719(after drying, 0.1 g, N,N-dimethylformamide, 10 mL, 100 mm). Purity Heavy metals <1.07>—Proceed with 0.5 g of Triam-cinolone according to Method 2, and perform the test. Pre-pare the control solution with 1.5 mL of Standard Lead So-lution (not more than 30 ppm). Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu- ＝ 31.03 mg of C 16H 22O6 um, phosphorus (V) oxide, 609C, 3 hours).
Triamcinolone Acetonide, when dried, contains not less than 97.
Triamcinolone Acetonide occurs as a white crystalline powder. It is sparingly soluble in ethanol (99.5) and in acetone, slightly soluble in methanol, and practically insoluble in water. Melting point: about 2909C (with decomposition). It shows crystal polymorphism.
(1) Dissolve 2 mg of Triamcinolone Acetonide in 40 mL of ethanol (95), add 5 mL of 2,6-di-tert-butylcresol TS and 5 mL of sodium hydroxide TS, and heat on a water bath under a reflux condenser for 20 minutes: a green color develops. Add 5 mL of water and 1 mL of Fehling's TS to 0.01 g of Triamcinolone Acetonide, and heat: a red precipi-tate is produced. Proceed with 0.01 g of Triamcinolone Acetonide as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid: the solution responds to Qualitative Tests <1.09> for fluoride. Determine the absorption spectrum of a solution of Triamcinolone Acetonide in ethanol (95) (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Triamcinolone Acetonide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Triamcinolone Acetonide, previously dried, as directed in the potassium bromide disk method under the Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Triamcinolone Acetonide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve 0.1 g each of Triamcinolone Acetonide and Triamcinolone Acetonide RS in 20 mL of ethanol (95), respectively, then evaporate the ethanol to dryness, and repeat the test on the dried residue. D Optical rotation <2.49> [a]25: ＋110 – ＋1209(after drying, 0.1 g, ethanol (99.5), 10 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 0.5 g of Triamcinolone Acetonide according to Method 2, and per-form the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 30 ppm). (2) Related substances—Dissolve 40 mg of Triamcino-lone Acetonide in 4 mL of acetone, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add acetone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and methanol (93:7) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z (0.5 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g, platinum crucible).
Dissolve about 20 mg each of Triamcinolone Acetonide and Triamcinolone Acetonide RS, previously dried and accurately weighed, in methanol to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL each of the internal standard solution, then add the mo-bile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 20 mL each of these solutions as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of triamcinolone acetonide to that of the internal standard. Amount (mg) of triamcinolone acetonide (C24H31FO6) ＝ MS × QT/QS MS: Amount (mg) of Triamcinolone Acetonide RS taken Internal standard solution—A solution of prednisolone in methanol (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. ‌Mobile phase: A mixture of water and acetonitrile (3:1). Flow rate: Adjust so that the retention time of triamcino-‌ lone acetonide is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and triamcinolone acetonide are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak height of triamcinolone acetonide to that of the internal standard is not more than 1.0z.
Triamcinolone, when dried, contains not less than 97.
Triamcinolone occurs as a white crystalline powder. Residue on ignition <2.44> Not more than 0.3z (0.5 g, platinum crucible).
Dissolve about 20 mg each of Triamcinolone and Triamcinolone RS, previously dried and accurately weighed, in a solution of L-ascorbic acid in methanol (1 in 1000) to make exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL each of the internal standard solution, add a solution of L-ascorbic acid in methanol (1 in 1000) to make 20 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of triamcinolone to that of the internal standard. Amount (mg) of triamcinolone (C21H27FO6) ＝ MS × QT/QS MS: Amount (mg) of Triamcinolone RS taken Internal standard solution—Dissolve 15 mg of methyl par-ahydroxybenzoate in a solution of L-ascorbic acid in metha-nol (1 in 1000) to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame- ‌ter and 15 cm in length, packed with octadecylsilanized silica gel (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (3:1). Flow rate: Adjust so that the retention time of triamcino- lone is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, triamcinolone and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak height of triamcinolone to that of the internal standard is not more than 1.5z.
Triamterene, when dried, contains not less than 98.
Triamterene occurs as a yellow crystalline powder. It is odorless, and tasteless. It is sparingly soluble in dimethylsulfoxide, very slightly soluble in acetic acid (100), and practically insoluble in water, in ethanol (95), and in diethyl ether. It dissolves in nitric acid and in sulfuric acid, but does not dissolve in dilute nitric acid, in dilute sulfuric acid and in di-lute hydrochloric acid.
(1) To 0.01 g of Triamterene add 10 mL of water, heat, and filter after cooling: the filtrate shows a pur-ple fluorescence. To 2 mL of the filtrate add 0.5 mL of hy-drochloric acid: the fluorescence disappears. The filtrate obtained in (1) responds to Qualitative Tests <1.09> for primary aromatic amines. Dissolve 0.01 g of Triamterene in 100 mL of acetic acid (100), and to 10 mL of the solution add water to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Triamterene according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Triamterene according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Triamterene in 20 mL of dimethylsulfoxide. To 2 mL of this solution add methanol to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28) and methanol (9:1:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 365 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.10z (1 g).
Weigh accurately about 0.15 g of Triamterene, pre-viously dried, and dissolve in 100 mL of acetic acid (100) by warming. Titrate <2.50> with 0.05 mol/L perchloric acid VS (indicator: 2 drops of crystal violet TS). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 12.66 mg of C12H11N7
Triazolam, when dried, contains not less than 98.
Triazolam occurs as a white crystalline pow-der. It is sparingly soluble in N,N-dimethylformamide, slightly soluble in ethanol (95), and practically insoluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Triazolam in ethanol (95) (1 in 200,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Triazolam RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tria-zolam, previously dried, as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Triazolam RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Triazolam as directed under Flame Coloration Test <1.04> (2): a green to blue-green color appears.
(1) Chloride <1.03>—To 1.0 g of Triazolam add 50 mL of water, and allow to stand for 1 hour while occasional shaking, and filter. Discard the first 10 mL of the filtrate, pipet 25 mL of the subsequent filtrate, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.028z). Heavy metals—Being specified separately when the drug is granted approval based on the Law. Related substances—Dissolve 0.14 g of Triazolam in 10 mL of N,N-dimethylformamide, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add N,N-dimethylformamide to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 12 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of peak other than triazolam obtained from the sample solution is not larger than 1/5 times the peak area of triazolam from the standard solution, and the total area of the peaks other than triazolam is not larger than the peak area of triazolam from the standard solution. For the areas of the peaks, related substance A having the relative retention time of about 0.7 to triazolam, related substance B having the relative reten-tion time of about 1.5, and related substance C having the relative retention time of about 2.4, multiply their correction factors, 1.8, 0.6 and 4.3, respectively. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 39 minutes after injec- tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add N,N-dimethylformamide to make exactly 10 mL. Confirm that the peak area of triazolam obtained with 12 mL of this solution is equivalent to 7 to 13z of that with 12 mL of the standard solution. System performance: When the procedure is run with 12 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of triazolam are not less than 4500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 12 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of triazolam is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 55 mg each of Triazolam and Triazolam RS, previously dried, dissolve each in N,N-dimethylformamide to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 12 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of triazolam in each solution. Amount (mg) of triazolam (C17H12Cl2N4) ＝ MS × AT/AS MS: Amount (mg) of Triazolam RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of methanol and diluted acetic acid-ammonium acetate buffer solution (pH 4.5) (1 in 10) (14:11). Mobile phase B: A mixture of methanol and diluted acetic acid-ammonium acetate buffer solution (pH 4.5) (1 in 10) (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 14 14 – 34 98 → 1 2 → 99 34 – 39 Flow rate: 2.0 mL per minute. System suitability— System performance: When the procedure is run with 12 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of triazolam are not less than 4500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 12 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of triazolam is not more than 1.0z.
Trichlormethiazide, when dried, contains not less than 97.
Trichlormethiazide occurs as a white powder. It is freely soluble in N,N-dimethylformamide and in ace- tone, slightly soluble in acetonitrile and in ethanol (95), and practically insoluble in water. A solution of Trichlormethiazide in acetone (1 in 50) shows no optical rotation. Melting point: about 2709C (with decomposition).
(1) Determine the absorption spectrum of a solution of Trichlormethiazide in ethanol (95) (3 in 250,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Trichlormethiazide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Trichlormethiazide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Trichlormethiazide RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Perform the test with Trichlormethiazide as directed under Flame Coloration Test <1.04> (2): a green color ap-pears.
(1) Chloride <1.03>—Dissolve 1.0 g of Trichlor-methiazide in 30 mL of acetone, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.036z). Sulfate <1.14>—Dissolve 1.0 g of Trichlormethiazide in 30 mL of acetone, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this so- lution as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.005 mol/L sulfuric acid VS add 30 mL of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Trichlor- methiazide according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.6 g of Trichlormethiazide according to Method 5, using 20 mL of N,N-dimethylformamide, and perform the test (not more than 3.3 ppm). Related substances—Dissolve 25 mg of Trichlor-methiazide in 50 mL of acetonitrile, and use the solution as the sample solution. Perform the test with 10 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late the amount of related substances by the area percentage method: the amount of 4-amino-6-chlorobenzene-1,3-disul-fonamide, having the relative retention time of about 0.3 to trichlormethiazide, is not more than 2.0z, and the total amount of the related substances is not more than 2.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:1). Mobile phase B: A mixture of acetonitrile and diluted phosphoric acid (1 in 1000) (3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 10 10 – 20 100 → 0 0 → 100 Flow rate: 1.5 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of trichlormethiazide, beginning after the sol-vent peak. System suitability— Test for required detectability: To exactly 1 mL of the sample solution add acetonitrile to make exactly 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add acetonitrile to make exactly 20 mL. Confirm that the peak area of trichlormethiazide obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solution for system suitability test. System performance: To 5 mL of the solution for system suitability test add 5 mL of water, and warm in a water bath at 609C for 30 minutes. When the procedure is run with 10 mL of this solution, after cooling, under the above operating conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide and trichlormethiazide are eluted in this order, the relative reten-tion time of 4-amino-6-chlorobenzene-1,3-disulfonamide to trichlormethiazide is about 0.3, and the number of theoreti-cal plates and the symmetry factor of the peak of trichlor- ‌methiazide are not less than 5000 and not more than 1.2, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of trichlormethiazide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 25 mg of Trichlormethiazide and Trichlormethiazide RS, previously dried, and dissolve separately in exactly 20 mL of the internal standard solution. To 1 mL of these solutions add acetonitrile to make 20 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of trichlormethiazide to that of the internal standard. Amount (mg) of trichlormethiazide (C H Cl N O S ) Identification To an amount of powdered Trichlor-methiazide Tablets, equivalent to 4 mg of Trichlor-methiazide, add 10 mL of acetone, shake vigorously for 5 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 4 mg of Trichlor-methiazide RS in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and methanol (10:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spots from the sample solution and the standard solution show the same Rf value. Purity Related substances—Powder a suitable amount of Trichlormethiazide Tablets in an agate mortar. Take an amount of the powder, equivalent to 10 mg of Trichlor-methiazide, add 20 mL of acetonitrile, shake vigorously for 15 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with 10 mL of the sample ＝ MS × QT/QS 8 8 3 3 4 2 solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak MS: Amount (mg) of Trichlormethiazide RS taken Internal standard solution—A solution of 3-nitrophenol in acetonitrile (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:1). Flow rate: Adjust so that the retention time of trichlor-methiazide is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the internal standard and trichlormethiazide are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times area by the automatic integration method, and calculate the amount of each related substance by the area percentage method: the amount of 4-amino-6-chlorobenzene-1,3-disul-foneamide, having the relative retention time of about 0.3 to trichlormethiazide, is not more than 4.0z, and the total amount of the peaks other than trichlormethiazide is not more than 5.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 268 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:1). Mobile phase B: A mixture of acetonitrile and diluted phosphoric acid (1 in 1000) (3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) the peak area of trichlormethiazide to that of the internal standard is not more than 1.0z.
Trichomycin occurs as a yellow to yellow-brown powder. It is practically insoluble in water, in ethanol (99.5) and in tetrahydrofuran. It dissolves in dilute sodium hydroxide TS. It is hygroscopic.
(1) To 2 mg of Trichomycin add 2 mL of sulfuric acid: a blue color appears, and the color is changed to a blue-purple after allowing to stand. (2) Dissolve 1 mg of Trichomycin in 50 mL of a solution of sodium hydroxide (1 in 200). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 359 nm and 365 nm, between 378 nm and 384 nm, and between 400 nm and 406 nm. Content ratio of the active principle Conduct this proce-dure without exposure to light, using light-resistant vessels. Dissolve 10 mg of Trichomycin in 50 mL of a mixture of tetrahydrofuran for liquid chromatography and water (3:1), and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, determine the peak areas by the automatic integration method, and calculate the amount of trichomycin A and trichomycin B by the area percentage method: the amount of trichomycin A is between 20z and 40z, and that of trichomycin B is between 15z and 25z. The relative reten-tion time of trichomycin B to trichomycin A is about 1.2. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 360 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate and 1.7 g of sodium lauryl sulfate in a mixture of 600 mL of water and 400 mL of acetonitrile for liquid chro-matography. Flow rate: Adjust so that the retention time of trichomy-cin A is about 8 minutes. Time span of measurement: About 4 times as long as the retention time of trichomycin A. System suitability— Test for required detectability: Measure 5 mL of the sam-ple solution, add a mixture of tetrahydrofuran for liquid chromatography and water (3:1) to make 50 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add a mixture of tetrahydrofuran for liquid chromatography and water (3:1) to make exactly 30 mL. Confirm that the peak area of trichomycin A obtained with 5 mL of this solution is equivalent to 12 to 22z of that with 5 mL of the solution for system suitability test. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above op-erating conditions, trichomycin A and trichomycin B are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of trichomycin A is not more than 2.0z. Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, 609C, 3 hours).
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately an amount of Trichomycin and Trichomycin RS, equivalent to about 150,000 units, dissolve them separately in a mixture of tetra-hydrofuran for liquid chromatography and water (3:1) to make exactly 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of trichomycin in each solution. Amount (unit) of trichomycin ＝ MS × AT/AS MS: Amount (unit) of Trichomycin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 360 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 15 g of ammonium acetate in 120 mL of water, and add 1000 mL of acetonitrile for liquid chromatography and 700 mL of methanol. Flow rate: Adjust so that the retention time of trichomy-cin is about 6 minutes. ‌System suitability— System performance: Dissolve 5 mg of Trichomycin and 1 mg of berberine chloride hydrate in 100 mL of a mixture of tetrahydrofuran for liquid chromatography and water (3:1). When the procedure is run with 20 mL of this solution under the above operating conditions, berberine and trichomycin are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of trichomycin is not more than 2.0z.
Trientine Hydrochloride occurs as white to pale yellow, crystals or crystalline powder. It is odorless or has slightly an ammonia-like odor. It is freely soluble in water, soluble in methanol, and slightly soluble in ethanol (99.5). It is hygroscopic. Melting point: about 1219C.
(1) Determine the infrared absorption spec-trum of Trientine Hydrochloride, previously dried, as di-rected in the paste method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. (2) A solution of Trientine Hydrochloride (1 in 100) re-sponds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> The pH of a solution obtained by dissolving 1 g of Trientine Hydrochloride in 100 mL of water is between 7.0 and 8.5.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Trientine Hydrochloride according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead solution (not more than 10 ppm). (2) Related substances—Dissolve 0.30 g of Trientine Hy-drochloride in 100 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 3 mL each of the sample solution and standard solution on two plates of silica gel for thin-layer chromatography. Develop the one plate with a mixture of 2-propanol and ammonia so-lution (28) (3:2) to a distance of about 6 cm, and air-dry the plate. Spray evenly ninhydrin-butanol TS on the plate, and heat the plate at 1309C for 5 minutes: the spots other than the principal spot and the spot nearby the starting point ob-tained from the sample solution is not more intense than the spot from the standard solution. Develop another plate with a mixture of ammonia solution (28), diethylether, aceto-nitrile, and ethanol (99.5) (10:4:3:3) to a distance of about 6 cm, and air-dry the plate. Spray evenly ninhydrin-butanol TS on the plate, and heat the plate at 1309C for 5 minutes: the spot nearby the starting point from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 2.0z (1 g, reduced pressure not exceeding 0.67 kPa, 409C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.22 g of Trientine Hydro- ‌chloride, and dissolve in 10 mL of 0.1 mol/L hydrochloric acid VS, 2 mL of a solution of sodium nitrate (9 in 20), 10 mL of acetic acid-ammonium acetate buffer solution (pH 4.8) and 50 mL of water. Titrate <2.50> with 0.1 mol/L cop-per (II) nitrate VS (potentiometric titration) using a copper electrode as the indicator electrode, a complex type silver-sil-ver chloride electrode as the reference electrode, and potas-sium chloride solution (1 in 4) as the inner solution. Perform a blank determination in the same manner, and make any necessary correction.‌ Each mL of 0.1 mol/L copper (II) nitrate VS ＝ 21.92 mg of C6H18N4.2HCl
Containers—Tight containers. Storage—Light-resistant, substituted by argon gas, at 2 –
Trihexyphenidyl Hydrochloride, when dried, con-tains not less than 98.5z of trihexyphenidyl hydro-chloride (C20H31NO.HCl). Description Trihexyphenidyl Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is soluble in ethanol (95), sparingly soluble in acetic acid (100), slightly soluble in water, very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: about 2509C (with decomposition). Identification (1) Dissolve 1 g of Trihexyphenidyl Hydro- ‌chloride in 100 mL of water by warming, and cool. Use this solution as the sample solution. To 5 mL of the sample solu-tion add 1 mL of a solution of 2,4,6-trinitrophenol in chlo-roform (1 in 50), and shake vigorously: a yellow precipitate is formed. To 20 mL of the sample solution obtained in (1) add 2 mL of sodium hydroxide TS: a white precipitate is formed. Collect the precipitate, wash with a small amount of water, recrystallize from methanol, and dry in a desiccator (in vacu-um, silica gel) for 2 hours: the crystals so obtained melt <2.60> between 1139C and 1179C. The sample solution obtained in (1) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Trihexyphenidyl Hydrochlo-ride in 100 mL of water by warming, and cool: the pH of this solution is between 5.0 and 6.0. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Trihexyphenidyl Hydrochloride in 100 mL of water by warming: the solution is clear and colorless. Heavy metals <1.07>—Dissolve 1.5 g of Trihexy- phenidyl Hydrochloride in 60 mL of water by warming on a water bath at 809C, cool, and filter. To 40 mL of the filtrate add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Piperidylpropiophenone—Dissolve 0.10 g of Trihexy-phenidyl Hydrochloride in 40 mL of water and 1 mL of 1 mol/L hydrochloric acid TS by warming, cool, and add water to make exactly 100 mL. Determine the absorbance of this solution at 247 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.50. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Trihexyphenidyl Hydrochloride, previously dried, dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 mol/L perchloric acid-1,4-dioxane VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid-dioxane VS Trihexyphenidyl Hydrochloride, add 30 mL of chloroform, shake, and filter. Evaporate the filtrate on a water bath to dryness. Dissolve the residue in 10 mL of water by warming, cool, and use this solution as the sample solution. With 5 mL of the sample solution, proceed as directed in the Identifica-tion (1) under Trihexyphenidyl Hydrochloride. Shake a quantity of powdered Trihexyphenidyl Hy-drochloride Tablets, equivalent to 0.01 g of Trihexyphenidyl Hydrochloride, with 5 mL of chloroform, filter, and use the filtrate as the sample solution. Dissolve 0.02 g of Trihex-yphenidyl Hydrochloride RS in 10 mL of chloroform, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and methanol (9:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate: the spots from the sample solution and the standard solution show a blue-pur-ple color and the same Rf value. The sample solution obtained in (1) responds to Qualitative Tests <1.09> (2) for chloride. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Trihexyphenidyl Hydrochloride Tablets add 2 mL of dilute hydrochloric acid and 60 mL of water, disin-tegrate by vigorous shaking for 10 minutes, and warm on a water bath with occasional shaking for 10 minutes. Cool, add 2 mL of methanol, and add water to make exactly V mL so that each mL contains about 20 mg of trihexyphenidyl hy-drochloride (C20H31NO.
Trihexyphenidyl Hydrochloride occurs as a white crystalline powder. It is odorless, and has a bitter taste. It is soluble in ethanol (95), sparingly soluble in acetic acid (100), slightly soluble in water, very slightly soluble in acetic anhydride, and practically insoluble in diethyl ether. Melting point: about 2509C (with decomposition).
(1) Dissolve 1 g of Trihexyphenidyl Hydro- ‌chloride in 100 mL of water by warming, and cool. Use this solution as the sample solution. To 5 mL of the sample solu-tion add 1 mL of a solution of 2,4,6-trinitrophenol in chlo-roform (1 in 50), and shake vigorously: a yellow precipitate is formed. To 20 mL of the sample solution obtained in (1) add 2 mL of sodium hydroxide TS: a white precipitate is formed. Collect the precipitate, wash with a small amount of water, recrystallize from methanol, and dry in a desiccator (in vacu-um, silica gel) for 2 hours: the crystals so obtained melt <2.60> between 1139C and 1179C. The sample solution obtained in (1) responds to Qualitative Tests <1.09> (2) for chloride. pH <2.54> Dissolve 1.0 g of Trihexyphenidyl Hydrochlo-ride in 100 mL of water by warming, and cool: the pH of this solution is between 5.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Trihexyphenidyl Hydrochloride in 100 mL of water by warming: the solution is clear and colorless. Heavy metals <1.07>—Dissolve 1.5 g of Trihexy- phenidyl Hydrochloride in 60 mL of water by warming on a water bath at 809C, cool, and filter. To 40 mL of the filtrate add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Piperidylpropiophenone—Dissolve 0.10 g of Trihexy-phenidyl Hydrochloride in 40 mL of water and 1 mL of 1 mol/L hydrochloric acid TS by warming, cool, and add water to make exactly 100 mL. Determine the absorbance of this solution at 247 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.50. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Trihexyphenidyl Hydrochloride, previously dried, dissolve in 50 mL of a mix-ture of acetic anhydride and acetic acid (100) (1:1), and titrate <2.50> with 0.1 mol/L perchloric acid-1,4-dioxane VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid-dioxane VS Trihexyphenidyl Hydrochloride, add 30 mL of chloroform, shake, and filter. Evaporate the filtrate on a water bath to dryness. Dissolve the residue in 10 mL of water by warming, cool, and use this solution as the sample solution. With 5 mL of the sample solution, proceed as directed in the Identifica-tion (1) under Trihexyphenidyl Hydrochloride. Shake a quantity of powdered Trihexyphenidyl Hy-drochloride Tablets, equivalent to 0.01 g of Trihexyphenidyl Hydrochloride, with 5 mL of chloroform, filter, and use the filtrate as the sample solution. Dissolve 0.02 g of Trihex-yphenidyl Hydrochloride RS in 10 mL of chloroform, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chlo-roform and methanol (9:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate: the spots from the sample solution and the standard solution show a blue-pur-ple color and the same Rf value. The sample solution obtained in (1) responds to Qualitative Tests <1.09> (2) for chloride. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Trihexyphenidyl Hydrochloride Tablets add 2 mL of dilute hydrochloric acid and 60 mL of water, disin-tegrate by vigorous shaking for 10 minutes, and warm on a water bath with occasional shaking for 10 minutes. Cool, add 2 mL of methanol, and add water to make exactly V mL so that each mL contains about 20 mg of trihexyphenidyl hy-drochloride (C20H31NO.HCl). Centrifuge, if necessary, and use the supernatant liquid as the sample solution. Separately, dissolve about 20 mg of Trihexyphenidyl Hydrochloride RS (determine previously its loss on drying <2.41> under the same conditions as Trihexyphenidyl Hydrochloride) in methanol to make exactly 20 mL. Pipet 2 mL of this solu-tion, and add 2 mL of dilute hydrochloric acid and water to make exactly 100 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample solution and stand-ard solution, transfer to glass-stoppered centrifuge tubes, add exactly 10 mL of bromocresol purple-dipotassium hydrogenphosphate-citric acid TS and 15 mL of chloroform, stopper tightly, shake well, and centrifuge. Pipet 10 mL each of the chloroform layers, add chloroform to make exactly 50 ＝ 33.79 mg of C 20H 31NO.HCl mL. Determine the absorbances, AT and AS, of the subse-quent solutions of the sample solution and standard solution
Trimebutine Maleate, when dried, contains not less than 98.
Trimebutine Maleate occurs as white, crystals or crystalline powder. It is freely soluble in N,N-dimethylformamide and in ace-tic acid (100), soluble in acetonitrile, and slightly soluble in water and in ethanol (99.5). It dissolves in 0.01 mol/L hydrochloric acid TS. A solution of Trimebutine Maleate in N,N-dimethyl-formamide (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Trimebutine Maleate in 0.01 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Trimebutine Maleate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Trimebutine Maleate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Trimebutine Maleate according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.10 g of Trimebutine Maleate in 100 mL of a mixture of 0.01 mol/L hydrochloric acid TS and acetonitrile (13:7), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of 0.01 mol/L hydrochloric acid TS and acetonitrile (13:7) to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than maleic acid and trimebutine obtained from the sample solu-tion is not larger than 1/2 times the peak area of trimebutine from the standard solution, and the total area of the peaks other than maleic acid and trimebutine is not larger than the peak area of trimebutine from the standard solution. ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 650 mL of diluted perchloric acid (17 in 20,000), previously adjusted the pH to 3.0 with a solution of ammonium acetate (1 in 1000), add 1 g of sodium 1-pen-tanesulfonate to dissolve. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of trimebutine is about 9 minutes. Time span of measurement: About 2 times as long as the retention time of trimebutine, beginning after the peak of maleic acid. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add a mixture of 0.01 mol/L hy-drochloric acid TS and acetonitrile (13:7) to make exactly 20 mL. Confirm that the peak area of trimebutine obtained with 20 mL of this solution is equivalent to 20 to 30z of that with 20 mL of the standard solution. System performance: Dissolve 40 mg of Trimebutine Maleate and 20 mg of imipramine hydrochloride in 100 mL of a mixture of 0.01 mol/L hydrochloric acid TS and aceto-nitrile (13:7). When the procedure is run with 20mL of this solution under the above operating conditions, trimebutine and imipramine are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of trimebutine is not more than 5z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.8 g of Trimebutine Male-ate, previously dried, dissolve in 70 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple through blue to blue-green (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS Trimetazidine Hydrochloride
Trimetazidine Hydrochloride contains not less than 98.
Trimetazidine Hydrochloride occurs as a white crystalline powder. It is very soluble in water and in formic acid, sparingly soluble in methanol, and slightly soluble in ethanol (99.5). The pH of a solution of 1.0 g of Trimetazidine Hydrochlo-ride in 20 mL of water is between 2.3 and 3.3. Melting point: about 2279C (with decomposition).
(1) Determine the absorption spectrum of a solution of Trimetazidine Hydrochloride in 0.1 mol/L hy-drochloric acid TS (1 in 6250) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Trimetazidine Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Trimetazidine Hydrochloride (1 in 50) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Trimetazidine Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.2 g of Trimetazidine Hydrochloride in 50 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 20 mL. Pipet 2 mL of this solution, ＝ 50.35 mg of C22H29 NO5.C4H4O4 add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL
Weigh accurately about 0.12 g of Trimetazidine Hy-drochloride, dissolve in 5 mL of formic acid, add exactly 15 mL of 0.1 mol/L perchloric acid VS, and heat at 90 – 1009C for 30 minutes. After cooling, add 45 mL of acetic acid (100), and titrate <2.50> the excess perchloric acid with 0.1 mol/L sodium acetate VS (potentiometric titration). Per-form a blank determination in the same manner. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.96 mg of C14H22N2O3.2HCl
Trimethadione, when dried, contains not less than 98.
Trimethadione occurs as white, crystals or crystalline powder. It has a camphor-like odor. It is very soluble in ethanol (95) and in chloroform, freely soluble in diethyl ether, and soluble in water.
(1) To 5 mL of a solution of Trimetha-dione (1 in 50) add 2 mL of barium hydroxide TS: a precipi-tate is formed immediately. (2) Determine the infrared absorption spectrum of a so-lution of Trimethadione in chloroform (1 in 50) as directed in the solution method under Infrared Spectrophotometry <2.25>, using a 0.1-mm fixed sodium chloride cell, and com- pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
Heavy metals <1.07>—Proceed with 2.0 g of Trimethadione according to Method 1, and perform the test. 14 22 2 3 S T S Prepare the control solution with 2.0 mL of Standard Lead MS: Amount (mg) of trimetazidine hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of parahy-droxybenzoic acid in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) (7 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0) and methanol (17:3). Flow rate: Adjust so that the retention time of trimetazi-dine is about 7 minutes. Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Trimethadione, pre-viously dried, in a glass-stoppered conical flask, dissolve in 5 mL of ethanol (95), add exactly measured 50 mL of 0.1 mol/L sodium hydroxide VS, stopper, and allow to stand for 15 minutes with occasional shaking. Titrate <2.50> the ex-cess sodium hydroxide with 0.1 mol/L hydrochloric acid VS (indicator: 4 drops of cresol red TS). Perform a blank deter-mination in the same manner. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 14.31 mg of C6H9NO3
Trimetoquinol Hydrochloride Hydrate occurs as white, crystals or crystalline powder. It is freely soluble in methanol, and sparingly soluble in water and in ethanol (99.5). Melting point: about 1519C (with decomposition, after drying in vacuum, 1059C, 4 hours).
(1) Determine the absorption spectrum of a solution of Trimetoquinol Hydrochloride Hydrate in 0.01 mol/L hydrochloric acid TS (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Trimetoquinol Hydrochloride Hydrate as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Trimetoquinol Hydrochloride Hydrate (1 in 50) responds to Qualitative Tests <1.09> (1) for chloride. D Optical rotation <2.49> [a]20: －16 – －199(0.25 g calcu-lated on the anhydrous basis, water, after warming and cool-ing, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Trimetoquinol Hydrochloride Hydrate in 100 mL of water by warming, and cool: the pH of this solution is between 4.5 and 5.5.
(1) Clarity and color of solution—Dissolve 0.10 g of Trimetoquinol Hydrochloride Hydrate in 10 mL of water by warming: the solution is clear and colorless. Sulfate <1.14>—Perform the test with 0.5 g of Trimetoquinol Hydrochloride Hydrate. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Heavy metals <1.07>—Proceed with 1.0 g of Trimeto- quinol Hydrochloride Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Trimeto-quinol Hydrochloride Hydrate in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the total area of the peaks other than trimetoquinol obtained from the sample solution is not larger than the peak area of trimetoquinol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2 g of potassium dihydrogen phos-phate and 2 g of sodium 1-pentane sulfonate in 1000 mL of water. Adjust with phosphoric acid to a pH between 2.8 and 3.2, and filter through a membrane filter with a pore size of 0.4 mm. Add 200 mL of acetonitrile to 800 mL of the filtrate. Flow rate: Adjust so that the retention time of trimeto- quinol is about 7 minutes. Time span of measurement: About twice as long as the retention time of trimetoquinol, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of trimetoquinol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 5 mg of Trimetoquinol Hy-drochloride Hydrate and 1 mg of procaine hydrochloride in 50 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, procaine and trimetoquinol are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of trimetoquinol is not more than 2.0z. Water <2.48> 3.5 – 5.5z (0.3 g, volumetric titration, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Trimetoquinol Hy-drochloride Hydrate, dissolve in 2 mL of 0.1 mol/L hydro-chloric acid VS and 70 mL of ethanol (99.5) with thorough shaking, and titrate <2.50> with 0.1 mol/L potassium hy-droxide-ethanol VS (potentiometric titration). Calculate the consumed volume of 0.1 mol/L potassium hydroxide-ethanol VS between the first inflection point and of the second inflection point. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 38.19 mg of C19H23NO5.HCl
Tropicamide, when dried, contains not less than 98.
Tropicamide occurs as a white crystalline pow-der. It is odorless, and has a bitter taste. It is freely soluble in ethanol (95) and in chloroform, slightly soluble in water and in diethyl ether, and practically insoluble in petroleum ether. It dissolves in dilute hydrochloric acid. The pH of a solution of 1.0 g of Tropicamide in 500 mL of water is between 6.5 and 8.0.
(1) To 5 mg of Tropicamide add 0.5 mL of a solution of ammonium vanadate (V) in sulfuric acid, (1 in 200), and heat: a blue-purple color develops. 5 mg, 2 mol/L hydrochloric acid TS, 200 mL).
(1) Chloride <1.03>—Dissolve 1.0 g of Tropicamide in 30 mL of ethanol (95), add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS, 30 mL of ethanol (95), 6 mL of dilute nitric acid, and add water to make 50 mL (not more than 0.016z). Heavy metals <1.07>—Dissolve 1.0 g of Tropicamide in 30 mL of ethanol (95), add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solu-tion as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution, 30 mL of ethanol (95), 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). N-Ethyl-g-picolylamine—Dissolve 0.10 g of Tropica-mide in 5 mL of water by heating, add 1 mL of a solution of acetaldehyde (1 in 20), and shake well. Add 1 to 2 drops of sodium pentacyanonitrosylferrate (III) TS and 1 to 2 drops of sodium hydrogen carbonate TS, and shake: no blue color develops. Tropic acid—To 10 mg of Tropicamide add 5 mg of sodium tetraborate decahydrate and 7 drops of 4-dimethylaminobenzaldehyde TS, and heat in a water bath for 3 minutes. Cool in ice water, and add 5 mL of acetic an-hydride: no red-purple color develops. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Tropicamide, previ-ously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 28.44 mg of C17H20N2O2
Troxipide occurs as a white crystalline powder. It is freely soluble in acetic acid (100), soluble in methanol, sparingly soluble in ethanol (99.5) and slightly soluble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. A solution of Troxipide in 1 mol/L hydrochloric acid TS (1 in 5) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Troxipide in 0.1 mol/L hydrochloric acid TS (1 in 62,500) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Troxipide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Trox-ipide as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Troxipide RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Troxipide in 30 mL of methanol, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 0.25 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of methanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.009z). Heavy metals <1.07>—Moisten 2.0 g of Troxipide with 1 mL of sulfuric acid, and gently heat until charred. After cooling, add 2 mL of nitric acid, carefully heat until white fumes are no longer evolved, and perform the test according to Method 2. Prepare the control solution as follows: evaporate 1 mL of sulfuric acid, 2 mL of nitric acid and 2 mL of hydrochloric acid on a water bath and then on a sand bath to dryness, and moisten the residue with 3 drops of hydrochloric acid. Proceed in the same manner for the preparation of the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 10 ppm). Related substances—Dissolve 0.20 g of Troxipide in 10 mL of methanol, and use this solution as the sample solu-tion. Pipet 2 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of metha-nol, ethyl acetate, water, hexane and ammonia water (28) (20:20:5:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the number of the spots other than the principal spot obtained from the sample solution is not more than three, and they are not more intense than the spot from the stand-ard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.6 g of Troxipide, previ-ously dried, dissolve in 40 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 29.44 mg of C15H22N2O4
Tulobuterol Hydrochloride occurs as white, crystals or crystalline powder. It is very soluble in methanol, freely soluble in water, in ethanol (95) and in acetic acid (100), sparingly soluble in ace-tic anhydride. A solution of Tulobuterol Hydrochloride (1 in 20) shows no optical rotation. Melting point: about 1639C.
(1) Determine the absorption spectrum of a solution of Tulobuterol Hydrochloride (1 in 2500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Tulobuterol Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Tulobuterol Hydrochloride (1 in 20) responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Tulobuterol Hydrochloride in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Tulobuterol Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 30 mg of Tulobuterol Hydrochloride in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than tulobuterol obtained from the sample solution is not larger than the peak area of tulobuterol from the standard solution, and the total area of the peaks other than the peak of tulobuterol from the sample solution is not larger than 5 times the peak area of tulobuterol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). add the mobile phase to make 10 mL, and use this solution as the solution for system suitability test. When the proce-dure is run with 25 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of tulobuterol are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of tulobuterol is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, 609C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Tulobuterol Hydro-chloride, previously dried, dissolve in 80 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.42 mg of C12H18ClNO.HCl
ツロブテロール経皮吸収型テープ
Tulobuterol Transdermal Tape contains not less than 90.
After removing the liner from an amount of Tulobuterol Transdermal Tape, equivalent to 20 mg of Tulobuterol, shake with 10 mL of hexane. Take the superna-tant liquid to an another vessel, shake with 10 mL of 0.1 mol/L hydrochloric acid TS, centrifuge, and take the aque-ous layer. To 3 mL of the layer add 0.1 mol/L hydrochloric acid TS to make 10 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 261 nm and 263 nm and between 265 nm and 267 nm, and a shoulder between 271 nm and 273 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. After removing the liner from 1 tape of Tulobuterol Transdermal Tape, add exactly V mL of the internal stand-ard solution so that each mL contains about 0.25 mg of tulobuterol (C12H18ClNO), shake, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of tulobuterol for assay (separately determine the water <2.48> in the same manner as Tulobuterol), and dissolve in the internal standard solution to make exactly 20 mL. Pipet 5 mL of this solution, add the internal standard solution to make exactly 20 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of tulobuterol (C12H18ClNO) ＝ MS × QT/QS × V/80 MS: Amount (mg) of tulobuterol for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of benzyl benzoate in hexane (1 in 4000). Adhesive strength Being specified separately when the drug standard solution, shake, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of tulobuterol for assay (separately determine the water <2.48> in the same manner as Tulobuterol), and dissolve in hexane to make exactly 100 mL. Pipet 10 mL of this solu-tion, add exactly 1 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tulobuterol to that of the internal standard. Amount (mg) of tulobuterol (C12H18ClNO) in 1 tape ＝ MS × QT/QS × V/1000 MS: Amount (mg) of tulobuterol for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of benzyl benzoate in hexane (1 in 200). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused-silica column 0.53 mm in inside diame- ter and 30 m in length, coated the inside surface with methyl silicon polymer for gas chromatography in 1.5 mm thickness. Column temperature: A constant temperature of about 1809C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of tulobuterol is about 3 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, tulobuterol and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of tulobuterol is not more than 2.0z.
Tulobuterol occurs as white, crystals or crys-talline powder. It is very soluble in methanol, freely soluble in ethanol (99.5) and in acetic acid (100), and practically insoluble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. It gradually sublimes at 409C. A solution of Tulobuterol in methanol (1 in 20) shows no optical rotation.
(1) Determine the absorption spectrum of a solution of Tulobuterol in 0.1 mol/L hydrochloric acid TS (3 in 5000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Tulobuterol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Tulobuterol according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 5 ppm). Related substances—Dissolve 25 mg of Tulobuterol in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL. Pipet 5 mL of this so-lution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: each peak area other than tulobuterol obtained from the sample solu-tion is not larger than the peak area of tulobuterol from the standard solution, and the total area of the peaks other than tulobuterol from the sample solution is not larger than 5 times the peak area of tulobuterol from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3 g of sodium 1-octanesulfonate in 900 mL of water, and add 5 mL of diluted phosphoric acid (1 in 150). To 650 mL of this solution add 350 mL of aceto-nitrile for liquid chromatography. Flow rate: Adjust so that the retention time of tulobuterol is about 7 minutes. Time span of measurement: About 5 times as long as the retention time of tulobuterol, beginning after the solvent peak. System suitability— System performance: To 1 mL of the sample solution add the mobile phase to make 100 mL. To 5 mL of this solution add the mobile phase to make 10 mL, and use this solution as the solution for system suitability test. When the proce-dure is run with 25 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of tulobuterol are not less than 5000 and not more than 2.0, re-spectively. System repeatability: When the test is repeated 6 times with 25 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of tulobuterol is not more than 2.0z. Boron—Put 50 mg of Tulobuterol and 3.0 mL of Standard Boron Solution separately in platinum crucibles, and add 5 mL of potassium carbonate-sodium carbonate TS to them. After evaporating to dryness on a water bath, dry them at 1209C for 1 hour, and immediately incinerate by ignition. After cooling, add 0.5 mL of water and 3 mL of curcumin TS to the residue in the crucibles, and warm gently on a water bath for 5 minutes. After cooling, add 3 mL of acetic acid-sulfuric acid TS, and allow to stand for 30 minutes. Then add ethanol (95) to make them exactly 100 mL, filter, discard the first 10 mL of the filtrate, and use these subsequent filtrates as the sample solution and the ‌standard solution, respectively. Perform the test with these solutions as directed under Ultraviolet-visible Spectropho-tometry <2.24> using methanol (95) as a blank: the absor-bance at 555 nm of the sample solution is not more than that of the standard solution. Water <2.48> Not more than 0.2z (2 g, volumetric titra-tion, direct titration). is granted approval based on the Law. Drug release Being specified separately when the drug is granted approval based on the Law.
After removing the liner from 10 tapes of Tulobuterol Transdermal Tape, add V mL of hexane so that each mL contains 0.5 mg of tulobuterol (C12H18ClNO), then add exactly V/10 mL of the internal Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Tulobuterol, dis-solve in 20 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from purple to blue-green through blue (indicator: 2 drops of crystal violet TS ). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 22.77 mg of C12H18ClNO
Ubenimex, when dried, contains not less than 98.
Ubenimex occurs as a white crystalline pow-der. It is freely soluble in acetic acid (100), slightly soluble in water, and very slightly soluble in ethanol (99.5). It dissolves in 1 mol/L hydrochloric acid TS. Melting point: about 2309C (with decomposition).
(1) Determine the absorption spectrum of a solution of Ubenimex (1 in 2000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －15.5 – －17.59(after dry-ing, 0.5 g, 1 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ubenimex according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Ubenimex in 10 mL of the mobile phase A, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase A to make exactly 200 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ubenimex obtained from the sample solution is not larger than 1/2 times the peak area of ubenimex from the standard solution. Furthermore, the total area of the peaks other than ubenimex from the sample solution is not larger than the peak area of ubenimex from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of diluted 0.1 mol/L potas-sium dihydrogen phosphate TS (13 in 20) and acetonitrile for liquid chromatography (17:3). Mobile phase B: A mixture of acetonitrile for liquid chro-matography and diluted 0.1 mol/L potassium dihydrogen phosphate TS (13 in 20) (2:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Ubenimex Capsules
Ubidecarenone contains not less than 98.
Ubidecarenone occurs as a yellow to orange crystalline powder. It is odorless and has no taste. It is soluble in diethyl ether, very slightly soluble in ethanol (99.5), and practically insoluble in water. It is gradually decomposed and colored by light. Melting point: about 489C.
(1) Dissolve 0.05 g of Ubidecarenone in 1 mL of diethyl ether, and add 10 mL of ethanol (99.5). To 2 mL of this solution add 3 mL of ethanol (99.5) and 2 mL of dimethyl malonate, then add dropwise 1 mL of a solution of potassium hydroxide (1 in 5), and mix: a blue color appears. (2) Determine the infrared absorption spectrum of Amount (mg) of ubenimex (C H N O ) Ubidecarenone as directed in the potassium bromide disk ＝ MS × QT/QS × 1/4 16 24 2 4 method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the ‌spectrum of Ubidecarenone RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Ubidecarenone according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.05 g of Ubidecare-none in 50 mL of ethanol (99.5) by warming at about 509C for 2 minutes, and after cooling use this solution as the sam-ple solution. To exactly 1 mL of the sample solution add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the total area of the peaks other than ubidecarenone obtained from the sam-ple solution is not larger than the peak area of ubidecare-none from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, flow rate, and selection of column: Proceed as directed in the operating conditions in the Assay. Detection sensitivity: Adjust so that the peak height of ubidecarenone obtained from 5 mL of the standard solution is between 20 mm and 40 mm. Time span of measurement: About 2 times of the retention time of ubidecarenone, beginning after the solvent peak. Water <2.48> Not more than 0.20z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Ubidecare-none and Ubidecarenone RS (separately determined the water <2.48> in the same manner as Ubidecarenone) dissolve each in 40 mL of ethanol (99.5) by warming at about 509C for 2 minutes, and after cooling add ethanol (99.5) to make exactly 50 mL each, and use these solutions as the sample solution and standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine peak areas, AT and AS, of ubidecarenone in each solution. and calculate the resolution. Use a column giving elution of ubiquinone-9 and ubidecarenone in this order with the reso-lution between these peaks being not less than 4. System repeatability: Repeat the test 5 times with the standard solution under the above operating conditions: the relative standard deviation of the peak areas of ubidecare-none is not more than 0.8z.
Ulinastatin is a solution of a glycoprotein having trypsin inhibiting activity, which is separated and puri-fied from human urine. It contains ulinastatin of not less than 45,000 Units per mL and not less than 2500 Units per mg protein.
Ulinastatin occurs as a light brown to brown, clear liquid.
(1) Dilute a suitable volume of Ulinastatin with water to make a solution containing 4000 Units of ulinastatin per mL. To 1 mL of this solution add 1 mL of a solution of phenol (1 in 20), then carefully add 5 mL of sul-furic acid, and mix: an orange to red-orange color develops. Dilute a suitable volume of Ulinastatin with water to make a solution containing 2000 units per mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Dilute a suitable volume of Ulinastatin with 2,2?,2!- nitrilotriethanol buffer solution (pH 7.8) to make a solution containing 500 Units of ulinastatin per mL, and use this solution as the sample solution. Use the same buffer solution as the control solution. To 0.1 mL each of the sample solution and control solution add 1.6 mL of the same buffer solution and 0.2 mL of trypsin TS for test of ulinastatin, mix, and allow them to stand in a water bath at 259C Amount (mg) of ubidecarenone (C ＝ MS × AT/AS 59H 90O4) for 1 minute. Then add 1 mL of N-a-benzoyl-L-arginine-4-nitroanilide TS, mix, and allow them to stand at 259C for 2 minutes: the solution obtained with the sample solution de- MS: Amount (mg) of Ubidecarenone RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column about 5 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and ethanol (99.5) (13:7). Flow rate: Adjust so that the retention time of ubidecare-none is about 10 minutes. Selection of column: Dissolve 0.01 g each of Ubidecare-none and ubiquinone–9 in 20 mL of ethanol (99.5) by warm-ing at about 509C for 2 minutes. After cooling, proceed with 5 mL of this solution under the above operating conditions, velops no color while that obtained with the control solution develops a yellow color. To 1.5 g of Powdered Agar add 100 mL of boric acid-sodium hydroxide buffer solution (pH 8.4), dissolve by warming in a water bath, then pour immediately into a Petri dish placed horizontally so that the agar layer is about 2 mm in thickness. After the agar becomes hard, bore two wells about 2.5 mm in diameter with a separation of 6 mm from each other. In one of the wells place 10 mL of a solution of Ulinastatin containing 500 Units per mL in boric acid-sodi-um hydroxide buffer solution (pH 8.4), and in the other well place 10 mL of anti-ulinastatin rabbit serum, cover the dish to avoid drying of the agar, and allow to stand for overnight at a room temperature: a clear precipitin line appears be-tween the wells. pH <2.54> 6.0 – 8.0 Specific activity When calculated from the results obtained by the Assay and the following method, the specific activity is not less than 2500 Units per 1 mg protein. Sample solution—To an exactly measured volume of Ulinastatin, equivalent to about 10,000 Units, add water to make exactly 20 mL. Standard solutions—Weigh accurately about 10 mg of bovine serum albumin for test of ulinastatin, and dissolve in water to make exactly 20 mL. To a suitable volume of this solution add water to make four solutions containing exactly 300, 200, 100 and 50 mg of the bovine serum albumin for test of ulinastatin per mL, respectively. Procedure—Pipet 0.5 mL each of the sample solution and standard solutions, put them in glass test tubes about 18 mm in internal diameter and about 130 mm in length, add exactly 5 mL of alkaline copper TS, mix, and allow the tubes to stand in a water bath at 309C for 10 minutes. Then add exactly 0.5 mL of diluted Folin's TS (1 in 2), mix, and warm in the water bath for 20 minutes. Determine the absorbances of these solutions at 750 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution obtained in the same manner with 0.5 mL of water as the blank. Plot the absorbances of the standard solutions on the ver-tical axis and their protein concentrations on the horizontal axis to prepare a calibration curve, and calculate the protein content of the sample solution from its absorbance by using this curve. Then calculate the amount of protein per mL of Ulinastatin.
(1) Heavy metals <1.07>—Proceed with 10 mL of Ulinastatin according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 1 ppm). Related substances—To a suitable volume of Ulinastatin add water to make a solution containing exactly 12,500 Units per mL, and use this solution as the sample stock solution. To exactly 0.25 mL of the sample stock solu-tion add exactly 0.2 mL of glycerin and exactly 0.05 mL of 0.05z bromophenol blue TS, mix, and use this solution as the sample solution. Separately, to exactly 1 mL of the sam-ple stock solution add water to make exactly 100 mL. To ex-actly 0.25 mL of this solution add exactly 0.2 mL of glycerin and exactly 0.05 mL of 0.05z bromophenol blue TS, mix, and use this solution as the standard solution. Perform the following test with the sample solution and standard solu-tion: the bands other than the principal band obtained from the sample solution are not more intense than the band ob-tained from the standard solution in the electrophoretogram. Tris buffer solution A for polyacrylamide gel electro-phoresis Dissolve 18.2 g of 2-amino-2-hydroxymethyl-1,3-propanediol in 80 mL of water, adjust to pH 8.8 with 6 mol/L hydrochloric acid TS, and add water to make 100 mL. Tris buffer solution B for polyacrylamide gel electro-phoresis Dissolve 6.0 g of 2-amino-2-hydroxymethyl-1,3-propanediol in 80 mL of water, adjust to pH 8.8 with 6 mol/L hydrochloric acid TS, and add water to make 100 mL. Tris buffer solution C for polyacrylamide gel elec-trophoresis Dissolve 3.0 g of 2-amino-2-hydroxymethyl-1,3-propanediol and 14.4 g of glycine in water to make 1000 mL. Acrylamide solution for polyacrylamide gel electro-phoresis Dissolve 30 g of acrylamide and 0.8 g of N,N?-methylenebisacrylamide in water to make 100 mL. Gel for separation Mix gently 15 mL of tris buffer solution A for polyacrylamide gel electrophoresis, 20 mL of acrylamide solution for polyacrylamide gel electrophoresis, 24.5 mL of water, 0.022 mL of N,N,N?,N?-tetramethyl- ethylenediamine, 0.32 mL of 10z ammonium peroxodisul- fate TS and 0.3 mL of 1 mol/L sodium sulfite TS, pour into a plate for slab gel preparation, then cover the gel mixture with a layer of water, and allow to set for 1 hour. Gel for concentration Remove the water layer on the gel for separation, and pour a mixture of 2.5 mL of tris buffer solution B for polyacrylamide gel electrophoresis, 2.66 mL of acrylamide solution for polyacrylamide gel elec-trophoresis, 14.6 mL of water, 0.01 mL of N,N,N?,N?-tetra-methylethylenediamine, 0.2 mL of 10z ammonium perox-odisulfate TS and 0.04 mL of 1 mol/L sodium sulfite TS on the gel. Then position a plastic sample well former so that the height of the gel for concentration is about 15 mm, and allow to set for 2 hours. Procedure Electrophoresis Set the gel in an apparatus for slab gel electrophoresis, and fill the upper and lower reservoirs with tris buffer solution C for polyacrylamide gel electrophoresis. Introduce carefully 10 mL each of the sample solution and standard solution into the wells using a different well for each solution, and allow electrophoresis to proceed using the electrode of the lower reservoir as the anode. Switch off the power supply when the bromophenol blue band has migrated to about 10 mm from the bottom of the gel. Staining Dissolve 2.0 g of Coomassie brilliant blue R-250 in a mixture of 400 mL of methanol and 100 mL of acetic acid (100), add water to make 1000 mL, and use this solution as the staining solution. Stain the gel for 2 hours in the stain-ing solution warmed to 409C. Decolorization To 100 mL of methanol and 75 mL of acetic acid (100) add water to make 1000 mL, and use this solution as the rinsing solution. Immerse the gel removed from the staining solution in the rinsing solution to decolorise. Kallidinogenase—Dilute a suitable volume of Ulina-statin with water so that each mL of the solution contains about 50,000 Units, and use this solution as the sample solu-tion. Take exactly 0.4 mL of the sample solution into a test tube, add exactly 0.5 mL of tris buffer solution (pH 8.2), mix, and allow the tube to stand in a water bath at 37 ± 0.29C for 5 minutes. Add exactly 0.1 mL of substrate TS for kallidinogenase assay (4), mix, allow the tube to stand in the water bath of 37 ± 0.29C for exactly 30 minutes, then add exactly 0.1 mL of diluted acetic acid (100) (1 in 2), mix, and use this solution as the test solution. Separately, take exactly 0.4 mL of the sample solution in a test tube, add exactly 0.5 mL of tris buffer solution (pH 8.2), mix, and allow the tube to stand in the water bath of 37 ± 0.29C for 35 minutes. Then add exactly 0.1 mL of diluted acetic acid (100) (1 in 2), mix, add exactly 0.1 mL of substrate TS for kallidinogenase assay (4), mix, and use this solution as the control solution. Determine the absorbances of the test solution and the con-trol solution at 405 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the blank, and cal-culate the difference between them: the difference is not more than 0.050. Molecular mass Dilute a suitable volume of Ulinastatin with the mobile phase so that each mL of the solution con-tains about 6500 Units, and use this solution as the sample solution. Separately, dissolve 1.0 mg each of g-globulin (molecular mass: 160,000), bovine serum albumin for test of ulinastatin (molecular mass: 67,000), and myoglobin (molecular mass: 17,000) in about 1 mL of the mobile phase, and use this solution as the molecular mass reference solu-tion. Perform the test with 50 mL each of the sample solution and molecular mass reference solution as directed under Liq-uid Chromatography <2.01> according to the following con- ‌ditions. Prepare a calibration curve by plotting the logarithm of molecular masses on the vertical axis and the retention times (minute) of the molecular mass reference substances on the horizontal axis, and determine the molecular mass of the sample using the calibration curve and the retention time ob-tained with the sample solution: the molecular mass is 67,000 ± 5000. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column about 7 mm in inside di-ameter and about 60 cm in length, packed with porous silica gel for liquid chromatography (10 – 12 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 16.33 g of potassium dihydrogen-phosphate and 124.15 g of ethylene glycol in water to make 1000 mL. If necessary, adjust to pH 4.0 with phosphoric acid. Flow rate: Adjust so that the retention time of bovine se-rum albumin is about 36 minutes. Selection of column: Proceed with 50 mL of the molecular mass reference solution according to the above operating conditions, and calculate the resolution. Use a column from which g-globulin, bovine serum albumin and myoglobin are eluted in this order with the resolution between their peaks being not less than 1.5, respectively. Antigenicity Dilute a suitable volume of Ulinastatin with isotonic sodium chloride solution so that each mL of the so-lution contains 15,000 Units, and use this solution as the sample solution. Inject 0.10 mL of the sample solution on 3 occasions at intervals of 2 days intraperitoneally to each of 4 well-nourished, healthy guinea pigs weighing 250 to 300 g. Inject 0.10 mL of horse serum intraperitoneally to each of 4 guinea pigs of another group as a control. Inject 0.20 mL of the sample solution intravenously into each of 2 guinea pigs of the first group 14 days after the first intraperitoneal injec-tion and into each of the remaining 2 guinea pigs 21 days after the injection, and inject 0.20 mL of horse serum intra-venously in the same manner into each guinea pig of the second group. Observe the signs of respiratory distress, col-lapse or death of the animals for 30 minutes after each in-travenous injection and 24 hours later: the animals of the first group exhibit none of the signs mentioned above, and all the animals of the second group exhibit symptoms of respiratory distress or collapse and not less than 3 animals are killed. Toxicity Inject intravenously 0.50 mL of Ulinastatin into each of five well-fed, healthy albino mice weighing 18 to 25 g: no mouse dies within 48 hours after injection. If any mouse dies within 48 hours, repeat the test using 5 albino mice weighing 19 to 21 g: all the animals survive for 48 hours.
Measure exactly a suitable volume of Ulinastatin, dilute with 2,2?,2!-nitrilotriethanol buffer solution (pH 7.8) so that each mL of the solution contains about 150 Units, and use this solution as the sample solution. Separately, di-lute a suitable volume of Ulinastatin RS with 2,2?,2!-nitrilotriethanol buffer solution (pH 7.8) so that each mL of the solution contains exactly 300, 200, 100, 50 or 0 Units, and use these solutions as the standard solutions. 2,2?,2!-Nitrilotriethanol buffer solution (pH 7.8) and N-a-benzoyl-L-arginine-4-nitroanilide TS are warmed in a water bath of 25 ± 19C for use as described below. Take exactly 0.1 mL each of the sample solution and the standard solutions in test tubes, add exactly 1.6 mL of 2,2?,2!-nitrilotriethanol buffer solution (pH 7.8) mix, and put the tubes in the water bath of 25 ± 19C. One minute after addition of the buffer solution add exactly 0.2 mL of ice-cooled trypsin TS for test of ulinastatin, mix, and put the tubes again in the water bath. One minute later add exactly 1 mL of N-a-benzoyl-L-arginine-4-nitroanilide TS, mix, and then put the tubes in the water bath. Exactly 2 minutes later add exactly 0.1 mL of di-luted acetic acid (100) (1 in 2) to stop the enzyme reaction, and determine the absorbances of the solutions so obtained at 405 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24> using water as the blank. Prepare a calibra-tion curve using the absorbances obtained with the standard solutions, and calculate ulinastatin Units in the sample solu-tion from its absorbance by using this curve.
Containers—Tight containers. Storage—Not exceeding at －209C.
Uncaria Hook is uncinate hook or short stem with opposite or single hook; the hook, 1 – 4 cm in length, curved and acuminate; externally red-brown to dark brown or grayish brown, some one with hairs, the transverse section oblong to elliptical, light brown; stem thin and prismatic square to cylindrical, 2 – 5 mm in diameter, externally, red-brown to dark brown or grayish brown; the transverse section, square to elliptical; the pith light brown, square to elliptical; hard in texture. Odorless and practically tasteless. Under a microscope <5.01>, a transverse section of the hook reveals vascular bundles in the cortex, unevenly dis-tributed and arranged in a ring. Parenchyma cells in the sec-ondary cortex containing sand crystals of calcium oxalate.
To 1.0 g of pulverized Uncaria Hook add 10 mL of water and 1 mL of ammonia TS, shake, then add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the diethyl ether layer as the sample solution. Separately, dissolve 1 mg each of rhyncophylline for thin-layer chroma-tography and hirsutine for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (7:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: one of the several spots obtained from the sample so-lution has the same color tone and Rf value with one of the spots from the standard solution. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.5z.
Weigh accurately about 0.2 g of moderately fine powder of Uncaria Hook, transfer into a glass-stoppered centrifuge tube, add 30 mL of a mixture of methanol and dilute acetic acid (7:3), shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add two 10-mL portions of a mixture of methanol and dilute acetic acid (7:3), proceed in the same manner, and combine all of the supernatant liquid. To the combined liquid add a mix-ture of methanol and dilute acetic acid (7:3) to make exactly 50 mL, and use this as the sample solution. Separately, weigh accurately about 5 mg of rhynchophylline for assay, previously dried in a desiccator (silica gel) for 24 hours, and dissolve in a mixture of methanol and dilute acetic acid (7:3) to make exactly 100 mL. Pipet 1 mL of this solution, add a mixture of methanol and dilute acetic acid (7:3) to make ex-actly 10 mL, and use this solution as the standard solution (1). Separately, dissolve 1 mg of hirsutine in 100 mL of a mixture of methanol and dilute acetic acid (7:3), and use this solution as the standard solution (2). Perform the test with exactly 20 mL each of the sample solution and standard solu-tions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATa and ATb, of rhynchophylline and hirsutine obtained from the sample solution, and the peak area, AS, of rhynchophylline from the standard solution (1). Amount (mg) of total alkaloids (rhynchophylline and hirsutine) ＝ MS × (ATa ＋ 1.405ATb)/AS × 1/20 MS: Amount (mg) of rhynchophylline for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). ‌JP XVIII Crude Drugs and Related Drugs / Unseiin Extract 2169 Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 200 mL of water, add 10 mL of acetic acid (100) and water to make 1000 mL, and add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of rhyn-chophylline is about 17 minutes. System suitability— System performance: Dissolve 5 mg of rhynchophylline for assay in 100 mL of a mixture of methanol and dilute acetic acid (7:3). To 5 mL of this solution add 1 mL of am-monia solution (28), and reflux for 10 minutes or warm at about 509C for 2 hours. After cooling, to 1 mL of the solu-tion so obtained add a mixture of methanol and dilute acetic acid (7:3) to make 5 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the peak of isorhynchophylline is appears in addition to the peak of rhynchophylline, and the resolution between these peaks is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution (1) under the above op-erating conditions, the relative standard deviation of the peak areas of rhynchophylline is not more than 1.5z.
Unseiin Extract contains not less than 24 mg and not more than 72 mg (for preparation prescribed 3 g of Peony Root) or not less than 32 mg and not more than 96 mg (for preparation prescribed 4 g of Peony lowed by a pungent taste.
Unseiin Extract occurs as a yellow-brown to very dark brown powder or black-brown viscous extract. It has a slight odor, and has a slightly sweet taste at first, fol- ple solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown to grayish brown spot from the standard solution (Scutellaria Root). To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, and heat the plate at 1059C for 1 minute: one of the several spots ob- Unseiin Extract / Crude Drugs and Related Drugs tained from the sample solution has the same color tone and Rf value with the purple to dark purple spot from the stand-ard solution (Gardenia Fruit). To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of coptisine chloride for thin-layer chromatogra-phy in 5 mL of methanol, and use this solution as the stand- Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C H O ) ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 2 mL ＝ MS × AT/AS × 5/8 23 28 11 each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28) and methanol (15:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Cop-tis Rhizome). To 1.0 g of dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of limonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sul-furic acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the purple to dark purple spot from the standard solution (Phellodendron Bark).
(1) To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 15 mL of water and 5 mL of 0.1 mol/L hydrochloric acid TS, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of diethyl ether to the residue, and use this solution as the sample solution. Separately, use (Z )-ligustilide TS for thin-layer chromatography as the standard solution. Per-form the test with these solutions as directed under Thin layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of butyl acetate and hexane (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu-tion (Japanese Angelica Root and Cnidium Rhizome). To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 1 minute: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red-purple to pur-ple spot from the standard solution (Peony Root). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate Root) of paeoniflorin (C23H 28O11 : 480.46), not less the solvent under low pressure (in vacuo), then add 2 mL of diethyl ether to the residue, and use this solution as the sam- than 39 mg and not more than 117 mg (for preparation prescribed 1.5 g of Scutellaria Root), or not less than 78 mg and not more than 234 mg (for preparation prescribed 3 g of Scutellaria Root) of baicalin (C21H18O11: 446.36), and not less than 20 mg and not more than 60 mg of berberine [as berberine chloride (C20H18ClNO4: 371.81)], per extract prepared with the amount specified in the Method of preparation. Method of preparation 1) 2) 3) Japanese Angelica Root 4 g 4 g 3 g Rehmannia Root 4 g 4 g 3 g Peony Root 3 g 4 g 3 g Cnidium Rhizome 3 g 4 g 3 g Scutellaria Root 3 g 3 g 1.5 g Gardenia Fruit 2 g 2 g 1.5 g Coptis Rhizome 1.5 g 1.5 g 1.5 g Phellodendron Bark 1.5 g 1.5 g 1.5 g Prepare a dry extract or viscous extract as directed under Extracts, according to the prescription 1) to 3), using the crude drugs shown above. Description Unseiin Extract occurs as a yellow-brown to very dark brown powder or black-brown viscous extract. It has a slight odor, and has a slightly sweet taste at first, fol- ple solution. Separately, dissolve 1 mg of wogonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown to grayish brown spot from the standard solution (Scutellaria Root). To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfric acid TS on the plate, and heat the plate at 1059C for 1 minute: one of the several spots ob- Unseiin Extract / Crude Drugs and Related Drugs tained from the sample solution has the same color tone and Rf value with the purple to dark purple spot from the stand-ard solution (Gardenia Fruit). To 0.5 g of the dry extract (or 1.5 g of the viscous ex-tract) add 10 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of coptisine chloride for thin-layer chromatogra-phy in 5 mL of methanol, and use this solution as the stand- Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C H O ) ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 2 mL ＝ MS × AT/AS × 5/8 23 28 11 each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ammonia solution (28) and methanol (15:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow fluorescent spot from the standard solution (Cop-tis Rhizome). To 1.0 g of dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), then add 2 mL of diethyl ether to the residue, and use this solution as the sam-ple solution. Separately, dissolve 1 mg of limonin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, hexane and acetic acid (100) (10:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sul-furic acid-ethanol TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the purple to dark purple spot from the standard solution (Phellodendron Bark).
(1) Paeoniflorin—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add exactly 50 mL of diluted methanol (1 in 2), shake for 15 minutes, and filter. Pipet 5 mL of the filtrate, flow through in a column packed with 2 g of polyamide for column chromatography, elute with 20 mL of water, then add 1 mL of acetic acid (100) to the eluate, add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dis-solve in diluted methanol (1 in 2) to make exactly 100 mL. MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: 1.0 mL per minute. System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z. Baicalin—Weigh accurately about 0.1 g of the dry ex-tract (or an amount of the viscous extract, equivalent to about 0.1 g of the dried substance), add exactly 50 mL of diluted methanol (7 in 10), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of baicalin in each solu-tion. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 200) and acetonitrile (19:6). Flow rate: 1.0 mL per minute. System suitability— ‌JP XVIII Crude Drugs and Related Drugs / Wood Creosote 2171 System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of baicalin are not less than 5000 and not more than 1.5, respectively. ‌System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z.‌ Berberine—Weigh accurately about 0.2 g of the dry extract (or an amount of the viscous extract, equivalent to about 0.2 g of the dried substance), add exactly 50 mL of the mobile phase, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform Containers to adjust the percentage. It may contain an ap-propriate quantity of Ethanol. Description Uva Ursi Fluidextract is a yellow-brown to dark red-brown liquid, and has a bitter and astringent taste. It is miscible with water and with ethanol (95). Identification To 1 mL of Uva Ursi Fluidextract add 30 mL of a mixture of ethanol (95) and water (7:3), shake, filter, and use the filtrate as the sample solution. Proceed as directed in the Identification (2) under Bearberry Leaf. Purity Heavy metals <1.07>—Prepare the test solution with 1.0 g of Uva Ursi Fluidextract as direct under the Fluidex-tracts (4), and perform the test (not more than 30 ppm). Assay Pipet 1 mL of Uva Ursi Fluidextract, add water to make exactly 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay under Bearberry Leaf. the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography Amount (mg) of arbutin ＝ MS × AT/AS <2.01> according to the following conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine chloride (C20H18ClNO4) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Berberine Chloride RS taken, calcu- MS: Amount (mg) of arbutin for assay taken
Urapidil, when dried, contains not less than 98.
Urapidil occurs as white to pale yellow-white, crystals or crystalline powder. It has a bitter taste. It is freely soluble in acetic acid (100), sparingly soluble in ethanol (95) and in acetone, and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Urapidil in ethanol (95) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Ura-pidil as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 3.0 g of Urapidil in 40 mL of acetone and 6 mL of dilute nitric acid, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows. To 0.25 mL of 0.01 mol/L hydrochloric acid VS add 40 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.003z). Heavy metals <1.07>—Proceed with 1.0 g of Urapidil according to Method 4, and perform the test. Prepare the ‌control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).‌ Related substances—Dissolve 40 mg of Urapidil in 5 mL of ethanol (95), and use this solution as the sample solu-tion. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy, develop with a mixture of ethyl acetate, ethanol (95) and ammonia water (28) (22:13:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution appears not more than one and it is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 70 mg of Urapidil, previ-ously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 12.92 mg of C20H29N5O3
Urea [57-13-6] Urea contains not less than 99.
Urea occurs as colorless to white, crystals or crystalline powder. It is odorless, and has a cooling, saline taste. It is very soluble in water, freely soluble in boiling ethanol (95), soluble in ethanol (95), and very slightly soluble in diethyl ether. A solution of Urea (1 in 100) is neutral.
(1) Heat 0.5 g of Urea: it liquefies and the odor of ammonia is perceptible. Continue heating until the liquid becomes turbid, then cool. Dissolve the resulting lump in a mixture of 10 mL of water and 2 mL of sodium hydrox-ide TS, and add 1 drop of copper (II) sulfate TS: a reddish purple color develops. (2) Dissolve 0.1 g of Urea in 1 mL of water, and add 1 mL of nitric acid: a white, crystalline precipitate is formed. Melting point <2.60> 132.5 – 134.59C
(1) Chloride <1.03>—Perform the test with 2.0 g of Urea. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.007z). Sulfate <1.14>—Perform the test with 2.0 g of Urea. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Heavy metals <1.07>—Proceed with 1.0 g of Urea ac- cording to Method 1, and perform the test. Prepare the con-trol solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Ethanol-insoluble substances—Dissolve 5.0 g of Urea in 50 mL of warm ethanol (95), filter through a tared glass filter (G4), wash the residue with 20 mL of warm ethanol (95), and dry at 1059C for 1 hour: the mass of the residue is not more than 2.0 mg. Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Urea, and dissolve in water to make exactly 200 mL. Measure exactly 5 mL of this solution into a Kjeldahl flask, and proceed as directed under Nitrogen Determination <1.08>. Each mL of 0.005 mol/L sulfuric acid VS ＝ 0.3003 mg of CH4N2O
Urokinase is an enzyme, obtained from human urine, that activates plasminogen, and has the molecu-lar mass of about 54,000. It is a solution using a suitable buffer solution as the solvent. It contains not less than 60,000 Units per mL, and not less than 120,000 Units per mg of protein.
Urokinase is a clear and colorless liquid. The pH is between 5.5 and 7.5.
(1) Dissolve 0.07 g of fibrinogen in 10 mL of phosphate buffer solution (pH 7.4). To this solution add 1 mL of a solution of thrombin containing 10 Units per mL in isotonic sodium chloride solution, mix, place in a Petri dish about 90 mm in inside diameter, and keep horizontally until the solution is coagulated. On the surface drop 10 mL of a solution of Urokinase containing 100 Units per mL in gelatin-tris buffer solution, and stand for overnight: lysis circle appears. (2) Dissolve 1.0 g of Powdered Agar in 100 mL of boric acid-sodium hydroxide buffer solution (pH 8.4) by warming, and pour the solution into a Petri dish until the height come to about 2 mm. After cooling, make two wells of 2.5 mm in diameter with the space of 6 mm. To each well place sepa-rately 10 mL of a solution of Urokinase containing 30,000 Units per mL in isotonic sodium chloride solution and 10 mL of anti-urokinase serum, and stand for overnight: a clear precipitin line appears.
(1) Heavy metals <1.07>—Proceed with 2.0 mL of Urokinase according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). (2) Blood group substances—Dilute Urokinase with iso-tonic sodium chloride solution so that each mL of the solu-tion contains 12,000 Units, and use this solution as the sam- ‌ple solution. To anti-A type antibody for blood typing add isotonic sodium chloride solution to dilute each 32, 64, 128, 256, 512 and 1024 times, place separately 25 mL each of these solutions in six wells on the first and second lane of a V-shaped 96-wells microplate. Next, add 25 mL of the sample solution into the six wells on the first lane and 25 mL of iso-tonic sodium chloride solution into the six wells of the second lane, mix, and allow to stand for 30 minutes. To each well add 50 mL of A-type erythrocyte suspension, mix, allow to stand for 2 hours, and compare the agglutination of erythrocyte in both lanes: dilution factor of anti-A type antibody of the wells which show the agglutination is equal in both lanes. Perform the same test by using anti-B type antibody for blood typing and B-type erythrocyte suspension. Abnormal toxicity Dilute Urokinase with isotonic sodium chloride solution so that each mL of the solution contains 12,000 Units, and use this solution as the sample solution. Inject 5.0 mL of the sample solution into the peritoneal cavity of each of 2 or more of well-nourished, healthy guinea pigs weighing about 350 g. Observe the conditions of the animals for more than 7 days: all the animals exhibit no abnormalities. High-molecular mass urokinase Dilute Urokinase with gelatin-phosphate buffer solution so that each mL of the solution contains 10,000 Units, and use this solution as the sample solution. Perform the test with 100 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas of two peaks eluted closely at about 35 minutes having smaller retention time, Aa, and larger retention time, Ab, by the automatic integration method: the value, Aa/(Aa ＋ Ab), is not less than 0.85. Operating conditions— Apparatus: Use a pumping system for the mobile phase, a sample injection port, a column, a pumping system for the reaction reagent, a reaction coil, a reaction chamber, a spec-trofluorometer and a recorder. Attach a 3-way tube to the outlet for the mobile phase of the column, connect the pumping system for the reaction reagent and the reaction coil, and join outlet of the reaction coil to the spec-trofluorometer. Detector: Spectrofluorometer (excitation wavelength: 365 nm, fluorescence wavelength: 460 nm). Column: A stainless steel column 7.5 mm in inside diame-ter and 60 cm in length, packed with porous silica gel for liq-uid chromatography (10 to 12 mm in particle diameter). Column temperature: A constant temperature of about 209C. Reaction coil: A stainless steel column 0.25 mm in inside diameter and 150 cm in length. Reaction coil temperature: 379C. Mobile phase: Gelatin-phosphate buffer solution. Flow rate of mobile phase: 0.5 mL per minute. Reaction reagent: 7-(Glutarylglycyl-L-arginylamino)-4-methylcoumarin TS. Flow rate of reaction reagent: 0.75 mL per minute. Selection of column: Adjust the pH of Urokinase to 7.5 with sodium hydroxide TS, allow to stand at 379C for over 24 hours, and add gelatin-phosphate buffer solution to make the solution containing 20,000 Units per mL. Proceed with 100 mL of this solution under the above operating condi-tions, and calculate the resolution. Use a column giving elu-tion of high molecular mass urokinase (molecular mass: 54,000) and low molecular mass urokinase (molecular mass: 33,000) in this order with the resolution between these peaks being not less than 1.0.
(1) Urokinase—Pipet 1 mL of Urokinase, dilute exactly with gelatin-tris buffer solution so that each mL of the solution contains about 30 Units, and use this solution as the sample solution. Add exactly 2 mL of gelatin-tris buffer solution to contents of one ampoule of High-Molecular Mass Urokinase RS to dissolve, pipet 1 mL of this solution, dilute exactly with gelatin-tris buffer solution so that each mL of the solution contains about 30 Units, and use this solution as the standard solution. Place 1.0 mL of L-pyro-glutamylglycyl-L-arginine-p-nitroaniline hydrochloride TS in two silicon-coated test tubes about 10 mm in inside diameter, warm them in a water bath at 35 ± 0.29C for 5 minutes, add separately 0.50 mL each of the sample solution and standard solution, warm in a water bath at 35 ± 0.29C for exactly 30 minutes, then add 0.50 mL of diluted acetic acid (100) (2 in 5). Determine the absorbances, AT and AS, of these solutions at 405 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank. Separately place 1.0 mL of L-pyroglutamylglycyl-L-arginine-p-nitroaniline hydrochloride TS in two test tubes, add 0.50 mL of diluted acetic acid (100) (2 in 5), and 0.50 mL each of the sample solution and standard solution. Determine the absorbances, AT0 and AS0, of these solutions at 405 nm as the same man-ner, using water as the blank. Amount (Units) of Urokinase ＝ (AT － AT0)/(AS － AS0) × a × b a: Amount (Units) of urokinase in 1 mL of the standard solution b: Total volume (mL) of the sample solution (2) Protein—Measure exactly a volume of Urokinase, equivalent to about 15 mg of protein, and perform the test as directed under Nitrogen Determination <1.08>. Each mL of 0.005 mol/L sulfuric acid VS ＝ 0.8754 mg of protein
Ursodeoxycholic Acid, when dried, contains not less than 98.
Ursodeoxycholic Acid occurs as a white, crys-tal or powder, with bitter taste. It is freely soluble in methanol, in ethanol (99.5) and in acetic acid (100), and practically insoluble in water.
Determine the infrared absorption spectrum of Ursodeoxycholic Acid as directed in the potassium bro- ‌mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋59.0 – ＋62.09(after dry-ing, 1 g, ethanol (99.5), 25 mL, 100 mm).
(1) Sulfate <1.14>—Dissolve 2.0 g of Ursodeoxy-cholic Acid in 20 mL of acetic acid (100), add water to make 200 mL, and allow to stand for 10 minutes. Filter this solu-tion, discard the first 10 mL of the filtrate, and use the sub-sequent filtrate as the sample solution. To 40 mL of the sam- which is calculated by the comparison with the spots from the standard solutions (A) and (B), is not more than 0.25z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.5 g of Ursodeoxycholic Acid, previously dried, dissolve in 40 mL of ethanol (95) and 20 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank de-termination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ple solution add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the ＝ 39.26 mg of C24 H40O4 test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS by adding 4 mL of acetic acid (100), 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 1.0 g of Ursodeoxycholic Acid according to Method 2 and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Barium—To 2.0 g of Ursodeoxycholic Acid add 100 mL of water and 2 mL of hydrochloric acid, boil for 2
Uva Ursi Fluidextract contains not less than 3.
Valaciclovir Hydrochloride contains not less than 95.
Valaciclovir Hydrochloride occurs as a white to pale yellow-white crystalline powder. It is freely soluble in water, and very slightly soluble in ethanol (99.5). It dissolves in 0.05 mol/L hydrochloric acid TS. D Optical rotation [a]20: － 7.1 – － 11.19(1 g, water, 20 the sample solution. Separately, weigh accurately about 0.1 g of ursodeoxycholic acid for assay, previously dried at 1059C for 2 hours, dissolve in exactly 20 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ursodeoxycholic acid to that of the internal standard. Amount (mg) of ursodeoxycholic acid (C24H40O4) ＝ MS × QT/QS MS: Amount (mg) of ursodeoxycholic acid for assay taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (4 in 5) (7 in 200,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica mL, 100 mm). It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Valaciclovir Hydrochloride in 0.05 mol/L hydro-chloric acid TS (3 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Valaciclovir Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Valaciclovir Hydrochloride as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Valaciclovir Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, suspend Valaciclovir Hydrochloride in a mixture of ethanol (99.5) and water (45:2), and heat under reflux for 24 hours while stirring. After cooling to room temperature, col- lect the obtained solid by filtration, dry at 609C for 1 hour under reduced pressure, and perform the same test with the solid. A solution of Valaciclovir Hydrochloride (1 in 25) responds to Qualitative Tests <1.09> for chloride.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Valaciclovir Hydrochloride according to Method 4, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 20 ppm). Palladium—Dissolve exactly 0.100 g of Valaciclovir Hydrochloride in a solution of hydrochloric acid in dimethylsulfoxide (1 in 50) to make exactly 10 mL, and use this solution as the sample solution. Separately, to exactly 6 mL of Standard Palladium Solution for ICP Analysis add a solution of hydrochloric acid in dimethylsulfoxide (1 in 50) to make exactly 100 mL. Pipet 5 mL of this solution, add a solution of hydrochloric acid in dimethylsulfoxide (1 in 50) to make exactly 50 mL. Pipet 5 mL of this solution, add a solution of hydrochloric acid in dimethylsulfoxide (1 in 50) to make exactly 50 mL. Pipet 5 mL of this solution, add a solution of hydrochloric acid in dimethylsulfoxide (1 in 50) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Inductively Coupled Plasma-Atomic Emission Spectrometry <2.63> according to and calculate their amount by the area percentage method: the amount of the peaks, having a relative retention time of about 0.54, about 1.06, about 1.17, about 1.61, about 1.66 and about 1.98 to valaciclovir, is not more than 0.1z, 0.2z, 0.5z, 0.8z, 0.2z and 0.3z, respectively, and the amount of the peaks other than valaciclovir, the peaks mentioned above, guanine (relative retention time is about 0.31), aciclovir (relative retention time is about 0.42) and the peak (relative retention time is about 1.09) is not more than 0.05z, and their total amount is not more than 0.2z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 159C. Mobile phase A: Dissolve 3 g of trifluoroacetic acid in water to make 1000 mL. Mobile phase B: Dissolve 3 g of trifluoroacetic acid in methanol to make 1000 mL. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. the following conditions: the emission intensity obtained from the sample solution is not more than that from the standard solution (not more than 6 ppm). Operating conditions—Wavelength: 340.458 nm. Related substances—(i) To 0.25 g of Valaciclovir Time after injection 0 – 5 5 – 35 90 → 60 10 → 40 of sample (min) Mobile phase A (volz) Mobile phase B (volz) Hydrochloride add 2 mL of water, and sonicate for 20 minutes. After cooling, add methanol to make exactly 10 mL, if necessary, filter, through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard stock solution. Pipet 1 mL and 0.5 mL of the standard stock solution, add methanol to make them exactly 10 mL, and use these solutions as the standard solution (1) and the standard solution (2), respectively. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 4 mL each of the sample solution and the standard solu-tions (1) and (2) on a plate of silica gel with fluorescent indi-cator for thin-layer chromatography, develop the plate with a mixture of chloroform, methanol, tetrahydrofuran, dichloromethane and ammonia solution (28) (46:34:12:8:3) to a distance of about 8 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot, having an Rf value of about 0.47, obtained from the sample solution is not more intense than the spot from the standard solution (1), and the spot, having an Rf value of about 0.67, from the sample solution is not more intense than the spot from the standard solution (2). Furthermore, when spray evenly a solution of fluorescamine in acetone (1 in 10,000) on the plate, and examine under ultraviolet light (main wave-length: 366 nm): the spot, having an Rf value of about 0.63, from the sample solution is not more intense than the spot from the standard solution (1). Dissolve 40 mg of Valaciclovir Hydrochloride in 100 mL of a mixture of water and ethanol (95) (4:1), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, deter-mine each peak area by the automatic integration method, Flow rate: 0.8 mL per minute. Time span of measurement: 35 minutes, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water and ethanol (95) (4:1) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add a mixture of water and ethanol (95) (4:1) to make exactly 20 mL. Confirm that the peak area of valaciclovir obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of valaciclovir are not less than 25,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of valaciclovir is not more than 2.0z. Perform the test with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peaks, having a relative retention time of about 0.14 and about 0.42 to valaciclovir, is not more than 2.0z and not more than 0.2z, respectively. For the amounts of the peaks, having a relative retention time of about 0.14 and about 0.42 to valaciclovir, multiply their cor-rection factors, 0.66 and 0.89, respectively. Operating conditions— Proceed as directed in the operating conditions in the ‌Assay. System suitability— Test for required detectability: Pipet 1 mL of the sample solution add 0.05 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the solution for system suitability test. To exactly 5 mL of the solution for system suitability test add 0.05 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of valaciclovir obtained with 10 mL of this solution is equivalent to 0.07 to 0.13z of that with 10 mL of the sample solution. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of valaciclovir are not less than 700 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of valaciclovir is not more than 2.0z. The total amount of the related substances obtained in (i), (ii) and (iii) is not more than 2.0z. Enantiomer—When perform the test according to (3) (iii), the amount of the peak of the enantiomer, having the relative retention time of about 0.57 to valaciclovir, is not more than 3.0z. Water <2.48> Not more than 1.7z (0.2 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (2 g).
Weigh accurately about 25 mg each of Valaciclovir Hydrochloride and Valaciclovir Hydrochloride RS (separate-ly determine the water <2.48> and the residual solvent in the same manners as Valaciclovir Hydrochloride), dissolve them separately in 0.05 mol/L hydrochloric acid TS to make ex-actly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of valaciclovir in each solution. Amount (mg) of valaciclovir hydrochloride (C13H20N6O4.HCl) ＝ MS × AT/AS MS: Amount (mg) of Valaciclovir Hydrochloride RS taken, calculated on the anhydrous and residual sol-vent-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with 18-crown ether-immobi-lized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 109C. Mobile phase: To 950 mL of water add 5 mL of perchloric acid and 30 mL of methanol. Flow rate: Adjust so that the retention time of valaciclovir is about 21 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of valaciclovir are not less than 700 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valaciclovir is not more than 1.0z.
Valsartan occurs as a white powder. It is very soluble in methanol and in ethanol (99.5), and practically insoluble in water.
(1) Determine the absorption spectrum of a solution of Valsartan in methanol (1 in 62,500) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Valsartan RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Val-sartan as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Valsartan RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. D Optical rotation <2.49> [a]20: －64 – －699(0.5 g calculated on the anhydrous and residual solvent-free basis, methanol, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Valsartan according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 50 mg of Valsartan in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having the relative retention time of about 0.8 to valsartan, obtained from the sample solution is not larger than 1/5 times the peak area of valsartan from the standard solution, the area of the peak other than valsartan and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of valsartan from the standard solution, and the total area of the peaks other than valsartan from the sample solution is not larger than 3/10 times the peak area of valsartan from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 6 times as long as the retention time of valsartan, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of valsartan obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of valsartan are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valsartan is not more than 2.0z. Enantiomer—Dissolve 75 mg of Valsartan in 100 mL of the mobile phase. To 5 mL of this solution add the mobile phase to make 25 mL, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the peak area of the enan-tiomer, having the relative retention time of about 0.6 to val-sartan, obtained from the sample solution is not larger than the peak area of valsartan from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 227 nm). Column: A stainless steel column 4 mm in inside diameter and 10 cm in length, packed with a1-acid glycoprotein bind-ing silica gel for liquid chromatography (5 mm in particle di-ameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 14.68 g of disodium hydrogen phosphate dodecahydrate and 3.81 g of potassium dihydro-gen phosphate in 1000 mL of water. To 490 mL of this solu-tion add 10 mL of 2-propanol. Flow rate: Adjust so that the retention time of valsartan is about 10 minutes. ‌System suitability— System performance: Dissolve about 75 mg of Valsartan, previously allowed to stand at 1059C for 30 minutes, in the mobile phase to make 100 mL. To 5 mL of this solution add the mobile phase to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, the enantiomer and valsartan are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valsartan is not more than 5z. Water <2.48> Not more than 2.0z (0.1 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Valsartan and Valsartan RS (separately determine the water <2.48> and the residual solvent in the same manner as Valsartan), and dis-solve them separately in the mobile phase to make exactly 100 mL. Pipet 5 mL each of these solutions, add exactly 3 mL of the internal standard solution, and add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of valsartan to that of the inter-nal standard. Amount (mg) of valsartan (C24H29N5O3) ＝ MS × QT/QS MS: Amount (mg) of Valsartan RS taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of diclofenac sodium in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 3 mm in inside diameter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile, and acetic acid (100) (500:500:1). Flow rate: Adjust so that the retention time of valsartan is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, valsartan and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valsartan to that of the internal standard is not more than 1.0z.
Vancomycin Hydrochloride is the hydrochloride of a glycopeptide substance having antibacterial activity produced by the growth of Streptomyces orientalis. It contains not less than 1000 mg (potency) and not more than 1200 mg (potency) per mg, calculated on the anhydrous basis.
Vancomycin Hydrochloride occurs as a white powder. It is freely soluble in water, soluble in formamide, slightly soluble in methanol, very slightly soluble in ethanol (95), and practically insoluble in acetonitrile. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Vancomycin Hydrochloride (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Vancomycin Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Van-comycin Hydrochloride as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Vancomycin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 20 mg of Vancomycin Hydrochloride in 10 mL of water, and add 1 drop of silver nitrate TS: a white turbidity is produced. D Optical rotation <2.49> [a]20: －30 – －409(0.2 g calculated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.25 g of Vancomycin Hydrochloride in 5 mL of water is between 2.5 and 4.5.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Vancomycin Hydrochloride according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Vancomycin Hydrochloride in 10 mL of the mobile phase A, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase A to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under the Liquid Chromatography <2.01> according to the following conditions. If necessary, proceed with 20 mL of the mobile phase A in the same man-ner to compensate for the base line. Determine each peak area by the automatic integration method: the area of each peak other than vancomycin obtained from the sample solu-tion is not larger than the peak area of vancomycin from the standard solution, and the total area of the peaks other than vancomycin from the sample solution is not larger than 3 times of the peak area of vancomycin from the standard so-lution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of triethylamine buffer solu- tion (pH 3.2), acetonitrile and tetrahydrofuran (92:7:1). Adjust the amount of acetonitrile so that the retention time of vancomycin is 7.5 to 10.5 minutes. Mobile phase B: A mixture of triethylamine buffer solu-tion (pH 3.2), acetonitrile and tetrahydrofuran (70:29:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 12 12 – 20 100 → 0 0 → 100 20 – 22 Flow rate: 1.5 mL per minute. Time span of measurement: As long as about 2.5 times of the retention time of vancomycin, beginning after the solvent peak. System suitability— Test for required detectability: Confirm that the peak area of vancomycin obtained with 20 mL of the standard solution is equivalent to 3 to 5z of that with 20 mL of the sample so-lution. System performance: Dissolve 5 mg of Vancomycin Hy-drochloride in 10 mL of water, heat at 659C for 48 hours, and cool to the ordinal temperature. When the procedure is run with 20 mL of this solution under the above operating conditions, related substance 1, vancomycin and related sub-stance 2 are eluted in this order, the resolution between the peaks of the related substance 1 and vancomycin is not less than 3, the number of theoretical plates of the peak of van-comycin is not less than 1500, and the related substance 2 is eluted between 15 minutes and 18 minutes. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vancomycin is not more than 2.0z. Water <2.48> Not more than 5.0z (0.1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (3:1)). Residue on ignition <2.44> Not more than 1.0z (1 g).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 6.2 to 6.4 after sterilization. Standard solutions—Weigh accurately an amount of Vancomycin Hydrochloride RS, equivalent to about 25 mg (potency), dissolve in water to make exactly 25 mL, and use this solution as the standard stock solution. Keep the stand-ard stock solution at 59C or below, and use within 7 days. Take exactly a suitable amount of the standard stock solu-tion before use, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make solutions so that each mL contains 100 mg (potency) and 25 mg (potency), and use these solutions as the high concentration standard solution and the low concentra-tion standard solution, respectively. Sample solutions—Weigh accurately an amount of Vancomycin Hydrochloride, equivalent to about 25 mg (po-tency), and dissolve in water to make exactly 25 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make solutions so that ‌each mL contains 100 mg (potency) and 25 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.‌
Verapamil Hydrochloride, when dried, contains not less than 98.
Verapamil Hydrochloride occurs as a white crystalline powder. It is freely soluble in methanol and in acetic acid (100), soluble in ethanol (95) and in acetic anhydride, and sparingly soluble in water.
(1) To 2 mL of a solution of Verapamil Hydrochloride (1 in 50) add 5 drops of Reinecke salt TS: a light red precipitate is produced. Determine the absorption spectrum of a solution of Verapamil Hydrochloride in 0.01 mol/L hydrochloric acid TS (1 in 50,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ver-apamil Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Verapamil Hydrochloride (1 in 50) ‌responds to Qualitative Tests <1.09> for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Verapamil Hydrochloride in 20 mL of water by warming: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Ver- apamil Hydrochloride according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Verapamil Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Verapamil Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Separately, pipet 5 mL of the standard stock solution, add methanol to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on two plates of silica gel for thin-layer chromatography. With the one plate, develop the plate with a mixture of cyclo-hexane and diethylamine (17:3) to a distance of about 15 cm, air-dry the plate, heat at 1109C for 1 hour, and cool. Exa-mine immediately after spraying evenly iron (III) chloride-iodine TS on the plate: the spots other than the principal spot and the spot on the original point obtained from the sample solution, are not more intense than the spot from the standard solution (2), and the number of them which are more intense than the spot from the standard solution (1) is not more than 3. With another plate, develop the plate with a mixture of toluene, methanol, acetone and acetic acid (100) (14:4:1:1), and perform the test in the same manner. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.7 g of Verapamil Hydro-chloride, previously dried, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 49.11 mg of C27H38N2O4.HCl
Vinblastine Sulfate contains not less than 96.
Vinblastine Sulfate occurs as a white to pale yellow powder. It is soluble in water, sparingly soluble in methanol, and ‌practically insoluble in ethanol (99.5). It is hygroscopic. D Optical rotation [a]20: －28 – －359(20 mg calculated on the dried basis, methanol, 10 mL, 100 mm).
(1) Determine the absorption spectrum of a solution of Vinblastine Sulfate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Vinblastine Sulfate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Vin- Atmospheric gas: dried Nitrogen. Flow rate of atmospheric gas: 40 mL per minute.
(1) Clarity and color of solution—Dissolve 50 mg of Vinblastine Sulfate in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve about 4 mg of Vin-blastine Sulfate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area of these so-lutions by the automatic integration method: the area of the peak other than vinblastine obtained from sample solution is not larger than 1/4 times the peak area of vinblastine from the standard solution, and the total area of the peaks other than vinblastine from the sample solution is not larger than 3/4 times the peak area of vinblastine from the standard so-lution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of vinblastine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2.5 mL of the standard solution add water to make exactly 100 mL. Con-firm that the peak area of vinblastine obtained with 200 mL of this solution is equivalent to 1.7 to 3.3z of that with 200 mL of the standard solution. System repeatability: When the test is repeated 6 times with 200 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vinblastine is not more than 1.5z. Loss on drying Perform the test with about 10 mg of Vin-blastine Sulfate as directed in Thermogravimetry under Thermal Analysis <2.52> according to the following condi-tions: not more than 15.0z. Operating conditions— Heating rate: 59C per minute. Temperature range: room temperature to 2009C. MS: Amount (mg) of Vinblastine Sulfate RS taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 262 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 7 mL of diethylamine add water to make 500 mL, and adjust the pH to 7.5 with phosphoric acid. To 380 mL of this solution add 620 mL of a mixture of methanol and acetonitrile (4:1). Flow rate: Adjust so that the retention time of vinblastine is about 8 minutes. System suitability— System performance: Dissolve 10 mg each of Vinblastine Sulfate and vincristine sulfate in 25 mL of water. When the procedure is run with 20 mL of this solution under the above operating conditions, vincristine and vinblastine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vinblastine is not more than 1.0z.
Weigh accurately about 10 mg each of Vinblastine Sulfate and Vinblastine Sulfate RS (previously determine the loss on drying under the same conditions as Vinblastine Sul-fate), dissolve in water to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of vin-blastine in each solution. blastine Sulfate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec- Amount (mg) of vinblastine sulfate (C ＝ MS × AT/AS 46H58N4O9.H2SO4) trum of Vinblastine Sulfate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Vinblastine Sulfate (1 in 100) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 15 mg of Vinblastine Sulfate in 10 mL of water: the pH of this solution is between 3.5 and 5.0. Purity (1) Clarity and color of solution—Dissolve 50 mg of Vinblastine Sulfate in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve about 4 mg of Vin-blastine Sulfate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area of these so-lutions by the automatic integration method: the area of the peak other than vinblastine obtained from sample solution is not larger than 1/4 times the peak area of vinblastine from the standard solution, and the total area of the peaks other than vinblastine from the sample solution is not larger than 3/4 times the peak area of vinblastine from the standard so-lution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of vinblastine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2.5 mL of the standard solution add water to make exactly 100 mL. Con-firm that the peak area of vinblastine obtained with 200 mL of this solution is equivalent to 1.7 to 3.3z of that with 200 mL of the standard solution. System repeatability: When the test is repeated 6 times with 200 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vinblastine is not more than 1.5z. Loss on drying Perform the test with about 10 mg of Vin-blastine Sulfate as directed in Thermogravimetry under Thermal Analysis <2.52> according to the following condi-tions: not more than 15.0z. Operating conditions— Heating rate: 59C per minute. Temperature range: room temperature to 2009C. MS: Amount (mg) of Vinblastine Sulfate RS taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 262 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 7 mL of diethylamine add water to make 500 mL, and adjust the pH to 7.5 with phosphoric acid. To 380 mL of this solution add 620 mL of a mixture of methanol and acetonitrile (4:1). Flow rate: Adjust so that the retention time of vinblastine is about 8 minutes. System suitability— System performance: Dissolve 10 mg each of Vinblastine Sulfate and vincristine sulfate in 25 mL of water. When the procedure is run with 20 mL of this solution under the above operating conditions, vincristine and vinblastine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vinblastine is not more than 1.0z.
Containers—Tight containers. Storage—Light-resistant, at not exceeding －209C.
Vincristine Sulfate contains not less than 95.
Vincristine Sulfate occurs as a white to light yellow-white powder. It is very soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic. D Optical rotation [a]20: ＋28.5 – ＋35.59(0.2 g calculated on the dried basis, water, 10 mL, 100 mm).
(1) Determine the absorption spectrum of a solution of Vincristine Sulfate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Vincristine Sulfate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Vin-cristine Sulfate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Vincristine Sulfate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Vincristine Sulfate (1 in 100) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 10 mg of Vincristine Sulfate in 10 mL of water: the pH of this solution is between 3.5 and 4.5.
(1) Clarity and color of solution—Dissolve 50 mg of Vincristine Sulfate in 10 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve 10 mg of Vincristine Sulfate in 10 mL of water, and use this solution as the sam-ple solution. Pipet 2 mL of the sample solution, add water to Amount (mg) of vinblastine sulfate (C ＝ MS × AT/AS × 10 46H58N4O9.H2SO4) make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard solution as directed under MS: Amount (mg) of Vinblastine Sulfate RS taken, calcu-lated on the dried basis
Weigh accurately about 10 mg each of Vincristine Sulfate and Vincristine Sulfate RS (separately determine the loss on drying under the same conditions as Vincristine Sul-fate), dissolve each in water to make exactly 10 mL, and use these solutions as the sample solution and the standard solu- procedure is run with 10 mL of this solution under the above operating conditions, vincristine and vinblastine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vincristine is not more than 1.0z.
Voglibose occurs as white, crystals or crystal-line powder. It is very slightly soluble in water, freely soluble in acetic acid (100), slightly soluble in methanol, and very slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS.
(1) Determine the infrared absorption spec-trum of Voglibose as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Determine the 1H spectrum of a solution of Vogli-bose in heavy water for nuclear magnetic resonance spectros-copy (3 in 70) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpro-pionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits 2 doublet signals A at about d 1.5 ppm, 2 doublet signals B at about d 2.1 ppm, a multiplet signal C at about d 2.9 ppm, and a multi-plet signal D between d 3.4 ppm and d 3.9 ppm. The area in-tensity ratio of each signal, A:B:C:D, is about 1:1:1:10. D Optical rotation <2.49> [a]20: ＋45 – ＋489(0.2 g calculated on the anhydrous basis, 0.1 mol/L hydrochloric acid TS, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Voglibose in 10 mL of water: the pH of the solution is between 9.8 and 10.4. side diameter and 2 m in length. Mobile phase: To 1.56 g of sodium dihydrogen phosphate dihydrate add water to make 500 mL. To this solution add a solution, prepared by dissolving 3.58 g of disodium hydro-gen phosphate dodecahydrate in water to make 500 mL, to adjust to pH 6.5. To 370 mL of this solution add 630 mL of acetonitrile. Reaction reagent: Dissolve 6.25 g of taurine and 2.56 g of sodium periodate in water to make 1000 mL. Reaction temperature: A constant temperature of about 1009C. Cooling temperature: A constant temperature of about 159C. Flow rate of mobile phase: Adjust so that the retention time of voglibose is about 20 minutes. Flow rate of reaction reagent: Same as the flow rate of the mobile phase. Time span of measurement: About 2.5 times as long as the retention time of voglibose, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of voglibose obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry ‌factor of the peak of voglibose are not less than 7000 and be-tween 0.8 and 1.2, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voglibose is not more than 3.0z. Water <2.48> Not more than 0.2z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Voglibose according to Method 1, and perform the test. Adjust the pH of the test solution between 3.0 and 3.5 with dilute hydrochloric acid instead of dilute acetic acid. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Voglibose in 50 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the total area of the peaks other than voglibose obtained from sample solution is not larger than 1/5 times the peak area of voglibose from the standard solution. For the area of the peaks, having the relative reten-tion time of about 1.7, about 2.0 and about 2.3 to voglibose, multiply their correction factors, 2, 2 and 2.5, respectively. Operating conditions— Apparatus: Use an apparatus consisting of 2 pumps for the mobile phase and reaction reagent transportation, sam-ple injection port, column, reaction coil, cooling coil, detec-tor and recording device, and the reaction coil and cooling coil maintained at a constant temperature. Detector: Fluorophotometer (excitation wavelength: 350 nm, fluorescence wavelength: 430 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with pentaethylenehexaami-nated polyvinyl alcohol polymer bead for liquid chromatog-raphy (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. more than 101.0z of voglibose (C10 lated on the anhydrous basis. H21 NO7), calcu- Reaction coil: A polytetrafluoroethylene tube 0.5 mm in inside diameter and 20 m in length. Cooling coil: A polytetrafluoroethylene tube 0.3 mm in in- Description Voglibose occurs as white, crystals or crystal-line powder. It is very slightly soluble in water, freely soluble in acetic acid (100), slightly soluble in methanol, and very slightly soluble in ethanol (99.5). It dissolves in 0.1 mol/L hydrochloric acid TS. Identification (1) Determine the infrared absorption spec-trum of Voglibose as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Determine the 1H spectrum of a solution of Vogli-bose in heavy water for nuclear magnetic resonance spectros-copy (3 in 70) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpro-pionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits 2 doublet signals A at about d 1.5 ppm, 2 doublet signals B at about d 2.1 ppm, a multiplet signal C at about d 2.9 ppm, and a multi-plet signal D between d 3.4 ppm and d 3.9 ppm. The area in-tensity ratio of each signal, A:B:C:D, is about 1:1:1:10. D Optical rotation <2.49> [a]20: ＋45 – ＋489(0.2 g calculated on the anhydrous basis, 0.1 mol/L hydrochloric acid TS, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Voglibose in 10 mL of water: the pH of the solution is between 9.8 and 10.4. side diameter and 2 m in length. Mobile phase: To 1.56 g of sodium dihydrogen phosphate dihydrate add water to make 500 mL. To this solution add a solution, prepared by dissolving 3.58 g of disodium hydro-gen phosphate dodecahydrate in water to make 500 mL, to adjust to pH 6.5. To 370 mL of this solution add 630 mL of acetonitrile. Reaction reagent: Dissolve 6.25 g of taurine and 2.56 g of sodium periodate in water to make 1000 mL. Reaction temperature: A constant temperature of about 1009C. Cooling temperature: A constant temperature of about 159C. Flow rate of mobile phase: Adjust so that the retention time of voglibose is about 20 minutes. Flow rate of reaction reagent: Same as the flow rate of the mobile phase. Time span of measurement: About 2.5 times as long as the retention time of voglibose, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of voglibose obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry ‌factor of the peak of voglibose are not less than 7000 and be-tween 0.8 and 1.2, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voglibose is not more than 3.0z. Water <2.48> Not more than 0.2z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Voglibose, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 26.73 mg of C10H21NO7
Voriconazole is a white crystalline powder. It is freely soluble in methanol and in acetonitrile, soluble in ethanol (99.5), and very slightly soluble in water. It dissolves in 1 mol/L hydrochloric acid TS. Optical rotation [a]25 : － 374 – － 4049(50 mg calculated on the anhydrous basis, methanol, 25 mL, 100 mm).
(1) Determine the absorption spectrum of a solution of Voriconazole in methanol (1 in 40,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Voriconazole RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Voriconazole as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Voriconazole RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers.
(1) Heavy metals—Take 2.0 g of Voriconazole in a porcelain crucible, moisten with an appropriate amount of sulfuric acid, cover the crucible loosely, and ignite at a low temperature until charred. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid to the content of the cruci-ble, heat gently the crucible until white fumes are no longer evolved, then ignite at 500 – 6009C. After cooling, add 4 mL of 6 mol/L hydrochloric acid TS, evaporate to dryness on a water bath, moisten the residue with 1 drop of hydrochloric acid, add 10 mL of boiling water, and heat for 2 minutes. After cooling, add appropriate drops of ammonia TS until litmus paper changes to blue, add water to make 15 mL, and adjust the pH between 3.0 and 4.0 with dilute acetic acid. Filter if necessary, wash the crucible and the filter paper with 10 mL of water, put the filtrate and washings to a Nessler tube, add water to make 40 mL, and use this solution as the sample solution. Separately, put 2.0 mL of Standard Lead Solution in another Nessler tube, add water to make 25 mL, adjust the pH between 3.0 and 4.0 with dilute acetic acid or ammonia TS, then add water to make 40 mL, and use this solution as the control solution. To each of the sample solu-tion and control solution add 2 mL of acetate buffer solution (pH 3.5), then add 1.2 mL of thioacetamide-alkaline glycerin TS, and add water to make 50 mL. After allowing to stand for 2 minutes, observe vertically both tubes against a white background: the color obtained with the test solution is not more intense than that with the control solution (not more than 10 ppm). Related substances—Dissolve 50 mg of Voriconazole in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks other than voriconazole obtained from the sample solution is not larger than the peak area of voriconazole from the standard solution. The total area of the peaks other than voriconazole from the sample solution is not larger than 4 times the peak area of voriconazole from the standard solution. For the area of the peak, having a relative retention time of about 0.26, about 0.32, and about 0.61 to voricona-zole, multiply the correction factor, 0.7, 0.7 and 2.1, respec-tively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.7 times as long as the retention time of voriconazole. System suitability— System performance: Suspend 0.1 g of Voriconazole in 10 mL of sodium hydroxide solution (1 in 25), add the mobile phase to make 20 mL, and allow to stand for 30 minutes. To 1 mL of this solution add the mobile phase to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution between the peaks, having the relative retention times of about 0.26 and about 0.32 to voriconazole, is not less than 1.7. System repeatability: When the test is repeated 6 times with 20 mL of the solution prepared by adding the mobile phase to 5 mL of the standard solution to make 10 mL under the above operating conditions, the relative standard devia-tion of the peak area of voriconazole is not more than 10.0z. Enantiomer—Dissolve 25 mg of Voriconazole in 2 mL of acetonitrile, add the mobile phase to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make ex-actly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the area of the peak of the enantiomer, having the relative retention time of about 1.3 to voriconazole, obtained from the sample solution is not larger than 1.2 times the peak area of voriconazole from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 256 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with 2-hydroxypropyl-b-cyclodextrinized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 0.77 g of ammonium acetate in 1000 mL of water, and adjust to pH 5.0 with acetic acid (100). To 820 mL of this solution add 180 mL of acetonitrile. Flow rate: Adjust so that the retention time of voricona- zole is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of voriconazole are not less than 2500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of a solution, prepared by adding the mobile phase to 5 mL of the standard solution to make 10 mL, under the above operating conditions, the relative standard deviation of the peak area of voriconazole is not more than 5.0z. Water <2.48> Not more than 0.2z (1.0 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g, a platinum crucible).
Weigh accurately about 50 mg each of Voriconazole and Voriconazole RS (separately determine the water <2.48> in the same manner as Voriconazole), dissolve each in the mobile phase to make exactly 100 mL. Pipet 5 mL each of these solutions, add each the mobile phase to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of voriconazole in each solution. Amount (mg) of voriconazole (C16H14F3N5O) ＝ MS × AT/AS MS: Amount (mg) of Voriconazole RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 256 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.9 g of ammonium formate in 1000 mL of water, and adjust to pH 4.0 with formic acid. To 550 mL of this solution add 300 mL of methanol and 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of voricona-zole is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of voriconazole are not less than 3500 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voriconazole is not more than 1.0z. ‌Containers and storage Containers—Well-closed contain-ers. Voriconazole for Injection
Ammonium <1.02>—Perform the test with 30 mL of Water as directed under Ammonium Limit Test. Prepare the control solution as follows: to 0.15 mL of Standard Ammo-nium Solution add water for ammonium limit test to make 30 mL (not more than 0.05 mg/L). Purified Water
Wheat Starch occurs as white masses or pow-der. It is practically insoluble in water and in ethanol (99.5).◆
(1) Examine under a microscope <5.01> using a mixture of water and glycerin (1:1), Wheat Starch presents large and small granules, and, very rarely, inter-mediate sizes. The large granules, usually 10 – 60 mm in di-ameter, are discoid or, more rarely, reniform when seen face-on. The central hilum and striations are invisible or barely visible and the granules sometimes show cracks on the edges. Seen in profile, the granules are elliptical and fusiform and the hilum appears as a slit along the main axis. The small granules, rounded or polyhedral, are 2 – 10 mm in diameter. Between orthogonally oriented polarizing plates or prisms, the granules show a distinct black cross intersecting at the hilum. To 1 g of Wheat Starch add 50 mL of water, boil for 1 minute, and allow to cool: a thin, cloudy mucilage is formed. To 1 mL of the mucilage obtained in (2) add 0.05 mL of diluted iodine TS (1 in 10): a deep blue color is formed, and the color disappears by heating. pH <2.54> Put 5.0 g of Wheat Starch in a non-metal vessel, add 25.0 mL of freshly boiled and cooled water, mix gently for 1 minute, and allow to stand for 15 minutes: the pH of the solution is between 4.5 and 7.0.
(1) Iron—To 1.5 g of Wheat Starch add 15 mL of 2 mol/L hydrochloric acid TS, mix, filter, and use the fil-trate as the test solution. To 2.0 mL of Standard Iron Solu-tion add water to make 20 mL, and use this solution as the control solution. Put 10 mL each of the test solution and the control solution in test tubes, add 2 mL of a solution of citric acid (2 in 10) and 0.1 mL of mercapto acetic acid, and mix. Alkalize with ammonia solution (28) to litmus paper, add water to make 20 mL, and mix. Transfer 10 mL each of these solutions into test tubes, allow to stand for 5 minutes, and compare the color of these solutions against a white background: the color of the test solution is not more intense than that of the control solution (not more than 10 ppm). Oxidizing substances—To 4.0 g of Wheat Starch add 50 mL of water, shake for 5 minutes, and centrifuge. To 30 mL of the supernatant liquid add 1 mL of acetic acid (100) and 0.5 to 1.0 g of potassium iodide, shake, and allow to stand for 25 to 30 minutes at a dark place. Add 1 mL of starch TS, and titrate <2.50> with 0.002 mol/L sodium thio-sulfate VS until the color of the solution disappears. Perform a blank determination in the same manner, and make any necessary correction: the volume of 0.002 mol/L sodium thiosulfate VS consumed is not more than 1.4 mL (not more than 20 ppm, calculated as hydrogen peroxide). Sulfur dioxide— Apparatus Use as shown in the figure. A: Three-necked round-bottom flask (500 mL) B: Cylindrical dropping funnel (100 mL) C: Condenser D: Test tube E: Tap Procedure Introduce 150 mL of water into the three-necked round-bottom flask, close the tap of the cylin-drical dropping funnel, and pass carbon dioxide through the whole system at a rate of 100 ± 5 mL per minute. Pass cool-ing water through the condenser, and place 10 mL of hydro-gen peroxide-sodium hydroxide TS in the test tube. After 15 minutes, remove the funnel without interrupting the stream of carbon dioxide, and introduce through the opening into the flask about 25 g of Wheat Starch, accurately weighed, with the aid of 100 mL of water. Apply tap grease to the out-side of the connection part of the funnel, and load the fun-nel. Close the tap of the funnel, pour 80 mL of 2 mol/L hy-drochloric acid TS into the funnel, open the tap to introduce the hydrochloric acid into the flask, and close the tap while several mL of the hydrochloric acid remains, in order to avoid losing sulfur dioxide. Place the flask in a water bath, and heat the mixture for 1 hour. Transfer the contents of the test tube with the aid of a little water to a wide-necked coni-cal flask. Heat in a water bath for 15 minutes, and cool. Add 0.1 mL of bromophenol blue TS, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to violet-blue lasting for at least 20 seconds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the amount of sulfur dioxide by applying the following formula: it is not more than 50 ppm. Amount (ppm) of sulfur dioxide ＝ V/M × 1000 × 3.203 M: Amount (g) of Wheat Starch taken V: Amount (mL) of 0.1 mol/L sodium hydroxide VS consumed (4) Foreign matter. Under a microscope <5.01>, Wheat Starch does not contain starch granules of any other origin. It may contain a minute quantity, if any of fragments of the tissue of the original plant.◆ (5) Total protein—Weigh accurately about 3 g of Wheat Starch, place it in a Kjeldahl flask, add 4 g of a decomposi-tion accelerator (a powdered mixture of 100 g of potassium sulfate, 3 g of copper (II) sulfate pentahydrate and 3 g of titanium (IV) oxide), ◇wash down any adhering substances from the neck of the flask with a fine jet of water.◇ Add 25 mL of sulfuric acid allowing to flow down the inside wall of the flask, and mix the contents. Close the mouth of the flask loosely, for example by means of a glass bulb with a short stem, to avoid excessive loss of the sulfuric acid. Heat the flask gradually at first, then increase the temperature until there is vigorous boiling with condensation of sulfuric acid in the neck of the flask, preventing the upper part of the flask from becoming overheated. Continue the heating until the solution become clear, ◇and the inside wall of the flask is free from a carbonaceous material.◇ After cooling, dissolve the solid material by adding cautiously 25 mL of water, cool again, and place in a steam-distillation apparatus previously washed by passing steam. Add exactly 25 mL of 0.01 mol/L hydrochloric acid VS and a suitable amount of water into the receiver, and immerse the tip of the condenser in this acid so-lution. Add the same quantity of a solution of sodium hy-droxide (21 in 50) as used for a blank determination through the funnel, and distill immediately by passing steam through the mixture. Collect about 40 mL of distillate, lower the receiver so that the tip of the condenser is above the surface of the acid solution, ◇then continue the distillation for a while, and rinse the end part of the condenser with a small amount of water.◇ Titrate <2.50> the excessive hydrochloric acid with 0.025 mol/L sodium hydroxide VS until the color of the solution changes from red-purple through grayish blue to green (indicator: 3 drops of methyl red-methylene blue TS). Perform a blank determination in the same manner. The amount of a solution of sodium hydroxide (21 in 50) to be added from the funnel is sufficient to change the color of the solution in the flask from bluish green to dark brown or black. ‌Amount (z) of nitrogen ＝ (a － b) × 0.035/M M: Amount (g) of Wheat Starch taken a: Volume (mL) of 0.025 mol/L sodium hydroxide VS consumed in a blank determination b: Volume (mL) of 0.025 mol/L sodium hydroxide VS consumed in the sample determination The amount of total protein is not more than 0.3z [0.048z as nitrogen (N:14.01) (using conversion factor of nitrogen to protein, 6.25)]. Loss on drying <2.41> Not more than 15.0z (1 g, 1309C, 90 minutes). Residue on ignition <2.44> Not more than 0.6z (1 g).
White Petrolatum is a white to pale yellow, ho-mogeneous, unctuous mass. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in ethanol (99.5). It dissolves in diethyl ether making a clear liquid or producing slight insoluble substances. It becomes a clear liquid when warmed.
(1) Color—Melt White Petrolatum by warming, and pour 5 mL of it into a test tube, and keep the content in a liquid condition: the liquid has no more color than the fol-lowing control solution, when observed transversely from side against a white background. Control solution: Add 3.4 mL of water to 1.6 mL of Iron (III) Chloride CS. Acidity or alkalinity—To 35.0 g of White Petrolatum add 100 mL of hot water, shake vigorously for 5 minutes, and then draw off the aqueous layer. Treat the White ‌Petrolatum layer in the same manner using two 50-mL por-tions of hot water. To the combined aqueous layer add 1 drop of phenolphthalein TS, and boil: no red color is pro-duced. Further add 2 drops of methyl orange TS: no red color is produced.‌ Heavy metals <1.07>—Proceed with 1.0 g of White Petrolatum according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of White Petrolatum, according to Method 3, and perform the test. Add 10 mL of a solution of magnesium nitrate hex-ahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydro-gen peroxide (30), and fire to burn (not more than 2 ppm). Sulfur compound—To 4.0 g of White Petrolatum add 2 mL of ethanol (99.5) and 2 drops of sodium hydroxide solution (1 in 5) saturated with lead (II) oxide, warm the mixture for 10 minutes at about 709C with frequent shaking, and allow to cool: no dark color is produced. Organic acids—To 100 mL of dilute ethanol add 1 drop of phenolphthalein TS, and titrate with 0.01 mol/L so-dium hydroxide VS, until the color of the solution changes to light red. Mix this solution with 20.0 g of White Petrola-tum, and boil for 10 minutes under a reflux condenser. Add 2 to 3 drops of phenolphthalein TS to the mixture and 0.40 mL of 0.1 mol/L sodium hydroxide VS with vigorous shak-ing: the color of the solution remains red. Fats and fatty oils or resins—To 10.0 g of White Petrolatum add 50 mL of sodium hydroxide solution (1 in 5), and boil for 30 minutes under a reflux condenser. Cool the mixture, separate the aqueous layer, and filter, if neces-sary. To the aqueous layer add 200 mL of dilute sulfuric acid: neither oily matter nor precipitate is produced. Residue on ignition <2.44> Not more than 0.05z (2 g).
White Shellac occurs as yellow-white to light yellow, hard, brittle granules. It is odorless or has a faint, characteristic odor. It is sparingly soluble in ethanol (95), and practically in-soluble in water. It dissolves in sodium hydroxide TS. Acid value <1.13> 65 – 90 Weigh accurately about 0.5 g of White Shellac, add 50 mL of neutralized ethanol, and dis-solve by warming. After cooling, perform the test as directed in the Acid value under Purified Shellac.
(1) Chloride <1.03>—Shake and dissolve 0.40 g of White Shellac in 5 mL of ethanol (95) while warming, add 40 mL of water, and cool. Add 12 mL of dilute nitric acid and water to make 100 mL, and filter. Perform the test using 50 mL of the filtrate as the test solution. Prepare the control solution as follows: to 0.80 mL of 0.01 mol/L hydrochloric acid VS add 2.5 mL of ethanol (95), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.140z). Sulfate <1.14>—Shake and dissolve 0.40 g of White Shellac in 5 mL of ethanol (95) by warming, add 40 mL of water, and cool. Add 2 mL of dilute hydrochloric acid and water to make 100 mL, and filter. Perform the test using 50 mL of the filtrate as the test solution. Prepare the control so-lution as follows: to 0.45 mL of 0.005 mol/L sulfuric acid VS add 2.5 mL of ethanol (95), 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.110z). Heavy metals <1.07>—Proceed with 2.0 g of White Shellac according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm.) Arsenic <1.11>—Prepare the test solution with 0.40 g of White Shellac according to Method 3, and perform the test. Add 10 mL of a solution of magnesium nitrate hexahy-drate in ethanol (95) (1 in 50), then add 1.5 mL of hydrogen peroxide (30), and fire to burn (not more than 5 ppm). Ethanol-insoluble substances—Dissolve about 5 g of White Shellac, accurately weighed, in 50 mL of ethanol (95) on a water bath while shaking. Pour the ethanol solution into a tared extraction thimble, previously dried at 1059C for 2 hours, in a Soxhlet extractor, and extract with ethanol (95) for 3 hours. Dry the extraction thimble at 1059C for 3 hours: the mass of the residue is not more than 2.0z. Use a cylin-drical weighing bottle for taring the extraction thimble. Rosin—To 2.0 g of White Shellac add 10 mL of ethanol (99.5), shake thoroughly to dissolve, and add gradu-ally 50 mL of hexane while shaking. Transfer the solution to a separator, wash with two 50-mL portions of water, filter the upper layer, and evaporate the filtrate on a water bath to dryness. To the residue add 5 mL of acetic anhydride, and heat on a water bath to dissolve, if necessary. Transfer this solution to a Nessler tube, and add 1 drop of sulfuric acid: the color of the solution does not change from red-purple through purple to reddish yellow. Wax—Dissolve 10.0 g of White Shellac in 150 mL of ‌a solution of sodium carbonate decahydrate (9 in 200) with shaking on a water bath, and continue the heating for 2 hours. After cooling, collect the floating wax by filtration, wash the wax and the filter paper with water, transfer to a beaker, and dry at 659C until the water is almost evaporated. Transfer the wax together with the filter paper to an ex-traction thimble in a Soxhlet extractor. Dissolve the wax remaining in the beaker with a suitable quantity of chlo-roform by warming. Pour the solution into the thimble, and extract with chloroform for 2 hours. Evaporate the chlo-roform solution to dryness, and dry the residue at 1059C for 3 hours: the mass of the residue is not more than 20 mg. Loss on drying Not more than 6.0z. Weigh accurately about 1 g of moderately fine powder of White Shellac, and dry at 409C for 4 hours, then for 15 hours in a desiccator (calcium chloride for drying). Total ash <5.01> Not more than 1.0z (1 g).
White Soft Sugar is colorless or white, crystals or crystalline powder. It is odorless and has a sweet taste. It is very soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. A solution of White Soft Sugar (1 in 10) is neutral.
(1) When 1 g of White Soft Sugar is ignited, it melts and swells, and decomposes, emitting an odor of caramel, to bulky charcoal. (2) To 0.1 g of White Soft Sugar add 2 mL of dilute sul-furic acid, boil, add 4 mL of sodium hydroxide TS and 3 mL of Fehling's TS, and heat to boiling: a red to dark red pre-cipitate is produced. beaker with a watch glass, and boil. Immediately add 50.0 mL of the sample solution, boil the mixture exactly for 5 minutes, add at once 50 mL of freshly boiled and cooled water, dip it in a water bath of a temperature below 109C for 5 minutes, and collect the precipitate in a tared glass filter (G4). Wash the residue on the filter with water until the last washing is neutral, then wash with 10 mL of ethanol (95) and 10 mL of diethyl ether, and dry at 1059C for 30 minutes: the mass of the residual precipitate is not more than 0.120 g. Loss on drying <2.41> Not more than 1.30z (15 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (2 g).
Wine is an alcoholic liquid obtained by fermenting the juice of the fruits of Vitis vinifera Linneá(Vitaceae) or allied plants. It contains not less than 11.
Wine is a light yellow or reddish purple to red-purple liquid. It has a characteristic and aromatic odor. It has a slightly astringent and faintly irritating taste. Optical rotation <2.49> Boil 160 mL of Wine, neutralize with potassium hydroxide TS, and concentrate to 80 mL on a water bath. Cool, dilute with water to 160 mL, add 16 mL of lead subacetate TS, shake well, and filter. To 100 mL of the filtrate add 10 mL of a saturated solution of sodium sul-fate decahydrate, shake well, filter, and use the filtrate as the sample solution. Allow 20 mL of the sample solution to stand for 24 hours, add 0.5 g of activated charcoal, shake, stopper, and allow to stand for 10 minutes. Filter, and ob-serve the optical rotation of the filtrate in a 200-mm cell. Multiply the optical rotation observed by 1.21, and designate as the optical rotation of Wine: it is between －0.39and ＋ 0.39. Specific gravity <2.56> d 20: 0.990 – 1.010
(1) Total acid [as L-tartaric acid (C4H6O6)]—To exactly 10 mL of Wine add 250 mL of freshly boiled and cooled water, and titrate <2.50> with 0.1 mol/L sodium hy-droxide VS (indicator: 1 mL of phenolphthalein TS). Each mL of 0.1 mol/L sodium hydroxide VS ＝ 7.504 mg of C4H6O6 Total acid is not less than 0.40 w/vz and not more than 0.80 w/vz. Volatile acid [as acetic acid (C2H4O2: 60.05)]—Trans-fer 100 mL of Wine to a beaker, add 1 mL of 1 mol/L sodi-um hydroxide VS and the same volume of 1 mol/L sodium hydroxide VS as that of 0.1 mol/L sodium hydroxide VS titrated in (1) to make the solution alkaline, and concentrate to 50 mL on a water bath. Cool, add water to make 100 mL, transfer to a 1000-mL distillation flask, containing previ-ously added 100 g of sodium chloride. Wash the beaker with 100 mL of water, and combine the washings in the distilla-tion flask. Add 5 mL of a solution of L-tartaric acid (3 in 20), and distil with steam cautiously to maintain the volume of the solution in the flask until 450 mL of the distillate is obtained for 45 minutes. Dilute the distillate to exactly 500 mL with water, and use this solution as the sample solution. Titrate <2.50> a 250-mL portion of the sample solution with 0.1 mol/L sodium hydroxide VS (indicator: 5 drops of phe-nolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 6.005 mg of C2H4O2 The volatile acid is not more than 0.15 w/vz. Sulfur dioxide—Stopper a 750-mL round-bottomed flask with a stopper having two holes. Through one hole, in-sert a glass tube A extending nearly to the bottom of the flask. Through the other hole, insert a glass tube B ending to the neck of the flask. Connect the tube B to a Liebig's con-denser, and the end of the condenser to a joint of which in-ner diameter is 5 mm at the lower end. Connect the other end of the joint with a holed rubber stopper to a U tube having three bulbs as shown in the Figure. Pass carbon dioxide washed with a solution of potassium permanganate (3 in 100) through the tube A. Displace the air in the apparatus by carbon dioxide, and place 50 mL of a freshly prepared and diluted starch TS (1 in 5) and 1 g of potassium iodide in the U tube. From the other end of the U tube, add 1 to 2 drops of 0.01 mol/L iodine VS from a burette. While passing car-bon dioxide, remove the stopper of the flask a little, add ex-actly 25 mL of Wine, 180 mL of freshly boiled and cooled water, 0.2 g of tannic acid, and 30 mL of phosphoric acid, and stopper again. Pass carbon dioxide for further 15 minutes, heat the distillation flask with caution so that 40 to 50 drops of the distillate may be obtained in 1 minute. When the color of starch TS in the U tube is discharged, add 0.01 mol/L iodine VS dropwise from a burette so that the color of the starch TS remains light blue to blue during the distilla-tion. Read the volume of 0.01 mol/L iodine VS consumed when exactly 60 minutes have passed after the beginning of distillation. In this case, however, the coloration of starch TS produced by 1 drop of 0.01 mol/L iodine VS should per-sist at least for 1 minute. Each mL of 0.01 mol/L iodine VS ＝ 0.6406 mg of SO2 The amount of sulfur dioxide (SO2: 64.06) does not exceed 7.5 mg. Total sulfuric acid—Transfer 10 mL of Wine to a beaker, boil, and add 50 mL of a solution prepared by dis-solving 5.608 g of barium chloride dihydrate in 50 mL of hydrochloric acid and water to make 1000 mL. Cover the beaker, and heat on a water bath for 2 hours, supplying the water lost by distillation. Cool, centrifuge, and decant the supernatant liquid in another beaker. To this solution add 1 to 2 drops of dilute sulfuric acid, and allow to stand for 1 hour: a white precipitate is formed. Arsenic <1.11>—Evaporate 10 mL of Wine on a water bath to dryness. Prepare the test solution with the residue according to Method 3, and perform the test (not more than 0.2 ppm). Glycerin—Pipet 100 mL of Wine into a 150-mL por-celain dish, and concentrate on a water bath to 10 mL. Add 1 g of sea sand (No. 1), and make the solution strongly alka-line by adding a solution prepared by dissolving 4 g of cal-cium hydroxide in 6 mL of water. Heat on a water bath with constant stirring and pushing down any material adhering to the wall of the dish until the contents of the dish become soft masses. Cool, add 5 mL of ethanol (99.5), and grind to a gruel-like substance. Heat on a water bath, add 10 to 20 mL of ethanol (99.5) while agitating, boil, and transfer to a 100-mL volumetric flask. Wash the dish with seven 10-mL portions of hot ethanol (99.5), combine the washings with the contents of the flask, cool, and add ethanol (99.5) to make exactly 100 mL. Filter through a dry filter paper, evaporate 90 mL of the filtrate on a water bath, taking care not to boil the solution during the evaporation. Dissolve the residue in a small amount of ethanol (99.5), transfer to a 50-mL glass-stoppered volumetric cylinder, wash with sever-al portions of ethanol (99.5), and add the washings to the solution in the cylinder to make 15 mL. Add three 7.5-mL portions of dehydrated diethyl ether, shake vigorously each time, and allow to stand. When the solution becomes quite clear, transfer to a tared, flat weighing bottle. Wash the volumetric cylinder with 5 mL of a mixture of dehydrated diethyl ether and ethanol (99.5) (3:2). Transfer the washings to the weighing bottle, and evaporate carefully on a water bath. When the liquid becomes sticky, dry at 1059C for 1 hour, and cool in a desiccator (silica gel), and weigh: the mass of the residue is not less than 0.45 g and not more than 0.90 g. Reducing sugars—To a 25-mL portion of the sample solution obtained in the Optical rotation add 50 mL of boil-ing Fehling's TS, and heat for exactly 2 minutes. Filter the separated precipitates by a tared glass filter by suction, wash successively with hot water, with ethanol (95) and with diethyl ether, and continue to dry the precipitates by suction. Heat the filter gently at first, and then strongly until the pre-cipitates become completely black. Cool the precipitates in a desiccator (silica gel), and weigh as copper (II) oxide: the mass of cupric oxide does not exceed 0.325 g. Sucrose—Transfer a 50-mL portion of the sample so-lution obtained in the Optical rotation to a 100-mL flask, neutralize with diluted hydrochloric acid (1 in 30), followed by further addition of 5 mL of diluted hydrochloric acid (1 in 30). Heat in a water bath for 30 minutes, cool, neutralize with a solution of potassium hydroxide (1 in 100), add 4 drops of sodium carbonate TS, filter into a 100-mL volumet-ric flask, wash with water, combine the washings with the filtrate, and add water to make 100 mL. To 25 mL of this solution add 50 mL of boiling Fehling's TS, and proceed as directed in (7), and weigh as copper (II) oxide. From the number obtained by multiplying the mass (g) of copper (II) oxide by 2, deduct the amount (g) of copper (II) oxide deter-mined in (7), and multiply again the number so obtained by 1.2: the number obtained does not exceed 0.104 (g). Benzoic acid, cinnamic acid and salicylic acid—Tran-sfer exactly 50 mL of the sample solution obtained in (2) to a separator, add 10 g of sodium chloride and 2 mL of dilute hydrochloric acid, and extract with three 10-mL portions of diethyl ether. Combine the diethyl ether extracts, wash with two 5-mL portions of water, and extract with three 10-mL portions of 0.1 mol/L sodium hydroxide VS. Combine the alkaline extracts, evaporate the diethyl ether by warming on a water bath, cool, neutralize with 1 mol/L hydrochloric acid VS, and add 5 mL of potassium chloride-hydrochloric acid buffer solution and water to make exactly 50 mL. Per-form the test as directed under Ultraviolet-visible Spectro-photometry <2.24> with this solution, using a solution pre-pared in the same manner instead of the sample solution as the blank: the absorbance does not exceed 0.15 at a wave-length between 220 nm and 340 nm. Boric acid—Transfer 50 mL of Wine to a porcelain dish, add 5 mL of sodium carbonate TS, evaporate on a water bath to dryness, and ignite: a half portion of the residue does not respond to Qualitative Tests <1.09> (1) for borate. Dissolve another half portion of the residue in 5 mL of hydrochloric acid: it does not respond to Qualitative Tests <1.09> (2) for borate. Methanol—Wine meets the requirements of Methanol Test <1.12>, when proceeding with exactly 1 mL of ethanol layer obtained by Method 1 of Alcohol Number Determination <1.01> and distilling without adding water after shaking with 0.5 g of calcium carbonate. Formaldehyde—To 25 mL of Wine add 5 g of sodi-um chloride and 0.2 g of L-tartaric acid, distil, and obtain 15 mL of the distillate. To 5 mL of the distillate add 5 mL of acetyl acetone TS, mix, and heat on a water bath for 10 minutes: the solution has no more color than that of the fol-lowing control solution. Control solution: Using 5 mL of water instead of the dis-tillate, perform the test in the same manner. Extract content 1.9 – 3.5 w/vz Pipet 25 mL of Wine to a 200-mL tared beaker containing 10 g of sea sand (No. 1), previously dried at 1059C for 2.5 hours, and evaporate to dryness on a water bath. Dry the residue at 1059C for 2 hours, cool in a desiccator (silica gel), and weigh. Total ash 0.13 – 0.40 w/vz Pipet 50 mL of Wine to a tared porcelain dish, and evaporate to dryness on a water bath. Ignite the residue to the constant mass, cool, and weigh.
(1) Ethanol—Pipet Wine into a 100-mL volumetric flask at 159C, transfer to a 300- to 500-mL flask, and wash this volumetric flask with two 15-mL portions of water. Add the washings to the sample in the flask, connect the flask to a ‌distillation tube having a trap, and distil using the volumetric flask as a receiver. When about 80 mL of the distillate is ob-tained (it takes about 20 minutes), stop the distillation, allow to stand in water at 159C for 30 minutes, and add water to make exactly 100 mL. Shake well, and determine the specific gravity at 159C under Specific Gravity <2.56> (Method 3 may be used): the specific gravity d 15 is between 0.98217 and 0.98547.‌ (2) L-Tartaric acid—Pipet 100 mL of Wine, add 2 mL of acetic acid (100), 0.5 mL of a solution of potassium acetate (1 in 5) and 15 g of powdered potassium chloride, and shake vigorously to dissolve as much as possible. Add 10 mL of ethanol (95), rub the inner wall of the beaker strongly for 1 minute to induce the crystallization, and allow to stand be-tween 09C and 59C for more than 15 hours. Filter the crys-tals by suction, wash successively the beaker and the crystals with 3-mL portions of a solution prepared by dissolving 15 g of powdered potassium chloride in 120 mL of diluted ethanol (1 in 6), and repeat the washings five times. Transfer the crystals together with the filter paper to a beaker, wash the filter with 50 mL of hot water, combine the washings in the beaker, and dissolve the crystals by heating. Titrate <2.50> the solution with 0.2 mol/L sodium hydroxide VS immediately (indicator: 1 mL of phenolphthalein TS). The number obtained by adding 0.75 to the amount (mL) of 0.2 mol/L sodium hydroxide VS consumed represents the amount (mL) of 0.2 mol/L sodium hydroxide VS consumed. Each mL of 0.2 mol/L sodium hydroxide VS ＝ 30.02 mg of C4H6O6
Xylitol, when dried, contains not less than 98.
Xylitol occurs as white, crystals or powder. It is odorless and has a sweet taste. It is very soluble in water, slightly soluble in ethanol (95).
(1) To 1 mL of a solution of Xylitol (1 in 2) add 2 mL of iron (II) sulfate TS and 1 mL of a solution of sodium hydroxide (1 in 5): blue-green color is produced without turbidity. (2) Determine the infrared absorption spectrum of Xylitol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 5.0 g of Xylitol in 10 mL of freshly boiled and cooled water: the pH of this solution is between 5.0 and 7.0. Melting point <2.60> 93.0 – 95.09C
(1) Clarity and color of solution—Dissolve 5 g of Xylitol in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 2.0 g of Xylitol. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.005z). Sulfate <1.14>—Perform the test with 4.0 g of Xylitol. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.006z). Heavy metals <1.07>—Proceed with 4.0 g of Xylitol according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 5 ppm). Nickel—Dissolve 0.5 g of Xylitol in 5 mL of water, add 3 drops of dimethylglyoxime TS and 3 drops of ammo-nia TS, and allow to stand for 5 minutes: no red color is pro-duced. Arsenic <1.11>—Prepare the test solution with 1.5 g of Xylitol according to Method 1, and perform the test (not more than 1.3 ppm). Sugars—Dissolve 5.0 g of Xylitol in 15 mL of water, add 4.0 mL of dilute hydrochloric acid, and heat in a water bath for 3 hours under a reflux condenser. After cooling, neutralize with sodium hydroxide TS (indicator: 2 drops of methyl orange TS). Then add water to make 50 mL, transfer 10 mL of this solution to a flask, add 10 mL of water and 40 mL of Fehling's TS, boil gently for 3 minutes, and allow to stand to precipitate copper (I) oxide. Remove the superna-tant liquid through a glass filter (G4), and wash the precipi-tate with warm water until the last washing does not show alkalinity. Filter these washings through the glass filter men-tioned above. Dissolve the precipitate in the flask in 20 mL of iron (III) sulfate TS, filter the solution through the glass filter mentioned above, wash with water, combine the wash-ings with the filtrate, heat at 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS: not more than 1.0 mL of 0.02 mol/L potassium permanganate VS is consumed. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Xylitol, previously dried, dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution into an iodine flask, add 50 mL of potas-sium periodate TS exactly, and heat in a water bath for 15 minutes. After cooling, add 2.5 g of potassium iodide, stop-per, shake well, allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (in-dicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium thiosulfate VS It is hygroscopic. ＝ 1.902 mg of C5H 12O5 Identification (1) To 1 mL of a solution of Xylitol (1 in 2) add 2 mL of iron (II) sulfate TS and 1 mL of a solution of sodium hydroxide (1 in 5): blue-green color is produced without turbidity. (2) Determine the infrared absorption spectrum of Xylitol, previously dried, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Dissolve 5.0 g of Xylitol in 10 mL of freshly boiled and cooled water: the pH of this solution is between 5.0 and 7.0. Melting point <2.60> 93.0 – 95.09C
Yellow Petrolatum occurs as a yellow, homo-geneous, unctuous mass, It is odorless and tasteless. It is slightly soluble in ethanol (95), and practically insolu-ble in water. It dissolves in diethyl ether, in petroleum benzine and in turpentine oil, making a clear liquid or producing slight in-soluble substances. It becomes a yellow, clear liquid with slight fluorescence when warmed.
(1) Color—Melt Yellow Petrolatum by warming, and pour 5 mL of it into a test tube, and keep the content in a liquid condition: the liquid has no more color than the fol-lowing control solution, when observed transversely from side against a white background. Control solution: To 3.8 mL of Iron (III) Chloride CS add 1.2 mL of Cobalt (II) Chloride CS. Acidity or alkalinity—To 35.0 g of Yellow Petrola-tum add 100 mL of hot water, shake vigorously for 5 minutes, and then draw off the aqueous layer. Treat the Yel-low Petrolatum layer in the same manner using two 50-mL portions of hot water. To the combined aqueous layer add 1 drop of phenolphthalein TS, and boil: no red color is pro-duced. Further add 2 drops of methyl orange TS: no red color is produced. Heavy metals <1.07>—Proceed with 1.0 g of Yellow Petrolatum according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Yellow Petrolatum, according to Method 3, and perform the test. Add 10 mL of a solution of magnesium nitrate hex-ahydrate in ethanol (95) (1 in 50), then add 1.5 mL of hydro-gen peroxide (30), and fire to burn (not more than 2 ppm). Sulfur compound—To 4.0 g of Yellow Petrolatum add 2 mL of ethanol (99.5) and 2 drops of sodium hydroxide solution (1 in 5) saturated with lead (II) oxide, warm the mixture for 10 minutes at about 709C with frequent shaking, and allow to cool: no dark color is produced. Organic acids—To 100 mL of dilute ethanol add 1 drop of phenolphthalein TS, and titrate with 0.01 mol/L so-dium hydroxide VS, until the color of the solution changes to light red. Mix this solution with 20.0 g of Yellow Petrola-tum, and boil for 10 minutes under a reflux condenser. Add 2 to 3 drops of phenolphthalein TS to the mixture and 0.40 mL of 0.1 mol/L sodium hydroxide VS with vigorous shak-ing: the color of the solution remains red. Fats and fatty oils or resins—To 10.0 g of Yellow Petrolatum add 50 mL of sodium hydroxide solution (1 in 5), and boil for 30 minutes under a reflux condenser. Cool the mixture, separate the aqueous layer, and filter, if neces-sary. To the aqueous layer add 200 mL of dilute sulfuric acid: neither oily matter nor precipitate is produced. Residue on ignition <2.44> Not more than 0.05z (2 g).
Yokukansan Extract contains not less than 0.
Yokukansan Extract is a light brown to grayish brown powder or a black-brown viscous extract. It has a slightly odor, and a slightly bitter and acid taste. Identiˆcation (1) To 2.0 g of the dry extract (or 6.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, and centrifuge. Separate the diethyl ether layer, add 10 mL of sodium hydroxide TS, shake, centrifuge, separate the diethyl ether layer, and use this layer as the sample solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of butyl acetate and hexane (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultrav-iolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white ‰uorescent spot from the standard solution (Japanese Angelica Root; Cnidium Rhizome). To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 20 mL of water and 2 mL of ammonia TS, shake, then add 20 mL of diethyl ether, shake, and separate the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, and use the solution as the sample solution. Separately, dissolve 1 mg each of rhyncophyllin for thin-layer chromatography and hirsutine for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, 1-propanol, water and acetic acid (100) (7:5:4:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): at least one of the several spots obtained from the sample solution has the same color tone and Rf value with one of the two dark violet spots from the standard solution (Uncaria Hook). For preparation prescribed Atractylodes Rhizome—To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 25 mL of diethyl ether, and shake. Separate the diethyl ether layer, evaporate the solvent under low pressure (in vacuo), dissolve the residue in 2 mL of diethyl ether, and use this solution as the sample so-lution. Separately, dissolve 1 mg of atractylenoide III for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution has the same color tone and Rf value with the red to purple-red spot from the standard solution (Atractylodes Rhizome). For preparation prescribed Atractylodes Lancea Rhi-zome—To 2.0 g of the dry extract (or 6.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 25 mL of hexane, and shake. Separate the hexane layer, evaporate the solvent under low pressure (in vacuo), add 2 mL of hexane to the residue, and use this solution as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a dark purple spot is observed at an Rf value of about 0.5. The spot shows a greenish brown color after being sprayed evenly 4-dimethylamino-benzaldehyde TS for spraying on the plate, heated at 1059C for 5 minutes, and allowed to cool (Atractylodes Lancea Rhizome). Yokukansan Extract / Crude Drugs and Related Drugs To 1.0 g of the dry extract (or 3.0 g of the viscous extract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of saikosaponin b2 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and examine under ul- mL, and use this solution as the sample solution. Separately, weigh accurately about 5 mg each of rhyncophylline for assay and hirsutine for assay, dissolve in a mixture of metha-nol and diluted acetic acid (7:3) to make exactly 100 mL. Pipet 10 mL of this solution, add the mixture of methanol and diluted acetic acid (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, ATR and ATH, and ASR and ASH, of rhyncophylline and hirsutine in each solution. Amount (mg) of total alkaloids (rhyncophylline and hirsutine) traviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color ＝ (MSR × ATR/ASR ＋ MSH × ATH/ASH ) × 1/50 tone and Rf value with the yellow fluorescent spot from the standard solution (Bupleurum Root). To 1.0 g of the dry extract (or 3.0 g of the viscous ex-tract) add 10 mL of water, shake, then add 10 mL of 1-butanol, shake, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of liquiritin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-green fluorescent spot from the standard solution (Glycyrrhiza).
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of the dry extract (or an amount of the viscous ex-tract, equivalent to 1.0 g of the dried substance) as directed under Extracts (4), and perform the test (not more than 30 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.67 g of the dry extract (or an amount of the viscous extract, equivalent to 0.67 g of the dried substance) according to Method 3, and perform the test (not more than 3 ppm). Loss on drying <2.41> The dry extract: Not more than 10.0z (1 g, 1059C, 5 hours). The viscous extract: Not more than 66.7z (1 g, 1059C, 5 hours). Total ash <5.01> Not less than 10.0z, calculated on the dried basis.
(1) Total alkaloids (rhyncophylline and hirsutine) —Weigh accurately about 1 g of the dry extract (or an amount of the viscous extract, equivalent to about 1 g of the dried substance), add 20 mL of diethyl ether, shake, then add 3 mL of 1 mol/L hydrochloric acid TS and 7 mL of water, and shake for 10 minutes, centrifuge, and remove the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, and proceed in the same manner as above. To the aqueous layer add 10 mL of sodium hydroxide TS and 20 mL of diethyl ether, shake for 10 minutes, centrifuge, and separate the diethyl ether layer. To the aqueous layer add 20 mL of diethyl ether, proceed in the same manner, and repeat MSR: Amount (mg) of rhyncophylline for assay taken MSH: Amount (mg) of hirsutine for assay taken Operation conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 1 g of sodium lauryl sulfate add 600 mL of methanol, shake, then add 400 mL of water and 1 mL of phosphoric acid. Flow rate: 1.0 mL per minute (the retention times of rhyn-cophylline and hirsutine are about 17 minutes and about 47 minutes, respectively). Systemic suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peaks of rhyncophylline and hirsutine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rhyncophylline and hirsutine is not more than 1.5z, respectively. Saikosaponin b2—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, separate the supernatant liquid, combine all the supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, use saikosaponin b2 standard TS for assay as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of sai-kosaponin b2 in each solution. Amount (mg) of saikosaponin b2 ＝ CS × AT/AS × 50 this procedure twice. Combine all the extracts, evaporate the solvent under low pressure (in vacuo) at not more than 409C, dissolve the residue in the mobile phase to make exactly 10 CS: Concentration (mg/mL) of saikosaponin b2 kosaponin b2 standard TS for assay in sai- JP XVIII Crude Drugs and Related Drugs / Yokukansan Extract 2175 Operation conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (5:3). Flow rate: 1.0 mL per minute (the retention time of sai-kosaponin b2 is about 12 minutes). Systemic suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of saikosaponin b2 are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saikosaponin b2 is not more than 1.5z. Glycyrrhizic acid—Weigh accurately about 0.5 g of the dry extract (or an amount of the viscous extract, equiva-lent to about 0.5 g of the dried substance), add 20 mL of diethyl ether and 10 mL of water, and shake for 10 minutes. After centrifugation, remove the diethyl ether layer, add 20 mL of diethyl ether, proceed in the same manner as de-scribed above, and remove the diethyl ether layer. To the aqueous layer add 10 mL of methanol, shake for 30 minutes, centrifuge, and take the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), shake for 5 minutes, centrifuge, and take the supernatant liquid. Combine these supernatant liquids, add diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of glycyr-rhizic acid is about 15 minutes). System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu- tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Zaltoprofen, when dried, contains not less than 99.
Zaltoprofen occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in acetone, soluble in methanol and in ethanol (99.5), and practically insoluble in water. It is gradually decomposed by light. A solution of Zaltoprofen in acetone (1 in 10) shows no optical rotation.
(1) To 0.2 g of Zaltoprofen add 0.5 g of so-dium hydroxide, heat gradually to melt, and then carbonize. After cooling, add 5 mL of diluted hydrochloric acid (1 in 2): the gas evolved darkens moisten lead (II) acetate paper. Determine the absorption spectrum of a solution of Zaltoprofen in ethanol (99.5) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Zalto-profen as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Zaltoprofen according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Zaltoprofen according to Method 3, using 10 mL of a so-lution of magnesium nitrate hexahydrate in ethanol (95) (2 in 25), and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Zaltoprofen in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than zaltoprofen and the peak having the relative retention time of about 0.7 to zaltoprofen obtained from the sample solution is not larger than the peak area of zaltoprofen from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (300:200:1). Flow rate: Adjust so that the retention time of zaltoprofen is about 4 minutes. Time span of measurement: About 15 times as long as the retention time of zaltoprofen, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of zaltoprofen obtained with 20 mL of this solution is equivalent to 8 to 12z of that with 20 mL of the standard solution. System performance: Dissolve 25 mg of zaltoprofen and 50 mg of isopropyl benzoate in 100 mL of ethanol (99.5). Pipet 1 mL of this solution, and add the mobile phase to make exactly 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, zal-toprofen and isopropyl benzoate are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌area of zaltoprofen is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Zaltoprofen, previ-ously dried, dissolve in 50 mL of methanol, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 29.84 mg of C17H14O3S
Zanthoxyli Piperiti Pericarpium Pulveratum
Powdered Japanese Zanthoxylum Peel occurs as a dark yellow-brown powder. It has a strong, characteris-tic aroma and an acrid taste, leaving a sensation of numb-ness on the tongue. Under a microscope <5.01>, Powdered Japanese Zanthox- ylum Peel reveals fragments of inner tissue of endocarp consisting of stone cells with cell walls about 2.5 mm in thick-ness; fragments of spiral and ring vessels 10 – 15 mm in diam-eter; fragments of oil sacs containing essential oil or resin; fragments of epidermal cells, polygonal in surface view, con-taining tannin; numerous oil drops; masses of tannin, colored red by adding vanillin-hydrochloric acid TS.
To 2 g of Powdered Japanese Zanthoxylum Peel add 10 mL of water, shake for 5 minutes, add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, hexane, methanol and acetic acid (100) (20:20:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot with an Rf value of about 0.3 is observed. Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 30.0 g of Powdered Japanese Zanthoxylum Peel: the volume of essen-tial oil is not less than 0.8 mL.
Pericarps of capsules of 2 or 3 flattened spheroidal mericarps, which are dehiscent in 2 pieces about 5 mm in diameter; the outer surface of pericarp, dark yellow-red to dark red-brown, with numerous dented spots originat-ed from oil sacs; the inner surface, light yellow-white. Odor, characteristically aromatic; taste, acrid, which gives numbing sensation to the tongue. Under a microscope <5.01>, a transverse section of Japanese Zanthoxylum Peel reveals the external epidermis and the adjoined unicellular layer containing red-brown tan-nin; the mesocarp holds oil sacs being up to approximately 500 mm in diameter and sporadically vascular bundles con-sisting mainly of spiral vessels; the endocarp consists of stone cell layers; inner epidermal cells very small.
To 2 g of pulverized Japanese Zanthoxylum Peel add 10 mL of water, shake for 5 minutes, add 5 mL of diethyl ether, shake, centrifuge, and use the diethyl ether layer as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, hexane, methanol and acetic acid (100) (20:20:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot with an Rf value of about 0.3 is observed.
(1) Seed—When perform the test of foreign matter <5.01>, the amount of the seeds contained in Japanese Zan-thoxylum Peel does not exceed 20.0z. Peduncle and twig—The amount of the peduncles and twigs contained in Japanese Zanthoxylum Peel does not exceed 5.0z. Foreign matter <5.01>—The amount of foreign mat- ter other than peduncles and twigs contained in Japanese Zanthoxylum Peel does not exceed 1.0z. Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 30.0 g of pulverized Japanese Zanthoxylum Peel: the volume of essen-tial oil is not less than 1.0 mL.
Zidovudine occurs as a white to pale yellow-white powder. It is freely soluble in methanol, soluble in ethanol (99.5), and sparingly soluble in water. It gradually turns yellow-brown on exposure to light. Melting point: about 1249C. It shows crystal polymorphism.
Determine the infrared absorption spectrum of Zidovudine as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Zidovudine RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Zidovudine and Zidovudine RS separately in a small amount of water and dry them in a desiccator (in vacuum, phosphorus (V) oxide), and perform the test with the residues. D Optical rotation <2.49> [a]25: ＋60.5 – ＋63.09(0.5 g calcu-lated on the anhydrous basis, ethanol (99.5), 50 mL, 100 mm).
(1) Heavy metals <1.07>— Proceed with 1.0 g of Zidovudine according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) 1-[(2R,5S )-2,5-Dihydro-5-(hydroxymethyl)-2-furyl] thymine, triphenylmethanol, and other related sub-stances—Dissolve 0.20 g of Zidovudine in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, add 1 mL of the sample solution to 20 mg each of thymine for liquid chromatography, 1-[(2R,5S )-2,5-di-hydro-5-(hydroxymethyl)-2-furyl]thymine for thin-layer chromatography, and triphenylmethanol for thin-layer chro-matography, and add methanol to dissolve to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of chloroform and methanol (9:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of thymine in each solution, and calculate the amount of thy-mine using the following formula: the amount is not more than 2.0z. Also, determine the peak area of each peak ob-tained from the sample solution by the automatic integration method, and calculate the amounts of related substances other than thymine by the area percentage method: the amount of 3?-chloro-3?-deoxythymidine, whose relative retention time to zidovudine is 1.2, is not more than 1.0z, and is not more than 0.5z for all other related substances. Finally, the total amount of thymine, 3?-chloro-3?-deox-ythymidine, and all related substances obtained above is not more than 3.0z. Amount (z) of thymine ＝ MS/MT × AT/AS × 10 MS: Amount (mg) of thymine for liquid chromatography taken MT: Amount (mg) of Zidovudine taken Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of zidovudine, beginning after the solvent peak. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of zidovudine obtained with 10 mL of this solu-tion is equivalent to 3.5 to 6.5z of that with 10 mL of the so-lution for system suitability test. Water <2.48> Not more than 1.0z (0.25 g, coulometric titration). Residue on ignition <2.44> Not more than 0.2z (0.5 g). ‌Assay Weigh accurately about 50 mg of Zidovudine and Zidovudine RS (separately determine the water <2.48> in the same manner as Zidovudine), and dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of each solution, add the mobile phase to make them exactly 50 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of zidovudine in each solution.‌ Purity (1) Clarity and color of solution—Dissolve 1.0 g of Zinc Chloride in 10 mL of water and 2 drops of hydro-chloric acid: the solution has no color, and is clear. Sulfate <1.14>—Perform the test with 2.0 g of Zinc Chloride. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Ammonium—Dissolve 0.5 g of Zinc Chloride in 5 mL of water, and warm with 10 mL of a solution of sodium hydroxide (1 in 6): the evolving gas does not change moistened red litmus paper to blue. Heavy metals—Dissolve 0.5 g of Zinc Chloride in 5 Amount (mg) of zidovudine (C10H13N5O4) ＝ M S × AT/AS mL of water in a Nessler tube, shake thoroughly with 15 mL of potassium cyanide TS, add 1 drop of sodium sulfide TS, MS: Amount (mg) of Zidovudine RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (particle diameter: 5 mm). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (4:1). Flow rate: Adjust so that the retention time of zidovudine is about 15 minutes. System suitability— System performance: Dissolve 50 mg of Zidovudine in 50 mL of the mobile phase. Separately, dissolve 5 mg of 3?-chloro-3?-deoxythymidine for liquid chromatography in 50 mL of the mobile phase. Mix 10 mL and 1 mL of these solu-tions, respectively, and add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above conditions, zidovudine and 3?-chloro-3?-deoxythymidine are eluted in this order with the resolution between these peaks being not less than 1.4, and the symme-try factor of the peak of zidovudine is not more than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above condi-tions, the relative standard deviation of the peak area of zidovudine is not more than 2.0z.
Zolpidem Tartrate contains not less than 98.
Zolpidem Tartrate occurs as a white crystalline powder. It is freely soluble in acetic acid (100), soluble in N,N-dimethylformamide and in methanol, sparingly soluble in water, and slightly soluble in ethanol (99.5) and in acetic an-hydride. It dissolves in 0.1 mol/L hydrochloric acid TS. It gradually changes to yellow in color on exposure to light. D Optical rotation [a]20: about ＋1.89(1 g, N,N-dimethyl- formamide, 20 mL, 100 mm).
(1) Dissolve 50 mg of Zolpidem Tartrate in 5 mL of acetic acid (100) and add 3 drops of Dragendorff's TS: an orange precipitate is formed. Determine the absorption spectrum of a solution of Zolpidem Tartrate in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Zol-pidem Tartrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of 1.0 g of Zolpidem Tartrate in 10 mL of methanol by warming, responds to Qualitative Tests <1.09> (3) for tartrate.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Zolpidem Tartrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 10 mg of Zolpidem Tartrate in 20 mL of methanol and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 20 mL, and use this solu-tion as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: each area of ‌the peak other than zolpidem obtained from the sample solu-tion is not larger than the peak area of zolpidem from the standard solution. Operating conditions— Detector: A ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel tube 4.6 mm in inside diameter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 4.9 g of phosphoric acid add 1000 mL of water, and adjust the pH to 5.5 with triethylamine. To 11 volumes of this solution add 5 volumes of methanol and 4 volumes of acetonitrile. Flow rate: Adjust so that the retention time of zolpidem is about 5 minutes. Time span of measurement: About 5 times as long as the retention time of zolpidem. System suitability— System performance: Dissolve 10 mg each of Zolpidem Tartrate and benzyl parahydroxybenzoate in 100 mL of methanol. When the procedure is run with 5 mL of this solu-tion under the above operating conditions, zolpidem and benzyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of zolpidem is not more than 5.0z. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.4 g of Zolpidem Tartrate, dissolve in 100 mL of a mixture of acetic anhydride and ace-tic acid (100) (7:3) and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.24 mg of (C19H21N3O)2.C4H6O6
Containers—Tight containers. Storage conditions—Light-resistant.
Zonisamide, when dried, contains not less than 98.
Zonisamide occurs as white to pale yellow, crystals or crystalline powder. It is freely soluble in acetone and in tetrahydrofuran, spar-ingly soluble in methanol, slightly soluble in ethanol (99.5), and very slightly soluble in water.
(1) Determine the absorption spectrum of a solution of Zonisamide in methanol (3 in 200,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Zonisamide RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Zonisamide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of dried Zonisamide RS: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—Dissolve 1.0 g of Zonisamide in 30 mL of acetone, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.01 mol/L hydrochloric acid VS add 30 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.036z). Sulfate <1.14>—Dissolve 1.0 g of Zonisamide in 30 mL of acetone, and add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 1.0 mL of 0.005 mol/L sulfuric acid VS add 30 mL of acetone, 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.0 g of Zonisa- mide according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 25 mg of Zonisamide in 8 mL of tetrahydrofuran, add water to make 50 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than zonisamide obtained from the sample ‌solution is not larger than 1/5 times the peak area of zonisamide from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of zonisamide, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 3 mL of the standard solution, add the mobile phase to make exactly 50 mL. Con-firm that the peak area of zonisamide obtained with 10 mL of this solution is equivalent to 4.2 to 7.8z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of zonisamide are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of zonisamide is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.1 g of Zonisamide, previ-ously dried, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Zonisamide RS, previously dried, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of zonisamide to that of the internal standard is not more than 1.0z.
Zopiclone occurs as a white to pale yellow crystalline powder. It is slightly soluble in ethanol (99.5), and practically in-soluble in water. It dissolves in 0.1 mol/L hydrochloric acid TS. It is gradually colored to pale brown by light. A solution of Zopiclone in 0.1 mol/L hydrochloric acid TS (1 in 40) shows no optical rotation. Melting point: 175 – 1789C Zopiclone shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Zopiclone in 0.1 mol/L hydrochloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectropho- Time span of measurement: About 1.5 times as long as the retention time of zopiclone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of zopiclone obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of zopiclone are not less than 7500 and not more than 1.5, respectively. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of zopiclone is not more than 3.0z. Loss on drying <2.41> Not more than 0.5z (2 g, in vacu-um, 1009C, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Zopiclone, dissolve in 50 mL of a mixture of acetic anhydride and acetic acid (100) (4:1), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 38.88 mg of C17H17ClN6O3
D-galactose, L-arabinose and L-rhamnose monohydrate
Atropine Sulfate RS or atropine sulfate hydrate
Cnidium Rhizome 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Gardenia Fruit 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Forsythia Fruit 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Mentha Herb 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Schizonepeta Spike 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Ephedra Herb 1.2 g 1.2 g 1.2 g 1.2 g 1.2 g
Atractylodes Rhizome 3 g 3 g Atractylodes Lancea Rhizome - -
Berberine Chloride RS or berberin chloride hydrate
carbazochrome sodium sulfonate trihydrate
Glycyrrhizic Acid RS or glycyrrhizic acid
Polyporus Sclerotium 3 g 4.5 g 4.5 g 3 g
Processed Ginger 2.5 g 3 g Ginger - -
Rhizom - 4 g - 4 g - Astragalus Root 4 g 4 g 4 g 4 g 3 g
Glycyrrhiza 1.5 g 1.5 g 1.5 g 1.5 g 1.5 g
Ginger 0.5 g 0.5 g 0.5 g 0.5 g 0.5 g Processed Ginger - - - - -
Cimicifuga Rhizome 1 g 1 g 0.5 g 0.5 g 1 g
Atractylodes Lancea Rhizome - 3 g - Poria Sclerotium 3 g 3 g 3.5 g
Rhizome - 3 g - 3 g - Poria Sclerotium 3 g 3 g 3 g 3 g 3 g
powder of Nux Vomica with hexane, extract with the percolation method, using a mixture of 750 mL of Ethanol,
Berberine Chloride RS or berberine chloride hydrate
Atractylodes Rhizome 4 g Atractylodes Lancea Rhizome -
Atractylodes Rhizome 3 g Atractylodes Lancea Rhizome -
Atractylodes Rhizome 4.5 g Atractylodes Lancea Rhizome -
Atractylodes Rhizome 3 g - 3 g Atractylodes Lancea Rhizome - 3 g -
Atractylodes Rhizome 4 g 4 g 4 g Atractylodes Lancea Rhizome - - -
observed at the position corresponding to the spot from the standard solution
Main root or main root with some lateral roots, with or without short remains of rhizome at the crown; main root, long cylindrical and sometimes somewhat tortuous, 15 – 90 cm in length, 0.3 – 0.7 cm in diameter; externally grayish yellow to yellow-brown, with numerous longitudinal wrinkles, and with scattering scars of lateral roots. Fractured surface is flat; grayish white to light brown on the circumference, and with yellow-white xylem in the center. Hard and brittle, or flexible. Odor, slight; taste, slightly sweet, and mucilaginous. Under a microscope <5.01>, a transverse section reveals a rather distinct cambium separating the cortex from the xylem; small protoxylem located at the center of the xylem, and surrounded by numerous vascular bundles arranged on several concentric circles; parenchyma cells containing sand crystals of calcium oxalate; starch grains absent.
Shake vigorously 0.5 g of pulverized Achyranthes Root with 10 mL of water: a lasting fine foam is produced.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Achyranthes Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Achyranthes Root according to Method 4, and perform the test (not more than 5 ppm). Stem—When perform the test of foreign matter <5.01>, the amount of stems contained in Achyranthes Root does not exceed 5.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than stems contained in Achyranthes Root does not exceed 1.0z. Loss on drying <5.01> Not more than 17.0z (6 hours). Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
1) Powdered Processed Aconite Root 1: Powdered Processed Aconite Root 1 occurs as a light grayish brown powder. It has a characteristic odor. Under a microscope <5.01>, Powered Processed Aconite Root 1 reveals gelatinized starch masses or starch grains and parenchymatous cells containing them, fragments of red-brown metaderm, fragments of pitted, scaraliform, reticu-late and spiral vessels; also square to oblong-square scleren-chymatous cells, 30 – 150 mm in diameter, 100 – 250 mm in length, cell wall of sclerenchymatous cells, 6 – 12 mm in thickness; starch grains of Aconitum carmichaeli Debeaux or Aconitum japonicum Thunberg (Ranunculaceae) origin, simple, spherical or ellipsoid, 2 – 25 mm in diameter, or 2- to a dozen or so- compound, hilum of starch grain distinct. 2) Powdered Processed Aconite Root 2: Powdered Processed Aconite Root 2 occurs as a light yellow-white layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia water (28) (40:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, air-dry the plate, and spray evenly sodium nitrite TS: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the stand-ard solution.
To 3 g of Powdered Processed Aconite Root in a glass-stoppered centrifuge tube add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes, centrifuge, and take the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), dissolve the residue in 1 mL of diethyl ether, and use this solution as the sample so-lution. Separately, dissolve 1 mg of benzoylmesaconine hy-drochloride for thin-layer chromatography in 5 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- CSJ: Concentration ( mg/mL) of jesaconitine for purity in aconitum diester alkaloids standard solution for puri-ty CSH: Concentration ( mg/mL) of hypaconitine for purity in aconitum diester alkaloids standard solution for purity CSM: Concentration ( mg/mL) of mesaconitine for purity in aconitum diester alkaloids standard solution for purity M: Amount (g) of Powdered Processed Aconitine Root taken, calculated on the dried basis ‌JP XVIII Crude Drugs and Related Drugs / Processed Ginger 2109 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: Adjust so that the retention time of mesaconi-tine is about 31 minutes. System suitability— System performance: When the procedure is run with 20 mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between their peaks is not less than 1.5, respectively. System repeatability: To 1 mL of aconitum diester alkaloids standard solution for purity add a mixture of phos-phate buffer solution for processed aconite root and aceto-nitrile (1:1) to make 10 mL. When the test is repeated 6 times with 20 mL of this solution under the above operating condi-tions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5z. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Powdered Processed Aconite Root 1: Not more than 4.0z. Powdered Processed Aconite Root 2: Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 0.7z.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Processed Aconite Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Processed Aconite Root according to Method 4, and perform the test (not more than 5 ppm). Aconitum diester alkaloids (aconitine, jesaconitine, hypaconitine and mesaconitine)—Weigh accurately about 0.5 g of Powdered Processed Aconite Root, put in a glass-stoppered centrifuge tube, suspend in 3.0 mL of water by shaking, and add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 1.0 mL of ammo-nia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, evaporate the solvent under low pressure (in vacuo), and dissolve the residue with exactly 10 mL of a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1). Centrifuge this solution, and use the supernatant liquid as the sample solution. Perform the test with exactly 20 mL each of the sample solution and aconitum diester alkaloids stand-ard solution for purity as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine the heights of the peaks corresponding to aconitine, HTA and HSA, jesaconitine, HTJ and HSJ, hypaconitine, HTH and HSH, and mesaconitine, HTM and HSM, respectively, and calculate the amounts of them by the following formulae: the amounts of aconitine, jesaconitine, hypaconitine and mesaconitine per g calculated on the dried basis are not more than 55 mg, 40 mg, 55 mg and 120 mg, respectively, and the total amount of them is not more than 230 mg. Amount ( mg) of aconitine ＝ CSA/M × HTA/HSA × 10 Amount ( mg) of jesaconitine ＝ CSJ/M × HTJ/HSJ × 10 Amount ( mg) of hypaconitine powder. It has a characteristic odor. Under a microscope <5.01>, Powered Processed Aconite ＝ CSH/M × H TH/HSH × 10 Root 2 reveals gelatinized starch masses and parenchyma-tous cells containing them, fragments of red-brown metaderm, fragments of pitted, scaraliform, reticulate and spiral vessels; also square to oblong-square sclerenchyma-tous cells, 30 – 150 mm in diameter, 100 – 250 mm in length, cell wall of sclerenchymatous cells, 6 – 12 mm in thickness. CSA Amount ( mg) of mesaconitine ＝ CSM/M × HTM/HSM × 10 : Concentration ( mg/mL) of aconitine for purity in aconitum diester alkaloids standard solution for purity Identification To 3 g of Powdered Processed Aconite Root in a glass-stoppered centrifuge tube add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes, centrifuge, and take the diethyl ether layer. Evaporate the solvent under low pressure (in vacuo), dissolve the residue in 1 mL of diethyl ether, and use this solution as the sample so-lution. Separately, dissolve 1 mg of benzoylmesaconine hy-drochloride for thin-layer chromatography in 5 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin- CSJ: Concentration ( mg/mL) of jesaconitine for purity in aconitum diester alkaloids standard solution for puri-ty CSH: Concentration ( mg/mL) of hypaconitine for purity in aconitum diester alkaloids standard solution for purity CSM: Concentration ( mg/mL) of mesaconitine for purity in aconitum diester alkaloids standard solution for purity M: Amount (g) of Powdered Processed Aconitine Root taken, calculated on the dried basis ‌JP XVIII Crude Drugs and Related Drugs / Processed Ginger 2109 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: Adjust so that the retention time of mesaconi-tine is about 31 minutes. System suitability— System performance: When the procedure is run with 20 mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between their peaks is not less than 1.5, respectively. System repeatability: To 1 mL of aconitum diester alkaloids standard solution for purity add a mixture of phos-phate buffer solution for processed aconite root and aceto-nitrile (1:1) to make 10 mL. When the test is repeated 6 times with 20 mL of this solution under the above operating condi-tions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5z. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Powdered Processed Aconite Root 1: Not more than 4.0z. Powdered Processed Aconite Root 2: Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 0.7z.
Weigh accurately about 2 g of Powdered Processed Aconite Root, put in a glass-stoppered centrifuge tube, and add 1.6 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 0.8 mL of ammonia TS and 20 mL of diethyl ether, and proceed as above. Repeat this process more three times. Combine all the extracts, evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in 5 mL of ethanol (99.5), add 30 mL of freshly boiled and cooled water, and titrate <2.50> with 0.01 mol/L hydrochloric acid VS until the color of the solution changes from green to gray-blue through blue-green (indicator: 3 drops of methyl red-methylene blue TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.01 mol/L hydrochloric acid VS ＝ 6.037 mg of total alkaloid [as benzoylaconine (C32H45NO10)]
1) Processed Aconite Root 1: Cut pieces ir-regularly polygonal, less than 10 mm in diameter; externally dark grayish brown to black-brown; hard in texture; cut sur-face flat, light brown to dark brown, usually horny and lus-trous. Odor, weak and characteristic. Under a microscope <5.01>, transverse and longitudinal sections reveal pitted, scaraliform, reticulate and spiral vessels; starch grains in parenchymatous cells usually gelatinized but sometimes not gelatinized; starch grains, simple, spherical or ellipsoid, 2 – 25 mm in diameter, or 2- to a dozen or so- compound, hilum of starch grain distinct. Processed Aconite Root 2: Nearly obconical root, 15 – 30 mm in length, 12 – 16 mm in diameter, slices cut longitu-dinally or transversely, 20 – 60 mm in length, 15 – 40 mm in width, and 200 – 700 mm in thickness, or cut pieces irregular-ly polygonal, less than 12 mm in diameter; externally light brown to dark brown or yellow-brown; hard in texture, usu-ally without wrinkles; cut surface flat, light brown to dark brown or yellow-white to light yellow-brown, usually horny, semi-transparent and lustrous. Odor, weak and characteristic. Under a microscope <5.01>, transverse and longitudinal sections reveal metaderm, primary cortex, endodermis, secondary cortex, cambium, and xylem; primary cortex con-tains oblong to oblong-square sclerenchymatous cells, 30 – 75 mm in short axis, 60 – 150 mm in long axis; endodermis single layered cell, endodermal cells elongated in tangential direction; cambium, star shaped or irregular polygons to orbicular; a group of vessel in xylem v-shaped; sometimes isolated ring of cambium appears in secondary cortex or in pith; vessels, pitted, scaraliform, reticulate and spiral; starch grains in parenchymatous cells gelatinized. Processed Aconite Root 3: Cut pieces irregularly poly-gonal, less than 5 mm in diameter; externally grayish brown; hard in texture; cut surface flat, light grayish brown to grayish white, not lustrous. Odor, weak and characteristic. Under a microscope <5.01>, transverse and longitudinal sections reveal pitted, scaraliform, reticulate and spiral ves- JP XVIII Crude Drugs and Related Drugs / Processed Aconite Root 2107 sels; starch grains, simple, spherical or ellipsoid, 2 – 25 mm in diameter, or 2- to a dozen or so- compound, hilum of starch grain distinct.
To 3 g of pulverized Processed Aconite Root in a glass-stoppered centrifuge tube add 20 mL of diethyl ether and 2 mL of ammonia TS, shake for 10 minutes, cen-trifuge, and take the diethyl ether layer. Evaporate the sol-vent under low pressure (in vacuo), dissolve the residue in 1 mL of diethyl ether, and use this solution as the sample solu-tion. Separately, dissolve 1 mg of benzoylmesaconine hydro-chloride for thin-layer chromatography in 5 mL of ethanol (99.5), and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia water (28) (40:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, air-dry the plate, and spray evenly sodium nitrite TS: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the stand-ard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Processed Aconite Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Processed Aconite Root according to Method 4, and perform the test (not more than 5 ppm). Aconitum diester alkaloids (aconitine, jesaconitine, hypaconitine and mesaconitine)—Weigh accurately about 0.5 g of pulverized Processed Aconite Root, put in a glass-stoppered centrifuge tube, suspend in 3.0 mL of water by shaking, and add 1.0 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 1.0 mL of ammo-nia TS and 20 mL of diethyl ether, and repeat the above process twice more. Combine all the extracts, evaporate under low pressure (in vacuo), and dissolve the residue with exactly 10 mL of a mixture of phosphate buffer solution for processed aconite root and acetonitrile (1:1). Centrifuge and use the supernatant liquid as the sample solution. Perform the test with exactly 20 mL each of the sample solution and aconitum diester alkaloids standard solution for purity as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine the heights of the peaks corresponding to aconitine, HTA and HSA, jesaconitine, HTJ and HSJ, hypaconitine, HTH and HSH, and mesaconitine, HTM and HSM, respectively, and calculate the amounts of them by the following formulae: the amounts of aconitine, jesaconitine, hypaconitine and mesaconitine per g calculated on the dried basis are not more than 60 mg, 60 mg, 280 mg and 140 mg, respectively, and the total amount of them is not more than 450 mg. Amount ( mg) of aconitine ＝ CSA/M × HTA/HSA × 10 Amount ( mg) of jesaconitine CSA: Concentration ( mg/mL) of aconitine for purity in aconitum diester alkaloids standard solution for purity CSJ: Concentration ( mg/mL) of jesaconitine for purity in aconitum diester alkaloids standard solution for puri-ty CSH: Concentration ( mg/mL) of hypaconitine for purity in aconitum diester alkaloids standard solution for purity CSM: Concentration ( mg/mL) of mesaconitine for purity in aconitum diester alkaloids standard solution for purity M: Amount (g) of Processed Aconite Root taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 231 nm for aconitine, hypaconitine and mesaconi-tine; 254 nm for jesaconitine). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of phosphate buffer solution for processed aconite root and tetrahydrofuran (183:17). Flow rate: Adjust so that the retention time of mesaconi-tine is about 31 minutes. System suitability— System performance: When the procedure is run with 20 mL of aconitum diester alkaloids standard solution for purity under the above operating conditions, using 254 nm, mesaconitine, hypaconitine, aconitine and jesaconitine are eluted in this order, and each resolution between their peaks is not less than 1.5, respectively. System repeatability: To 1 mL of aconitum diester alkaloids standard solution for purity add a mixture of phos-phate buffer solution for processed aconite root and aceto-nitrile (1:1) to make 10 mL. When the test is repeated 6 times with 20 mL of this solution under the above operating condi-tions, using 231 nm, the relative standard deviation of the peak height of mesaconitine is not more than 1.5z. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Processed Aconite Root 1: Not more than 4.0z. Processed Aconite Root 2: Not more than 12.0z. Processed Aconite Root 3: Not more than 19.0z. Acid-insoluble ash <5.01> Not more than 0.9z.
Weigh accurately about 2 g of pulverized Processed Aconite Root, put in a glass-stoppered centrifuge tube, and add 1.6 mL of ammonia TS and 20 mL of diethyl ether, shake for 30 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 0.8 mL of ammonia TS and 20 mL of diethyl ether, and proceed as above. Repeat this process more three times. Combine all the extracts, evapo-rate the solvent under low pressure (in vacuo), dissolve the residue in 5 mL of ethanol (99.5), add 30 mL of freshly boiled and cooled water, and titrate <2.50> with 0.01 mol/L ＝ CSJ/M × H TJ/HSJ × 10 hydrochloric acid VS until the color of the solution changes from green to gray-blue through blue-green (indicator: 3 Amount ( mg) of hypaconitine ＝ CSH/M × HTH/HSH × 10 Amount ( mg) of mesaconitine ＝ CSM/M × HTM/HSM × 10 drops of methyl red-methylene blue TS). Perform a blank determination in the same manner, and make any necessary correction. ‌2108 Powdered Processed Aconite Root / Crude Drugs and Related Drugs Each mL of 0.01 mol/L hydrochloric acid VS ＝ 6.037 mg of total alkaloid [as benzoylaconine (C32H45NO10)]
Agar is the solid residue obtained by freezing
Circular or ellipsoidal sections 0.2 – 0.3 cm in thickness, and 1 – 3 cm in diameter; phloem on both frac-tured surfaces is dark grayish brown; xylem reveals light brown vessel portions and grayish white medullary rays lined alternately and radially; pith light grayish yellow, and dis-tinct; flank grayish brown, and with circular or transversely elongated elliptical lenticels. Almost odorless; slightly acrid taste. Under a microscope <5.01>, a transverse section reveals ring layers mainly consisting of fiber bundles with crystal cells and stone cell groups and surrounding the outside of the phloem in arc shape. Medullary rays of the cortex consisting of sclerenchyma cells containing solitary crystals; portion near cambium is distinct; cells around the pith remarkably thick-walled; xylem medullary rays and parenchyma cells around the pith contain solitary crystals of calcium oxalate and starch grains less than 8 mm in diameter.
To 0.5 g of pulverized Akebia Stem add 10 mL of water, boil, allow to cool, and shake vigorously: lasting fine foams are produced. Total ash <5.01> Not more than 10.0z.
Powdered Alisma Tuber occurs as a light grayish brown powder, and has a slight odor and a slightly bitter taste. Under a microscope <5.01>, Powdered Alisma Tuber re- veals mainly starch grains, fragments of parenchyma con-taining them, parenchyma cells containing yellow contents, and fragments of vascular bundles. Starch grains, spheroidal to ellipsoidal simple grains, 3 – 15 mm in diameter.
To 1.0 g of Powdered Alisma Tuber add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Use alisma tuber triterpenes TS for identification as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 1 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with a spot among the three spots from the standard solution.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Powdered Alisma Tuber according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Alisma Tuber according to Method 4, and per-form the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Spherical or conical tubers, 3 – 8 cm in length, 3 – 5 cm in diameter, sometimes a 2- to 4-branched irregular tuber; externally light grayish brown to light yellow-brown, and slightly annulate; many remains of root appearing as small warty protrusions; fractured surface nearly dense, the outer portion grayish brown, and the inner part white to light yellow-brown in color; rather light in texture and difficult to break. Slight odor and slightly bitter taste.
To 1.0 g of pulverized Alisma Tuber add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Use alisma tuber triterpenes TS for identification as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 1 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with a spot among the three spots from the standard solution.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Alisma Tuber according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Alisma Tuber according to Method 4, and per-form the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Powdered Aloe occurs as a dark brown to yel- phy. Develop the plate with a mixture of ethyl acetate, ace-tone, water and acetic acid (100) (20:5:2:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and the same Rf value with the red fluorescent spot from the standard solution.
(1) Dissolve 0.5 g of Powdered Aloe in 50 mL of water by warming. After cooling, add 0.5 g of siliceous earth, and filter. Perform the following tests with the filtrate as the sample solution. Dissolve 0.2 g of sodium tetraborate decahydrate in 5 mL of the sample solution by warming in a water bath. Add a few drops of this solution into 30 mL of water, and shake: a green fluorescence is produced. Shake 2 mL of the sample solution with 2 mL of nitric acid: a yellow-brown color which changes gradually to green is produced. Then warm this colored solution in a water bath: the color of the solution changes to red-brown. (2) To 0.2 g of Powdered Aloe add 10 mL of methanol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of barbaloin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra- MS: Amount (mg) of barbaloin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 360 nm). Column: A stainless steel column about 6 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (74:26:1). Flow rate: Adjust so that the retention time of barbaloin is about 12 minutes. System suitability— System performance: To about 10 mg of barbaloin for assay add 40 mg of oxalic acid dihydrate, and dissolve in methanol to make 100 mL. To 5 mL of the solution add 1 mL of a solution of ethenzamide in methanol (1 in 2000) and methanol to make 10 mL. When the procedure is run with 5 mL of this solution under the above operating conditions except the wavelength of 300 nm, barbaloin and ethenzamide are eluted in this order with the resolution between these ‌1944‌ Alpinia Officinarum Rhizome / Crude Drugs and Related Drugs peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of barbaloin is not more than 1.5z.
(1) Resin—Warm 0.5 of Powdered Aloe with 10 mL of diethyl ether on a water bath, and filter. Wash the residue and the filter paper with 3 mL of diethyl ether. Com-bine the filtrate and the washing, and evaporate the solvent: the mass of the residue does not exceed 5.0 mg. Ethanol-insoluble substances—Heat 1.0 g of Pow-dered Aloe with 50 mL of ethanol (95) for 30 minutes under a reflux condenser. Filter the warm mixture through a tared glass filter (G4), and wash the residue on the filter with ethanol (95) until the last washing becomes colorless. Dry the residue at 1059C for 5 hours, and weigh: the mass of the residue is not more than 0.10 g. Loss on drying <5.01> Not more than 12.0z. Total ash <5.01> Not more than 2.0z. Extract content <5.01> Water-soluble extract: not less than 40.0z.
Weigh accurately about 0.1 g of Powdered Aloe, add 40 mL of methanol, and heat under a reflex condenser for 30 minutes. After cooling, filter, and add methanol to the filtrate to make exactly 50 mL. Pipet 5 mL of the solu-tion, add methanol to make exactly 10 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of barbaloin for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 24 hours, add 40 mg of oxalic acid dihydrate, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of the solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of bar-baloin in each solution. lowish dark brown powder. It has a characteristic odor and an extremely bitter taste. Amount (mg) of barbaloin ＝ MS × AT/AS × 1/2 Under a microscope <5.01>, Powdered Aloe, immersed in olive oil or liquid paraffin, reveals greenish yellow to reddish brown, angular or rather irregular fragments. Identification (1) Dissolve 0.5 g of Powdered Aloe in 50 mL of water by warming. After cooling, add 0.5 g of siliceous earth, and filter. Perform the following tests with the filtrate as the sample solution. Dissolve 0.2 g of sodium tetraborate decahydrate in 5 mL of the sample solution by warming in a water bath. Add a few drops of this solution into 30 mL of water, and shake: a green fluorescence is produced. Shake 2 mL of the sample solution with 2 mL of nitric acid: a yellow-brown color which changes gradually to green is produced. Then warm this colored solution in a water bath: the color of the solution changes to red-brown. (2) To 0.2 g of Powdered Aloe add 10 mL of methanol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of barbaloin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra- MS: Amount (mg) of barbaloin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 360 nm). Column: A stainless steel column about 6 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (74:26:1). Flow rate: Adjust so that the retention time of barbaloin is about 12 minutes. System suitability— System performance: To about 10 mg of barbaloin for assay add 40 mg of oxalic acid dihydrate, and dissolve in methanol to make 100 mL. To 5 mL of the solution add 1 mL of a solution of ethenzamide in methanol (1 in 2000) and methanol to make 10 mL. When the procedure is run with 5 mL of this solution under the above operating conditions except the wavelength of 300 nm, barbaloin and ethenzamide are eluted in this order with the resolution between these ‌1944‌ Alpinia Officinarum Rhizome / Crude Drugs and Related Drugs peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of barbaloin is not more than 1.5z.
Aloe is the dried juice of the leaves mainly of Aloe
Spherical to fusiform fruit, with both ends somewhat pointed; 1 – 2 cm in length, 0.7 – 1 cm in width; externally brown to dark brown, with numerous longitudi-nal, knob-like protruding lines; pericarp 0.3 – 0.5 mm in thickness, closely adhering to the seed mass, and difficult to separate; inside divided vertically into three loculi by thin membranes, each loculus containing 5 to 8 seeds adhering by aril; seeds irregularly polygonal, about 3.5 mm in diameter, brown to dark brown in color, and hard in texture. Odor, characteristic; taste, slightly bitter. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 2.5z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Bitter Cardamon: the volume of essential oil is not less than 0.4 mL.
Alpinia Officinarum Rhizome is a slightly curved and cylindrical rhizome, sometimes branched; 2 – 8 cm in length, 0.6 – 1.5 cm in diameter; externally red-brown to dark brown with fine striped lines, grayish white nodes and several traces of rootlet; hard to break; fracture surface, light brown in color and thickness of cortex is approximately the same as that of stele. Odor, characteristic; taste, extremely pungent. Under a microscope <5.01>, a transverse section reveals epidermal cells often containing oil-like substances; cortex, endodermis and stele present beneath the epidermis; cortex and stele divided by endodermis; vascular bundles sur-rounded by fibers, scattered throughout the cortex and stele, cortex and stele composed of parenchyma interspersed with oil cells; parenchyma cells containing solitary crystals of calcium oxalate and starch grains, starch grains generally simple (sometimes 2- to 8-compound), narrowly ovate, ellip-soidal or ovate, 10 – 40 mm in diameter and with an eccentric navel.
To 0.5 g of pulverized Alpinia Officinarum Rhizome add 5 mL of acetone, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of cyclo-hexane, ethyl acetate and acetic acid (100) (12:8:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots ap-pear at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Alpinia Officinarum Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Alpinia Officinarum Rhizome according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 7.5z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-extract: not less than 14.0z.
Aluminum Silicate Hydrate with Silicon Dioxide
Powdered Amomum Seed occurs as a grayish brown powder, and has a characteristic aroma and an acrid taste. Under a microscope <5.01>, Powdered Amomum Seed reveals fragments of wavy perisperm cells filled with starch grains and containing in each cell a calcium oxalate crystal; yellow and long epidermal cells of seed coat and fragments of thin-walled tissue perpendicular to them; fragments of groups of brown, thick-walled polygonal stone cells.
To 2.0 g of Powdered Amomum Seed add 20 mL of hexane, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, use a mixture of hexane and borneol acetate (1000:1) as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, diethyl ether and metha-nol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Essential oil content <5.01> Perform the test with 30.0 g of Powdered Amomum Seed: the volume of essential oil is not less than 0.4 mL.
Approximately spherical or ellipsoidal mass, 1 – 1.5 cm in length, 0.8 – 1 cm in diameter; externally grayish brown to dark brown, and with white powder in those dried by spreading lime over the seeds; the seed mass is divided into three loculi by thin membranes, and each locu-lus contains 10 to 20 seeds joining by aril; each seed is poly-gonal and spherical, 0.3 – 0.5 cm in length, about 0.3 cm in diameter, externally dark brown, with numerous, fine pro-trusions; hard tissue; under a magnifying glass, a longitu-dinal section along the raphe reveals oblong section, with deeply indented hilum and with slightly indented chalaza; white perisperm covering light yellow endosperm and long embryo. Characteristic aroma when cracked, and taste acrid.
To 1.0 g of coarse powdered Amomum Seed add 20 mL of hexane, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, use a mixture of hexane and borneol acetate (1000:1) as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane, diethyl ether and methanol (15:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Essential oil content <5.01> Perform the test with 30.0 g of pulverized Amomum Seed: the volume of essential oil is not less than 0.6 mL.
Rather flat and cord-like rhizome, 3 – 15 cm in length, 0.5 – 1.5 cm in diameter, slightly bent and branched; externally yellow-brown to brown; on the upper surface, a longitudinal furrow and hair-like remains or scars of leaf sheath forming fine ring-nodes; on the lower surface, scars of root appearing as numerous round spot-like hollows; light and easily broken. Under a magnifying glass, a light yellow-brown transverse section reveals an extremely narrow cortex; stele porous, with many irregularly scattered vascular bun-dles. Odor, slight; taste, slightly sweet and mucous, followed by bitterness.
(1) Shake vigorously 0.5 g of pulverized Anemarrhena Rhizome with 10 mL of water in a test tube: a lasting fine foam is produced. Filter the mixture, and to 2 mL of the filtrate add 1 drop of iron (III) chloride TS: a dark green precipitate is produced. (2) To 1 g of pulverized Anemarrhena Rhizome add 10 mL of 1 mol/L hydrochloric acid TS, and heat under a reflux condenser for 30 minutes. After cooling, centrifuge, and remove the supernatant liquid. To the residue add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of sarsasapogenin for thin-layer chromatogra-phy in 1 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate ‌1946‌ Angelica Dahurica Root / Crude Drugs and Related Drugs of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (7:3) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Anemarrhena Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Anemarrhena Rhizome according to Method 4, and perform the test (not more than 5 ppm). Foreign matter <5.01>—The amount of fiber, originating from the dead leaves, and other foreign matters contained in Anemarrhena Rhizome is not more than 3.0z. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 2.5z.
Powdered Japanese Angelica Root occurs as a light grayish brown powder. It has a characteristic odor and a slight, sweet taste with a slightly pungent aftertaste. Under a microscope <5.01>, Powdered Japanese Angelica Root reveals starch grains or masses of gelatinized starch, and fragments of parenchyma containing them; fragments of light yellow-brown cork tissue; fragments of rather thick-walled collenchyma and phloem tissue; fragments of oil canal surrounded by secretory cells; fragments, 20 – 60 mm in diameter, of scalariform and reticulate vessels with simple perforation; starch grains composed of simple grains not more than 20 mm in diameter, and rarely 2- to 5-compound grains, sometimes comes up to 25 mm. Identiˆcation To 1.0 g of Powdered Japanese Angelica Root add 5 mL of methanol, shake for 10 minutes, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, use (Z)-ligustilide TS for thin-layer chro-matography as the standard solution (1). Dissolve 1 mg of scopoletin for thin-layer chromatography in 10 mL of methanol, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL each of the standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (30:25:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): two of the several spots obtained from the sample solu-tion have the same color tones and Rf values with the corre-sponding blue-white ‰uorescent spots from the standard so-lutions (1) and (2).
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Japanese Angelica Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Japanese Angelica Root according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Japanese Angelica Root does not show remarkably lignified sclerenchymatous cells. ‌2040‌‌ Japanese Gentian / Crude Drugs and Related Drugs Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 35.0z.
Thick and short main root, with numerous branched roots, nearly fusiform; 10 – 25 cm in length; exter-nally dark brown to red-brown, with longitudinal wrinkles and horizontal protrusions composed of numerous scars of fine rootlets; fractured surface is dark brown to yellow-brown in color, and smooth; and with a little remains of leaf sheath at the crown. Odor, characteristic; taste, slightly sweet, followed by slight pungency. Under a microscope <5.01>, a transverse section reveals to 10 cellular layers of cork, with several cellular layers of collenchyma inside of the layer; the cortex exhibits many oil canals surrounded by secretory cells and often large hollows appear; boundary of cortex and xylem is distinct; in the xylem, numerous vessels radiate alternately with medullary rays; vessels in the outer part of the xylem are singly or in several groups, and disposed rather densely in a cuneiform pattern, but vessels in the region of the center are scattered very sparsely; starch grains are simple grains, not more than 20 mm in diameter, and rarely 2- to 5-compound grains, some times up to 25 mm in diameter; starch grains often gelatinized. Identiˆcation To 1.0 g of pulverized Japanese Angelica Root add 5 mL of methanol, shake for 10 minutes, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, use (Z)-ligustilide TS for thin-layer chro-matography as the standard solution (1). Dissolve 1 mg of scopoletin for thin-layer chromatography in 10 mL of methanol, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL each of the standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (30:25:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): two of the several spots obtained from the sample solu-tion have the same color tones and Rf values with the corre-sponding blue-white ‰uorescent spots from the standard so-lutions (1) and (2).
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Japanese Angelica Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Japanese Angelica Root according to Method 4, and perform the test (not more than 5 ppm). Leaf sheath—When perform the test of foreign mat- ter <5.01>, the amount of leaf sheath contained in Japanese Angelica Root does not exceed 3.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than leaf sheath contained in Japanese Angelica Root does not exceed 1.0z. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 35.0z.
Main root from which many long roots are branched out and nearly fusiform and conical in whole shape, 10 – 25 cm in length; externally grayish brown to dark brown, with longitudinal wrinkles, and with numerous scars of rootlets laterally elongated and protruded. A few remains of leaf sheath at the crown and ring-nodes closely protruded near the crown. In a transverse section, the outer region is grayish white in color, and the central region is sometimes dark brown in color. Odor, characteristic; taste, slightly bitter.
To 0.2 g of pulverized Angelica Dahurica Root add 5 mL of ethanol (95), shake for 5 minutes, and filter. Examine the filtrate under ultraviolet light (main wavelength: 365 nm): a blue to blue-purple fluorescence develops.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Angelica Dahurica Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Angelica Dahurica Root according to Method 4, and perform the test (not more than 5 ppm). Leaf sheath—When perform the test of foreign mat-ter <5.01>, the amount of leaf sheath contained in Angelica Dahurica Root does not exceed 3.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than leaf sheath contained in Angelica Dahurica Root is not more than 1.0z. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
Aralia Rhizome is curved, irregular cylindrical to masses occasionally with remains of short roots. 4 – 12 cm in length, 2.5 – 7 cm in diameter, often cut crosswise or ‌1948‌ Areca / Crude Drugs and Related Drugs lengthwise. 1 to several, enlarged dents by remains of stems on the upper part or rarely 1.5 – 2.5 cm in diameter, remains of short stem. The outer surface is dark brown to yellow-brown, with longitudinally wrinkles, bases or dents of root. The transverse section of rhizome reveals dark brown to yel-low-brown, scattered brownish small spots with oil canals, and with numerous splits. Odor, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section of rhi-zome reveals the outermost layer to be cork layer, rarely composed of cork stone cells, followed these appeared several cellular layers of collenchyma. Vascular bundle and medullary rays is distinct, pith broad. Phloem fibre bundles are sometimes observed at the outer portion of phloem. Oil canals composed of schizogenous intercellular space in cor-tex and pith. Cortex composed of vessels, xylem fibres, and occasionally thick-wall xylem parenchyma. Vascular bundles scattered on the pith. And, parenchymatous cells observed rosette aggregates of calcium oxalate. Starch grains com-posed of simple grains, 2- to 6- compound grains.
To 1 g of pulverized Aralia Rhizome add 10 mL of methanol, shake for 5 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (30:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a purple spot appears at an Rf value of about 0.5.
Heavy metals <1.07>—Proceed with 1.0 g of pulver-ized Aralia Rhizome according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Loss on drying <5.01> Not more than 12.0z. Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 15.0z.
Burdock Fruit is slightly curved, long obovate achene, 5 – 7 mm in length, 2.0 – 3.2 mm in width, 0.8 to 1.5 mm in thickness; externally grayish brown to brown, with black spots; hollow about 1 mm in diameter at one broad end; flat, indistinct, longitudinal ridge at the other narrow end. 100 fruits weigh 1.0 – 1.5 g. Practically odorless; taste, bitter and oily. Under a microscope <5.01>, transverse section reveals an exocarp composed of an epidermis, mesocarp of slightly sclerified parenchyma, and endocarp of a single cellular layer of stone cells; seed coat composed of radially elongat-ed, sclerified epidermis, and parenchyma of several cellular layers; parenchymatous cells of the mesocarp contain a brown substance; stone cells of endocarp contain solitary, discrete crystals of calcium oxalate; cotyledons with starch grains, oil drops, aleurone grains, and minute crystals of cal-cium oxalate.
To 0.5 g of pulverized Burdock Fruit add 20 mL of methanol, shake for 10 minutes, filter, and use filtrate as the sample solution. Perform the test with the sample so-lution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of acetone, ethyl acetate and water (15:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfu-ric acid on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.4. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-extract: not less than 15.0z.
Rounded-conical or flattened nearly spherical seed 1.5 – 3.5 cm high and 1.5 – 3 cm in diameter; hilum at the center of its base and usually forming a dent; externally grayish red-brown to grayish yellow-brown, with a network of pale lines; hard in texture; cross section dense in texture, exhibiting a marbly appearance of grayish brown seed coat alternating with white albumen; center of the seed often hollow. Odor, slight; taste, astringent and slightly bitter.
To 1.0 g of pulverized Areca add 5 mL of 0.01 mol/L hydrochloric acid TS and 5 mL of ethyl acetate, shake for 15 minutes, centrifuge, and remove the ethyl acetate layer. To the aqueous layer add 1 mL of sodium hy-droxide TS and 5 mL of ethyl acetate, shake for 15 minutes, centrifuge, and use the ethyl acetate layer as the sample solu-tion. Separately, dissolve 1 mg of arecoline hydrobromide for thin-layer chromatography in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of acetone, water and acetic acid (100) (10:6:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS, air-dry, then spray evenly sodium nitrite TS: one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. The color of this spot fades immediately and then disappears after air-drying.
(1) Pericarp—When perform the test of foreign matter <5.01>, the amount of pericarp contained in Areca is not more than 2.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than the pericarp contained in Areca does not exceed 1.0z. Total ash <5.01> Not more than 2.5z.
Flattened, somewhat asymmetric ovoid seed, 1.1 – 1.8 cm in length, 0.8 – 1.3 cm in width, 0.4 – 0.7 cm in thickness; sharp at one end and rounded at the other end where chalaza situated; seed coat brown and its surface being powdery with rubbing easily detachable stone cells of epidermis; numerous vascular bundles running from chalaza throughout the seed coat, appearing as thin vertical furrows; seed coat and thin semitransparent white albumen easily separate from cotyledon when soaked in boiling water; cotyledon, white in color. Almost odorless; taste, bitter and oily. Under a microscope <5.01>, surface of epidermis reveals stone cells on veins protruded by vascular bundles, forming round polygon to ellipse and approximately uniform in shape, with uniformly thickened cell walls, and 60 – 90 mm in diameter; in lateral view, stone cell appearing obtusely trian-gular and its cell wall extremely thickened at the apex.
(1) When Apricot Kernel is knocked and ground together with water, the odor of benzaldehyde is produced. (2) To 1.0 g of ground Apricot Kernel add 10 mL of methanol, immediately heat under a reflux condenser for 10 minutes, cool, filter, and use the filtrate as the sample solu-tion. Separately, dissolve 2 mg of amygdalin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:5:4) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a spot with a bluish white fluorescence appears at an Rf value of about 0.7. Spray evenly thymol-sulfuric acid-methanol TS for spraying upon the plate, and heat the plate at 1059C for 5 minutes: one of the several spots from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Rancidity—Grind Apricot Kernel with hot water: no unpleasant odor of rancid oil is perceptible. (2) Foreign matter <5.01>—When perform the test with not less than 250 g of Apricot Kernel, it contains not more than 0.10z of fragments of endocarp. Loss on drying <5.01> Not more than 7.0z (6 hours).
Weigh accurately 0.5 g of ground Apricot Kernel, add 40 mL of diluted methanol (9 in 10), heat immediately ‌JP XVIII Crude Drugs and Related Drugs / Aralia Rhizome 1947 ‌under a reflux condenser for 30 minutes, and cool. Filter the mixture, add diluted methanol (9 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. Containers and Ethanol (3:1), mix well, and filter. Deter-mine the amount of hydrogen cyanide in the solution as directed in the Assay, and, if the amount is more than that specified above, dilute the solution to the specified concen-tration by the addition of the mixture of Purified Water or Purified Water in Containers and Ethanol (3:1). Description Apricot Kernel Water is a clear, colorless or pale yellow liquid. It has an odor of benzaldehyde and a characteristic taste. pH: 3.5 – 5.0 Identification To 2 mL of Apricot Kernel Water add 1 mL of ammonia TS, and allow to stand for 10 minutes: a slight Amount (mg) of amygdalin ＝ MS × AT/AS × 2 turbidity is produced. Allow to stand for 20 minutes: the tur-bidity is intensified. MS: Amount (mg) of amygdalin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihy-drogen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amygdalin is not more than 1.5z.
Capitulum, of ovoid to spherical, about 1.5 – 2 mm in length, about 2 mm in diameter, with linear leaves and pedicels. Outer surface of capitulum, light green to light yellow-brown in color; outer surface of leaf, green to green-brown; outer surface of pedicel, green-brown to dark brown. Under a magnifying glass, at the capitulum, involucral scale in 3 – 4 succubous rows; outer scale, of ovate with obtuse; inner scale, of elliptical, 1.5 mm in length, longer than outer one, with keel midrib and thin membranous margin. Floret, tubular; marginal ‰ower, of female; disk ‰ower, of her-maphrodite. Achene, of obovoid, 0.8 mm in length. Light in texture. Odor, characteristic, slight; taste, slightly acrid, which gives slightly numbing sensation to the tongue.
To 0.5 g of pulverized Artemisia Capillaris Flower add 10 mL of methanol, shake for 3 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone and hexane (1:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a principal spot with a blue fluorescence appears at an Rf value of about 0.5.
Stem—When perform the test of foreign matter <5.01>, Artemisia Capillaris Flower does not contain any ‌JP XVIII Crude Drugs and Related Drugs / Asiasarum Root 1949‌ stem more than 2 mm in diameter. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 15.0z.
Wrinkled leaves and their fragments, fre-quently with thin stems. The upper surface of leaf dark green, the lower surface covered densely with grayish white cotton-like hairs. When smoothed by immersion in water, unfolded laminas 4 – 15 cm long, 4 – 12 cm wide, 1- to 2-pinnately cleft or pinnately parted. Segments in 2 to 4 pairs, oblong-lanceolate to oblong, apex acuminate sometimes obtuse, margins irregularly lobed or entire. Small sized leaves tri-cleft or entire, lanceolate. Order, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section of leaf re-veals several-cells-layered collenchyma beneath epidermis of midvein; vascular bundles at the central portion of midvein, occasionally fiber bundles adjacent to phloem and xylem; laminas composed of upper epidermis, palisade tissue, spongy tissue and lower epidermis, long soft hairs, T-shaped hairs and glandular hairs on epidermis of laminas; epidermal cells contain tannin-like substances, parenchyma cells con-tain oil-like substances and tannin-like substances.
To 0.5 g of pulverized Artemisia Leaf (the parts like a floccose substance which are not easily pulver-ized may be removed) add 5 mL of a mixture of methanol and water (3:2), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg each of umbelliferone for thin-layer chroma-tography and scopoletin for thin-layer chromatography in 10 mL each of methanol, and use these solutions as the stand-ard solution (1) and the standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample so-lution and 5 mL each of the standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (20:10:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): two of the several spots obtained from the sample solution have the same color tone and Rf value with the cor-responding blue-white fluorescent spots from the standard solutions (1) and (2). System suitability—(Ultraviolet light (main wavelength: 365 nm)). To 1 mL of the standard solution (1) add methanol to make 10 mL. Confirm that when perform the test with 1 mL of this solution under the above conditions, a bluish white fluorescent spot is detectable.
Artemisia argyi—To 0.5 g of powdered Artemisia Leaf (the parts like a floccose substance which are not easily pulverized may be removed) add 5 mL of a mixture of meth-anol and water (3:2), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, to 0.5 g of artemisia･argyi for purity test add 5 mL of a mix-ture of methanol and water (3:2), shake for 10 minutes, cen-trifuge, and use the supernatant liquid as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (20:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): no spot appears from the sam-ple solution at the position of the green fluorescent spot (Rf value of about 0.5) obtained from the standard solution. Loss on drying <5.01> Not more than 14.0z. Total ash <5.01> Not more than 13.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 16.0z.
Asiasarum Root is a nearly cylindrical rhizome with numerous thin and long roots, externally light brown to dark brown. The root, about 15 cm in length, about 0.1 cm in diameter, with shallow longitudinal wrinkles on the sur-face, and brittle. The rhizome, 2 – 4 cm in length, 0.2 – 0.3 cm in diameter, often branched, with longitudinal wrinkles on the surface; internode short; each node has several scars of petiole and peduncle, and several thin and long roots. Odor, characteristic; taste, acrid, which gives slightly numbing sensation to the tongue.
To 1 g of pulverized Asiasarum Root add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of asarinin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sul-furic acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample ‌1950‌ Asparagus Root / Crude Drugs and Related Drugs solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Asiasarum Root according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Asiasarum Root according to Method 4, and perform the test (not more than 5 ppm). Terrestrial part—When perform the test of foreign matter <5.01>, any terrestrial parts are not found. Foreign matter <5.01>—The amount of foreign mat- ter other than terrestrial part contained in Asiasarum Root is not more than 1.0z. Aristolochic acid I—To exactly 2.0 g of pulverized Asiasarum Root add exactly 50 mL of diluted methanol (3 in 4), shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve exactly 1.0 mg of aristolochic acid I for crude drugs purity test in diluted meth-anol (3 in 4) to make exactly 100 mL. Pipet 1 mL of this so-lution, add diluted methanol (3 in 4) to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the following conditions: the sample so- lution shows no peak at the retention time corresponding to aristolochic acid I from the standard solution. If the sample solution shows such a peak, repeat the test under different conditions to confirm that the peak in question is not aristolochic acid I. Operating conditions— Detector: An ultraviolet or visible absorption photometer (wavelength: 400 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution prepared by dis-solving 7.8 g of sodium dihydrogen phosphate dihydrate and 2 mL of phosphoric acid in water to make 1000 mL and acetonitrile (11:9). Flow rate: Adjust so that the retention time of aristolochic acid I is about 15 minutes. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add diluted methanol (3 in 4) to make exactly 10 mL. Confirm that the ratio, S/N, of the signal (S) and noise (N) of aristolochic acid I obtained with 20 mL of this solution is not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of aristolochic acid I is not more than 5.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Essential oil content <5.01> Perform the test with 30.0 g of pulverized Asiasarum Root: the volume of essential oil is not less than 0.6 mL.
Fusiform to cylindrical tubers, 5 – 15 cm in length, 5 – 20 mm in diameter; externally light yellow-brown to light brown, translucent and often with longitudinal wrin-kles; flexible, or hard and easily broken in texture; fractured surface, grayish yellow, glossy and horny. Odor, characteristic; taste, sweet at first, followed by a slightly bitter aftertaste. Under a microscope <5.01>, a transverse section reveals stone cells and their groups scattered on outer layer of cor-tex; mucilaginous cells containing raphides of calcium oxa-late in the parenchyma cells of cortex and stele; no starch grains.
To 1 g of the coarse cutting of Asparagus Root add 5 mL of a mixture of 1-butanol and water (40:7), shake for 30 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of 1-butanol, water and acetic acid (100) (10:6:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfu-ric acid on the plate, and heat the plate at 1059C for 2 minutes: the spot of a red-brown at first then changes to brown color appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Asparagus Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Asparagus Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 18.0z (6 hours). Total ash <5.01> Not more than 3.0z.
Merrill (Liliaceae), from which most of the
Nearly cylindrical root, 30 – 100 cm in length, 0.7 – 2 cm in diameter, with small bases of lateral root dis-persed on the surface, twisted near the crown; externally light grayish yellow to light yellow-brown, and covered with irregular, dispersed longitudinal wrinkles and horizontal JP XVIII Crude Drugs and Related Drugs / Powdered Atractylodes Lancea Rhizome 1951 ‌lenticel-like patterns; difficult to break; fractured surface fibrous. Under a magnifying glass, a transverse section reveals an outer layer composed of periderm; cortex light yellowish white, xylem light yellow, and zone near the cam-bium somewhat brown in color; thickness of cortex from about one-third to one-half of the diameter of xylem; white medullary ray from xylem to cortex in thin root, but often appearing as radiating cracks in thick root; usually pith unobservable.‌ Odor, slight; taste, sweet.
To 1 g of pulverized Astragalus Root add 5 mL of potassium hydroxide TS and 5 mL of acetonitrile in a glass-stoppered centrifuge tube. After shaking this for 10 minutes, centrifuge, and use the acetonitrile layer as the sam-ple solution. Separately, dissolve 1 mg of astragaloside IV for thin-layer chromatography in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, methanol and water (20:5:4) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and exa-mine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the yellow-brown fluorescent spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Astragalus Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Astragalus Root according to Method 4, and perform the test (not more than 5 ppm). Root of Hedysarum species and others—Under a microscope <5.01>, a vertical section of Astragalus Root re-veals no crystal fiber containing solitary crystals of calcium oxalate outside the fiber bundle. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Powdered Atractylodes Lancea Rhizome oc-curs as a yellow-brown powder. It has a characteristic odor, and a slightly bitter taste. Under a microscope <5.01>, Powdered Atractylodes Lan- cea Rhizome reveals mainly parenchyma cells, spherocrystals of inulin, fragments of parenchyma cells containing small needle crystals of calcium oxalate as their contents; and fur-ther fragments of light yellow thick-walled fibers, stone cells and cork cells; a few fragments of reticulate and scalariform vessels, and small yellow-brown secreted masses or oil drops; starch grains absent.
To 2.0 g of Powdered Atractylodes Lancea Rhizome add 5 mL of hexane, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test ‌1952‌ Atractylodes Rhizome / Crude Drugs and Related Drugs with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and acetic acid (100) (10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a grayish green spot appears at an Rf value of about 0.5.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Atractylodes Lancea Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Atractylodes Lancea Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Atractylodes Lancea Rhizome: the volume of essential oil is not less than 0.5 mL.
Irregularly curved, cylindrical rhizome, 3 – 10 cm in length, 1 – 2.5 cm in diameter; externally dark grayish brown to dark yellow-brown; a transverse section nearly orbicular, with light brown to red-brown secretes as fine points. Often white cotton-like crystals produced on its surface. Odor, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section usually reveals periderm with stone cells; parenchyma of cortex, usually without any fiber bundle; oil sacs, containing light brown to yellow-brown substances, located at the end region of medullary rays; xylem exhibits vessels surrounded by fiber bundles and arranged radially on the region adjoining the cambium; pith and medullary rays exhibit the same oil sacs as in the cortex; parenchyma cells contain spherocrystals of inulin and small needle crystals of calcium oxalate.
To 2.0 g of pulverized Atractylodes Lancea Rhizome add 5 mL of hexane, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and acetic acid (100) (10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a grayish green spot appears at an Rf value of about 0.5.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Atractylodes Lancea Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Atractylodes Lancea Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Atractylodes Lancea Rhizome: the volume of essential oil is not less than 0.7 mL.
Powdered Atractylodes Rhizome occurs as a light brown to yellow-brown powder, and has a characteris-tic odor and a slightly bitter or slightly sweet taste, followed by a slightly bitter aftertaste. Under a microscope <5.01>, Powdered Atractylodes Rhi- zome reveals mainly parenchyma cells, crystals of inulin and fragments of parenchyma cells containing small needle crystals of calcium oxalate; fragments of light yellow thick-walled fibers, stone cells and cork cells; a few fragments of reticulate and scalariform vessels; small yellow-brown se-crete masses or oil droplets; starch grains absent.
To 2.0 g of Powdered Atractylodes Rhizome add 5 mL of hexane, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra- ‌JP XVIII Crude Drugs and Related Drugs / Bakumondoto Extract 1953‌ phy <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetic acid (100) (10:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.6.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Powdered Atractylodes Rhizome according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Atractylodes Rhizome according to Method 4, and perform the test (not more than 5 ppm). Atractylodes lancea rhizome—When proceed as di-rected in the Identification, using exactly 5 mL of hexane, any grayish green spot does not appear at an Rf value of about 0.5, immediately below the red-purple spot appeared at an Rf value of about 0.6. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Atractylodes Rhizome: the volume of essential oil is not less than 0.4 mL.
1) Wa-byakujutsu—Periderm-removed rhi-zome is irregular masses or irregularly curved cylinder, 3 – 8 cm in length, 2 – 3 cm in diameter; externally light grayish yellow to light yellowish white, with scattered grayish brown parts. The rhizome covered with periderm is externally grayish brown, often with node-like protuberances and coarse wrinkles. Difficult to break, and the fractured surface is fibrous. A transverse section, with fine dots of light yel-low-brown to brown secrete. Odor, characteristic; taste, somewhat bitter. Under a microscope <5.01>, a transverse section reveals periderm with stone cell layers; fiber bundles in the paren-chyma of the cortex, often adjoined to the outside of the phloem; oil sacs containing light brown to brown substances, situated at the outer end of medullary rays; in the xylem, radially lined vessels, surrounding large pith, and distinct fiber bundle surrounding the vessels; in pith and in medul-lary rays, oil sacs similar to those in cortex, and in paren-chyma, crystals of inulin and small needle crystals of calcium oxalate. 2) Kara-byakujutsu—Irregularly enlarged mass, 4 – 8 cm in length, 2 – 5 cm in diameter; externally grayish yellow to dark brown, having sporadic, knob-like small protrusions. Difficult to break; fractured surface has a light brown to dark brown xylem remarkably fibrous. Odor, characteristic; taste, somewhat sweet, but followed by slight bitterness. Under a microscope <5.01>, a transverse section usually reveals periderm with stone cells, absence of fibers in the cortex; oil sacs containing yellow-brown contents in phloem ray and also at the outer end of it; xylem with radially lined vessels surrounding large pith, and distinct fiber bundle sur-rounding the vessels; pith and medullary ray exhibit oil sacs as in cortex; parenchyma contains crystals of inulin and small needle crystals of calcium oxalate.
To 2.0 g of pulverized Atractylodes Rhizome add 5 mL of hexane, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetic acid (100) (10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.6.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Atractylodes Rhizome according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Atractylodes Rhizome according to Method 4, and perform the test (not more than 5 ppm). Atractylodes lancea rhizome—When proceed as di-rected in the Identification, using exactly 5 mL of hexane, any grayish green spot does not appear at an Rf value of about 0.5, immediately below the red-purple spot appeared at an Rf value of about 0.6. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Atractylodes Rhizome: the volume of essential oil is not less than 0.5 mL.
Nearly spherical fruit, 1 – 2 cm in diameter, or semispherical, 1.5 – 4.5 cm in diameter; external surface, deep green-brown to brown, and without luster, with numerous small dents associated with oil sacs; the outer por-tion of transverse section exhibits pericarp and mesocarp about 0.4 cm in thickness, yellow-brown in color in the region contacting epidermis, and light grayish brown color in the other parts; the central portion is radially divided into 8 to 16 small loculi; each loculus is brown and indented, often containing immature seeds. Odor, characteristic; taste, bitter.
To 0.5 g of pulverized Immature Orange add 10 mL of methanol, boil gently for 2 minutes, and filter. To 5 mL of the filtrate add 0.1 g of magnesium ribbon and 1 mL of hydrochloric acid, and allow to stand: a red-purple color develops. Total ash <5.01> Not more than 7.0z.
Belladonna Total Alkaloids contains not less than
Cylindrical root, usually 10 – 30 cm in length, 0.5 – 4 cm in diameter; often cut crosswise or lengthwise; ex- Amount (mg) of hyoscyamine (C 17H 23NO3) ternally grayish brown to grayish yellow-brown, with longitudinal wrinkles; periderm often removed; fractured surface is light yellow to light yellow-brown in color and is powdery. Almost odorless.
Place 2.0 g of pulverized Belladonna Root in a glass-stoppered centrifuge tube, add 30 mL of ammonia TS, and centrifuge after irradiation of ultrasonic waves for 5 minutes. Transfer the supernatant liquid to a separator, add 40 mL of ethyl acetate, and shake. Drain off the ethyl ace-tate layer, add 3 g of anhydrous sodium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent of the filtrate to dryness under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. Separately, dissolve 2 mg of Atropine Sulfate RS or atropine sulfate hydrate for thin-layer chromatography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, water and ammonia water (28) (90:7:3) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS on the plate: the principal spot obtained from the sample solution is the same in color tone and Rf value with the spot from the standard solution.
(1) Stem and crown—When perform the test of foreign matter <5.01>, the amount of stem and crowns con-tained in Belladonna Root does not exceed 10.0z. (2) Foreign matter <5.01>—The amount of foreign ＝ MS × QT/QS × 1/5 × 0.855 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption spectrometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 900 mL of water, add 10 mL of triethylamine, adjust with phosphoric acid to pH 3.5, and add water to make 1000 mL, and mix this solution with acetonitrile (9:1). Flow rate: Adjust so that the retention time of atropine is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, atropine and the internal standard are eluted in this order with the resolution between these peaks being not less than 4.
Weigh accurately about 0.7 g of pulverized Bella-donna Root, previously dried at 609C for 8 hours, place in a glass-stoppered centrifuge tube, and moisten with 15 mL of ammonia TS. To this add 25 mL of diethyl ether, stopper the centrifuge tube tightly, shake for 15 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 25 mL of diethyl ether, proceed in the same manner, and repeat this procedure twice. Combine all the extracts, and evaporate the solvent on a water bath. Dissolve the residue in 5 mL of the mobile phase, add exactly 3 mL of the internal standard so-lution, and add the mobile phase to make 25 mL. Filter this solution through a filter of a porosity of not more than 0.8 mm, discard the first 2 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately about 25 mg of Atropine Sulfate RS (previously deter-mine the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, add exactly 3 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Calculate the ratios, QT and QS, of the hyoscyamine (C 17H23 NO3: 289.37). peak area of hyoscyamine (atropine), to that of the internal standard. Description Cylindrical root, usually 10 – 30 cm in length, 0.5 – 4 cm in diameter; often cut crosswise or lengthwise; ex- Amount (mg) of hyoscyamine (C 17H 23NO3) ternally grayish brown to grayish yellow-brown, with longitudinal wrinkles; periderm often removed; fractured surface is light yellow to light yellow-brown in color and is powdery. Almost odorless. Identification Place 2.0 g of pulverized Belladonna Root in a glass-stoppered centrifuge tube, add 30 mL of ammonia TS, and centrifuge after irradiation of ultrasonic waves for 5 minutes. Transfer the supernatant liquid to a separator, add 40 mL of ethyl acetate, and shake. Drain off the ethyl ace-tate layer, add 3 g of anhydrous sodium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent of the filtrate to dryness under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. Separately, dissolve 2 mg of Atropine Sulfate RS or atropine sulfate hydrate for thin-layer chromatography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, water and ammonia water (28) (90:7:3) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS on the plate: the principal spot obtained from the sample solution is the same in color tone and Rf value with the spot from the standard solution. Purity (1) Stem and crown—When perform the test of foreign matter <5.01>, the amount of stem and crowns con-tained in Belladonna Root does not exceed 10.0z. (2) Foreign matter <5.01>—The amount of foreign ＝ MS × QT/QS × 1/5 × 0.855 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption spectrometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 900 mL of water, add 10 mL of triethylamine, adjust with phosphoric acid to pH 3.5, and add water to make 1000 mL, and mix this solution with acetonitrile (9:1). Flow rate: Adjust so that the retention time of atropine is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, atropine and the internal standard are eluted in this order with the resolution between these peaks being not less than 4.
1) Benincasa cerifera origin—Flattened, ovate to orbicular ovate seed, 10 – 13 mm in length, 6 – 7 The amount (mg) of hyoscyamine (C ＝ MSA × QTA/QSA × 0.855 17H23NO3) mm in width, about 2 mm in thickness; slightly acute at base; hilum and germ pore form two protrusions; externally light grayish yellow to light yellowish brown; prominent The amount (mg) of scopolamine (C17H21NO4) ＝ MSS × QTS/QSS × 6/125 × 0.789 MSA: The amount (mg) of Atropine Sulfate RS taken, cal-culated on the dried basis MSS: The amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution: A solution of brucine n-hydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of around 209C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 900 mL of water, add 10 mL of triethylamine, adjust to pH 3.5 with phosphoric acid, and add water to make 1000 mL. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of atropine is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, scopolamine, atropine and the internal standard are eluted in this order, and the resolutions between scopolamine and atropine, and atropine and the internal standard are not less than 11 and not less than 4, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of scopolamine to that of the internal standard is not more than 1.5z.
To 0.5 g of pulverized Benincasa Seed add 10 mL of a mixture of methanol and water (4:1), shake for 10 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sam-ple solution on a plate of silica gel for thin-layer chromatog-raphy, develop the plate with a mixture of 1-butanol, water and acetic acid (100) (8:6:3) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): two blue-white fluorescent spots ap-pear at an Rf value of about 0.4, and the spot having the smaller Rf value shows more intense fluorescence.
Foreign matter <5.01>—It contains not more than 2.0z. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 5.0z. ‌1960‌‌ Benzoin / Crude Drugs and Related Drugs Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 3.0z.
Long cone or column shape, single or bran-ched root, 10 – 20 cm in length, 0.5 – 1.5 cm in diameter; oc-casionally with remains of stem on the crown; externally light brown to brown and sometimes with deep wrinkles; easily broken, and fractured surface somewhat fibrous. Un-der a magnifying glass, a transverse section reveals the thick-ness of cortex reaching 1/3 – 1/2 of the radius and tangen-tially extended clefts in cortex. Odor, characteristic, and taste, slightly bitter. Under a microscope <5.01>, a transverse section reveals the cortex scattered with a good many oil canals 15 – 35 mm in diameter; in xylem, vessels lined radially or in a staircase pat-tern, and fiber bundles scattered; in the pith at the crown, the same oil canals as in the cortex; parenchyma cells con-taining starch grains and oil droplets. Starch grains com-posed of simple grains, 2 – 10 mm in diameter, or compound grains.
(1) Shake vigorously 0.5 g of pulverized Bupleurum Root with 10 mL of water: lasting fine foams are produced. (2) To 1.0 g of the pulverized Bupleurum Root, add 10 mL of methanol, and boil gently under a reflux condenser for 15 minutes. After cooling, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of saikosapo-nin a for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the spot from the standard solution, accompa-nied by the adjacent yellow-red spot above.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Bupleurum Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Bupleurum Root according to Method 4, and perform the test (not more than 5 ppm). Stem and leaf—When perform the test of foreign matter <5.01>, the amount of the stems and leaves contained in Bupleurum Root does not exceed 10.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than stems and leaves contained in Bupleurum Root does not exceed 1.0z. Loss on drying <5.01> Not more than 12.5z (6 hours). Total ash <5.01> Not more than 6.5z. ‌JP XVIII Crude Drugs and Related Drugs / Byakkokaninjinto Extract 1969‌ Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 11.0z.
Weigh accurately about 1 g of pulverized Bupleurum Root, transfer in a glass-stoppered centrifuge tube, add 20 mL of diluted methanol (9 in 10), shake for 15 minutes, cen-trifuge, and separate the supernatant liquid. Perform the same procedure with the residue using two 15-mL potions of diluted methanol (9 in 10), combine all the extracts, and add diluted methanol (9 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add 2.5 mL of dilute sodium hydroxide TS, heat in a water bath at 509C for 1 hour, and add 7.5 mL of phosphate buffer solution for assay of bupleurum root. Allow this solution to flow through a chromatographic column [about 10 mm inside diameter containing 0.36 g of octadecylsilanized silica gel for pretreatment (55 to 105 mm in particle diameter), conditioned with 10 mL of methanol then 10 mL of water just before use]. Wash the column with 10 mL of diluted methanol (7 in 20), then flow with metha-nol to get exactly 10 mL of effluent solution, and use this as the sample solution. Use saikosamponins a and d standard TS for assay as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, ATA and ASA, of saikosaponin a and ATD and ASD, of saikosaponin d in each solusion. Calculate the amount of saikosaponin a and saikosaponin d by the following equa-tion, and designate the total as the amount of total saponin. Amount (mg) of saikosaponin a ＝ MSA × ATA/ASA × 1/2 MSA: Amount (mg) of saikosaponin a for assay taken Amount (mg) of saikosaponin d ＝ MSD × ATD/ASD × 1/2 MSD: Amount (mg) of saikosaponin d for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 206 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water and acetonitrile (3:2). Flow rate: Adjust so that the retention time of saikosapo- nin a is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, saikosaponin a and saikosaponin d are eluted in this order, and the numbers of theoretical plates and the symme-try factors of their peaks are not less than 4000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of saikosaponin a and saikosaponin d are not more than 1.5z, respectively.
Powdered Calumba occurs as a grayish yellow powder, and has a characteristic odor and a bitter taste. Under a microscope <5.01>, Powdered Calumba reveals numerous starch grains, fragments of parenchyma cells con-taining them; fragments of cork cells, stone cells, fibers, sub-stitute fibers, vessels, tracheids, and also solitary crystals of calcium oxalate; starch grains consisting of solitary grains or 2- to 3-compound grains; hilum, unevenly scattered, usually 25 – 50 mm, but up to 90 mm in diameter.
To 3 g of Powdered Calumba add 30 mL of water, allow to stand for 5 minutes with occasional shaking, and filter. To 2 mL of the filtrate add gently 1 mL of sulfuric acid, and after cooling, add carefully chlorine TS to make two layers: a light red to red color develops at the zone of contact.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Powdered Calumba according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Calumba according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.5z.
Disk-like slices, 0.5 – 2 cm in thickness, 3 – 8 cm in diameter; mostly with concave center and slightly waved; side surface grayish brown in color, with irregular wrinkles; cut surface light yellow and powdery, with pale and dark radiating stripes; cortex rather yellowish; cambium and its neighborhood light grayish brown, warty protrusions in the center; hard in texture, but brittle. Odor characteristic; taste, bitter.
To 3 g of pulverized Calumba add 30 mL of water, allow to stand for 5 minutes with occasional shaking, and filter. To 2 mL of the filtrate add gently 1 mL of sulfuric acid, and after cooling, add carefully chlorine TS to make two layers: a light red to red color develops at the zone of contact.
(1) Heavy metals <1.07>—Proceed with 2.0 g of pulverized Calumba according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Calumba according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.5z.
Hemp Fruit is a slightly compressed void fruit, 4 – 5 mm in length, 3 – 4 mm in diameter; externally grayish green to grayish brown; pointed at one end, a scar of gyno-phore at the other end, and ridge on both sides; outer sur-face lustrous with white mesh-like pattern; slightly hard pericarp; seed, slightly green in color and internally has grayish white albumen; 100 fruits weigh 1.6 – 2.7 g. Practically odorless, aromatic on chewing; taste, mild and oily. Under a microscope <5.01>, a transverse section reveals the exocarp composed of an epidermis; mesocarp composed of parenchyma, a pigment cell layer and rows of short, small cells; endocarp made up of a single cellular layer of radially elongated stone cells; seed coat comprises a tubular cellular layer and spongy tissue. Inside of the seed; exosperm con- ‌2032 Hochuekkito Extract / Crude Drugs and Related Drugs sists of one cellular layer of parenchymatous cells, en-dosperm of one to several cellular layers of parenchymatous cells; most of the embryo composed of parenchyma, vascu-lar bundles occurring in the center of hypocotyls and cotyledons; embryo parenchyma contains aleurone grains and oil drops.
To 0.3 g of pulverized Hemp Fruit add 3 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and ethyl acetate (9:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a dark blue-purple spot appears at an Rf value of about 0.6.
Bract—When perform the test of foreign matter <5.01>, Hemp Fruit does not contain bract. Loss on drying <5.01> Not more than 9.0z (6 hours). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Powdered Capsicum occurs as a yellow-red powder. It has a slight, characteristic odor and a hot, acrid taste. Under a microscope <5.01>, Powdered Capsicum reveals fragments of parenchyma containing oil droplets and yellow-red chromoplasts; fragments of epidermis from outer sur-face of pericarp with thick cuticle; fragments of stone cells from inner surface of pericarp, with wavy curved side walls; fragments of thin vessels; fragments of seed coat with thick wall, and fragments of parenchyma consisting of small cells of endosperm containing fixed oil and aleuron grains.
To 1.0 g of Powdered Capsicum add 5 mL of ethanol (95), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of (E)-capsaicin for thin-layer chromatography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each ‌1974 Capsicum Tincture / Crude Drugs and Related Drugs of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, ethyl acetate and formic acid (10:9:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and expose to an ammonia vapor: a spot obtained from the sample solution and blue spot obtained from the standard solution show the same in color tone and Rf value. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 1.2z.
Weigh accurately about 0.5 g of Powdered Capsi-cum in a glass-stoppered centrifuge tube, add 30 mL of methanol, shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 10 mL of methanol, shake for 5 minutes, centrifuge, and separate the superna-tant liquid. Repeat this procedure again, combine all the ex-tracts, add methanol to make exactly 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 10 mg of (E)-capsaicin for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 409C) for 5 hours, and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add methanol to make exactly 25 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, ATC and ATD, of (E)-capsaicin and dihydrocapsaicin (the relative retention time to (E)-capsaicin is about 1.3) obtained with the sample solution, and the peak area, AS, of (E)-capsaicin obtained with the standard solution. Amount (mg) of total capsaicins ＝ MS × (ATC ＋ ATD)/AS × 0.08 MS: Amount (mg) of (E)-capsaicin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 281 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of (E)-capsai-cin is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of (E)-capsaicin for assay and 4-hydroxy-3-methoxybenzyl nonylic acid amide in methanol to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-hydroxy-3-methoxybenzyl nonylic acid amide and (E)-capsaicin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of (E)-capsaicin is not more than 1.5z.
Elongated conical to fusiform fruit, often bent, 3 – 10 cm in length, about 0.8 cm in width; outer sur-face lustrous and dark red to dark yellow-red; interior of pericarp hollow and usually divided into two loculi, contain-ing numerous seeds nearly circular and compressed, light yellow-red, about 0.5 cm in diameter. Usually it remains of calyx and peduncle. Odor, slight and characteristic; taste, hot and acrid.
To 1.0 g of pulverized Capsicum add 5 mL of ethanol (95), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 1 mg of (E)-capsaicin for thin-layer chromatography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, ethyl acetate and formic acid (10:9:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and expose to an ammonia vapor: a spot obtained from the sample solution and a blue spot from the standard solution show the same color tone and the same Rf value.
Foreign matter <5.01>—The amount of foreign mat-ter contained in Capsicum does not exceed 1.0z. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 1.2z.
Weigh accurately about 0.5 g of moderately fine powder of Capsicum in a glass-stoppered centrifuge tube, add 30 mL of methanol, shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 10 mL of methanol, shake for 5 minutes, centrifuge, and sepa-rate the supernatant liquid. Repeat this procedure again, combine all the extracts, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of (E)-capsaicin for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 409C) for 5 hours, and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add methanol to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, ATC and ATD, of (E)- the standard solution. Amount (mg) of total capsaicins ＝ MS × (ATC ＋ ATD)/AS × 0.08 MS: Amount (mg) of (E)-capsaicin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 281 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of (E)-capsai-cin is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of (E)-capsaicin for assay and 4-hydroxy-3-methoxybenzyl nonylic acid amide in methanol to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 4-hydroxy-3-methoxybenzyl nonylic acid amide and (E)-capsaicin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of (E)-capsaicin is not more than 1.5z.
Nearly ellipsoidal, 1 – 2 cm in length, 0.5 – 1 cm in diameter; externally, light yellow with three blunt ridges and many longitudinal lines; 0.1 – 0.2-cm beak at one end; pericarp thin, light and fibrous; interior longitudinally divided into three loculi by thin membranes, each loculus containing 3 to 7 seeds joining by aril; seed irregularly angu-lar ovoid, 0.3 – 0.4 cm in length, dark brown to blackish brown; the dorsal side convex, the ventral side longitudinally grooved; external surface coarsely tuberculated. Seed has a characteristic aroma, and pungent, slightly bit-ter taste; pericarp, slight characteristic odor and practically tasteless. Total ash <5.01> Not more than 6.0z (seed). Acid-insoluble ash <5.01> Not more than 4.0z (seed). Essential oil content <5.01> Perform the test with 30.0 g of the pulverized seeds of Cardamon: the volume of essential oil is not less than 1.0 mL.
Red to red-brown corolla, yellow style and sta-men, rarely mixed with immature ovary; total length about 1 cm; corolla, tubular and with 5 lobes; 5 stamens surrounding long pistil; pollen grains yellow and approximately spherical, about 50 mm in diameter, with fine protrusions on the sur-face. The pressed slab, about 0.5 cm in thickness, consists of a collection of numerous corollas. Odor, characteristic; taste, slightly bitter. ‌2122‌ Saffron / Crude Drugs and Related Drugs Identiˆcation To 1.0 g of pulverized SaŠlower add 10 mL of a mixture of acetone and water (4:1), shake for 10 minutes, ˆlter, and use the ˆltrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl acetate, water, formic acid and methanol (35:15:10:2) to a distance of about 7 cm, and air-dry the plate: a red spot ap-pears at an Rf value of about 0.5.
Foreign matter <5.01>—The amount of ovaries, stems, leaves and other foreign matter contained in Safflow-er does not exceed 2.0z. Total ash <5.01> Not more than 18.0z.
Powdered Clove occurs as a dark brown pow-der. It has a strong, characteristic odor and a pungent taste, leaving a sensation of numbness on the tongue. Under a microscope <5.01>, Powdered Clove reveals epidermal tissue with stomata, collenchyma, parenchyma with oil sacs, and spongy parenchyma or its fragments; fur-thermore, a few fusiform thick-walled fibers, spiral vessels 6 – 10 mm in diameter, anther and pollen grains, and rosette aggregates of calcium oxalate 10 – 15 mm in diameter. Epidermis of anther shows characteristically reticulated walls; pollen grains tetrahedral 10 – 20 mm in diameter; rosette aggregates of calcium oxalate arranged in crystal cell rows, or contained in collenchyma cells and parenchyma cells.
To 1.5 g of Powdered Clove add 20 mL of ethyl acetate, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 5 mg of eugenol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of hexane and ace-tone (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
Foreign matter <5.01>—Under a microscope, Pow-dered Clove does not contain stone cells or starch grains. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 0.5z. Essential oil content <5.01> Perform the test with 10.0 g of Powdered Clove: the volume of essential oil is not less than 1.3 mL.
Dark brown to dark red buds, 1 – 1.8 cm in length, consisting of slightly compressed and four-sided receptacle, crowned by 4 thick sepals and 4 nearly spherical, membranous, imbricated petals, enclosing numerous sta-mens and a single style. Odor, strong and characteristic; taste, pungent, which gives numbing sensation to the tongue.
To 1.5 g of pulverized Clove add 20 mL of ethyl acetate, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 5 mg of eugenol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 1 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of hexane and ace-tone (2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Stem—When perform the test of foreign mat-ter <5.01>, the amount of the stem contained in Clove does not exceed 5.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than the stem contained in Clove does not exceed 1.0z. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 0.5z. Essential oil content <5.01> Perform the test with 10.0 g of pulverized Clove: the volume of essential oil is not less than 1.6 mL.
Short cylindrical seed, 3 – 6 mm in length, 2 – 3.5 mm in diameter; acuminate at one end and flat at the other; externally green-brown to brown and lustrous, with light yellow-brown longitudinal lines or bands on both sides; hard in texture; cross section round or obtuse polygonal; under a magnifying glass, albumen enclosing a bent, dark-colored cotyledon. When ground, characteristic odor and taste. Identiˆcation To 1.0 g of pulverized Cassia Seed add 10 mL of diluted methanol (4 in 5), heat on a water bath for 5 minutes, and ˆlter. Evaporate the solvent of the ˆltrate, dis-solve the residue in 5 mL of water, add 2 mL of ethyl acetate, and shake for 10 minutes. Centrifuge this solution, and use the ethyl acetate layer as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of diethyl ether, cyclohexane and formic acid (5:5:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly potassium hydroxide-ethanol TS on the plate: an orange to yellow-brown spot appears at an Rf value of about 0.35.
Foreign matter <5.01>—The amount of foreign mat-ter contained in Cassia Seed does not exceed 1.0z. Total ash <5.01> Not more than 5.0z.
Slender stick-like fruit, 30 – 40 cm in length and about 0.5 cm in diameter; externally, dark brown; inner part contains numerous seeds; seed compressed or semitubu-lar, about 3 cm in length and about 0.3 cm in width, exter-nally grayish brown; hairs, about 1 cm in length, attached to both ends of seed; pericarp, thin and brittle. Odor, slight; taste, slightly astringent.
To 1.0 g of pulverized Catalpa Fruit add 20 mL of water, warm on a water bath for 5 minutes, and filter immediately. Transfer the filtrate to a separator, and extract with two 20-mL portions of 1-butanol. Combine the ex-tracts, evaporate the solvent under low pressure (in vacuo) on a water bath, dissolve the residue in 1 mL of methanol, ‌JP XVIII Crude Drugs and Related Drugs / Chotosan Extract 1977 ‌and use this solution as the sample solution. Separately, dis-solve 1 mg of parahydroxybenzoic acid in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (20:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution and a dark purple spot from the standard solution show the same color tone and the same Rf value. Prescribe that the moving distance of the spot corresponding to parahydrox-ybenzoic acid from the sample solution is 1: a dark purple spot develops at the relative moving distance of about 0.3.
Peduncle—When perform the test of foreign matter <5.01>, the amount of peduncles contained in Catalpa Fruit does not exceed 5.0z. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 0.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
1) Chrysanthemum indicum origin—Capitu-lum, 3 – 10 mm in diameter; involucre, consisting of 3 to 5 rows of involucral scales, often with peduncle; the outer in-volucral scale, linear to lanceolate; inner involucral scale, narrow ovate to ovate; ligulate ‰ower, in a single circle, yel-low to light yellow-brown in color; tubular ‰owers, numerous, light yellow-brown; outer surface of involucre, yellow-brown to brown; light in texture and easy to break. Odor, characteristic; taste, slightly bitter. 2) Chrysanthemum morifolium origin—Capitulum, 15 – 40 mm in diameter; involucre, consisting of 3 to 4 rows of in-volucral scales, often with peduncle; the outer involucral scale, linear to lanceolate; inner involucral scale, narrow o-vate to ovate; ligulate ‰owers, numerous, white to yellow in color; tubular ‰owers, small in number, light yellow-brown, occasionally degenerate; outer surface of involucre, green-brown to brown; light in texture and easy to break. tion on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, 2-buta-none, water and formic acid (25:3:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 8.5z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Knotted, irregularly shaped rhizome, 6 – 18 cm in length, 1 – 2.5 cm in diameter; externally dark brown to blackish brown, with many remains of roots, often with scars of terrestrial stems; the center of the scar dented, and the circumference being pale in color and showing a radial pattern; fractured surface fibrous; pith dark brown in color and often hollow; light and hard in texture. Almost odorless; taste, bitter and slightly astringent.
Dissolve 1 g of pulverized Cimicifuga Rhi-zome add 5 mL of dilute hydrochloric acid and 5 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the diethy ether layer as the sample solution. Use (E)-isoferulic acid-(E)-ferulic acid TS for thin-layer chromatography as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (30:10:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the spot among the several spots obtained from the sample solution has the same color tone and Rf value with the blue fluorescent spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Cimicifuga Rhizome according to Method 3, and ‌JP XVIII Crude Drugs and Related Drugs / Powdered Cinnamon Bark 1981‌ perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Cimicifuga Rhizome according to Method 4, and perform the test (not more than 5 ppm). Rhizome of Astilbe thunbergii Miquel—Under a microscope <5.01>, pulverized Cimicifuga Rhizome does not contain crystal druses in the parenchyma. Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 18.0z.
Powdered Cinnamon Bark is red-brown to brown in color. It has a characteristic aroma and a sweet, pungent taste with a slightly mucilaginous and astringent aftertaste. Under a microscope <5.01>, Powdered Cinnamon Bark re- veals starch grains, fragments of parenchyma cells contain-ing them; fragments of fibers, oil cells containing yellow-brown oil droplets, stone cells, cork stone cells, cork tissue, and fine crystals of calcium oxalate. Starch grains are simple and compound grains 6 to 20 mm in diameter.
To 2.0 g of Powdered Cinnamon Bark add 10 mL of diethyl ether, shake for 3 minutes, filter, and use the filtrate as the sample solution. Perform the test with this solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a purple spot develops at an Rf value of about 0.4. Spray 2,4-dinitrophenylhydrazine TS upon the spot: a yellow orange color develops.
(1) Petiole—Under a microscope <5.01>, Pow-dered Cinnamon Bark does not reveal epidermal cells, hairs, cells containing chlorophyll granules, and fragments of vas-cular bundle. (2) Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 6.0z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Cinnamon Bark provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.35 mL.
Usually semi-tubular or tubularly rolled pieces of bark, 0.1 – 0.5 cm in thickness, 5 – 50 cm in length, 1.5 – 5 cm in diameter; the outer surface dark red-brown, and the inner surface red-brown and smooth; brittle; the fractured surface is slightly fibrous, red-brown, exhibiting a light brown, thin layer. Characteristic aroma; taste, sweet and pungent at first, later rather mucilaginous and slightly astringent. Under a microscope <5.01>, a transverse section of Cinna- mon Bark reveals a primary cortex and a secondary cortex divided by an almost continuous ring consisting of stone cells; nearly round bundles of fibers in the outer region of the ring; cell wall of each stone cell often thickened in a U-shape; secondary cortex lacking stone cells, and with a small number of sclerenchymatous fibers coarsely scattered; parenchyma scattered with oil cells, mucilage cells and cells containing starch grains; medullary rays with cells contain-ing fine needles of calcium oxalate.
To 2.0 g of pulverized Cinnamon Bark add 10 mL of diethyl ether, shake for 3 minutes, filter, and use the filtrate as the sample solution. Perform the test with this solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a purple spot develops at an Rf value of about 0.4. Spray evenly 2,4-dinitrophenylhydrazine TS upon the spot: a yellow-orange color develops.
Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 15.5z (6 hours). Total ash <5.01> Not more than 6.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Cinnamon Bark provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.5 mL.
1) Cistanche salsa origin—Flatly cylindrical, 5 – 25 cm in length, 1 – 2.5 cm in diameter; the one end mostly slightly narrow and curved; external surface brown to blackish brown, covered with thick scales; fleshy and solid, slightly soft and oily, hardly broken; fractured surface yellow-brown to brown, vascular bundles light brown and arranged in a wavy ring. Odor, characteristic; taste, slightly sweet, followed by slight bitterness. Under a microscope <5.01> a transverse section of middle part reveals the outermost part is an epidermis coated with cuticle; cortex composed of parenchyma; collateral vascular bundles fusiform or rhombic and arranged in a wavy ring in the inner portion of cortex; groups of cells with slightly thickened cell walls sometimes attached outside of phloem of collateral vascular bundles, and exhibit tail like form; pith composed of parenchyma; parenchyma contains starch grains or gelatinized starch. Cistanche deserticola origin—Flatly cylindrical, and approximate to 1), but large in size, 5 – 50 cm in length, 1 – 8 cm in diameter. Odor, characteristic; taste, slightly sweet, followed by slight bitterness. Under a microscope <5.01> a transverse section of middle part reveals, approximate to 1). Cistanche tubulosa origin—Flatly fusiform to cylin-drical, slightly curved, 5 – 25 cm in length, 2 – 9 cm in diam-eter; external surface brown to blackish brown, covered with thick scales; solid in texture and firm, hardly broken; frac-tured surface light grayish brown to yellow-brown, vascular bundles yellow-white and scattered throughout the surface. Odor, characteristic; taste, slightly sweet, followed by slight bitterness. Under a microscope <5.01> a transverse section of middle part reveals, approximate to 1) and 2), but collateral vascu-lar bundles distributed throughout the parenchyma from marginal region to the center of transverse section; cells with slightly thickened cell walls observed sometimes around col-lateral vascular bundles, but exhibit no tail like form;
To 1 g of pulverized Cistanche Herb add 5 mL of water and 5 mL of 1-butanol, shake for 15 minutes, centrifuge, and use the 1-butanol layer as the sample solu-tion. Separately, dissolve 1 mg of verbascoside for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution and 10 mL of the standard solution on a plate of silica gel for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 2,6-dibromo-N-chlolo-1,4-benzoquinone monoimine TS on the plate, and allow to stand in an ammonia gas: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Cistanche Herb according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Cistanche Herb according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 20.0z. Total ash <5.01> Not more than 11.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 35.0z.
Clematis Root consists of short rhizome and numerous slender roots. The root, 10 – 20 cm in length, 1 – 2 mm in diameter, externally brown to blackish brown, with fine longitudinal wrinkles, brittle. The cortex easily separa-ble from central cylinder; root, grayish white to light yellow-brown in the transverse section, light grayish yellow to yel-low in the central cylinder; under a magnifying glass, central cylinder almost round, slight 2 – 4 sinuses on xylem. The rhizome, 2 – 4 cm in length, 5 – 20 mm in diameter, exter-nally light grayish brown to grayish brown; cortex peeled off and fibrous, often with rising node; apex having the residue of lignified stem. Odor, slight; practically tasteless. Under a microscope, <5.01> transverse section of root reveals an epidermis in the outermost layer; with exodermis lying just inside of the epidermis; cortex and stele divided by endodermis; cortex composed of parenchymatous tissue; xylem with 2 – 4 small concavities where phloem is present; parenchymatous cells contain both simple and 2- to 8-com-pound starch grains.
(1) To 0.5 g of pulverized Clematis Root add 10 mL of water, and boil for 2 to 3 minutes. After cool-ing, shake vigorously: lasting fine foams appear. (2) To 0.5 g of pulverized Clematis Root add 3 mL of acetic anhydride, warm on a water bath for 2 minutes, and filter. To the filtrate add 1 mL of sulfuric acid gently: a brown color appears at the zone of contact.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Clematis Root according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Clematis Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 13.0z (6 hours). ‌1984‌‌ Clove / Crude Drugs and Related Drugs Total ash <5.01> Not more than 8.5z. Acid-insoluble ash <5.01> Not more than 3.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 15.0z.
Root according to Method 3, and perform the test
Elliptical cremocarp, often each mericarp separated; 2 – 3 mm in length, 1 – 2 mm in width; externally light brown to brown, each mericarp usually with five winged longitudinal ridges; inner surface of mericarp almost flat. Odor, characteristic; it gives characteristic aroma, later a slight sensation of numbness on chewing. Under a microscope <5.01>, a transverse section reveals one oil canal between longitudinal ridges, usually two oil canals in the inner part of mericarp facing to gynophore; longitudinal ridges composed of slightly lignified parenchy-matous cells, with vascular bundles in the base; epidermal cells and parenchymatous cells of longitudinal ridges contain solitary crystals of calcium oxalate; parenchymatous cells of albumen contain oil drops and aleurone grains, and occa-sionally starch grains.
To 1 g of pulverized Cnidium Monnieri Fruit add 10 mL of ethyl acetate, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of osthole for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and ethyl acetate (2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and the Rf value with the blue-white fluorescent spot from the standard solu-tion. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 17.0z. Acid-insoluble ash <5.01> Not more than 6.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
Powdered Cnidium Rhizome occurs as a gray to light grayish brown powder. It has a characteristic odor and a slightly bitter taste. Under a microscope <5.01>, Powdered Cnidium Rhizome reveals colorless and gelatinized starch masses, and frag-ments of parenchyma containing them; fragments of scalariform and reticulate vessels 15 – 30 mm in diameter; fragments of thick-walled and lignified xylem fibers 20 – 60 mm in diameter; fragments of yellow brown cork tissue; fragments of secretory tissue.
To 1 g of Powdered Cnidium Rhizome add 5 mL of methanol and 0.1 mL of sodium hydroxide TS, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, use (Z)-ligustilide TS for thin-layer chromatography as the standard solution (1). Dis-solve 1 mg of (E )-ferulic acid in 2 mL of methanol and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 20 mL of the sample solution and 5 mL each of the standard solution (1) and the standard solution (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (30:25:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu-tion (1). Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes and allow to cool: one of the several spots from the sample solution has the same color tone and Rf value with the spot from the standard solution (2).
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Cnidium Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Cnidium Rhizome according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow-dered Cnidium Rhizome does not contain a large quantity of starch grains, stone cells, crystals of calcium oxalate or other foreign matter. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Irregular massive rhizome, occasionally cut lengthwise; 5 – 10 cm in length, and 3 – 5 cm in diameter; ex-ternally grayish brown to dark brown, with gathered nodes, and with knobbed protrusions on the node; margin of the vertical section irregularly branched; internally grayish white to grayish brown, translucent and occasionally with hollows; dense and hard in texture. Odor, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section reveals cortex and pith with scattered oil canals; in the xylem, thick-walled and lignified xylem fibers appear in groups of various sizes; starch grains usually gelatinized, but rarely remaining as grains of 5 – 25 mm in diameter; crystals of calcium oxa-late not observable.
To 1 g of pulverized Cnidium Rhizome add 5 mL of methanol and 0.1 mL of sodium hydroxide TS, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, use (Z )-ligustilide TS for thin-layer chromatography as the standard solution (1). Dis-solve 1 mg of (E )-ferulic acid in 2 mL of methanol and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 20 mL of the sample solution and 5 mL each of the standard solution (1) and the standard solution (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, acetone and acetic acid (100) (30:25:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solu-tion (1). Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes ‌1986‌ Powdered Cnidium Rhizome / Crude Drugs and Related Drugs ‌and allow to cool: one of the several spots from the sample solution has the same color tone and Rf value with the spot from the standard solution (2).
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Cnidium Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Cnidium Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Codonopsis Root nearly cylindrical, 8 – 30 cm in length, 0.5 – 2.5 cm in diameter; gradually slender to the apex, often branched; outer surface light yellow to grayish brown; from the base to central part with ring-like wrinkles, and longitudinal wrinkles entirely obvious; numerous projec-tions composed of scars of stems at the crown, with a round dent at the distal end; blackish brown and tremellose secre-tion often at the scars of lateral roots; flexible and easily bendable or hard and easily breakable in texture; in cut sur-face yellow-white to light brown in cortex, light yellow in xylem, sometimes with slit in cortex. Odor, slight and characteristic; taste, slightly sweet. Under a microscope <5.01>, a transverse section reveals cork layer at the outermost portion, outer 1- to 10-layer con-sisting of cork stone cells; groups of laticifers containing light yellow substances arranged radially in phloem, intercel-lular spaces usually observed; vessels of xylem arranged radi-ally; starch grains and crystals of inulin usually contained in phloem parenchyma cells. ‌JP XVIII Crude Drugs and Related Drugs / Condurango 1987‌‌
To 2.0 g of pulverized Codonopsis Root add 50 mL of water, and heat in a water bath for 1 hour. After cooling, filter, and wash the filtrate with two 20-mL portions of ethyl acetate. Separate the aqueous layer, extract with two 30-mL portions of water saturated 1-butanol. Combine the 1-butanol layers, and evaporate the solvent in a water bath under low pressure (in vacuo). Dissolve the residue in 1 mL of methanol, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of 1-propanol, water and ethyl acetate (6:5:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly naphthoresorcin-phosphoric acid TS on the plate, and heat the plate at 1059C for 10 minutes: an orange to red-purple spot at an Rf value of about 0.5 is observed.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Codonopsis Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Codonopsis Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 23.0z (6 hours). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
Powdered Coix Seed occurs as a brownish, grayish white to grayish yellow-white powder, and has a slight odor and a slightly sweet taste. Under a microscope <5.01>, Powdered Coix Seed reveals starch grains, and fragments of endosperm containing them; fragments of tissue accompanied with epidermal cells of pericarp composed of yellowish and oblong cells, and frag-ments of parenchyma cells containing fixed oil, aleuron grains and starch grains; a very few fragments of spiral ves-sels. Starch grains are simple and 2-compound grains, simple grain nearly equidiameter to obtuse polygon, 10 – 20 mm in diameter, and have a stellate cleft-like hilum in the center. Spherical starch grains, coexisting with aleuron grains, are spherical simple grains, 3 – 7 mm in diameter.
Place a small amount of Powdered Coix Seed on a slide glass, add dropwise iodine TS, and examine under a microscope <5.01>: nearly equidiameter and obtuse polygonal simple starch grains, usually 10 – 15 mm in diame-ter, and compound starch grains have a reddish brown color. Small spheroidal starch grains, coexisting with fixed oil and with aleuron grains in parenchymatous cells, have a blue-purple color.
Foreign matter—Under a microscope <5.01>, Powdered Coix Seed reveals no fragments of tissue having silicified cell wall, no stone cells, no fragments of other thick-walled and lignified cells, no fragments of reticulate, scalariform and pitted vessels, no fragments of fibers and hairs, and no large starch grains, more than 10 mm in diame-ter, appearing blue-purple upon addition of iodine TS. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 3.0z.
Ovoid or broad ovoid seed, about 6 mm in length, and about 5 mm in width; with a slightly hollowed apex and base; dorsal side distended; ventral side longitudi-nally and deeply furrowed in the center; dorsal side mostly white in color and powdery; in the furrow on the ventral sur-face, attached brown, membranous pericarp and seed coat. Under a magnifying glass, the cross section reveals light yellow scutellum in the hollow of the ventral side. Hard in texture. Odor, slight; taste, slightly sweet; adheres to the teeth on chewing.
To a transverse section of Coix Seed add iodine TS dropwise: a dark red-brown color develops in the endosperm, and a dark gray color develops in the scutellum. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 3.0z.
Compound Phellodendron Powder for Cataplasm
Tubular or semi-tubular pieces of bark, 0.1 – 0.6 cm in thickness, 4 – 15 cm in length; outer surface grayish brown to dark brown, nearly smooth and with numerous lenticels, or more or less scaly and rough; inner surface light grayish brown and longitudinally striate; frac-tured surface fibrous on the outer region and generally granular in the inner region. Odor, slight; taste, bitter. Under a microscope <5.01>, a transverse section reveals a cork layer composed of several cellular layers of thin-walled cells; primary cortex with numerous stone cell groups; secon-dary cortex with phloem fiber bundles scattered inside the ‌1988‌ Condurango Fluidextract / Crude Drugs and Related Drugs starch sheath consisting of one-cellular layer; articulate latex tubes scattered in both cortices; parenchyma cells containing starch grains or rosette aggregates of calcium oxalate; starch grain 3 – 20 mm in diameter.
Digest 1 g of pulverized Condurango in 5 mL of water, and filter: the clear filtrate becomes turbid on heating, but becomes clear again upon cooling.
Foreign matter <5.01>—The xylem and other for-eign matter contained in Condurango do not exceed 2.0z. Total ash <5.01> Not more than 12.0z.
Powdered Coptis Rhizome occurs as a yellow-brown to grayish yellow-brown powder. It has a slight odor and an extremely bitter, lasting taste, and colors the saliva yellow on chewing. Under a microscope <5.01>, almost all elements are yellow in color; it reveals mainly fragments of vessels, tracheids and xylem fibers; parenchyma cells containing starch grains; polygonal cork cells. Usually, round to obtuse polygonal stone cells and their groups, and phloem fibers, 10 – 20 mm in diameter, and fragments of their bundles. Sometimes, po-lygonal and elongated epidermal cells, originated from the petiole, having characteristically thickened cell walls. Starch grains are single grains 1 – 7 mm in diameter.
(1) To 0.5 g of Powdered Coptis Rhizome add 10 mL of water, allow to stand for 10 minutes with occa-sional shaking, and filter. To 2 to 3 drops of the filtrate add 1 mL of hydrochloric acid and 1 to 2 drops of hydrogen peroxide TS, and shake: a red-purple color develops. (2) To 0.5 g of Powdered Coptis Rhizome add 20 mL of methanol, shake for 2 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Berberine Chloride RS or berberine chloride hydrate for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the spot among the several spots from the sam-ple solution and a yellow to yellow-green fluorescence spot from the standard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Powdered Coptis Rhizome according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). When the decision is difficult by this method, perform the test as di-rected under Atomic Absorption Spectrophotometry <2.23>. Put 5.0 g of Powdered Coptis Rhizome in a platinum, quartz or porcelain crucible, heat gently, and then incinerate by ig-nition between 4509C and 5509C. After cooling, add a small amount of 2 mol/L nitric acid TS, filter if necessary, and wash the crucible and filter several times with small portions of 2 mol/L nitric acid TS. Combine the filtrate and the washings, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sample solution. Separately, to 2.5 mL of Standard Lead Solution add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the stand-ard solution. Perform the test with the sample solution and the standard solution according to the following conditions: the absorbance of the sample solution is not more than that ‌1990‌ Corn Oil / Crude Drugs and Related Drugs of the standard solution (not more than 5 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: A lead hollow-cathode lamp. Wavelength: 283.3 nm. Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Coptis Rhizome according to Method 4, and perform the test (not more than 5 ppm). Phellodendron bark—Under a microscope <5.01>, crystal cell rows or mucilage masses are not observable. Stir 0.5 g of Powdered Coptis Rhizome with 2 mL of water: the solution does not become gelatinous. Curcuma—Place Powdered Coptis Rhizome on a filter paper, drop diethyl ether on it, and allow to stand. Re-move the powder from the filter paper, and drop 1 drop of potassium hydroxide TS: no red-purple color develops. Under a microscope <5.01>, Powdered Coptis Rhizome does not contain gelatinized starch or secretory cells containing yellow-red resin. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Weigh accurately about 0.5 g of Powdered Coptis Rhizome, add 30 mL of a mixture of methanol and dilute hydrochloric acid (100:1), heat under a reflux condenser for 30 minutes, cool, and filter. Repeat the above procedure twice with the residue, using 30-mL and 20-mL portions of a mixture of methanol and dilute hydrochloric acid (100:1). To the last residue add 10 mL of methanol, shake well, and filter. Combine the whole filtrates, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chlo-ride RS (previously determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in metha-nol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine [as berberine chloride (C20H18ClNO4)] ＝ MS × AT/AS MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen-phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in methanol to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, palmatine and berber- ine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Irregular, cylindrical rhizome, 2 – 4 cm, rarely up to 10 cm in length, 0.2 – 0.7 cm in diameter, slightly curved and often branched; externally grayish yellow-brown, with ring nodes, and with numerous remains of rootlets; generally remains of petiole at one end; fractured surface rather fibrous; cork layer light grayish brown, cortex and pith are yellow-brown to reddish yellow-brown, xylem is yellow to reddish yellow in color. Odor, slight; taste, extremely bitter and lasting; it colors the saliva yellow on chewing. Under a microscope <5.01>, a transverse section of Coptis Rhizome reveals a cork layer composed of thin-walled cork cells; cortex parenchyma usually exhibiting groups of stone cells near the cork layer and yellow phloem fibers near the cambium; xylem consisting chiefly of vessels, tracheids and xylem fibers; medullary ray distinct; pith large; in pith, stone cells or stone cells with thick-walled and lignified cells are sometimes recognized; parenchyma cells contain minute starch grains.
(1) To 0.5 g of pulverized Coptis Rhizome add 10 mL of water, allow to stand for 10 minutes with occa-sional shaking, and filter. To 2 to 3 drops of the filtrate add 1 mL of hydrochloric acid and 1 to 2 drops of hydrogen peroxide TS, and shake: a red-purple color develops. (2) To 0.5 g of pulverized Coptis Rhizome add 20 mL of methanol, shake for 2 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Berberine Chloride RS or berberin chloride hydrate for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion and a yellow to yellow-green fluorescence spot from the standard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Coptis Rhizome according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). When the decision is difficult by this method, perform the test as di-rected under Atomic Absorption Spectrophotometry <2.23>. Put 5.0 g of pulverized Coptis Rhizome in a platinum, quartz or porcelain crucible, heat gently, and then incinerate by ignition between 4509C and 5509C. After cooling, add a small amount of 2 mol/L nitric acid TS, filter if necessary, and wash the crucible and filter several times with small por-tions of 2 mol/L nitric acid TS. Combine the filtrate and the washings, add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the sample solution. Separately, to 2.5 mL of Standard Lead Solution add 2 mol/L nitric acid TS to make exactly 20 mL, and use this solution as the stand-ard solution. Perform the test with the sample solution and the standard solution according to the following conditions: the absorbance of the sample solution is not more than that of the standard solution (not more than 5 ppm). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: A lead hollow-cathode lamp. Wavelength: 283.3 nm. The procedure and permissible limit for Coptis Rhizome labeled to be used for extracts or infusions and decoctions are as follows. To 4.0 g of moderately fine cuttings of Coptis Rhizome add 80 mL of water, and heat until the amount becomes about 40 mL with occasional stirring. After cooling, filter, and proceed with the filtrate according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of ‌JP XVIII Crude Drugs and Related Drugs / Powdered Coptis Rhizome 1989 Standard Lead Solution (not more than 5 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Coptis Rhizome according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Weigh accurately about 0.5 g of pulverized Coptis Rhizome, add 30 mL of a mixture of methanol and dilute hydrochloric acid (100:1), heat under a reflux condenser for 30 minutes, cool, and filter. Repeat the above procedure twice with the residue, using 30-mL and 20-mL portions of a mixture of methanol and dilute hydrochloric acid (100:1). To the last residue add 10 mL of methanol, shake well, and filter. Combine the whole filtrates, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chlo-ride RS (previously determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in metha-nol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine [as berberine chloride (C20H18ClNO4)] ＝ MS × AT/AS MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen-phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in methanol to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, palmatine and berber-ine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Flattened oblong, 1.5 – 2 cm in length, about 1 cm in width; externally dark red-purple to dark purple, lus-trous, and with coarse wrinkles; a crack-like scar formed by removal of true fruits; a scar of calyx at one end, and a scar of peduncle at the other; soft in texture. Odor, slight; taste, acid and occasionally slightly sweet.
To 1 g of coarse cuttings of Cornus Fruit add 10 mL of methanol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of loganin for thin-layer chromatography in 2 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (6:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-metho-xybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ‌JP XVIII Crude Drugs and Related Drugs / Corydalis Tuber 1991 obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Further, a spot, slightly different in color tone from the above-mentioned spot, is found immediately below of the spot.
(1) Foreign matter <5.01>—The amount of its peduncles and other foreign matter contained in Cornus Fruit does no exceed 2.0z. (2) Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 35.0z.
Weigh accurately about 1 g of fine cuttings of Cor-nus Fruit (separately determine the loss on drying <5.01>), put in a glass-stoppered centrifuge tube, add 30 mL of diluted methanol (1 in 2), shake for 20 minutes, centrifuge, and take the supernatant liquid. To the residue add 30 mL of diluted methanol (1 in 2), and repeat the above process twice more. Combine all the extacts, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of loga-nin for assay, dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of loganin in each solution. Amount (mg) of loganin ＝ MS × AT/AS MS: Amount (mg) of loganin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, acetonitrile and metha-nol (55:4:1). Flow rate: Adjust so that the retention time of loganin is about 25 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of loganin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of loganin is not more than 1.5z.
Powdered Corydalis Tuber occurs as a green-ish yellow to grayish yellow powder. Almost odorless; taste, bitter. Under a microscope <5.01>, Powdered Corydalis Tuber reveals mainly, masses of gelatinized starch or light yellow to colorless parenchymatous cells containing starch grains, fragments of cork layers, light yellow stone cells, scleren-chymatous cells, reticulate vessels, spiral vessels and ring vessels; starch grains observed simple grains and 2- to 3-compound grains.
To 2 g of Powdered Corydalis Tuber add 10 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, dissolve 1 mg of de- Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 17.91 g of disodium hydrogen phosphate dodecahydrate in 970 mL of water, and adjust to pH 2.2 with phosphoric acid. To this solution add 14.05 g of sodium perchlorate, dissolve, and add water to make exactly 1000 mL. Add 450 mL of acetonitrile, and dissolve 0.20 g of sodium lauryl sulfate in this solution. Flow rate: Adjust so that the retention time of dehydro- ‌JP XVIII Crude Drugs and Related Drugs / Curcuma Rhizome 1993 corydaline is about 24 minutes. System suitability— ‌System performance: Dissolve 1 mg of dehydrocorydaline nitrate for assay and 1 mg of berberine chloride hydrate in 20 mL of a mixture of water and acetonitrile (20:9). When the procedure is run with 5 mL of this solution under the above operating conditions, berberine and dehydrocory-daline are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of dehydrocorydaline is not more than 1.5z.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Corydalis Tuber according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Corydalis Tuber according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 15.0z. Total ash <5.01> Not more than 3.0z.
Weigh accurately about 1 g of Powdered Corydalis Tuber, add 30 mL of a mixture of methanol and dilute hy-drochloric acid (3:1), heat under a reflux condenser for 30 minutes, and filter after cooling. To the residue add 15 mL of the mixture of methanol and dilute hydrochloric acid (3:1), and proceed in the same way as above. Combine all the filtrates, add the mixture of methanol and dilute hydro-chloric acid (3:1) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of dehydrocorydaline nitrate for assay, previ-ously dried in a desiccator (silica gel) for not less than 1 hour, dissolve in the mixture of methanol and dilute hydro-chloric acid (3:1) to make exactly 200 mL, and use this solu-tion as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of dehydrocorydaline in each solution. Amount (mg) of dehydrocorydaline [as dehydrocorydaline nitrate (C22H24N2O7)] ＝ MS × AT/AS × 1/4 MS: Amount (mg) of dehydrocorydaline nitrate for assay daline [as dehydrocorydaline nitrate (C22H calculated on the basis of dried material. 24N2O7)], taken Operating conditions— Description Powdered Corydalis Tuber occurs as a green-ish yellow to grayish yellow powder. Almost odorless; taste, bitter. Under a microscope <5.01>, Powdered Corydalis Tuber reveals mainly, masses of gelatinized starch or light yellow to colorless parenchymatous cells containing starch grains, fragments of cork layers, light yellow stone cells, scleren-chymatous cells, reticulate vessels, spiral vessels and ring vessels; starch grains observed simple grains and 2- to 3-compound grains. Identification To 2 g of Powdered Corydalis Tuber add 10 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Separately, dissolve 1 mg of de- Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 17.91 g of disodium hydrogen phosphate dodecahydrate in 970 mL of water, and adjust to pH 2.2 with phosphoric acid. To this solution add 14.05 g of sodium perchlorate, dissolve, and add water to make exactly 1000 mL. Add 450 mL of acetonitrile, and dissolve 0.20 g of sodium lauryl sulfate in this solution. Flow rate: Adjust so that the retention time of dehydro- ‌JP XVIII Crude Drugs and Related Drugs / Curcuma Rhizome 1993 corydaline is about 24 minutes. System suitability— ‌System performance: Dissolve 1 mg of dehydrocorydaline nitrate for assay and 1 mg of berberine chloride hydrate in 20 mL of a mixture of water and acetonitrile (20:9). When the procedure is run with 5 mL of this solution under the above operating conditions, berberine and dehydrocory-daline are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of dehydrocorydaline is not more than 1.5z.
Crataegus cuneata origin—Nearly spherical fruits, 8 – 14 mm in diameter; externally yellow-brown to grayish brown, with fine reticulated wrinkles, remained dent of 4 – 6 mm in diameter at one end, often the base of calyx around the dent, short peduncle or scar at the other end. True fruits, usually five loculus, often split five, mericarp, 5 – 8 mm in length, light brown, usually, containing one seed into each mericarp. Almost odorless; taste, slightly acid. Under a microscope <5.01>, a transverse section of central parts reveals in the outermost layer composed of epidermis to be covered with comparatively thick cuticle layer, cuticle intrude into lateral cell walls of epidermis, and reveal wedge-like. Cell of the epidermis or 2- to 3-cellular layer of paren-chyma cells beneath these observed contents of yellow-brown to red-brown in color followed these appeared paren-chyma. Vascular bundles and numerous stone cells appear single or gathered 2 to several cells scattered on the paren-chyma, and observed solitary crystals and clustera crystals of calcium oxalate. Pericarp of true fruits composed of mainly sclerenchyma cells, seed covered with seed coats, perisperm, endosperm, cotyledon observed inside seed coats; scleren-chyma cells of true fruits and cells of seed coats containing solitary crystals of calcium oxalate. Crataegus pinnatifida var. major origin—Approxi- mate to 1), but it is large in size, 17 – 23 mm in diameter, the outer surface red-brown and lustrous, spot-like scars of hairs are distinct. At one end remained dent, 7 – 9 mm in diame-ter, mericarp, 10 – 12 mm in length, yellow-brown in color, usually ripe seeds are absent. Odor, characteristic; taste, acid. Under a microscope <5.01>, a transverse section of the cen-tral parts approximate to 1), but it contains a few stone cells in parenchyma.
Crataegus cuneata origin—To 1.0 g of pulverized Crataegus Fruit add 5 mL of methanol, shake for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of rutin for thin-layer chromatography in 20 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy, develop the plate with a mixture of ethyl acetate, 2-butanone, water and formic acid (5:3:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly dilute sulfu-ric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the green fluorescent spot from the standard solution, and one or two similar green fluorescent spots are found at an Rf value of about 0.5. These spots disappear gradually by allowing to cool, and appear again by heating. Crataegus pinnatifida var. major origin—To 1 g of pulverized Crataegus Fruit add 5 mL of methanol, shake for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of hyperoside for thin-layer chromatography in 20 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography, develop the plate with a mixture of ethyl ace-tate, 2-butanone, water and formic acid (5:3:1:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wave-length: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the green fluorescent spot from the standard solution, and a similar fluorescent spot is found just above the spot. These spots disappear gradually by allowing to cool, and appear again by heating. Loss on drying <5.01> Not more than 17.0z. Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
Crude Glycyrrhiza Extract contains not less than 4.
Crude Glycyrrhiza Extract occurs as lustrous, dark yellow-red to black-brown plates, rods or masses. It is comparatively brittle when cold, and the fractured surface is dark yellow-red, shell-like, and lustrous. It softens when warmed. It has a characteristic odor and a sweet taste. It dissolves in water with turbidity.
To 0.6 g of Crude Glycyrrhiza Extract add 10 mL of a mixture of ethanol (95) and water (7:3), dissolve by warming if necessary, cool, centrifuge, and use the super-natant liquid as the sample solution. Proceed as directed in the Identification under Glycyrrhiza.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Crude Glycyrrhiza Extract as directed in the Extracts (4) under General Rules for Preparations, and per-form the test (not more than 30 ppm). Water-insoluble substances—Boil 5.0 g of pulverized Crude Glycyrrhiza Extract with 100 mL of water. After cool-ing, filter the mixture through tared filter paper, wash with water, and dry the residue at 1059C for 5 hours: the mass of the residue is not more than 1.25 g. Foreign matter—The filtrate obtained in (2) does not have a strong bitter taste. Starch—To about 1 g of pulverized Crude Glycyr-rhiza Extract add water to make 20 mL, shake the mixture thoroughly, and filter. Examine the insoluble substance on the filter paper under a microscope: the residue contains no starch grains. Total ash <5.01> Not more than 12.0z (1 g).
Weigh accurately about 0.15 g of Crude Glycyrrhiza Extract, place in a glass-stoppered centrifuge tube, add 25 mL of dilute ethanol, and heat at 509C for 30 minutes with occasional shaking. Cool, centrifuge, and take the superna-tant liquid. To the residue add 20 mL of dilute ethanol, and proceed in the same manner. Combine all the extracts, add dilute ethanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in dilute ethanol to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: Adjust so that the retention time of glycyrrhizic acid is about 15 minutes. System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak with the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
Powdered Turmeric occurs as a yellow-brown to dark yellow-brown powder. It has a characteristic odor and a bitter, stimulant taste, and colors the saliva yellow on chewing. Under a microscope <5.01>, all elements are yellow in color; it reveals parenchymatous cells containing mainly masses of gelatinized starch or yellow substances, also frag-ments of scalariform vessels; fragments of cork layers, epidermis, thick-walled xylem parenchymatous cells, and non-glandular hairs are occasionally observed.
(1) To 0.5 g of Powdered Turmeric, add 20 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (11:9:1) to a distance about 7 cm, and air-dry the plate: a yel-low spot appears at an Rf value of about 0.4. (2) To 0.2 g of Powdered Turmeric, add 25 mL of a mix- ture of methanol and acetic acid (100) (99:1), centrifuge after shaking for 20 minutes. Perform the test with the superna-tant liquid as directed in the Assay, and determine the peak areas of curcumin, demethoxycurcumin and bisdemethox-ycurcumin: the peak area of curcumin is larger than the peak area of demethoxycurcumin and is larger than 0.69 times the peak area of bisdemethoxycurcumin.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Turmeric according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Turmeric according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 17.0z (6 hours). Total ash <5.01> Not more than 7.5z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 9.0z.
Weigh accurately about 0.2 g of Powdered Turmer-ic, add 25 mL of a mixture of methanol and acetic acid (100) (99:1), shake for 20 minutes, centrifuge, and separate the supernatant liquid. To the residue, add 25 mL of a mixture of methanol and acetic acid (100) (99:1), and proceed in the same manner as described above. Combine all the extracts, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of curcumin for assay, and dissolve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, add meth-anol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as described under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, ATC, ATD and ATB of curcumin, demethoxycurcumin and bis-demethoxycurcumin in the sample solution as well as the peak area AS of curcumin in the standard solution. Amount (mg) of total curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin) ＝ MS × (ATC ＋ ATD ＋ ATB × 0.69)/AS × 1/5 MS: Amount (mg) of curcumin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilianized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (56:43:1). Flow rate: 1.0 mL/per minute (the retention time of cur-cumin is about 11 minutes). System suitability— System performance: Dissolve 1 mg each of curcumin for assay, demethoxycurcumin and bisdemethoxycurcumin in methanol to make 5 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, bisdemethoxycurcumin, demethoxycurcumin and curcumin are eluted in this order with the resolution among these peaks being not less than 1.5. ‌2168‌‌ Turpentine Oil / Crude Drugs and Related Drugs System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of curcumin is not more than 1.5z.
Turmeric is a main rhizome or a lateral rhi-zome; main rhizome, nearly ovoid, about 3 cm in diameter, about 4 cm in length; lateral rhizome, cylindrical, with round tips, curved, about 1 cm in diameter, 2 - 6 cm in length; both main and lateral rhizomes with cyclic nodes; rhizome with cork layer, yellow-brown, lustrous; rhizome without cork layer, dark yellow-red, with yellow-red pow-ders on surface; hard in texture, not easily broken; transver-sely cut surface yellow-brown to red-brown, lustrous like wax. Odor, characteristic; taste, slightly bitter and stimulant, it colors a saliva yellow on chewing. Under a microscope <5.01>, a transverse section reveals the outermost layer to be composed of a cork layer 4 – 10 cells thick; sometimes a cork layer partly remains; cortex and stele, divided by an endodermis, composed of parenchyma, vascular bundles scattered; oil cells scattered in parenchyma; parenchymatous cells contain yellow substances, sandy and solitary crystals of calcium oxalate, and gelatinized starch.
(1) To 0.5 g of pulverized Turmeric, add 20 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (11:9:1) to a distance about 7 cm, and air-dry the plate: a yel-low spot appears at an Rf value of about 0.4. (2) To 0.2 g of pulverized Turmeric, add 25 mL of a mix-ture of methanol and acetic acid (100) (99:1), centrifuge after shaking for 20 minutes. Perform the test with the superna-tant liquid as directed in the Assay, and determine the peak areas of curcumin, demethoxycurcumin and bisdemethox-ycurcumin: the peak area of curcumin is larger than the peak area of demethoxycurcumin and is larger than 0.69 times the peak area of bisdemethoxycurcumin.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Turmeric according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Turmeric according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 17.0z (6 hours). Total ash <5.01> Not more than 7.5z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 9.0z.
Weigh accurately about 0.2 g of pulverized Turmer-ic, add 25 mL of a mixture of methanol and acetic acid (100) (99:1), shake for 20 minutes, centrifuge, and separate the su-pernatant liquid. To the residue, add 25 mL of a mixture of methanol and acetic acid (100) (99:1), and proceed in the same manner as described above. Combine all the extracts, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of curcumin for assay, and dissolve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, add meth-anol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as described under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, ATC, ATD and ATD of curcumin, demethoxycurcumin and bis-demethoxycurcumin in the sample solution as well as the peak area AS of curcumin in the standard solution. ‌JP XVIII Crude Drugs and Related Drugs / Powdered Turmeric 2167 Amount (mg) of total curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin) ＝ MS × (ATC ＋ ATD ＋ ATB × 0.69)/AS × 1/5 MS: Amount (mg) of curcumin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilianized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (56:43:1). Flow rate: 1.0 mL per minute (the retention time of curcu-min is about 11 minutes). System suitability— System performance: Dissolve 1 mg each of curcumin for assay, demethoxycurcumin and bisdemethoxycurcumin in methanol to make 5 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, bisdemethoxycurcumin, demethoxycurcumin and curcumin are eluted in this order with the resolution among these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of curcumin is not more than 1.5z.
Nearly ovoid to oblong-ovoid or conical rhi-zome, 2 – 8 cm in length, 1.5 – 4 cm in diameter; externally grayish yellow-brown to grayish brown; nodes protruded as rings; internode of 0.3 – 0.8 cm, with scars of roots, and small protrusions consisting of scars of branched rhizomes; hard in texture; a transverse section reveals cortex and stele distinctly; cortex 2 – 5 mm in thickness; a transverse section, ‌1994‌ Cyperus Rhizome / Crude Drugs and Related Drugs grayish brown in rhizome of 1) Curcuma zedoria origin, light yellow to grayish yellow or light yellow-green to grayish yellow-green in 2) Curcuma phaeocaulis origin and purplish brown to dark purple-brown in 3) Curcuma kwangsiensis origin, and sometimes lustrous. Odor, characteristic; taste, pungent, bitter and cool feel-ing on chewing. Under a microscope <5.01>, a transverse section of central part reveals the outermost layer usually consisting of a cork layer 4 – 10 cells thick; cortex and stele divided by endoder-mis, composed of parenchyma cells, vascular bundles scat-tered; small sized vascular bundles line up beneath the endodermis; oil cells contain yellow-brown to dark brown oily substances, scattered in parenchyma; parenchyma contains gelatinized starch and rarely crystals of calcium oxalate.
To 2.0 g of pulverized Curcuma Rhizome add 5 mL of water, shake, then add 5 mL of hexane, shake for 10 minutes, centrifuge, and use the hexane layer as the sample solution. Perform the test with this solution as direct-ed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (4:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybezaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: a deep blue to dark brown spot and a red-brown to brown spot appear at Rf values of about 0.3 and about 0.2, respectively.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Curcuma Rhizome according to Method 3, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Curcuma Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Curcuma Rhizome, provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.5 mL.
Powdered Cyperus Rhizome occurs as a light red-brown powder, and has a characteristic odor and taste. Under a microscope <5.01>, Powdered Cyperus Rhizome reveals fragments of polygonal parenchyma cells, scalari-form vessels, and seta-like fibers; a large quantity of starch, mostly gelatinized; an extremely small number of stone cells.
To 2.0 g of Powdered Cyperus Rhizome add 10 mL of diethyl ether, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, cyclohexane and formic acid (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.35.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Cyperus Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Cyperus Rhizome according to Method 4, and ‌JP XVIII Crude Drugs and Related Drugs / Daiokanzoto Extract 1995‌ perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow-dered Cyperus Rhizome does not show extremely lignified cells, except stone cells, and crystals. Total ash <5.01> Not more than 3.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Cyperus Rhizome provided that 1 mL of silicon resin is previously added on the sample in the flask: the volume of essential oil is not less than 0.2 mL.
Fusiform rhizome, 1.5 – 2.5 cm in length, 0.5 – 1 cm in diameter; externally grayish brown to grayish blackish brown, with 5 to 8 irregular ring nodes, and with hair-like fiber bundles on each node; hard in texture. The transverse section red-brown to light yellow in color, with waxy luster; thickness of cortex approximately equal to or slightly smaller than the diameter of stele. Under a mag-nifying glass, a cut surface reveals fiber bundles as brown spots lined in rings along circumference; here and there in the cortex, vascular bundles appear as red-brown spots, and numerous secretory cells scattered as minute yellow-brown spots; in the stele, numerous vascular bundles scattered as spots or lines. Characteristic odor and taste.
To 2.0 g of pulverized Cyperus Rhizome add 10 mL of diethyl ether, shake for 5 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, cyclohexane and formic acid (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.35.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Cyperus Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Cyperus Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 3.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Cyperus Rhizome, provided that 1 mL of silicon resin is previously added on the sample in the flask: the volume of essential oil is not less than 0.3 mL.
Diluted Opium Powder contains not less than
solution as directed in Assay under Powdered Opium
Powdered Dioscorea Rhizome occurs as nearly yellowish white to white; odorless and tasteless. Under a microscope <5.01>, Dioscorea rhizome powder re- veals starch grains; fragments of parenchyma cells contain-ing starch grains; raphides of calcium oxalate, 100 to 200 mm in length and its containing mucilage cells; ring and scalariform vessels, 15 to 35 mm in diameter; starch grain isosceles deltoid or oblong, solitary, 18 to 35 mm, hilum and striation being distinct.
(1) To 0.2 g of Powdered Dioscorea Rhi-zome add 2 mL of acetic anhydride, warm on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add care-fully 0.5 mL of sulfuric acid to make two layers: a red-brown to purple-brown color develops at the zone of con-tact. (2) To 1 g of Powdered Dioscorea Rhizome add 4 mL of a mixture of methanol and water (4:1), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 1 mg of allantoin for thin-layer chromatography in 2 mL of a mixture of methanol and water (4:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solu-tion and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (7:3:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly a so-lution of 0.2 g of 4-dimethylaminocinnamaldehyde in 10 mL of 6 mol/L hydrochloric acid TS and 10 mL of ethanol (99.5) on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Dioscorea Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Dioscorea Rhizome according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Cylindrical or irregular cylindrical rhizome, 5 – 15 cm in length, 1 – 4 cm in diameter, occasionally longi-tudinally split or transversely cut; externally whitish to yel-lowish white; fractured surface, whitish, smooth and powd-ery; hard in texture but breakable. Practically odorless and tasteless.
(1) To the cut surface of Dioscorea Rhi-zome add diluted iodine TS (1 in 50) dropwise: a dark purple to dark blue color develops. To 0.2 g of pulverized Dioscorea Rhizome add 2 mL of acetic anhydride, warm on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add 0.5 mL of sulfuric acid carefully to make two layers: a red-brown to purple-brown color appears at the zone of contact. To 1 g of pulverized Dioscorea Rhizome add 4 mL of a mixture of methanol and water (4:1), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 1 mg of allantoin for thin-layer chromatography in 2 mL of a mixture of methanol and water (4:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solu-tion and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (7:3:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly a so-lution of 0.2 g of 4-dimethylaminocinnamaldehyde in 10 mL of 6 mol/L hydrochloric acid TS and 10 mL of ethanol (99.5) on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Dioscorea Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Dioscorea Rhizome according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Flattened ellipsoidal to flattened orbicular-ovate seed, 9 – 14 mm in length, 6 – 10 mm in width, 4 – 7 mm in thickness; externally light yellowish white to light yel-low, smooth and somewhat lustrous; caruncle white, like a half-moon, protrudent at one side; hard in texture. Almost odorless; taste, slightly sweet and acid. Under a microscope <5.01>, a transverse section reveals the outermost layer of seed coat composed of a single layer of palisade like epidermal cells coated with cuticle; beneath epidermis a single layer of sclerenchymatous and sandglass like cells; inside of the layer mentioned above parenchyma lie, the innermost portion of the parenchyma decayed; cotyledons occur inside of the seed coat; the outermost layer of cotyledon composed of a single layer of epidermal cells, inner part of cotyledon mainly parenchyma, containing aleu-rone grains and oil drops, and occasionally starch grains.
To 3 g of pulverized Dolichos Seed add 30 mL of methanol, shake for 10 minutes, centrifuge, and take the supernatant liquid. Evaporate the solvent of the superna-tant liquid, add 30 mL of water and 50 mL of ethyl acetate to the residue, shake, and take the ethyl acetate layer. To the ethyl acetate add 10 g of anhydrous sodium sulfate, shake, and filter. Evaporate the solvent of the filtrate, add 1 mL of ethyl acetate to the residue, and use this solution as the sam-ple solution. Perform the test with the sample solution as di-rected under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate and acetic acid (100) (100:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a blue-white fluorescent spot ap-pears at an Rf value of about 0.4. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 4.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 9.0z.
Slightly curved subcolumnar rhizome, 15 – 30 cm in length, 1 – 2.5 cm in diameter; externally grayish brown and slightly rough; transversely cut surface light brown, cortex thin, xylem thick with a pith in center; ex-tremely hard in texture. Odor, slightly characteristics; tasteless or slightly sweet, astringency. Under a microscope <5.01>, a transverse section reveals the outermost layer consisting of a cork layer 3 – 7 cells thick; oil canals scattered in parenchyma; fiber bundles lined stepwise in phloem; phloem and xylem separated clearly by cambium; xylem composed of vessels, xylem fibers and xylem paren-chyma; ray composed of 2 – 6 rows of cells; pith composed of parenchyma; parenchyma of cortex and ray contain ag-gregate crystals of calcium oxalate; occasionally starch grains in ray, parenchyma of cortex and xylem.
To 0.5 g of pulverized Eleutherococcus Senticosus Rhizome add 20 mL of diluted methanol (1 in 2), shake for 15 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of eleutheroside B for liquid chromatography in diluted metha-nol (1 in 2) to make 20 mL. To 2 mL of this solution add diluted methanol (1 in 2) to make 20 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions: the peak corresponding to eleutheroside B in the chromatogram obtained from the sample solution shows the same retention time with the peak of eleutheroside B in the chromatogram from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water and acetonitrile (9:1). Flow rate: Adjust so that the retention time of eleuthero- side B is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of eleutheroside B are not less than 5000 and not more than 1.5, respectively.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Eleutherococcus Senticosus Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Eleutherococcus Senticosus Rhizome accord-ing to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 2.5z.
Thin cylindrical or ellipsoidal cylinder, 0.1 – 0.2 cm in diameter; 3 – 5 cm in length of internode; light green to yellow-green; numerous parallel vertical furrows on the surface; scaly leaves at the node portion; leaves, 0.2 – 0.4 cm in length, light brown to brown in color, usually being opposite at every node, adhering at the base to form a tubu-lar sheath around the stem. Under a magnifying glass, the transverse section of the stem appears as circle and ellipse, the outer portion grayish green to yellow-green in color, and the center filled with a red-purple substance or hollow. When fractured at internode, the outer part is fibrous and easily split vertically. Odor, slight; taste, astringent and slightly bitter, giving a slight sensation of numbness on the tongue.
To 0.5 g of pulverized Ephedra Herb add 10 mL of methanol, shake for 2 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly ninhydrin-ethanol TS for spraying, and heat the plate at 1059C for 5 minutes: a red-purple spot appears at an Rf value of about 0.35.
(1) Woody stem—When perform the test of for-eign matter <5.01>, the amount of the woody stems con-tained in Ephedra Herb does not exceed 5.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than woody stems contained in Ephedra Herb does not exceed 1.0z. Loss on drying <5.01> Not more than 12.5z (6 hours). Total ash <5.01> Not more than 11.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of moderately fine powder of Ephedra Herb, place in a glass-stoppered centri-fuge tube, add 20 mL of diluted methanol (1 in 2), shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 20 mL of diluted methanol (1 in 2), pro-ceed in the same manner, and repeat this procedure twice. Combine all the extracts, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of ephe-drine hydrochloride for assay of crude drugs, previously dried at 1059C for 3 hours, and dissolve in diluted methanol (1 in 2) to make exactly 20 mL. Pipet 2 mL of the solution, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATE and ATP, of ephedrine and pseudoephedrine (the relative retention time to ephedrine is about 0.9) obtained from the sample solution, and the peak area, AS, of ephe-drine obtained from the standard solution. Amount (mg) of total alkaloids (ephedrine and pseudoe-phedrine) ＝ MS × (ATE ＋ ATP)/AS × 1/10 × 0.819 MS: Amount (mg) of ephedrine hydrochloride for assay of crude drugs taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 5 g of sodium lauryl sulfate add 350 mL of acetonitrile, shake, and add 650 mL of water and 1 mL of phosphoric acid to dissolve lauryl sulfate. Flow rate: Adjust so that the retention time of ephedrine is about 27 minutes. System suitability— System performance: Dissolve 1 mg of ephedrine hydro-chloride for assay of crude drugs and 1 mg of pseudoephed-rine hydrochloride in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, pseudoephedrine and ephedrine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 1.5z.
Epimedium Herb is composed of a stem and a ternate to triternate compound leaf; leaflet ovate to broadly ovate or ovate-lanceolate, 3 – 20 cm in length, 2 – 8 cm in width, petiolule 1.5 – 7 cm in length, apex of leaflet acuminate, needle hair on margin 0.1 – 0.2 cm in length, base of leaflet cordate to deeply cordate, lateral leaflet asymmetry; upper surface green to green-brown, sometimes lustrous, lower surface light green to grayish green-brown, often pilose, especially on vein densely pilose, papery or coriaceous; petiole and stem cylindrical, light yellowish brown to slightly purplish and light green-brown, easily broken. ‌2002‌ Eucalyptus Oil / Crude Drugs and Related Drugs Odor, slight; taste, slightly bitter. Under a microscope <5.01>, a transverse section of the leaf reveals 3 – 6 vascular bundles in midvein; mesophyll composed of upper epidermis, single-layered palisade, spongy tissue and lower epidermis; leaf margins orbicular or oblong, sclerenchymatous; multi-cellular hairs on epidermis; 8 – 20 vascular bundles in petiole and 6 – 15 vascular bundles in petiolule. Under a microscope <5.01>, a transverse section of the stem reveals a single to several-layered hypodermis, cortex of 4 – 10 cellular layers of sclerenchyma, vascular bundle 13 – 30 in number, oblong to obovate.
To 2 g of pulverized Epimedium Herb add 20 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of icariin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 12.5z (6 hours). Total ash <5.01> Not more than 8.5z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 17.0z.
Loquat Leaf is an oblong to wide lanceolate leaf, 12 – 30 cm in length, 4 – 9 cm in width; pointed at the apex and wedge-shaped at the base; roughly serrate leaf with short petiole; occasionally being cut into strips 5 – 10 mm in shorter diameter and several cm in longer diameter; upper surface green to green-brown in color, lower surface light green-brown with light brown woolly hairs; vein, light yel-low-brown in color, raised out on the lower surface of the leaf. Odor, slight; practically tasteless. Under a microscope <5.01>, a transverse section of Loquat Leaf reveals thick cuticle on both surfaces; palisade tissue, mostly 4 to 5 cellular layers with several large cells without chloroplast; at main vein, ring of collateral bundle partly cut by intruding fundamental tissue at xylem side, and group of fiber attaching to phloem; solitary and clustered crystals of calcium oxalate in mesophyll; woolly hair, unicellular and curved, about 25 mm in thickness, and up to 1.5 mm in length.
To 0.3 g of pulverized Loquat Leaf add 10 mL of methanol, warm on a water bath for 5 minutes with occasional shaking, cool, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of water and acetonitrile (3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: a red-pur-ple principal spot appears at an Rf value of about 0.5.
Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 10.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 16.0z.
Eucommia Bark is a semi-tubular or plate-like bark, 2 – 6 mm in thickness; externally pale grayish brown to grayish brown, and rough in texture, sometimes red-brown due to the cork layer falling off; internally dark violet, smooth and covered with a linear pattern that runs longitudi-nally, silk-like threads of gutta-percha (a thermoplastic rub-ber-like substance) appearing when broken. It has a faint but characteristic odor and taste. Under a microscope <5.01>, transverse section reveals parenchymatous cells containing gutta-percha; phloem with stone-cell and fiber layers; rays in rows of 2 – 3 cells; calcium oxalate crystals absent. ‌JP XVIII Crude Drugs and Related Drugs / Powdered Fennel 2003‌‌ ‌Identification Put 1 g of pulverized Eucommia Bark in a glass-stoppered centrifuge tube, add 10 mL of water and 20 mL of diethyl ether, shake for 15 minutes, and centrifuge. Take the diethyl ether layer so obtained, evaporate the solvent on a water bath, and add 1 mL of ethanol (99.5) to the residue: colloidal substances appear. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 7.0z.
Flattened spheroidal or globular fruit, 2 – 5 mm in diameter; externally dark brown to grayish brown, with many oil sacs appearing as hollow pits, and often with peduncle, 2 – 5 mm in length, covered densely with hairs; pericarp in matured split to five loculi, and each loculus containing obovoid or globular seeds of a lustrous brown to black-brown or bluish black color. Odor, characteristic; taste, acrid, followed by a lasting bitterness.
To 1.0 g of pulverized Euodia Fruit add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of acetone, 2-propanol, water and formic acid (7:7:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a blue-white fluorescent spot is ob-served at an Rf value of about 0.6. The spot shows a yellow-red color after being sprayed evenly Dragendorff's TS for spraying.
(1) Peduncle—The amount of peduncles contained in Euodia Fruit does not exceed 5.0z. (2) Foreign matter <5.01>—The amount of foreign mat- ter other than peduncles contained in Euodia Fruit does not exceed 1.0z. Total ash <5.01> Not more than 8.0z.
Powdered Fennel occurs as a greenish light brown to greenish brown, and is a characteristic odor and taste. Under a microscope <5.01>, Powdered Fennel reveals frag- ments of parenchyma cells of perisperm containing aleurone grain, fragments of parenchyma cells of endosperm contain-ing fatty oil, fragments of sclerenchyma with characteristic simple pits, fragments of oil canal within yellow-brown ma-terial, fragments of endocarp shown scalariform, spiral ves-sels, fragments of epidermis or epidermis with stomata. ‌2004‌‌ Fennel Oil / Crude Drugs and Related Drugs
To 0.5 g of Powdered Fennel add 10 mL of hexane, allow to stand for 5 minutes with occasional shak-ing, filter, and use the filtrate as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate prepared with silica gel with fluorescent indicator for thin-layer chromatography. Then develop the plate with a mixture of hexane and ethyl acetate (20:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot with dark purple color appears at an Rf value of about 0.4. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Fennel: the volume of essential oil is not less than 0.45 mL.
Cylindrical cremocarp, 3.5 – 8 mm in length, 1 – 2.5 mm in width; externally grayish yellow-green to grayish yellow; two mericarps closely attached with each other, and with five longitudinal ridges; cremocarp often with pedicel 2 – 10 mm in length. Characteristic odor and taste. Under a microscope <5.01>, ridges near the ventral side are far protruded than those on the dorsal side; one large oil canal between each ridge, and two oil canals on the ventral side.
To 0.5 g of pulverized Fennel add 10 mL of hexane, allow to stand for 5 minutes with occasional shak-ing, filter, and use the filtrate as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (20:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot with a dark purple color appears at an Rf value of about 0.4.
(1) Peduncle—When perform the test of foreign matter <5.01>, the amount of peduncles contained in Fennel does not exceed 3.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than the peduncle contained in Fennel does not exceed 1.0z. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Fennel: the volume of essential oil is not less than 0.7 mL.
Ovoid to long ovoid capsule, 1.5 – 2.5 cm in length, 0.5 – 1 cm in width, with acute apex, and sometimes with a peduncle at the base; externally light gray to dark brown, scattered with light gray and small ridged dots, and with two longitudinal furrows; a capsule dehiscing along the longitudinal furrows has the apexes bent backward; the inner surface of dehisced pericarp is yellow-brown in color, with a longitudinal partition-wall in the middle; seeds, slender and oblong, 0.5 – 0.7 cm in length, and usually with a wing. Odor, slight; taste, slightly bitter.
To 1.0 g of pulverized Forsythia Fruit add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (20:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: a red-purple to red-brown spot is observed at an Rf value of about 0.3.
(1) Branchlet—When perform the test of foreign matter <5.01>, the amount of branchlets contained in For-sythia Fruit does not exceed 5.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than branchlets contained in Forsythia Fruit does not exceed 1.0z. Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not ‌JP XVIII Crude Drugs and Related Drugs / Powdered Gambir 2005‌‌‌ less than 10.0z.
Fritillaria Bulb is a depressed spherical bulb, 2 – 3 cm in diameter, 1 – 2 cm in height, consisting of 2 thickened scaly leaves often separated; externally and inter-nally white to light yellow-brown in color; inside base is in a slightly dark color; the bulb sprinkled with lime before dry-ing is dusted with white powder; fractured surface, white in color and powdery. Odor, slight and characteristic; taste, bitter. ‌Under a microscope <5.01>, a transverse section reveals the outermost layer to be composed of an epidermis; numerous vascular bundles scattered throughout the parenchyma inside of the epidermis; parenchyma filled with starch grains; starch grains are mainly simple (rarely 2- to 3-compound), 5 – 60 mm in diameter, narrowly ovate to ovate or triangular to obovate, stratiform figure obvious; epidermal cells and parenchyma cells near the vessels contain solitary crystals of calcium oxalate.
Put 2 g of pulverized Fritillaria Bulb in a glass-stoppered centrifuge tube, add 10 mL of ammonia TS and 20 mL of a mixture of ethyl acetate and diethyl ether (1:1), shake for 20 minutes, and centrifuge. Take the upper layer, add 20 g of anhydrous sodium sulfate to the layer, shake, and filter. Evaporate the solvent of the filtrate, dis-solve the residue in 1 mL of ethanol (99.5), and use this solu-tion as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (17:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: spots of a yellow-red color appear at Rf values of about 0.4 and about 0.6.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Fritillaria Bulb according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Fritillaria Bulb according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 16.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
Oval caryopsis, 10 mm in length, 3 – 4 mm in width, furrowed on one surface; externally light yellow, sometimes with plumule at one end, with hairs and some-times with roots at the other end; cut surface of caryopsis white and powdery; easily broken and light in texture. Odor, slight; taste, slightly sweet. Under a microscope <5.01>, a transverse section of the caryopsis reveals glume, pericarp, seed coat and endosperm in this order from the outside; 2 – 4 cellular layered aleurone layers on the circumference of endosperm; endosperm filled with starch grains; starch grains as spheroidal or ellipsoidal, large grains about 20 mm and small grains about 2 mm in di-ameter mixed together.
To 3.0 g of pulverized Malt add 5 mL of methanol, shake for 15 minutes, centrifuge, and use the su-pernatant liquid as the sample solution. Perform the test with the sample solution as directed under Liquid Chroma-tography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, water and acetic acid (100) (8:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly a solution of 0.1 g of 2,3-indolinedione in 50 mL of acetone on the plate, and heat the plate at 1059C for 5 minutes: a blue-purple spot appears at an Rf value of about 0.4. Loss on drying <5.01> Not more than 11.0z. Total ash <5.01> Not more than 2.6z. Acid-insoluble ash <5.01> Not more than 0.8z. Extract content <5.01> Dilute ethanol-soluble extract: Not ‌2070 Maoto Extract / Crude Drugs and Related Drugs less than 15.0z.
Gambir is the dried aqueous extract prepared from
Powdered Gardenia Fruit occurs as a yellow-brown powder, and has a slight odor and a bitter taste. Under a microscope <5.01>, Powdered Gardenia Fruit reveals fragments of yellow-brown epidermis consisting of polygonal epidermal cells in surface view; unicellular hairs, spiral and ring vessels, stone cells often containing crystals of calcium oxalate; fragments of thin-walled parenchyma containing yellow pigments, oil drops and rosette aggregates of calcium oxalate (the above elements from fruit receptacle and pericarp); fragments of large and thick-walled epidermis of seed coat, containing a red-brown substance; fragments of endosperm filled with aleuron grains (the above elements from seed).
(1) To 1.0 g of Powdered Gardenia Fruit, previously dried in a desiccator (silica gel) for 24 hours, add 100 mL of hot water, warm the mixture between 609C and 709C for 30 minutes with frequent shaking, and filter after cooling. To 1.0 mL of the filtrate add water to make 10 mL: the color of the resulting solution is yellow and is not lighter ‌JP XVIII Crude Drugs and Related Drugs / Gentian 2007 than that of the following control solution. ‌Control solution: Dissolve 9.8 mg of carbazochrome so-dium sulfonate trihydrate in water to make exactly 10 mL. Pipet 1 mL of this solution, and add water to make exactly 50 mL. (2) To 1.0 g of Powdered Gardenia Fruit add 20 mL of methanol, warm for 3 minutes on a water bath, cool, filter, and use the filtrate as the sample solution. Separately, dis-solve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and methanol (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxy-benzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 13.0z. Total ash <5.01> Not more than 6.0z.
Weigh accurately about 0.5 g of Powdered Gardenia Fruit, transfer into a glass-stoppered centrifuge tube, add 40 mL of diluted methanol (1 in 2), shake for 15 minutes, cen-trifuge, and take the supernatant liquid. To the residue add 40 mL of diluted methanol (1 in 2), and repeat the same procedure as above. Combine the extracts so obtained, and add diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of the solution, add methanol to make exactly 20 mL, use this solution as the sample solution. Separately, weigh accurately about 10 mg of geniposide for assay, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of the solu-tion, add methanol to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. Amount (mg) of geniposide ＝ MS × AT/AS × 2 MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (22:3). Flow rate: Adjust so that the retention time of geniposide is about 15 minutes. System suitability— System performance: Dissolve 1 mg each of geniposide for assay and caffeine in methanol to make 15 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, caffeine and geniposide are eluted in this order with the resolution between these peaks being not less than 3.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z.
Nearly long ovoid to ovoid fruit, 1 – 5 cm in length, 1 – 1.5 cm in width; externally yellow-brown to yellow-red, usually having 6, rarely 5 or 7, markedly raised ridges; calyx or its scar at one end, and sometimes peduncle at the other end; inner surface of pericarp yellow-brown, smooth and lustrous; internally divided into two loculi, con-taining a mass of seeds in yellow-red to dark red placenta; seed nearly circular, flat, about 0.5 cm in major axis, black-brown or yellow-red. Odor, slight; taste, bitter.
(1) To 1.0 g of pulverized Gardenia Fruit, previously dried in a desiccator (silica gel) for 24 hours, add 100 mL of hot water, warm the mixture between 609C and 709C for 30 minutes with frequent shaking, and filter after cooling. To 1.0 mL of the filtrate add water to make 10 mL: the color of the resulting solution is yellow and is not lighter than that of the following control solution. Control solution: Dissolve 9.8 mg of carbazochrome so-dium sulfonate trihydrate in water to make exactly 10 mL. Pipet 1 mL of this solution, and add water to make exactly 50 mL. (2) To 1.0 g of pulverized Gardenia Fruit add 20 mL of methanol, warm for 3 minutes on a water bath, cool, filter, and use the filtrate as the sample solution. Separately, dis-solve 1 mg of geniposide for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and methanol (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxy-benzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 13.0z. Total ash <5.01> Not more than 6.0z.
Weigh accurately about 0.5 g of pulverized Gardenia Fruit, transfer into a glass-stoppered centrifuge tube, add 40 mL of diluted methanol (1 in 2), shake for 15 minutes, cen-trifuge, and take the supernatant liquid. To the residue add 40 mL of diluted methanol (1 in 2), and repeat the same procedure as above. Combine the extracts so obtained, and add diluted methanol (1 in 2) to make exactly 100 mL. Pipet 5 mL of the solution, add methanol to make exactly 20 mL, use this solution as the sample solution. Separately, weigh accurately about 10 mg of geniposide for assay, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of the solu-tion, add methanol to make exactly 10 mL, and use this solu- tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of geniposide in each solution. Amount (mg) of geniposide ＝ MS × AT/AS × 2 MS: Amount (mg) of geniposide for assay taken, calcu-lated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (22:3). Flow rate: Adjust so that the retention time of geniposide is about 15 minutes. System suitability— System performance: Dissolve 1 mg each of geniposide for assay and caffeine in methanol to make 15 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, caffeine and geniposide are eluted in this order with the resolution between these peaks being not less than 3.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of geniposide is not more than 1.5z.
Gastrodia Tuber is an irregularly curved and flattened cylindrical to flattened fusiform tuber, 5 – 15 cm in length, 2 – 5 cm in diameter, 1 – 2 cm in thickness; externally light yellow-brown to light yellow-white; with ring nodes, and irregular longitudinal wrinkles; hard in texture; frac-tured surface, dark brown to yellow-brown in color, with luster, horny and gluey. Odor, characteristic; practically tasteless. Under a microscope <5.01>, a transverse section reveals parenchyma cells containing raphides of calcium oxalate; starch grain absent.
To 1 g of pulverized Gastrodia Tuber add 5 mL of methanol, shake for 15 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 1 mL of metha-nol, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, de-velop the plate with a mixture of ethyl acetate, methanol and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: a red-purple to light brown spot appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Gastrodia Tuber according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Gastrodia Tuber according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 16.0z (6 hours). Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 16.0z.
Powdered Gentian occurs as a yellow-brown powder, and has a characteristic odor. It has a sweet taste at first, which later becomes persistently bitter. Under a microscope <5.01>, Powdered Gentian reveals parenchyma cells containing oil droplets and minute needle crystals, vessels, tracheids, cork tissues, and crystals of cal- cium oxalate. Vessels are chiefly reticulate vessels and scalariform vessels, 20 – 80 mm in diameter. Starch grains are observed very rarely, in simple grains about 10 – 20 mm in diameter.
(1) Place 0.1 g of Powdered Gentian, pre-viously dried in a desiccator (silica gel) for 48 hours, on a slide glass, put a glass ring 10 mm in both inside diameter and in height on it, then cover with another slide glass, and heat gently and gradually: light yellow crystals are sublimed on the upper glass. The crystals are insoluble in water and in ethanol (95), and soluble in potassium hydroxide TS. (2) To 0.5 g of Powdered Gentian add 10 mL of metha-nol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of gentiopicroside for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution and a spot from the standard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Powdered Gentian according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Gentian according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, stone cell and fiber are not observed. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 3.0z.
Nearly cylindrical pieces, 10 – 50 cm in length, 2 – 4 cm in diameter; externally dark brown; the rhizome short, with fine, transverse wrinkles, and sometimes with ‌2008‌ Powdered Gentian / Crude Drugs and Related Drugs buds and remains of leaves at the upper edge. The root lon-gitudinally and deeply wrinkled, and more or less twisted; fractured surface yellow-brown and not fibrous, and a cam-bium and its neighborhood tinged dark brown. Odor, characteristic; taste, sweet at first, later persistently bitter. Under a microscope <5.01>, a transverse section of the root reveals several cellular layers of collenchyma adjoined internally to 4 to 6 cellular layers of thin-walled cork; secondary cortex with irregularly distributed phloem; xylem consisting chiefly of parenchyma, with individual or clustered vessels and tracheids, and exhibiting some sieve tubes of xylem; parenchyma of the xylem and the cortex con-taining oil droplets, minute needle crystals of calcium oxa-late and very rarely starch grains 10 – 20 mm in diameter.
(1) Place 0.1 g of pulverized Gentian, pre-viously dried in a desiccator (silica gel) for 48 hours, on a slide glass, put a glass ring 10 mm in both inside diameter and in height on it, then cover with another slide, and heat gently and gradually: pale yellow crystals are sublimed on the upper slide. The crystals are insoluble in water and in ethanol (95), and soluble in potassium hydroxide TS. (2) To 0.5 g of pulverized Gentian add 10 mL of metha-nol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of gentiopicroside for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution and a spot from the standard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Gentian according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Gentian according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 3.0z.
Powdered Japanese Gentian occurs as a grayish yellow-brown powder. It has a slight odor and a lasting, extremely bitter taste. Under a microscope <5.01>, Powdered Japanese Gentian reveals fragments of parenchyma cells containing oil droplets and fine crystals, fragments of endodermis divided into daughter cells with suberized cell wall, exodermis, and vessels; vessels mainly reticulate vessels and scalariform ves-sels, 20 – 30 mm in diameter.
To 0.5 g of Powdered Japanese Gentian add 10 mL of methanol, shake for 20 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of gentiopicroside for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solu-tion and a spot from the standard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Japanese Gentian according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Japanese Gentian according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Japanese Gentian usually reveals no stone cells and fibers. No starch grains; if any, very few. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 3.0z.
Irregular, cylindrical, short rhizome with numerous, slender roots around, and externally yellow-brown to grayish yellow-brown. The root is 10 – 15 cm in length, about 0.3 cm in diameter, and has longitudinal, coarse wrinkles on the outer surface; flexible; fractured surface, smooth and yellow-brown in color. The rhizome is about 2 cm in length, about 0.7 cm in diameter, and has buds or short remains of stems at the top. Odor, slight; taste, extremely bitter and lasting. Under a microscope <5.01>, a transverse section of the young root reveals epidermis, exodermis and a few cellular layers of primary cortex; usually, the outermost layer is endodermis consisting of characteristic cells divided into a few daughter cells, often with collenchyma of 1 to 2 cellular layers contacting the inner side; secondary cortex having rents here and there, and irregularly scattered sieve tubes; vessels arranged rather radially in xylem, sieve tubes existing in xylem; the rhizome has a large pith, rarely with sieve tubes; parenchyma cells contain small needle, plate or sand crystals of calcium oxalate and oil drops; starch grains usu-ally absent.
To 0.5 g of pulverized Japanese Gentian add 10 mL of methanol, shake for 20 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of gentiopicroside for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solu-tion has the same color tone and the same Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Japanese Gentian according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Japanese Gentian according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 3.0z.
Powdered Geranium Herb occurs as a grayish green to light yellow-brown powder. It has a slight odor and an astringent taste. Under a microscope <5.01>, Powdered Geranium Herb reveals mainly fibers, spiral vessels, pitted vessels, and unicellular hairs; furthermore, multicellular glandular hairs, epidermis with stomata, fragments of palisade tissue, rosette aggregates of calcium oxalate, and starch grains. Fiber is thick-walled, with somewhat distinct pits; unicellular hair shows small point-like protrusions on the surface; palisade tissue consisting of circular parenchyma cells in surface view, each cell containing one rosette aggregate of calcium oxalate which is about 20 mm in diameter. Starch grains consisting of simple grains but rarely of 2-compound grains, ovoid to spherical, 5 – 30 mm in diameter, with distinct hilum.
Boil 0.1 g of Powdered Geranium Herb with 10 mL of water, filter, and to the filtrate add 1 drop of iron (III) chloride TS: a dark blue color develops.
Foreign matter—Under a microscope <5.01>, Pow-dered Geranium Herb reveals no stone cells. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 15.0z.
Stem with leaves opposite; stem, slender and long, green-brown; stem and leaf covered with soft hairs; leaf divided palmately into 3 to 5 lobes, and 2 – 4 cm in length, grayish yellow-green to grayish brown; each lobe oblong to obovate, and its upper margin crenate. Odor, slight; taste, astringent.
Boil 0.1 g of Geranium Herb with 10 mL of water, filter, and to the filtrate add 1 drop of iron (III) chlo-ride TS: a blackish blue color develops.
Foreign matter <5.01>—The amount of the root and other foreign matter contained in Geranium Herb does not exceed 2.0z. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 15.0z.
Powdered Ginseng occurs as a light yellow-white to light yellow-brown powder. It has characteristic odor and is a slight sweet taste followed by a slight bitter-ness. for spraying on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Powdered Ginseng according to Method 4, and perform the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Powdered Ginseng according to Method 4, and perform the test (not more than 2 ppm). Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 4.2z. Acid-insoluble ash <5.01> Not more than 0.5z. Extract content <5.01> Dilute ethanol-soluble extract; not less than 14.0z.
(1) Ginsenoside Rg1—Weigh accurately about 1.0 g of Powdered Ginseng, put in a glass-stoppered centri-fuge tube, add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. Repeat the procedure with the residue using 15 mL of diluted methanol (3 in 5), combine the supernatant liquids, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of dilute sodium hydroxide TS, allow to stand for 30 minutes, add 3 mL of 0.1 mol/L hydrochloric acid TS and diluted methanol (3 in 5) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rg1 RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ginsenoside Rg1 in each solution. ‌JP XVIII Crude Drugs and Related Drugs / Glycyrrhiza 2013‌ Amount (mg) of ginsenoside Rg1 (C42H72O14) ＝ MS × AT/AS MS: Amount (mg) of Ginsenoside Rg1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of ginsenoside Rg1 is about 25 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rg1 RS and ginsenoside Re in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rg1 and ginsenoside Re are eluted in this order with the reso-lution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rg1 is not more than 1.5z. (2) Ginsenoside Rb1—Use the sample solution obtained in (1) as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determined the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS MS: Amount (mg) of Ginsenoside Rb1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of ginsenoside Rb1 is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rb1 RS and ginsenoside Rc in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rb1 and ginsenoside Rc are eluted in this order with the reso-lution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z.
Thin and long cylindrical to fusiform root, often branching out into 2 to 5 lateral roots from the middle; 15 minutes, centrifuge, and separate the supernatant liquid. Repeat the procedure with the residue using 15 mL of diluted methanol (3 in 5), combine the supernatant liquids, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of dilute sodium hydroxide TS, allow to stand for 30 minutes, add 3 mL of 0.1 mol/L hydrochloric acid TS and diluted methanol (3 in 5) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rg1 RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg) dissolve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ginsenoside Rg1 in each solution. 5 – 25 cm in length, main root 0.5 – 3 cm in diameter; exter-nally light yellow-brown to red-brown, and translucent and with longitudinal wrinkles; crown somewhat constricted, and Amount (mg) of ginsenoside Rg1 ＝ MS × AT/AS (C42H 72O 14) sometimes with short remains of stem; fractured surface flat; horny and hard in texture. Odor, characteristic; taste, at first slightly sweet, followed by a slight bitterness.
(1) To 0.2 g of pulverized Red Ginseng add 2 mL of acetic anhydride, warm on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add gently 0.5 mL of sulfuric acid to make two layers: a red-brown color develops at the zone of contact. (2) To 2.0 g of pulverized Red Ginseng add 10 mL of water and 10 mL of 1-butanol, shake for 15 minutes, centri-fuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of ginsenoside Rg1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 2 mL of the standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol and water (14:5:4) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Red Ginseng according to Method 4, and perform the test. Prepare the control solution with 1.5 mL of Stand-ard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of pulverized Red Ginseng according to Method 4, and per-form the test (not more than 2 ppm). Foreign matter <5.01>—The amount of stems and other foreign matter contained in Red Ginseng does not exceed 2.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 15.5z (6 hours). Total ash <5.01> Not more than 4.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 18.0z.
(1) Ginsenoside Rg1—Weigh accurately about 1 g of pulverized Red Ginseng, put in a glass-stoppered centri-fuge tube, add 30 mL of diluted methanol (3 in 5), shake for MS: Amount (mg) of Ginsenoside Rg1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of ginsenoside Rg1 is about 25 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rg1 RS and ginsenoside Re in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rg1 and ginsenoside Re are eluted in this order with the reso-lution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rg1 is not more than 1.5z. (2) Ginsenoside Rb1—Use the sample solution obtained in (1) as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS MS: Amount (mg) of Ginsenoside Rb1 RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ‌JP XVIII Crude Drugs and Related Drugs / Rhubarb 2113‌ Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of ginsenoside Rb1 is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rb1 RS and ginsenoside Rc in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rb1 and ginsenoside Rc are eluted in this order with the reso-lution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z.
Thin and long cylindrical to fusiform root, often branching 2 to 5 lateral roots from the middle; 5 – 20 cm in length, main root 0.5 – 3 cm in diameter; externally light yellow-brown to light grayish brown, with longitudinal the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) On a section of Ginseng add dilute iodine TS dropwise: a dark blue color is produced on the surface. (2) To 2.0 g of pulverized Ginseng add 10 mL of water and 10 mL of 1-butanol, shake for 15 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Sepa-rately, dissolve 1 mg of ginsenoside Rg1 for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of ginsenoside Rg1 is about 25 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rg1 RS and ginsenoside Re in diluted methanol (3 in 5) to Powdered Ginseng / Crude Drugs and Related Drugs make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rg1 and ginsenoside Re are eluted in this order with the reso-lution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rg1 is not more than 1.5z. ‌(2) Ginsenoside Rb1—Use the sample solution obtained in (1) as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS Under a microscope <5.01>, Powdered Ginseng reveals round to rectangular parenchyma cells containing starch grains, occasionally gelatinized starch, vessels, secretory cell, sclerenchyma cell, big and thin-walled cork cell; crystals of calcium oxalate and starch. Vessels are reticulate vessel frag-ments, scalariform vessel and spiral vessel, 15 – 40 mm in di-ameter. Secretory cell containing a mass of yellow glistened contents; rosette aggregate of calcium oxalate, 20 – 60 mm in diameter, and 1 – 5 mm in diameter, rarely up to 30 mm in di-ameter of its single crystal; sclerenchymatous cells and thin-walled cork cells. Starch grains are observed in simple grain and 2 to 6-compound grain, simple grain, 3 – 20 mm in diam-eter. Identification To 2.0 g of Powdered Ginseng add 10 mL of water and 10 mL of 1-butanol, shake for 15 minutes, centri-fuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of ginsenoside Rg1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 2 mL of the standard MS: Amount (mg) of Ginsenoside Rb1 lated on the anhydrous basis Operating conditions— RS taken, calcu- solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol and water (14:5:4) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of ginsenoside Rb1 is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rb1 RS and ginsenoside Rc in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rb1 and ginsenoside Rc are eluted in this order with the reso-lution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Ginseng according to Method 4, and perform the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of pulverized Ginseng according to Method 4, and perform the test (not more than 2 ppm). Foreign matter <5.01>—The amount of stems and other foreign matter contained in Ginseng does not exceed 2.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 4.2z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 14.0z.
(1) Ginsenoside Rg1—Weigh accurately about 1.0 g of pulverized Ginseng, put in a glass-stoppered centri-fuge tube, add 30 mL of diluted methanol (3 in 5), shake for 15 minutes, centrifuge, and separate the supernatant liquid. Repeat the procedure with the residue using 15 mL of diluted methanol (3 in 5), combine the supernatant liquids, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 3 mL of dilute sodium hydroxide TS, allow to stand for 30 minutes, add 3 mL of 0.1 mol/L hydrochloric acid TS and diluted methanol (3 in 5) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rg1 RS (separately determine the water <2.48> by coulomet-ric titration, using 10 mg), dissolve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ginsenoside Rg1 in each solution. Amount (mg) of ginsenoside Rg1 (C42H72O14) ＝ MS × AT/AS wrinkles and scars of rootlets; sometimes crown somewhat constricted and with short remains of rhizome; fractured surface practically flat, light yellow-brown in color, and brown in the neighborhood of the cambium. MS: Amount (mg) of Ginsenoside Rg1 lated on the anhydrous basis Operating conditions— RS taken, calcu- Odor, characteristic; taste, at first slightly sweet, followed by a slight bitterness. Identification (1) On a section of Ginseng add dilute iodine TS dropwise: a dark blue color is produced on the surface. (2) To 2.0 g of pulverized Ginseng add 10 mL of water and 10 mL of 1-butanol, shake for 15 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Sepa-rately, dissolve 1 mg of ginsenoside Rg1 for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (4:1). Flow rate: Adjust so that the retention time of ginsenoside Rg1 is about 25 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rg1 RS and ginsenoside Re in diluted methanol (3 in 5) to Powdered Ginseng / Crude Drugs and Related Drugs make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rg1 and ginsenoside Re are eluted in this order with the reso-lution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rg1 is not more than 1.5z. ‌(2) Ginsenoside Rb1—Use the sample solution obtained in (1) as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ginsenoside Rb1 in each solu-tion. Amount (mg) of ginsenoside Rb1 (C54H92O23) ＝ MS × AT/AS Under a microscope <5.01>, Powdered Ginseng reveals round to rectangular parenchyma cells containing starch grains, occasionally gelatinized starch, vessels, secretory cell, sclerenchyma cell, big and thin-walled cork cell; crystals of calcium oxalate and starch. Vessels are reticulate vessel frag-ments, scalariform vessel and spiral vessel, 15 – 40 mm in di-ameter. Secretory cell containing a mass of yellow glistened contents; rosette aggregate of calcium oxalate, 20 – 60 mm in diameter, and 1 – 5 mm in diameter, rarely up to 30 mm in di-ameter of its single crystal; sclerenchymatous cells and thin-walled cork cells. Starch grains are observed in simple grain and 2 to 6-compound grain, simple grain, 3 – 20 mm in diam-eter. Identification To 2.0 g of Powdered Ginseng add 10 mL of water and 10 mL of 1-butanol, shake for 15 minutes, centri-fuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of ginsenoside Rg1 for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution and 2 mL of the standard MS: Amount (mg) of Ginsenoside Rb1 lated on the anhydrous basis Operating conditions— RS taken, calcu- solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol and water (14:5:4) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS Detector: An ultraviolet absorption photometer (wave-length: 203 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of ginsenoside Rb1 is about 20 minutes. System suitability— System performance: Dissolve 1 mg each of Ginsenoside Rb1 RS and ginsenoside Rc in diluted methanol (3 in 5) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ginsenoside Rb1 and ginsenoside Rc are eluted in this order with the reso-lution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ginsenoside Rb1 is not more than 1.5z.
Cylindrical to long conical root or rhizome, 10 – 20 cm in length, 0.5 – 1.5 cm in diameter; externally light yellow-brown to red-brown. Rhizome short, with fine ring nodes; roots having longitudinal wrinkles and numerous, dark red-brown, warty protrusions or transverse-ly elongated protuberances. Brittle and easily breakable. A transverse section white and powdery, and under a mag-nifying glass, oil canals scattered as brown dots. Odor, slight; taste, slightly sweet.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Glehnia Root and Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Glehnia Root and Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
Glycyrrhiza Extract contains not less than 3.
Glycyrrhiza Extract is a brown to blackish brown, viscous extract, and has a characteristic odor and a sweet taste. It dissolves in water, forming a clear solution, or with a slight turbidity.
To 0.8 g of Glycyrrhiza Extract add 10 mL of a mixture of ethanol (95) and water (7:3), shake for 2 minutes, centrifuge, and use the supernatant liquid as the sample solution. Proceed as directed in the Identification under Glycyrrhiza.
(1) Heavy metals <1.07>—Prepare the test solution with 1.0 g of Glycyrrhiza Extract as directed under the Extracts (4), and perform the test (not more than 30 ppm). (2) Insoluble matter—Dissolve 2.0 g of Glycyrrhiza Ex-tract in 18 mL of water, and filter. To 10 mL of the filtrate add 5 mL of ethanol (95): a clear solution results.
Weigh accurately about 0.5 g of Powdered Glycyr-rhiza in a glass-stoppered centrifuge tube, add 70 mL of dilute ethanol, shake for 15 minutes, centrifuge, and sepa-rate the supernatant liquid. To the residue add 25 mL of dilute ethanol, and proceed in the same manner. Combine all the extracts, add dilute ethanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Glycyrrhizic Acid RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in dilute ethanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glycyrrhizic acid in each solution. Amount (mg) of glycyrrhizic acid (C42H62O16) ＝ MS × AT/AS MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 720 mL of water, and add 5 mL of acetic acid (100) and 280 mL of acetonitrile. Flow rate: Adjust so that the retention time of glycyrrhizic acid is about 15 minutes. System suitability— System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak with the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glycyrrhizic acid is not more than 1.5z.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Poria Sclerotium according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Poria Sclerotium according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 1.0z.
Powdered Glycyrrhiza is light yellow-brown or light yellow to grayish yellow (powder of peeled Glycyrrhiza) in color. It has a slight odor and a sweet taste. Under a microscope <5.01>, Powdered Glycyrrhiza reveals mainly yellow sclerenchymatous fiber bundles accompanied with crystal cell rows; vessels, 80 – 200 mm in diameter, with pitted, reticulate and scalariform pits, and with round perfo-rations; parenchyma cells, containing starch grains and soli-tary crystals of calcium oxalate, their fragments, and cork tissues; but powder of peeled Glycyrrhiza shows no cork tissue; if any, a very few. Starch grains are simple grains, 2 – 20 mm in diameter; solitary crystals of calcium oxalate, 10 – 30 mm in a diameter.
To 2 g of Powdered Glycyrrhiza add 10 mL of a mixture of ethanol (95) and water (7:3), heat by shaking on a water bath for 5 minutes, cool, filter, and use the fil-trate as the sample solution. Separately, dissolve 5 mg of Glycyrrhizic Acid RS or glycyrrhizic acid for thin-layer chro-matography in 1 mL of a mixture of ethanol (95) and water (7:3), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solu-tion and a spot from the standard solution show the same color tone and the same Rf value. ‌JP XVIII Crude Drugs and Related Drugs / Glycyrrhiza Extract 2015
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Glycyrrhiza according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Glycyrrhiza according to Method 4, and per- Glycyrrhiza Extract
Licorice Root
Glycyrrhiza
甘草
kanzou
Glycyrrhizae Radix consists of the root and stolon of Glycyrrhiza glabra Linne, Glycyrrhiza uralensis Fisher, or related species (Leguminosae), from which the cork layer is removed, dried, and usually processed.
Collected in autumn to spring of 3-4 year old plants
Root: cylindrical, often laterally split, 30-100 cm long, 0.5-3.5 cm in diameter; Stolon: cylindrical, sometimes with branches, 1-3 m long, 0.5-1.5 cm in diameter
Fracture surface is fibrous, outer part reddish brown, inner part yellowish brown
Cork cells usually absent; phloem composed of fiber bundles (surrounded by crystal sheath containing calcium oxalate prisms), sieve tubes and parenchyma; xylem composed of vessels (arranged in radial rows), tracheids, wood fibers and parenchyma; vessels 30-180 μm in diameter with simple perforation plates; prisms of calcium oxalate crystals 15-30 μm in length scattered in the parenchyma cells near the fiber bundles
Powder shows fragments of fibers surrounded by crystal sheath containing prisms of calcium oxalate; fragments of vessels with simple perforation plates; parenchyma cells containing starch grains (simple or compound, 3-15 μm in diameter) and prisms of calcium oxalate
Fiber bundles surrounded by crystal sheath containing prisms of calcium oxalate; vessels with simple perforation plates; starch grains 3-15 μm in diameter; sweet taste
To 1.0 g of pulverized Glycyrrhizae Radix add 20 mL of ammonia TS and 20 mL of chloroform, shake for 15 minutes, filter, and use the chloroform layer as the sample solution. Separately, dissolve 1 mg of glycyrrhizin RS in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography. Spot 10 μL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ammonium acetate TS, ammonia solution and ethyl acetate (6:3:1:1) to a distance of about 10 cm, and dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, and heat at 80°C for 10 minutes: a spot from the sample solution and a spot from the standard solution show the same blue-purple color.
Perform the test as directed under Liquid Chromatography with the sample solution prepared in the Assay. The retention time of the peak corresponding to glycyrrhizic acid from the sample solution is the same as that from the standard solution.
Weigh accurately about 0.5 g of pulverized Glycyrrhizae Radix, add 50 mL of dilute ammonia TS, shake for 15 minutes, filter, and add dilute ammonia TS to make exactly 50 mL. Pipet 5 mL of this solution, add 2 mL of dilute hydrochloric acid (1 in 3), and extract with two 20-mL portions of 2-propanol. Combine the 2-propanol extracts, evaporate to dryness under reduced pressure, dissolve the residue in the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of glycyrrhizin RS (previously determine the water content), dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 μL each of the sample solution and standard solution as directed under Liquid Chromatography.
Powdered Acacia occurs as a white to light yel-lowish white powder. It is odorless, tasteless, but produces a mucilaginous sensation on the tongue. Under a microscope <5.01>, Powdered Acacia, immersed in olive oil or liquid paraffin, reveals colorless, angular fragments or nearly globular grains. Usually starch grains or vegetable tissues are not observed or very trace, if any. Powdered Acacia (1.0 g) dissolves almost completely in 2.0 mL of water, and the solution is acid. It is practically insoluble in ethanol (95).
To 1 g of Powdered Acacia add 25 mL of water and 1 mL of sulfuric acid, and heat under a reflux con-denser for 60 minutes. After cooling, add gently 2.0 g of an-hydrous sodium carbonate. To 1 mL of this solution add 9 mL of methanol, mix well, centrifuge, and use the superna-tant liquid as the sample solution. Separately, dissolve 10 mg each of D-galactose, L-arabinose and L-rhamnose monohy-drate in 1 mL water, add methanol to make 10 mL, and use these solutions as the standard solutions, (1), (2) and (3), re-spectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, acetic acid (100) and water (12:3:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, and heat the plate at 1059C for 2 minutes: the three spots obtained from the sample solution are the same with each spot from the standard solutions (1), (2) and (3) in the color tone and the Rf value, respectively.
(1) Insoluble residue—To 5.0 g of Powdered Aca-cia add 100 mL of water and 10 mL of dilute hydrochloric acid, and dissolve by gentle boiling for 15 minutes with swirling. Filter the warm mixture through a tared glass filter (G3), wash the residue thoroughly with hot water, and dry at 1059C for 5 hours: the mass of the residue does not exceed 10.0 mg. Tannin-bearing gums—To 10 mL of a solution of Powdered Acacia (1 in 50) add 3 drops of iron (III) chloride TS: no dark green color is produced. ‌1940‌ Achyranthes Root / Crude Drugs and Related Drugs Glucose—Use the sample solution obtained in the Identification as the sample solution. Separately, dissolve 10 mg of glucose in 1 mL of water, add methanol to make 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of acetone and water (9:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: no spot obtained from the sample solution is observed at the po-sition corresponding to the spot from the standard solution. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Colorless or light yellow-brown, translucent or somewhat opaque spheroidal tears, or angular fragments with numerous fissures on the surface; very brittle; the frac-tured surface glassy and occasionally iridescent. Odorless; tasteless, but produces a mucilaginous sensation on the tongue. Pulverized Acacia (1.0 g) dissolves almost completely in 2.0 mL of water, and the solution is acid. It is practically insoluble in ethanol (95).
To 1 g of pulverized Acacia add 25 mL of water and 1 mL of sulfuric acid, and heat under a reflux con-denser for 60 minutes. After cooling, add gently 2.0 g of an-hydrous sodium carbonate. To 1 mL of this solution add 9 mL of methanol, mix well, centrifuge, and use the superna-tant liquid as the sample solution. Separately, dissolve 10 mg each of D-galactose, L-arabinose and L-rhamnose monohy-drate in 1 mL water separately, add methanol to make 10 mL, and use these solutions as the standard solutions (1), (2) and (3), respectively. Perform the test with these solutions as directed under Thin-layer chromatography <2.03>. Spot 2 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol, acetic acid (100) and water (12:3:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, and heat the plate at 1059C for 2 minutes: the three spots obtained from the sample solution are the same with each spot from the standard solutions (1), (2) and (3) in the color tone and the Rf value, respectively.
(1) Insoluble residue—To 5.0 g of pulverized Aca-cia add 100 mL of water and 10 mL of dilute hydrochloric acid, and dissolve by gentle boiling for 15 minutes with swirling. Filter the warm mixture through a tared glass filter (G3), wash the residue thoroughly with hot water, and dry at 1059C for 5 hours: the mass of the residue does not exceed 10.0 mg. Tannin-bearing gums—To 10 mL of a solution of Acacia (1 in 50) add 3 drops of iron (III) chloride TS: no dark green color is produced. Glucose—Use the sample solution obtained in the Identification as the sample solution. Separately, dissolve 10 mg of glucose in 1 mL of water, add methanol to make 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of acetone and water (9:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 1-naphthol-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: no spot obtained from the sample solution is observed at the po-sition corresponding to the spot from the standard solution. Loss on drying <5.01> Not more than 17.0z (6 hours). Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Hedysarum Root is nearly cylindrical, 20 – 100 cm in length, 0.5 – 2.5 cm in diameter; outer surface yellow-brown to red-brown, with irregular longitudinal wrinkles; often horizontal lenticels and scars of lateral roots; periderm peeled easily, internally light yellow-brown to light red-brown; soft in texture, flexible and difficult to break; frac-tured surface fibrous, powdery; in transverse section nearly white in cortex, brownish around cambium, light yellow-brown in xylem; ray obvious. Odor, slightly characteristic; taste, slightly sweet. Under a microscope <5.01>, a transverse section reveals cork layer 6 – 8 cells layered, 2 – 4 cells layered parenchyma cells with sparingly thick wall inside the cork layer; ray obvi- ous in secondary cortex and often appearing cracked tissue in outer portion of secondary cortex; phloem fiber bungles arranged stepwise in phloem; ray obvious in xylem, reticu-late, scalariform, pitted, and spiral vessels; xylem tissues around vessels; thin walled cells containing solitary crystals of calcium oxalate in peripheral region of phloem fibers and xylem fibers and appearing as crystal cell rows in a longitudi-nal section; solitary crystals of calcium oxalate 7 – 20 mm in diameter, starch grains simple or 2- to 8-compound grains in parenchyma.
To 1.0 g of pulverized Hedysarum Root add 10 mL of methanol, shake for 10 minutes, and filter. Evaporate the solvent of the filtrate under low pressure (in vacuo), add 1 mL of methanol to the residue, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, 2-butanone and formic acid (60:40:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a blue-white fluorescent spot at an Rf value of about 0.4 is observed.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Hedysarum Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Hedysarum Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 16.0z (6 hours). Total ash <5.01> Not more than 5.5z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
root of Hedysarum polybotrys Handel-Mazzetti (Leguminosae)
Stem with alternate leaves and spikes; stem light brown, with longitudinal furrows and protruded nodes; when soaked in water and smoothed out, leaves wide ovate and cordate, 3 – 8 cm in length, 3 – 6 cm in width; light green-brown; margin entire, apex acuminate; petiole long, and membranous stipule at the base; spike, 1 – 3 cm in length, with numerous light yellow-brown achlamydeous florets, and the base enclosed by 4 long ovate, light yellow to light yellow-brown involucres. Odor, slight; taste, slight.
Heat 2 g of pulverized Houttuynia Herb with 20 mL of ethyl acetate under a reflux condenser for 15 minutes, and filter. Evaporate the filtrate to dryness, add 10 mL of water to the residue, warm the mixture on a water bath for 2 minutes, and, after cooling, filter. Shake well the filtrate with 20 mL of ethyl acetate in a separator, take 15 mL of ethyl acetate solution, and evaporate the solution on a water bath to dryness. Dissolve the residue in 5 mL of meth-anol, add 0.1 g of magnesium ribbon and 1 mL of hydro-chloric acid, and allow the mixture to stand: a light red to red color develops.
Foreign matter <5.01>—The amount of the rhizome, roots and other foreign matter contained in Houttuynia Herb is not more than 2.0z. Total ash <5.01> Not more than 14.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 10.0z. ‌2036‌‌‌ Immature Orange / Crude Drugs and Related Drugs
Powdered Sweet Hydrangea Leaf occurs as a dark yellow-green powder, and has a faint odor and a char-acteristic, sweet taste. Under a microscope <5.01>, Powdered Sweet Hydrangea Leaf reveals fragments of epidermis with wavy lateral cell wall; stomata with two subsidiary cells; unicellular and thin- ‌JP XVIII Crude Drugs and Related Drugs / Swertia Herb 2157 walled hair with numerous protrusions of the surface, 150 – 300 mm in length; fragments of palisade tissue and spongy tissue; fragments of vascular bundle and mucilage cells con-taining raphides of calcium oxalate 50 – 70 mm in length.
To 1.0 g of Powdered Sweet Hydrangea Leaf add 10 mL of methanol, shake for 10 minutes, centri-fuge, and use the supernatant liquid as the sample solution. Separately, dissolve 2 mg of sweet hydrangea leaf dihydro-isocoumarin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of diethyl ether, hexane and formic acid (5:5:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two of the several spots obtained from the sample solution have the same color tone and Rf value with the spots from the standard solution.
Foreign matter <5.01>—Under a microscope, Pow-dered Sweet Hydrangea Leaf does not show stone cells, a large quantity of fibers or starch grains. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.5z.
Usually wrinkled and contracted leaf, dark green to dark yellow-green in color. When soaked in water and smoothed out, it is lanceolate to acuminately ovate, 5 – 15 cm in length, 2 – 10 cm in width; margin serrated, base slightly wedged; coarse hair on both surfaces, especially on the veins; lateral veins not reaching the margin but curving upwards and connecting with each other; petiole short and less than one-fifth of the length of lamina. Odor, slight; taste, characteristically sweet.
To 1.0 g of pulverized Sweet Hydrangea Leaf add 10 mL of methanol, shake for 10 minutes, centri-fuge, and use the supernatant liquid as the sample solution. Separately, dissolve 2 mg of sweet hydrangea leaf di-hydroisocoumarin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of diethyl ether, hexane and formic acid (5:5:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two of the several spots obtained from the sample solution have the same color tone and Rf value with the spots from the standard solution.
(1) Stem—When perform the test of foreign mat-ter <5.01>, the amount of stems contained in Sweet Hydran-gea Leaf does not exceed 3.0z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than stems contained in Sweet Hydrangea Leaf does not exceed 1.0z. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.5z.
Hydrous Lanolin is Purified Lanolin to which water
Long and thin cylindrical rhizome, 0.3 – 0.5 cm in diameter; sometimes branched; externally yellow-white, with slight longitudinal wrinkles, and with nodes at 2 – 3 cm intervals; difficult to break; fractured surface fi-brous. Cross section irregularly round; thickness of cortex is slightly smaller than the diameter of the stele; pith often forms a hollow. Under a magnifying glass, a transverse sec-tion reveals cortex, yellow-white, and with scattered brown spots; stele, yellow-brown in color. Almost odorless, and tasteless at first, but later slightly sweet.
To 1 g of pulverized Imperata Rhizome add 20 mL of hexane, allow the mixture to stand for 30 minutes with occasional shaking, and filter. Evaporate the solvent of the filtrate under low pressure (in vacuo), dissolve the residue in 5 mL of acetic anhydride, place 0.5 mL of this so-lution in a test tube, and add carefully 0.5 mL of sulfuric acid to make two layers: a red-brown color develops at the zone of contact, and the upper layer acquires a blue-green to blue-purple color.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Imperata Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Imperata Rhizome according to Method 4, and perform the test (not more than 5 ppm). Rootlet and scaly leaf—When perform the test of foreign matter <5.01>, the amount of the rootlets and scaly leaves contained in Imperata Rhizome is not more than 3.0 z. Foreign matter <5.01>—The amount of foreign mat- ter other than rootlets and scaly leaves is not more than 1.0z. Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
Ipecac Syrup is a syrup containing not less than
Powdered Ipecac occurs as a light grayish yel-low to light brown powder. It has a slight odor, which is ir-ritating to the nasal mucosa, and has a somewhat bitter and unpleasant taste. Under a microscope <5.01>, Powdered Ipecac reveals starch grains and needle crystals of calcium oxalate; frag-ments of parenchyma cells containing starch grains or the needle crystals; substitute fibers,thin-walled cork tissue; ves-sels and tracheids with simple or bordered pits; a few wood fibers and wood parenchyma. Starch grains inherent in Ipecac, mainly 2 – 8-compound grains, rarely simple grains 4 – 22 mm in diameter; and needle crystals of calcium oxalate 25 – 60 mm in length.
To 0.5 g of Powdered Ipecac add 2.5 mL of hydrochloric acid, allow to stand for 1 hour with occasional shaking, and filter. Collect the filtrate into an evaporating dish, and add a small pieces of chlorinated lime: circumfer-ence of it turns red.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Ipecac according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Ipecac according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, groups of stone cells and sclerenchymatous fibers are not observed. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of Powdered Ipecac, transfer into a glass-stoppered centrifuge tube, add 30 mL of 0.01 mol/L hydrochloric acid TS, shake for 15 minutes, centrifuge, and separate the supernatant liquid. Repeat this procedure twice with the residue using 30-mL portions of 0.01 mol/L hydrochloric acid TS. Combine all the extracts, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of emetine hydrochloride for assay, previously dried in a desiccator (in vacuum, phospho-rus (V) oxide, 509C) for 5 hours, dissolve in 0.01 mol/L hy-drochloric acid TS to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATE and ATC, of emetine and cephaeline obtained with the sample so-lution, and the peak area, ASE, of emetine obtained with the standard solution. ‌2038 Ipecac Syrup / Crude Drugs and Related Drugs Amount (mg) of total alkaloids (emetine and cephaeline) ＝ MS ×{ATE ＋ (ATC × 0.971)}/ASE × 0.868 MS: Amount (mg) of emetine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 2.0 g of sodium 1-heptane sul-fonate in 500 mL of water, adjust the pH 4.0 with acetic acid (100), and add 500 mL of methanol. Flow rate: Adjust so that the retention time of emetine is about 14 minutes. System suitability— System performance: Dissolve 1 mg each of emetine hy-drochloride for assay and cephaeline hydrobromide in 10 mL of 0.01 mol/L hydrochloric acid TS. When the procedure is run with 10 mL of this solution under the above operating conditions, cephaeline and emetine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of emetine is not more than 1.5z.
Slender, curved, cylindrical root, 3 – 15 cm in length, 0.3 – 0.9 cm in diameter; mostly twisted, and some-times branched; outer surface gray, dark grayish brown, red-brown in color and irregularly annulated; when root frac-tured, cortex easily separable from the xylem; the cortex on the fractured surface is grayish brown, and the xylem is light brown in color: thickness of cortex up to about two-thirds of radius in thickened portion. Scales in rhizome opposite. Odor, slight; powder irritates the mucous membrane of the nose; taste, slightly bitter and unpleasant. Under a microscope <5.01>, the transverse section of Ipecac reveals a cork layer, consisting of brown thin-walled cork cells; in the cortex, sclerenchyma cells are absent; in the xylem, vessels and tracheids arranged alternately; paren-chyma cells filled with starch grains and sometimes with raphides of calcium oxalate.
To 0.5 g of pulverized Ipecac add 2.5 mL of hydrochloric acid, allow to stand for 1 hour with occasional shaking, and filter. Collect the filtrate into an evaporating dish, and add a small pieces of chlorinated lime: circumfer-ence of it turns red.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Ipecac according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Ipecac according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 12.0z (6 hours). ‌JP XVIII Crude Drugs and Related Drugs / Powdered Ipecac 2037 Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of pulverized Ipecac, in a glass-stoppered centrifuge tube, add 30 mL of 0.01 mol/L hydrochloric acid TS, shake for 15 minutes, centri-fuge, and separate the supernatant liquid. Repeat this proce-dure twice with the residue using 30-mL portions of 0.01 mol/L hydrochloric acid TS. Combine all the extracts, add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of emetine hydrochloride for assay, previously dried in a desiccator (in vacuum, phospho-rus (V) oxide, 509C) for 5 hours, dissolve in 0.01 mol/L hy-drochloric acid TS to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATE and ATC, of emetine and cephaeline obtained with the sample so-lution, and the peak area, ASE, of emetine obtained with the standard solution. Amount (mg) of total alkaloids (emetine and cephaeline) ＝ MS ×{ATE ＋ (ATC × 0.971)}/ASE × 0.868 MS: Amount (mg) of emetine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 2.0 g of sodium 1-heptane sul-fonate in 500 mL of water, adjust the pH 4.0 with acetic acid (100), and add 500 mL of methanol. Flow rate: Adjust so that the retention time of emetine is about 14 minutes. System suitability— System performance: Dissolve 1 mg each of emetine hy-drochloride for assay and cephaeline hydrobromide in 0.01 mol/L hydrochloric acid TS to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, cephaeline and emetine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of emetine is not more than 1.5z.
Stem, leaves, and flowers usually cross sec-tioned, stems square, 0.2 – 3 cm in diameter, yellow-green to green-brown in color, covered densely with white short hairs; the pith white, a great parts of central of cut surface. Light in texture. Leaves opposite, petiolated, 3-dissected to 3-incised, each lobe splits pinnately, and end lobes reveals linear-lanceolate, acute or acuminate, the upper surface light green, the lower surface bristle with white short hairs, grayish green. Flower, verticillate; sepal, tubular, and the upper end acerate with five lobes; light green to light green-brown in color, corolla labiate, light red-purple to light brown. Odor, slightly; taste, slightly bitter, astringent. Under a microscope <5.01>, a transverse section of stem reveals four ridges, a part of the ridge of Leonurus sibiricus Linneáprotruding knobby. Epidermis, observed non-glandu-lar hairs from 1 to 3 cells, glandular hairs with head of 1 to 4 celled or glandular scale with 8 cells. Each ridge parts, beneath epidermis, collenchyma developed, development of xylem fibers remarkably. Cortex composed of several cellu-lar layers parenchymatous cells. Collateral vascular bundle arranged in a circle. Phloem fibers observed at the outer por-tion of phloem. Parenchymatous cells of cortex and pith ob-serve needle crystals or plate-like crystals of calcium oxalate.
To 1 g of pulverized Leonurus Herb add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, de-velop the plate with a mixture of water and methanol (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS followed by immediate spraying of so-dium nitrite TS on the plate: a grayish brown spot appears at an Rf value of about 0.5, which color fades soon and then disappears after air-drying the plate. Loss on drying <5.01> Not more than 12.0z. Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 12.0z.
Lilium Bulb reveals oblong with narrowed apex, lanceolate, or narrowly triangular boat-shaped, trans-lucent, 1.3 – 6 cm in length, 0.5 – 2.0 cm in diameter, exter-nally milky white to light yellow-brown occasionally purplish in color, nearly smooth; central portion somewhat thick-ened, circumferential portion thin, slightly waved, occa-sionally rolled inside; usually several lines of vascular bun-dles longitudinally in parallel are seen through parenchyma; hard in texture, easy to break; fractured surface horny and flat. Almost odorless; taste, slightly acid and bitter. Under a microscope <5.01>, the surface reveals epidermal cells rectangular to almost square, stomata nearly circular, the cells adjacent to stomata mostly 4 in number. Under a microscope <5.01>, a transverse section reveals the outermost layer composed of epidermal cells covered with smooth cuti-cle; epidermis circular to quadrangular parenchymatous cells distributed evenly, palisade tissue not observed; in paren-chyma of mesophyll collateral vascular bundles extended from adaxial side to abaxial side of scaly leaves are arranged almost in a transverse line; starch grains contained in paren-chymatous cells, usually gelatinized.
To 3 g of pulverized Lilium Bulb add 10 mL of 1-butanol, shake, add 10 mL of water, shake for 30 minutes, and centrifuge. Evaporate the solvent under low pressure (in vacuo), add 1 mL of methanol to the residue, shake gently, and use the supernatant liquid so obtained as the sample solution. Perform the test with the sample solu-tion as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, methanol and water (12:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots appear at an Rf value of about 0.3. When ex-amine these spots under ultraviolet light (main wavelength: 365 nm) after spraying with sodium carbonate TS, they ap-pear as blue-purple fluorescent spots. Loss on drying <5.01> Not more than 16.0z. Total ash <5.01> Not more than 4.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
Fusiform or rosary-like root, 10 – 15 cm in length, 10 – 25 mm in diameter; externally yellow-brown to brown, with a few scars of rootlets; a transversely cut sur-face reveals cortex brown, xylem light yellow-brown, con-centric circles and radially arranged lines brown; dense and hard in texture. Odor, camphor-like; taste, bitter. Under a microscope <5.01>, a transverse section of the root with periderm reveals a cork layer several cells thick, partially consisting of cork stone cells; cortex parenchyma sometimes contains oil cells and fibers; in xylem, vessels-xylem fibers and rays are arranged alternately; parenchya-tous cells of cortex and xylem contain sandy and columnar crystals of calcium oxalate, simple starch grains 1 – 15 mm in diameter, and 2- to 4- compound starch grains.
To 3 g of pulverized Lindera Root add 40 mL of hexane, and heat under a reflux condenser for 30 minutes. After cooling, filter, to the residue add 10 mL of ammonia TS and 30 mL of a mixture of ethyl acetate and diethyl ether (1:1), shake vigorously for 20 minutes, and cen-trifuge. Separate the upper layer, add 10 g of anhydrous so-dium sulfate, shake, and filter. Evaporate the solvent of the filtrate, dissolve the residue with 0.5 mL of ethanol (99.5), and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, methanol and ammonia water (28) (10:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: a yellow-brown spot appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Lindera Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Lindera Root according to Method 4, and per-form the test (not more than 5 ppm). Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 2.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 6.0z.
Rather slender conical root, often branched, 6 – 10 cm in length, 0.5 – 1.5 cm in diameter; externally dark purple, coarse in texture, thin and easily peeled; mostly with twisted and deep longitudinal furrows, which sometimes reach to xylem; sometimes remains of stem at the crown; easily broken; fractured surface granular and with many clefts. Under a magnifying glass, a transverse section reveals a dark purple color at the outer portion of cortex, and light brown inner portion making irregular wave; xylem yellowish in color; the center of the crown is often cracked, and the surrounding part red-purple. Odor, slight; taste, slightly sweet.
(1) Heat 0.5 g of pulverized Lithospermum Root in a test tube: red vapor evolves, which condenses on the wall of the upper part of the tube into red-brown oil drops. Shake 0.5 g of pieces or powder of Lithospermum Root with 1 mL of ethanol (95), and to the red solution thereby obtained add 1 drop of sodium hydroxide TS: the red color changes to blue-purple. To this solution add 1 to 2 drops of dilute hydrochloric acid: the color turns red again. To 0.5 g of pulverized Lithospermum Root add 5 mL of ethanol (95), shake for 30 minutes, filter, and evaporate the solvent of the filtrate at a temperature not higher than 409C under low pressure (in vacuo). Add 1 mL of ethanol (95) to the residue, and use this solution as the sample solu-tion. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chroma-tography. Develop the plate with a mixture of ethyl acetate and ethanol (95) (3:1) to a distance of about 7 cm, and air-dry the plate: a red-purple spot appears at an Rf value of about 0.75.
(1) Heavy metals <1.07>—Proceed with 2.0 g of pulverized Lithospermum Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Lithospermum Root according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 11.0z. Acid-insoluble ash <5.01> Not more than 3.5z.
solution according to Method 3, and perform the test
Leaves and opposite leaves on short stem; leaf, ovate and entire, with short petiole, 3 – 7 cm in length, 1 – 3 cm in width; upper surface green-brown, lower surface light grayish green; under a magnifying glass, both surfaces pubescent. Stem, 1 – 4 mm in diameter; externally grayish yellow-brown to purplish brown, a transversely cut surface of stem, round and hollow. Almost odorless; taste, slightly astringent, followed by a litter bitterness. Under a microscope <5.01>, a transverse section of leaf re- veals the outermost layer of upper and lower surfaces to be composed of a single-layered epidermis, uni-cellular non-glandular hairs and multi-cellular glandular hairs on epider-mis; in midvein, several-cellular-layered collenchyma present beneath the epidermis and vascular bundles in the center; in mesophyll, palisade layer adjacent to upper epidermis, spongy tissue adjacent to lower epidermis; glandular hairs contain brown secretion, parenchymatous cells contain ag-gregate crystals of calcium oxalate, and occasionally starch grains.
To 1 g of pulverized Lonicera Leaf and Stem add 5 mL of methanol, shake for 5 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of chlorogenic acid for thin-layer chromatog-raphy in 2 mL of methanol, and use this solution as the standard solution (1). Separately, dissolve 1 mg of loganin for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (6:1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the stand-ard solution (1). Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution (2).
Stem—Lonicera Leaf and Stem does not contains the stems larger than 5 mm in diameter. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 9.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 12.0z.
Tubular to semitubular bark, 1 – 6 mm in thickness; externally light brown to light yellow-brown, periderm peeled easily as scale; internally grayish brown, longitudinally striate; brittle in texture; fractured surface, grayish white, not fibrous. Odor, weak and characteristic; taste, slightly sweet at first. Under a microscope <5.01>, a transverse section reveals periderm composed of a cork layer of several cellular layers of thin-walled cork cells; in cortex parenchyma cells contain-ing sandy crystals of calcium oxalate sparsely distributed, oc-casionally a few fibers observed; parenchyma cells contain starch grains, 1 – 10 mm in diameter; stone cells very rare.
To 1.0 g of pulverized Lycium Bark add 10 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, ammonium acetate solution (1 in 20) and acetic acid (100) (2:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, heat the plate at 1059C for 2 minutes, then spray evenly sodium nitrite TS, and allow to stand for 5 minutes: a dark brown principal spot appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 2.0 g of pulverized Lycium Bark according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Lycium Bark according to Method 4, and per-form the test (not more than 5 ppm). Loss on drying <5.01> Not more than 11.5z (6 hours). Total ash <5.01> Not more than 20.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 10.0z.
Fusiform fruit with acute apex, 6 – 20 mm in length, 3 – 8 mm in diameter, pericarp red to dark red, externally roughly wrinkled; under a magnifying glass, a transverse section of fruit reveals two locules containing numerous seeds; seed light brown to light yellow-brown, about 2 mm in a diameter, compressed reniform. Odor, characteristic; taste, sweet, occasionally slightly bit-ter.
To 1.0 g of pulverized Lycium Fruit add 5 mL of ethyl acetate, shake for 15 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and ethyl acetate (10:1) to a dis-tance of about 7 cm, and air-dry the plate: a yellow principal spot appears at an Rf value of about 0.6.
Foreign matter <5.01>—It contains not more than 2.0z of foreign matter such as peduncle or others. Total ash <5.01> Not more than 8.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 35.0z.
Powdered Magnolia Bark occurs as a yellow-brown powder, and has a slight odor and a bitter taste. Under a microscope <5.01>, Powdered Magnolia Bark re- veals starch grains and parenchyma cells containing them; stone cells of various sizes or its groups; fibers 12 to 25 mm in MS: Amount (mg) of magnolol for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 289 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (50:50:1). Flow rate: Adjust so that the retention time of magnolol is about 14 minutes. System suitability— System performance: Dissolve 1 mg each of magnolol for assay and honokiol in diluted methanol (7 in 10) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, honokiol and mag-nolol are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of magnolol is not more than 1.5z.
Plate-like or semi-tubular bark, 2 – 7 mm in thickness; externally grayish white to grayish brown, and rough, sometimes cork layer removed, and externally red-brown; internally light brown to dark purple-brown; cut surface extremely fibrous, and light red-brown to purple-brown. Odor, slight; taste, bitter. Under a microscope <5.01>, a transverse section reveals a thick cork layer or several thin cork layers, and internally adjoining the circular tissue of stone cells of approximately equal in diameter; primary cortex thin; fiber groups scat-tered in the pericycle; groups of phloem fibers lined alter-nately with the other tissue of phloem between medullary rays in the secondary cortex, and then these tissues show a latticework; oil cells scattered in the primary and secondary cortex, but sometimes observed in the narrow medullary rays.
To 1.0 g of pulverized Magnolia Bark add 10 mL of methanol, stir for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS on the plate: a yellow ‌2068 Powdered Magnolia Bark / Crude Drugs and Related Drugs spot appears at an Rf value of about 0.3. Total ash <5.01> Not more than 6.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 11.0z.
Weigh accurately about 0.5 g of pulverized Magno-lia Bark, add 40 mL of diluted methanol (7 in 10), heat under a reflux condenser for 20 minutes, cool, and filter. Repeat the above procedure with the residue, using 40 mL of diluted methanol (7 in 10). Combine the whole filtrates, add diluted methanol (7 in 10) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of magnolol for assay, dissolve in diluted methanol (7 in 10) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of magnolol in each solution. Amount (mg) of magnolol ＝ MS × AT/AS MS: Amount (mg) of magnolol for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 289 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and acetic diameter; yellow-red-brown cork tissue; oil cells containing a yellow-brown to red-brown substance. Simple starch grains about 10 mm in diameter and 2- to 4-compound starch grains. Identification To 1.0 g of Powdered Magnolia Bark add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS on the plate: a yellow spot appears at an Rf value of about 0.3. Total ash <5.01> Not more than 6.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 11.0z. Assay Weigh accurately about 0.5 g of Powdered Magnolia Bark, add 40 mL of diluted methanol (7 in 10), heat under a reflux condenser for 20 minutes, cool, and filter. Repeat the above procedure with the residue, using 40 mL of diluted methanol (7 in 10). Combine the whole filtrates, add diluted methanol (7 in 10) to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 10 mg of magnolol for assay, dissolve in diluted meth-anol (7 in 10) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of magnolol in each solution. acid (100) (50:50:1). Flow rate: Adjust so that the retention time of magnolol is Amount (mg) of magnolol ＝ MS × AT/AS about 14 minutes. System suitability— System performance: Dissolve 1 mg each of magnolol for assay and honokiol in diluted methanol (7 in 10) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, honokiol and mag-nolol are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of magnolol is not more than 1.5z.
Magnolia Flower is a fusiform flower bud, 15 – 45 mm in length, 6 – 20 mm in diameter at central part; often having ligneous peduncles on base; usually 3 bracts, externally with sparse hairs, brown to dark brown, or with dense hairs, grayish white to light yellow-brown, and the inner surface of 3 bracts smooth and dark brown in color; interior perianth of 9 pieces or 12 pieces, same size or outer three pieces are smaller; 50 – 100 stamens and numerous pistils. Brittle in texture. Odor, characteristic; taste, acrid and slightly bitter.
To 1 g of pulverized Magnolia Flower add 10 mL of methanol, shake for 15 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 20 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, acetone, water and formic acid (5:3:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: a yellow-red spot appears at an Rf value of about 0.3. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 5.5z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-extract: not less than 13.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Magnolia Flower: the volume of essential oil is not less than 0.5 mL.
Mallotus Bark is flat or semitubular pieces of bark, 1 – 3 mm in thickness; externally greenish gray to brownish gray brow in color, with a vertically striped shape gathering numerous lenticels; internal surface light yellow-brown to grayish brown in color, and smooth with numerous fine striped lines; easy to break; slightly fibrous at fractured surface. Mallotus Bark has a slight odor, a bitter taste and slightly astringent.
To 0.5 g pulverized Mallotus Bark add 10 mL of methanol, warm on a water bath for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dis-solve 1 mg of bergenin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (100:17:13) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 11.0z.
a clear, colorless liquid, having the odor of mentha oil
Stem with opposite leaves; stem, square, light brown to red-purple in color, and with fine hairs; when smoothed by immersing in water, leaf, ovate to oblong, with acute apex and base, 2 – 8 cm in length, 1 – 2.5 cm in width, margin irregularly serrated; the upper surface, light brown-yellow to light green-yellow, and the lower surface, light green to light green-yellow in color; petiole 0.3 – 1 cm in length. Under a magnifying glass, leaf reveals hairs, glandu-lar hairs and scales. It has a characteristic aroma and gives a cool feeling on keeping in the mouth.
To 1.0 g of pulverized Mentha Herb add 10 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of menthol in 1 mL of diethyl ether, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of hexane and acetone (7:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid-acetic acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
Foreign matter <5.01>—The amount of roots and other foreign matter contained in Mentha Herb does not exceed 2.0z. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.5z. Essential oil content <5.01> Perform the test with 50.0 g of Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) pulverized Mentha Herb after adding 1 mL of silicone resin to the sample in the flask: the volume of essential oil is not less than 0.4 mL. MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Tubular, semi-tubular or cord-like bark, 1 – 6 mm thick, often in fine lateral cuttings; externally, white to yellow-brown; in the case of the bark with periderm, its periderm is yellow-brown in color, easy to peel, with numerous longitudinal, fine wrinkles and numerous red-purple lenticels laterally elongated; inner surface, dark yel-low-brown in color and flat; cross section, white to light brown in color, and fibrous. Odor, slight; taste, slight. Under a microscope <5.01>, a transverse section of bark with periderm reveals 5 to 12 cellular layers of cork cells in the outer portion; phloem fibers or their bundles scattered in the cortex, arranged alternately and stepwise with phloem parenchyma; lactiferous tubes; solitary crystals of calcium oxalate; starch grains as spheroidal or ellipsoidal, simple or compound grains, simple grain 1 – 7 mm in diameter.
Heat 1 g of pulverized Mulberry Bark with 20 mL of hexane under a reflux condenser for 15 minutes, and filter. Evaporate the solvent of the filtrate under low pressure (in vacuo), dissolve the residue in 10 mL of acetic anhydride, place 0.5 mL of the solution in a test tube, and add carefully 0.5 mL of sulfuric acid to make two layers: a red-brown color develops at the zone of contact.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Mulberry Bark according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g Amount (mg) of [6]-shogaol ＝ MS × AT/AS × 1/10 of pulverized Mulberry Bark according to Method 4, and perform the test (not more than 5 ppm). MS: Amount (mg) of [6]-shogaol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 0.1 g of oxalic acid dihydrate in 600 mL of water, and add 400 mL of acetonitrile. Flow rate: 1.0 mL per minute (the retention time of [6]-shogaol is about 30 minutes). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-shogaol are not less than 5000 and not more than 1.5, respectively. Foreign matter <5.01>—The amount of the root xylem and other foreign matter is not more than 1.0z. Total ash <5.01> Not more than 11.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Powdered Moutan Bark occurs as a light grayish yellow-brown powder. It has a characteristic odor and a slight, pungent and bitter taste. tone and Rf value with the spot from the standard solution. (2) Evaporate the solvent to dryness 1 mL of the sample solution obtained in (1), dissolve the residue in 50 mL of ethanol (95), and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima at around 228 nm, 274 nm and 313 nm.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Moutan Bark according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Moutan Bark according to Method 4, and per-form the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, usually vessels and other sclerenchymatous cells are not observable. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Weigh accurately about 0.5 g of Powdered Moutan Bark, add 40 mL of methanol, heat under a reflux condenser for 30 minutes, cool, and filter. Repeat the above procedure with the residue, using 40 mL of methanol. Combine all the filtrates, add methanol to make exactly 100 mL, then pipet 10 mL of this solution, add methanol to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of paeonol for assay, dissolve in methanol to make exactly 100 mL, then pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of paeonol in each solution. Amount (mg) of paeonol ＝ MS × AT/AS × 1/2 MS: Amount (mg) of paeonol for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). ‌JP XVIII Crude Drugs and Related Drugs / Mukoi-Daikenchuto Extract 2075 Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile, and acetic acid (100) (65:35:2). Flow rate: Adjust so that the retention time of paeonol is about 14 minutes. System suitability— System performance: Dissolve 1 mg of paeonol for assay and 5 mg of butyl parahydroxybenzoate for resolution check in methanol to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, paeonol and butyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeonol is not more than 1.5z.
Tubular to semi-tubular bark, about 0.5 cm in thickness, 5 – 8 cm in length, 0.8 – 1.5 cm in diameter; exter-nally dark brown to purple-brown, with small and transver-sely elongated ellipsoidal scars of lateral roots, and with lon-gitudinal wrinkles; internally, light grayish brown to pur-plish brown and smooth; fractured surface coarse; white crystals often attached on the internal and fractured sur-faces. Odor, characteristic; taste, slightly pungent and bitter.
To 2.0 g of pulverized Moutan Bark add 10 mL of hexane, shake for 3 minutes, filter, and use the fil-trate as the sample solution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of hexane, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultravi-olet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Moutan Bark according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Moutan Bark according to Method 4, and perform the test (not more than 5 ppm). Xylem—When perform the test of foreign matter <5.01>, the amount of the xylem contained in Moutan Bark is not more than 5.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than xylem contained in Moutan Bark is not exceed 1.0z. Powdered Moutan Bark / Crude Drugs and Related Drugs Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 6.0z. ‌Acid-insoluble ash <5.01> Not more than 1.0z.
Weigh accurately about 0.3 g of pulverized Moutan Bark, add 40 mL of methanol, heat under a reflux condenser for 30 minutes, cool, and filter. Repeat the above procedure with the residue, using 40 mL of methanol. Combine all the filtrates, add methanol to make exactly 100 mL, then pipet 10 mL of this solution, add methanol to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of paeonol for assay, dissolve in methanol to make exactly 100 mL, then pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of paeonol in each solution. Under a microscope <5.01>, Powdered Moutan Bark re-veals starch grains and fragments of parenchyma containing them; fragments of cork tissue containing tannin; fragments of somewhat thick-walled collenchyma, medullary rays, and phloem parenchyma; rosette aggregates of calcium oxalate and also fragments of parenchyma cells containing them. Starch grains are simple or 2- to 10-compound grains, 10 – 25 mm in diameter; rosette aggregates are 20 – 30 mm in di-ameter. Identification (1) To 2.0 g of Powdered Moutan Bark add 10 mL of hexane, shake for 3 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of paeonol for thin-layer chromatography in 1 mL of hexane, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultravi- Amount (mg) of paeonol ＝ MS × AT/AS × 1/2 olet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color MS: Amount (mg) of paeonol for assay taken, calculated on the basis of the content obtained by qNMR Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4 to 6 mm in inside diameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile, and acetic acid (100) (65:35:2). Flow rate: Adjust so that the retention time of paeonol is about 14 minutes. System suitability— System performance: Dissolve 1 mg of paeonol for assay and 5 mg of butyl parahydroxybenzoate for resolution check in methanol to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, paeonol and butyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with the standard solution under the above operating condi-tions, the relative standard deviation of the peak area of paeonol is not more than 1.5z.
Ovoid-globose to ellipsoidal seeds, 1.5 – 3.0 cm in length, 1.3 – 2.0 cm in diameter; externally grayish brown, with wide and shallow longitudinal furrows and fine wrinkles; usually, reveals a hilum at one end, the hilum grayish white to grayish yellow and slightly protruding, and a chalaza at the other end, the chalaza grayish brown to dark brown and slightly concave; dark brown thin perisperm ex-tending irregularly into the light yellow-white to light brown endosperm, by which cut surface exhibiting a marble-like ap-pearance. Odor, characteristic and strong; taste, acrid and slightly bitter. Under a microscope <5.01>, a transverse section reveals perisperm composed of outer and inner layers; the outer layer composed of parenchyma containing red-brown to dark red-brown contents; the inner layer, composed of parenchyma filled with red-brown to dark red-brown con-tents, often extends into endosperm, where numerous oil cells are scattered: several vascular bundles present in the inner layer, the vessels spiral; in parenchyma cells of en-dosperm, simple or compound starch grains and aleurone grains observed; and in the parenchyma cells in the outer layer of perisperm and the marginal part of endosperm, numerous crystals of calcium oxalate observed.
To 1 g of pulverized Nutmeg add 5 mL of methanol, allow to stand for 10 minutes with occasional shaking, filter, and use the filtrate as the sample solution. Separately, dissolve 2 mg of myristicin for thin-layer chro-matography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (9:1) to a dis-tance of about 7 cm, and air-dry the plate. Spray evenly diluted sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution. Loss on drying <5.01> Not more than 16.0z (6 hours). Total ash <5.01> Not more than 2.5z. Essential oil content <5.01> When the test is performed with 10.0 g of pulverized Nutmeg, the essential oil content is not less than 0.5 mL.
Ovoid to ellipsoidal seed, at the base a papil-late protuberance surrounded with shallow depression, 1.0 – 1.7 cm in length, 0.5 – 1.2 cm in width; externally light red-brown to light yellow-brown; projection part dark reddish brown; endocarp not lustrous and hardly peeled off; en-dosperm yellow-white, a green embryo in the center. Almost odorless; taste, slightly sweet and oily, embryo is extremely bitter. Under a microscope <5.01>, a transverse section of the seed at central portion reveals endocarp composed of paren-chyma or endocarp occasionally left out; seed coat com-posed of epidermis and parenchyma of compressed cells; vascular bundles scattered in parenchyma; endosperm com-posed of epidermis and parenchyma; aggregate crystals of calcium oxalate and tannin-like substances contained in en-docarp remained; parenchymatous cells of seed coat contain tannin-like substances; parenchyma of endosperm contain starch grains.
To 0.5 g of pulverized Nelumbo Seed add 5 mL of water, shake for 5 minutes, and centrifuge. To 0.5 mL of the supernatant liquid add 1 drop of a solution of 1-naphthol in ethanol (99.5) (1 in 5), mix, then add gently 1 mL of sulfuric acid: the solution shows a purple color. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 14.5z.
Notopterygium is slightly curved, cylindrical to conical, 3 – 10 cm in length, 5 – 20 mm in diameter; rhizome occasionally branched; externally yellow-brown to dark brown. The rhizome with nearly orbicular, hollowed stem scars at the apex, sometimes having short residue of stem; externally node rising, internode short; root scars in warty processes on the node; externally root has coarse lon-gitudinal wrinkles and lateral root scars in warty processes; light and slightly brittle in texture, easy to break. The trans-verse section of the rhizome reveals numerous radial cracks; cortex yellow-brown to brown; xylem light yellow to light grayish yellow; pith grayish white to light brown. Under a magnifying glass, the rhizome reveals brown, fine points of resin canals in the cortex and pith. Odor, characteristic; taste, a slight characteristic taste at first, followed by a slightly pungent and slightly numbing aftertaste. Under a microscope <5.01>, transverse section shows the outermost layer to be composed of a cork layer several to a dozen or so cells thick; several-layered collenchyma just in-side of the cork layer; oil canals scattered in cortex, large ones more than 300 mm in diameter; intercellular space oc-curring in radial direction in cortex; oil canals scattered in pith, large ones more than 500 mm in diameter; parenchyma-tous cells contain simple and 2- to 3-compound starch grains.
To 0.3 g of pulverized Notopterygium add 3 mL of hexane in a glass-stoppered centrifuge tube, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of octadecylsilanized silica gel with fluorescent indicator for thin-layer chromatog-raphy, develop the plate with a mixture of methanol and water (9:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a blue-white fluorescent spot appears at an Rf value of about 0.5, which shows a dark purple color under ultraviolet light (main wavelength: 254 nm).
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Notopterygium according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Notopterygium according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 20.0z.
rhizome and root of Notopterygium incisum Ting ex H
Usually, longitudinally split irregular column, twisted, bent or somewhat pressed, 20 – 30 cm in length, about 2 cm in width; the outer surface, dark brown, and the cut surface, white to grayish white in color; one side shows nearly round to blunt triangular scars of petiole about 1 cm in diameter, and the other side numerous scars of roots less than 0.3 cm in diameter; light, spongy in texture, and easily broken; fractured surface flat and powdery. Under a mag-nifying glass, a transverse section reveals a black outer por- ‌2078‌ Nutmeg / Crude Drugs and Related Drugs tion, and porous tissue with scattered vascular bundles in the inner portion. Odor, slight; taste, slightly bitter and unpleasant.
To 1.0 g of pulverized Nuphar Rhizome add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography, de-velop the plate with a mixture of ethyl acetate, methanol and ammonia solution (28) (20:3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: a yellow-brown spot appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Nuphar Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Nuphar Rhizome according to Method 4, and perform the test (not more than 5 ppm). Petiole—When perform the test of foreign matter <5.01>, the amount of the petioles contained in Nuphar Rhi-zome does not exceed 3.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than petioles is not more than 1.0z. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Peanut Oil is a pale yellow, clear oil. It is odor-less or has a slight odor. It has a mild taste. It is miscible with diethyl ether and with petroleum ether. It is slightly soluble in ethanol (95). Specific gravity d 25: 0.909 – 0.916 Congealing point of the fatty acids: 22 – 339C
Saponify 5 g of Peanut Oil by boiling with 2.5 mL of sodium hydroxide solution (3 in 10) and 12.5 mL of ethanol (95). Evaporate the ethanol, dissolve the residue in 50 mL of hot water, and add dilute hydrochloric acid in excess until the free fatty acids separate as an oily layer. Cool the mixture, remove the separated fatty acids, and dis-solve them in 75 mL of diethyl ether. To the diethyl ether so-lution add a solution of 4 g of lead (II) acetate trihydrate in 40 mL of ethanol (95), and allow the mixture to stand for 18 hours. Filter the supernatant liquid, transfer the precipitate to the filter with the aid of diethyl ether, and filter by suc-tion. Place the precipitate in a beaker, heat it with 40 mL of dilute hydrochloric acid and 20 mL of water until the oily layer is entirely clear, cool, and decant the aqueous layer. Boil the fatty acids with 50 mL of diluted hydrochloric acid (1 in 100). When the solution prepared by dissolving 0.1 g of the fatty acids in 10 mL of ethanol (95) is not darkened by the addition of 2 drops of sodium sulfide TS, allow the fatty acids to solidify, and press them between dry filter papers to exclude moisture. Dissolve the solid fatty acid in 25 mL of diluted ethanol (9 in 10) with the aid of gentle heat, and then cool to 159C to crystallize the fatty acids. Recrystallize them from diluted ethanol (9 in 10) and dry in a desiccator (phosphorus (V) oxide, in vacuum) for 4 hours: the melting point <1.13> of the dried crystals is between 739C and 769C. material. Description Cylindrical root, 7 – 20 cm in length, 1 – 2.5 cm in diameter; externally brown to light grayish brown, with distinct longitudinal wrinkles, with warty scars of later-al roots, and with laterally elongated lenticels; fractured sur-face dense in texture, light grayish brown, and with light brown radial lines in xylem. Odor, characteristic; taste, slightly sweet at first, followed by an astringency and a slight bitterness. Identification (1) Shake 0.5 g of pulverized Peony Root with 30 mL of ethanol (95) for 15 minutes, and filter. Shake 3 mL of the filtrate with 1 drop of iron (III) chloride TS: a blue-purple to blue-green color is produced, and it changes to dark blue-purple to dark green. (2) To 2 g of pulverized Peony Root add 10 mL of meth-anol, warm on a water bath for 5 minutes, cool, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, ethyl acetate and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Peony Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Peony Root according to Method 4, and per-form the test (not more than 5 ppm). Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Weigh accurately about 0.5 g of pulverized Peony Root, add 50 mL of diluted methanol (1 in 2), heat under a reflux condenser for 30 minutes, cool, and filter. To the residue add 50 mL of diluted methanol (1 in 2), and proceed ‌2094 Powdered Peony Root / Crude Drugs and Related Drugs in the same manner. Combine the filtrates, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas, AT and AS, of paeoniflorin in each solution. Identification (1) Shake 0.5 g of Powdered Peony Root with 30 mL of ethanol (95) for 15 minutes, and filter. To 3 mL of the filtrate add 1 drop of iron (III) chloride TS, and mix: a blue-purple to blue-green color is produced, and thereafter it changes to dark blue-purple to dark green. (2) To 2 g of Powdered Peony Root add 10 mL of meth-anol, warm on a water bath for 5 minutes, cool, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Paeoniflorin RS or paeoniflorin for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions Amount (mg) of paeoniflorin (C23 ＝ MS × AT/AS H28O 11) as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). Flow rate: Adjust so that the retention time of paeoniflo-rin is about 10 minutes. System suitability— System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z.
Orange Oil is a yellow to yellow-brown liquid. It has a characteristic, aromatic odor, and a slightly bitter taste. It is miscible with an equal volume of ethanol (95) with turbidity. D Refractive index <2.45> n20: 1.472 – 1.474 D Optical rotation <2.49> a20: ＋43 – ＋509(50 mm). Specific gravity <1.13> d 20: 0.842 – 0.848
Heavy metals <1.07>—Proceed with 1.0 mL of Orange Oil according to Method 2, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 40 ppm).
Cacao Butter occurs as a yellow-white, hard, brittle mass. It has a slight, chocolate-like odor, and has no odor of rancidity. It is freely soluble in diethyl ether and in petroleum ether, soluble in boiling ethanol (99.5), and very slightly soluble in ethanol (95). Congealing point of the fatty acids: 45 – 509C Melting point 31 – 359C (Cram the sample into a capillary tube without melting the sample). Specific gravity <1.13> d 40: 0.895 – 0.904 Acid value <1.13> Not more than 3.0. Saponification value <1.13> 188 – 195 Iodine value <1.13> 35 – 43
Camellia Oil is a colorless or pale yellow, clear oil. It is nearly odorless and tasteless. It is miscible with diethyl ether and with petroleum ether. It is slightly soluble in ethanol (95). It congeals partly at －109C, and completely at －159C. Specific gravity d 25: 0.910 – 0.914
To 2 mL of Camellia Oil add dropwise 10 mL of a mixture of fuming nitric acid, sulfuric acid, and water (1:1:1), previously cooled to room temperature: a bluish green color develops at the zone of contact. Acid value <1.13> Not more than 2.8. Saponification value <1.13> 188 – 194 ‌JP XVIII Crude Drugs and Related Drugs / Powdered Capsicum 1973‌ Unsaponifiable matters <1.13> Not more than 1.0z. Iodine value <1.13> 78 – 83 capsaicin and dihydrocapsaicin (the relative retention time to (E)-capsaicin is about 1.3) obtained with the sample solu-tion, and the peak area, AS, of (E)-capsaicin obtained with
Clove Oil is a colorless or light yellow-brown, clear liquid. It has a characteristic aroma and a burning taste. It is miscible with ethanol (95) and with diethyl ether. ‌JP XVIII Crude Drugs and Related Drugs / Cnidium Rhizome 1985‌ It is slightly soluble in water. It acquires a brown color upon aging or by air.
(1) To 5 drops of Clove Oil add 10 mL of calcium hydroxide TS, and shake vigorously: the oil forms a flocculent mass, and a white to light yellow color develops. (2) Dissolve 2 drops of Clove Oil in 4 mL of ethanol (95), and add 1 to 2 drops of iron (III) chloride TS: a green color is produced. D Refractive index <2.45> n20: 1.527 – 1.537 Specific gravity <1.13> d 20: 1.040 – 1.068
(1) Clarity of solution—Dissolve 1.0 mL of Clove Oil in 2.0 mL of diluted ethanol (7 in 10): the solution is clear. Water-soluble phenols—To 1.0 mL of Clove Oil add 20 mL of boiling water, shake vigorously, filter the aqueous layer after cooling, and add 1 to 2 drops of iron (III) chloride TS: a yellow-green, but no blue or violet, color develops. Heavy metals <1.07>—Proceed with 1.0 mL of Clove Oil according to Method 2, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 40 ppm). D Optical rotation <2.49> a20: 0 – －1.59(100 mm).
Take 10.0 mL of Clove Oil in a Cassia flask, add 70 mL of sodium hydroxide TS, shake for 5 minutes and warm for 10 minutes in a water bath with occasional shaking, add sodium hydroxide TS to the volume after cooling, and allow to stand for 18 hours. Measure the volume (mL) of the sepa-rated oily layer. Total eugenol (volz) ＝ {10 － (volume of separated oily layer)}× 10
Cinnamon Oil is a yellow to brown liquid. It has a characteristic, aromatic odor and a sweet, pungent taste. It is clearly miscible with ethanol (95) and with diethyl ether. It is practically insoluble in water. It is weakly acidic. Upon aging or long exposure to air, it darkens and becomes viscous. Specific gravity d 20: 1.010 – 1.065
Shake 4 drops of Cinnamon Oil with 4 drops of nitric acid: the mixture forms white to light yellow crystals at a temperature below 59C.
(1) Rosin—Mix 1.0 mL of Cinnamon Oil with 5 mL of ethanol (95), then add 3 mL of freshly prepared, saturated ethanol solution of lead (II) acetate trihydrate: no precipitate is produced. (2) Heavy metals <1.07>—Proceed with 1.0 mL of Cinna- mon Oil according to Method 2, and perform the test. Pre-pare the control solution with 4.0 mL of Standard Lead So-lution (not more than 40 ppm).
Pipet 5.0 mL of Cinnamon Oil into a cassia flask, add 70 mL of sodium hydrogensulfite TS, and heat the mix-ture in a water bath with frequent shaking to dissolve com-pletely. To this solution add sodium hydrogensulfite TS to raise the lower level of the oily layer within the graduate por-tion of the neck. Allow to stand for 2 hours, and measure the volume (mL) of the separated oily layer. Total aldehydes (volz) ＝ {5.0 － (volume of separated oily layer)}× 20
Coconut Oil is a white to light yellow mass or a colorless or light yellow, clear oil. It has a slight, characteris-tic odor and a mild taste. It is freely soluble in diethyl ether and in petroleum ether. It is practically insoluble in water. At a temperature below 159C, it congeals to a hard and brittle solid. Melting point: 20 – 289C Acid value <1.13> Not more than 0.2. Saponification value <1.13> 246 – 264 Unsaponifiable matter <1.13> Not more than 1.0z. Iodine value <1.13> 7 – 11
Eucalyptus Oil is a clear, colorless or pale yel-low liquid. It has a characteristic, aromatic odor and a pun-gent taste. It is neutral.
Shake 1 mL of Eucalyptus Oil vigorously with 1 mL of phosphoric acid, and allow to stand: the solu-tion congeals within 30 minutes. D Refractive index <2.45> n20: 1.458 – 1.470 Specific gravity <1.13> d 20: 0.907 – 0.927
(1) Clarity of solution—Mix 1.0 mL of Eucalyptus Oil with 5 mL of diluted ethanol (7 in 10): the solution is clear. (2) Heavy metals <1.07>—Proceed with 1.0 mL of Eu- calyptus Oil according to Method 2, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 40 ppm).
Weigh accurately about 0.1 g each of Eucalyptus Oil and cineol for assay, and dissolve each in hexane to make ex-actly 25 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution to each, then add hexane to make 100 mL, and use these solutions as the sam-ple solution and the standard solution, respectively. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions. Calculate the ratios, QT and QS, of the peak area of cineol to that of the internal standard. Amount (mg) of cineol ＝ MS × QT/QS MS: Amount (mg) of cineol for assay taken Internal standard solution—A solution of anisole in hexane (1 in 250). Operating conditions— Detector: A hydrogen ‰ame-ionization detector. Column: A glass column 3 mm in inside diameter and 5 m in length, packed with silanized porous silica gel for gas chromatography coated in 5z with alkylene glycol phthalate ester for gas chromatography (150 to 180 mm in particle di-ameter). Column temperature: A constant temperature of about 1209C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of cineol is about 11 minutes. System suitability— System performance: Dissolve 0.1 g each of cineol for as-say and limonene in 25 mL of hexane. To 1 mL of this solu-tion add hexane to make 20 mL. When the procedure is run with 2 mL of this solution under the above operating condi-tions, limonene and cineol are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cineol to that of the internal standard is not more than 1.0z.
Fennel Oil is a colorless to pale yellow liquid. It has a characteristic, aromatic odor and a sweet taste with a slight, bitter aftertaste. It is miscible with ethanol (95) and with diethyl ether. It is practically insoluble in water. When cold, white crystals or crystalline masses may often separate from the oil.
Dissolve 0.30 g of Fennel Oil in 20 mL of hexane, pipet 1 mL of this solution, add hexane to make exactly 10 mL, and use this solution as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (20:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): a spot with a dark purple color appears at the Rf value of about 0.4. D Refractive index <2.45> n20: 1.528 – 1.560 Specific gravity <1.13> d 20: 0.955 – 0.995
(1) Clarity of solution—To 1.0 mL of Fennel Oil add 3 mL of ethanol (95): the solution is clear. To this solu-tion add 7 mL of ethanol (95): the solution remains clear. (2) Heavy metals <1.07>—Proceed with 1.0 mL of Fennel Oil according to Method 2, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 40 ppm).
Corn Oil is a clear, light yellow oil. It is odor-less or has a slight odor, and a mild taste. It is miscible with diethyl ether and with petroleum ether. It is slightly soluble in ethanol (95), and practically insolu- ble in water. At －79C, it congeals to an unguentary mass. Specific gravity d 25: 0.915 – 0.921 Acid value <1.13> Not more than 0.2. Saponification value <1.13> 187 – 195 Unsaponifiable matter <1.13> Not more than 1.5z. Iodine value <1.13> 103 – 130
Mentha Oil is a colorless or pale yellow, clear ‌JP XVIII Crude Drugs and Related Drugs / Moutan Bark 2073‌ liquid. It has a characteristic, pleasant aroma and has a pun-gent taste, followed by a cool aftertaste. It is miscible with ethanol (95), with ethanol (99.5), with warm ethanol (95), and with diethyl ether. It is practically insoluble in water. D Refractive index <2.45> n20: 1.455 – 1.467 D Optical rotation <2.49> a20: －17.0 – －36.09(100 mm). Specific gravity <1.13> d 25: 0.885 – 0.910 Acid value <1.13> Not more than 1.0.
(1) Clarity and color of solution—To 1.0 mL of Mentha Oil add 3.5 mL of diluted ethanol (7 in 10), and shake: Mentha Oil dissolves clearly. To the solution add 10 mL of ethanol (95): the solution is clear or has no more tur-bidity, if any, than the following control solution. Control solution: To 0.70 mL of 0.01 mol/L hydrochloric acid VS add 6 mL of dilute nitric acid and water to make 50 mL, add 1 mL of silver nitrate TS, and allow to stand for 5 minutes. (2) Heavy metals <1.07>—Proceed with 1.0 mL of Men- tha Oil according to Method 2, and perform the test. Pre-pare the control solution with 4.0 mL of Standard Lead So-lution (not more than 40 ppm).
Weigh accurately about 5 g of Mentha Oil, and dis-solve in ethanol (95) to make exactly 20 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard so-lution, and use this solution as the sample solution. Sepa-rately, weigh accurately about 10 g of l-menthol for assay, and dissolve in ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, and use this solution as the standard solu-tion. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chromatogra-phy <2.02> according to the following conditions. Calculate the ratios, QT and QS, of the peak area of menthol to that of the internal standard. Amount (mg) of menthol ＝ MS × QT/QS × 1/5 MS: Amount (mg) of l-menthol for assay taken Internal standard solution—A solution of n-ethyl caprylate in ethanol (95) (1 in 25). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column about 3 mm in inside diameter and about 2 m in length, packed with 25z of polyethylene glycol 6000 for gas chromatography supported on acid-washed 180 – 250 mm siliceous earth for gas chromatogra-phy. Column temperature: A constant temperature of about 1509C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of the internal standard is about 10 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, the internal standard and l-menthol are eluted in this order with the resolution between these peaks being not less than 5.
Olive Oil is a light yellow oil. It has a faint odor, which is not rancid, and has a bland taste. It is miscible with diethyl ether, with petroleum ether. It is slightly soluble in ethanol (95). The whole or a part of it congeals between 09C and 69C. Congealing point of the fatty acids: 17 – 269C Specific gravity <1.13> d 25: 0.908 – 0.914 Acid value <1.13> Not more than 1.0. Saponification value <1.13> 186 – 194 Unsaponifiable matters <1.13> Not more than 1.5z. Iodine value <1.13> 79 – 88
(1) Drying oil—Mix 2 mL of Olive Oil with 10 mL of diluted nitric acid (1 in 4), add 1 g of powdered sodium nitrite little by little with thorough shaking, and allow to stand in a cold place for 4 to 10 hours: the mixture congeals to a white solid. (2) Peanut oil—Weigh exactly 1.0 g of Olive Oil, dissolve in 60 mL of sulfuric acid-hexane-methanol TS, heat for 2.5 hours under a reflux condenser, cool, transfer to a separa-tor, and add 100 mL of water. Wash the vessel for the reflux extraction with 50 mL of petroleum ether, add the washing to the separator, shake, allow to stand, and separate the pe-troleum ether layer. Extract the aqueous layer with another 50 mL of petroleum ether, and combine the petroleum ether layer with the former petroleum ether solution. Wash the pe-troleum ether solution repeatedly with 20-mL portions of water until the washings show no more acidity to methyl orange TS. Then add 5 g of anhydrous sodium sulfate, shake, filter, wash anhydrous sodium sulfate with two 10-mL portions of petroleum ether, and filter the washings earth for gas chromatography (150 to 180 mm in particle di-ameter), coated with polyethylene glycol 20 mol/L in a ratio of 5z. Column temperature: A constant temperature of about 2209C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of methyl be-henate is about 18 minutes. System suitability— Test for required detectability: Adjust so that the peak height of methyl behenate obtained with 2 mL of the stand-ard solution is 5 to 10 mm.
Rape Seed Oil is a clear, pale yellow, slightly viscous oil. It is odorless or has a slight odor and a mild taste. It is miscible with diethyl ether and with petroleum diethyl ether. It is slightly soluble in ethanol (95). Specific gravity d 25: 0.906 – 0.920 Acid value <1.13> Not more than 0.2. Saponification value <1.13> 169 – 195 Unsaponifiable matters <1.13> Not more than 1.5z. Iodine value <1.13> 95 – 127
Castor Oil is a colorless or pale yellow, clear, viscous oil. It has a slight, characteristic odor, and has a bland at first, and afterwards slightly acrid taste. It is miscible with ethanol (99.5) and with diethyl ether. It is freely soluble in ethanol (95), and practically insoluble in water. When cooled to 09C, it becomes more viscous, and tur-bidity is gradually formed.
To 3 g of Castor Oil add 1 g of potassium hydroxide, and heat the mixture carefully to fuse: a charac-teristic odor is perceptible. Dissolve the fused matter in 30 mL of water, add an excess of magnesium oxide, and filter. Acidify the filtrate with hydrochloric acid: white crystals is produced. Specific gravity <1.13> d 25: 0.953 – 0.965 Acid value <1.13> Not more than 1.5. Saponification value <1.13> 176 – 187 Hydroxyl value <1.13> 155 – 177 Iodine value <1.13> 80 – 90
Adulteration—Shake to mix 1.0 g of Castor Oil with 4.0 mL of ethanol (95): it dissolves clearly. Add 15 mL of ethanol (95): no turbidity is produced.
Sesame Oil is a clear, pale yellow oil. It is odorless or has a faint, characteristic odor, and has a bland taste. It is miscible with diethyl ether and with petroleum ether. It is slightly soluble in ethanol (95). It congeals between 09C and －59C. Congealing point of the fatty acids: 20 – 259C
To 1 mL of Sesame Oil add 0.1 g of sucrose and 10 mL of hydrochloric acid, and shake for 30 seconds: the acid layer becomes light red and changes to red on stand-ing. Specific gravity <1.13> d 25: 0.914 – 0.921 Acid value <1.13> Not more than 0.2. Saponification value <1.13> 187 – 194 Unsaponifiable matters <1.13> Not more than 2.0z. Iodine value <1.13> 103 – 118
Soybean Oil is a clear, pale yellow oil. It is odorless or has a slight odor, and has a bland taste. It is miscible with diethyl ether and with petroleum ether. It is slightly soluble in ethanol (95), and practically insolu- ble in water. It congeals between －109C and －179C. Congealing point of the fatty acids: 22 – 279C Specific gravity <1.13> d 25: 0.916 – 0.922 Acid value <1.13> Not more than 0.2. Saponification value <1.13> 188 – 195 Unsaponifiable matter <1.13> Not more than 1.0z. Iodine value <1.13> 126 – 140
Turpentine Oil is a clear, colorless to pale yel-low liquid. It has a characteristic odor and a pungent, bitter taste. Turpentine Oil (1 mL) is miscible with 5 mL of ethanol (95) and this solution is neutral. D Refractive index <2.45> n20: 1.465 – 1.478 Specific gravity <1.13> d 20: 0.860 – 0.875
(1) Foreign matter—Turpentine Oil has no offen-sive odor. Shake 5 mL of Turpentine Oil with 5 mL of a so-lution of potassium hydroxide (1 in 6): the aqueous layer does not show a yellow-brown to dark brown color. Hydrochloric acid-coloring substances—Shake 5 mL of Turpentine Oil with 5 mL of hydrochloric acid, and allow to stand for 5 minutes: the hydrochloric acid layer is light yellow and not brown in color. Mineral oil—Place 5 mL of Turpentine Oil in a Cas-sia flask, cool to a temperature not exceeding 159C, add dropwise 25 mL of fuming sulfuric acid while shaking, warm between 609C and 659C for 10 minutes, and add sulfuric acid to raise the lower level of the oily layer to the graduated portion of the neck: not more than 0.1 mL of oil separates. Distilling range <2.57> 150 – 1709C, not less than 90 volz.
Fusiform root, 1 – 2.5 cm in length, 0.3 – 0.5 cm in diameter, somewhat sharp at one end, and somewhat rounded at the other; externally light yellow to light yellow-brown, with longitudinal wrinkles of various sizes; when fractured, cortex flexible and friable, stele strong; fractured surface of cortex light yellow-brown in color, slightly trans-lucent and viscous. Odor, slight; taste, slightly sweet and mucous. Under a microscope <5.01>, a transverse section reveals brown, 4- to 5-cellular layer velamen internally adjoining the epidermis; a single-layer exodermis inside the velamen, and cortex of parenchyma cells inside the exodermis; endodermis is distinct; about 20 protoxylems in actionstele; cortex paren-chyma contains columnar crystals and raphides of calcium oxalate; oil drops in the exodermis.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Ophiopogon Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Ophiopogon Root according to Method 4, and perform the test (not more than 5 ppm). Rootlets—When perform the test of foreign matter <5.01>, the amount of the rootlets contained in Ophiopogon Root is not exceed 1.0z. ‌2082‌ Powdered Opium / Crude Drugs and Related Drugs Total ash <5.01> Not more than 3.0z.
Brown Rice occurs as ellipsoidal, slightly flat-tened, 4 – 6 mm in length; externally translucent, light yel-lowish white to light brown. Slightly cave in and a white embryo at one end; a brown small dent of scar of style at the other end; few longitudinally striates on the surface. Odor, slight; taste, slightly sweet. Under a microscope <5.01>, a transverse section of the caryopsis reveals the outermost layer composed of pericarp; vascular bundles in the pericarp; seed coat adhering closely to the pericarp; in the interior, 1 or 2 cellular layered aleuron layers; parenchymatous cells of endosperm contain simple or compound starch grains.
(1) To 0.1 g of pulverized Brown Rice add 50 mL of water, and heat in a water bath for 5 minutes. After cooling, add 1 drops of iodine TS, and shake: a blue-purple color develops. (2) To 1 g of pulverized Brown Rice add 5 mL of ethyl acetate, shake for 10 minutes, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 1 mg of cycloartenyl ferulate for thin-layer chromatography in 1 mL of ethyl acetate, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer chromatography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and acetone (5:2) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-purple fluorescent spot obtained from the standard solution. Total ash <5.01> Not more than 1.5z.
Powdered Peony Root occurs as a light grayish brown powder, and has a characteristic odor and a slightly sweet taste at first, followed by an astringency and a slight bitterness. Under a microscope <5.01>, Powdered Peony Root reveals starch grains and fragments of parenchyma cells containing them; fragments of cork cells, vessels, tracheids and xylem fibers; rosette aggregates of calcium oxalate, and fragments of rows of crystal cells containing them. Starch grains con-sist of simple grains, 5 – 25 mm in diameter, occasionally 2-to 3-compound grains. MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). ‌JP XVIII Crude Drugs and Related Drugs / Peucedanum Root 2095 Flow rate: Adjust so that the retention time of paeoniflo-rin is about 10 minutes. System suitability— ‌System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Peony Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Peony Root according to Method 4, and per-form the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Peony Root does not show groups of light yellow stone cells and fibers. Loss on drying <5.01> Not less than 14.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Weigh accurately about 0.5 g of Powdered Peony Root, add 50 mL of diluted methanol (1 in 2), heat under a reflux condenser for 30 minutes, cool, and filter. To the residue add 50 mL of diluted methanol (1 in 2), and proceed in the same manner. Combine the filtrates, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Paeoniflorin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas, AT and AS, of paeoniflorin in each solution. Amount (mg) of paeoniflorin (C23H28O11) (C23H 28O11 : 480.46), calculated on the basis of dried ＝ MS × AT/AS material. Description Powdered Peony Root occurs as a light grayish brown powder, and has a characteristic odor and a slightly sweet taste at first, followed by an astringency and a slight bitterness. Under a microscope <5.01>, Powdered Peony Root reveals starch grains and fragments of parenchyma cells containing them; fragments of cork cells, vessels, tracheids and xylem fibers; rosette aggregates of calcium oxalate, and fragments of rows of crystal cells containing them. Starch grains con-sist of simple grains, 5 – 25 mm in diameter, occasionally 2-to 3-compound grains. MS: Amount (mg) of Paeoniflorin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (850:150:1). ‌JP XVIII Crude Drugs and Related Drugs / Peucedanum Root 2095 Flow rate: Adjust so that the retention time of paeoniflo-rin is about 10 minutes. System suitability— ‌System performance: Dissolve 1 mg each of Paeoniflorin RS and albiflorin in diluted methanol (1 in 2) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, albiflorin and paeoniflorin are eluted in this order with the resolution be-tween these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paeoniflorin is not more than 1.5z.
Powdered Panax Japonicus Rhizome occurs as a light grayish yellow-brown powder, and has a slight odor and a slightly bitter taste. Under a microscope <5.01>, Powdered Panax Japonicus Rhizome reveals mainly starch grains or gelatinized starch masses, and fragments of parenchyma cells containing them; also fragments of cork tissue, somewhat thick-walled collen-chyma, phloem tissue, and reticulate vessels; rarely frag-ments of scalariform vessels with a simple perforation, fibers and fiber bundles, rosette aggregates of calcium oxalate, and parenchyma cells containing them; yellow to orange-yellow resin; starch grains consisting of simple grains or 2- to 4-compound grains, simple grains, 3 – 18 mm in diameter; rosette aggregates of calcium oxalate are 20 – 60 mm in diam-eter.
Shake 0.5 g of Powdered Panax Japonicus Rhizome with 10 mL of methanol for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 2 mg of chikusetsusaponin IV for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (5:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1109C for 5 minutes: one of the several spots obtained from the sample solution shows the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Panax Japonicus Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Panax Japonicus Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Irregularly cylidrical rhizome with distinct nodes, 3 – 20 cm in length, 1 – 1.5 cm in diameter, internode 1 – 2 cm; externally light yellow-brown, with fine longitudi- ‌JP XVIII Crude Drugs and Related Drugs / Peach Kernel 2091‌ nal wrinkles; stem scars, hollowed at the center, protruding on the upper surface, and root scars protruding as knobs on internodes; easily broken; fractured surface approximately flat, and light yellow-brown in color; horny in texture. Odor, slight; taste, slightly bitter.
Shake 0.5 g of pulverized Panax Japonicus Rhizome with 10 mL of methanol for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 2 mg of chikusetsusaponin IV for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water and formic acid (5:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1109C for 5 minutes: one of the several spots obtained from the sample solution shows the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Panax Japonicus Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Panax Japonicus Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Usually, contracted and wrinkled leaves, often with thin stems. Both surfaces of the leaf are brownish pur-ple, or the upper surface is grayish green to brownish green, and the lower surface is brownish purple in color. When smoothed by immersion in water, the lamina is ovate to ob-cordate, 5 – 12 cm in length, 5 – 8 cm in width; the apex, acuminate; the margin, serrate; the base, broadly cuneate; petiole, 3 – 5 cm in length; cross sections of stem and petiole, square. Under a magnifying glass, hairs are observed on both surfaces of the leaf, but abundantly on the vein and sparsely on other parts; small glandular hairs are observed on the lower surface. Odor, characteristic; taste slightly bitter.
To 0.6 g of pulverized Perilla Herb, add 10 mL of diethyl ether, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of perillaldehyde for thin-layer chromatography in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane and ethyl acetate (3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxybenzaldehyde-sulfuric acid-acetic acid-ethanol TS for spray on the plate, and heat the plate at 1059C for 2 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Stem—When perform the test of foreign mat-ter <5.01>, Perilla Herb does not contain its stems equal to or greater than 3 mm in diameter. Foreign matter <5.01>—The amount of foreign mat-ter other than the stems contained in Perilla Herb does not exceed 1.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 16.0z. Acid-insoluble ash <5.01> Not more than 2.5z.
Weigh accurately about 0.2 g of freshly prepared pulverized Perilla Herb, put in a glass-stoppered centrifuge tube, add 20 mL of methanol, shake for 10 minutes, centri-fuge, and separate the supernatant liquid. To the residue, add 20 mL of methanol, and proceed in the same manner. Combine all the extracts, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of diphenyl sulfone for assay, and dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT of perillaldehyde and As of diphenyl sul-fone, in each solution. Amount (mg) of perillaldehyde ＝ MS × AT/AS × 1/20 × 0.700 MS: Amount (mg) of diphenyl sulfone for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 234 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (13:7). Flow rate: 1.0 mL per minute. System suitability— System performance: Dissolve 1 mg of (E )-asarone and 1 mg of perillaldehyde for thin-layer chromatography in the standard solution to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, diphenyl sulfone, perillaldehyde, and (E )-asarone are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diphenyl sulfone is not more than 1.5z.
Powdered Peach Kernel occurs as a reddish-light brown to light brown powder. It is almost odorless and is oily and has slightly a bitter taste. Under a microscope <5.01>, Powdered Peach Kernel frag- ments of outer seed coat epidermis; elliptical to ovoid, con-taining yellow-brown compound 50 to 80 mm in diameter and stone cell; cap-like shape to ovoid, yellow-brown in color. The stone cell is element of epidermis, 50 to 80 mm in diameter and 70 to 80 mm in height, cell wall of the top, 12 to 25 mm thickness, the base 4 mm in thickness, with obvious and numerous pits. Inner seed coat, yellow-brown, irregular and somewhat long polygon, 15 to 30 mm in diameter; and fragments of cotyledon and albumen containing aleurone grains and fatted oil, Aleurone grains are almost spherical grains, 5 to 10 mm in diameter.
To 1.0 g of Powdered Peach Kernel add 10 mL of methanol, and immediately heat under a reflux con- Amount (mg) of amygdalin ＝ MS × AT/AS × 2 denser for 10 minutes. After cooling, filter, and use the fil-trate as the sample solution. Separately, dissolve 2 mg of MS: Amount (mg) of amygdalin for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilianized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amygdalin is not more than 1.5z.
Weigh accurately 0.5 g of Powdered Peach Kernel, add 40 mL of diluted methanol (9 in 10), heat immediately under a reflux condenser for 30 minutes, and cool. Filter the mixture, add diluted methanol (9 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, dissolve in diluted methanol (1 in 2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test exactly with 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. Amount (mg) of amygdalin ＝ MS × AT/AS × 2 MS: Amount (mg) of amygdalin for assay taken ‌JP XVIII Crude Drugs and Related Drugs / Peony Root 2093‌ Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilianized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and methanol (5:1). Flow rate: 0.8 mL per minute (the retention time of amyg-dalin is about 12 minutes). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amygdalin are not less than 5000 and Acid value <1.13> Not more than 0.2. Saponification value <1.13> 188 – 196 Unsaponifiable matters <1.13> Not more than 1.5z. Iodine value <1.13> 84 – 103
Flattened, asymmetric ovoid seed, 1.2 – 2.0 cm in length, 0.6 – 1.2 cm in width, and 0.3 – 0.7 cm in thick-ness; somewhat sharp at one end, and round at the other end with chalaza; seed coat red-brown to light brown; externally, its surface being powdery by easily detachable stone cells of epidermis; numerous vascular bundles running and rarely branching from chalaza through the seed coat, and, appear-ing as dented longitudinal wrinkles; when soaked in boiling water and softened, the seed coat and thin, translucent, white albumen easily separated from the cotyledone; cotyle-done white in color. Almost odorless; taste, slightly bitter and oily. Under a microscope <5.01>, the outer surface of seed coat reveals polygonal, long polygonal, or obtuse triangular stone cells on the protrusion from vascular bundles, shape of which considerably different according to the position, and their cell walls almost equally thickened; in lateral view, ap-pearing as a square, rectangle or obtuse triangle.
To 1.0 g of ground Peach Kernel add 10 mL of methanol, immediately heat under a reflux condenser for 10 minutes, cool, filter, and use the filtrate as the sample so-lution. Separately, dissolve 2 mg of amygdalin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. ‌2092 Powdered Peach Kernel / Crude Drugs and Related Drugs Develop the plate with a mixture of ethyl acetate, methanol and water (20:5:4) to a distance of about 7 cm, and air-dry the plate. Spray evenly thymol-sulfuric acid-methanol TS for spraying upon the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Rancidity—Grind Peach Kernel with boiling water: no odor of rancid oil is perceptible. (2) Foreign matter <5.01>—When perform the test with not less than 250 g of Peach Kernel, it contains not more than 0.10z of broken pieces of endocarp. Loss on drying <5.01> Not more than 8.0z (6 hours).
Weigh accurately 0.5 g of ground Peach Kernel, add 40 mL of diluted methanol (9 in 10), heat immediately under a reflux condenser for 30 minutes, and cool. Filter the mix-ture, add diluted methanol (9 in 10) to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, filter, and use the filtrate as the sample solution. Sepa-rately, weigh accurately about 10 mg of amygdalin for assay, previously dried in a desiccator (silica gel) for not less than 24 hours, dissolve in diluted methanol (1 in 2) to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amygdalin in each solution. Powdered Peach Kernel Persicae Semen Pulveratum
1) Peucedanum Praeruptorum Root—Slen-der obconical to cylindrical root, occasionally dichotomized at the lower part, 3 – 15 cm in length, 0.8 – 1.8 cm in diame- ‌2096‌ Pharbitis Seed / Crude Drugs and Related Drugs ter at the crown; externally light brown to dark brown; ring-node-like wrinkles numerous at the crown, sometimes with hair-like remains of leaf sheath; the root having somewhat deep longitudinal wrinkles and scars of cutting off of lateral roots; transverse section surface light brown to whitish in color; brittle in texture. Odor, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section reveals the outermost layer composed of a cork layer, inner tangential walls of some cork cells thickened; collenchyma just inside of the cork layer; in cortex numerous oil canals scattered and intercellular air spaces observed; occasionally phloem fibers observed at the terminal portion of phloem; vessels and scat-tered oil canals in xylem; starch grains in parenchyma, 2 to 10 several-compound grains. 2) Angelica Decursiva Root—Similar to 1), but without hair-like remains of leaf sheath at the crown. Odor, characteristic; taste, slightly bitter. Under a microscope <5.01>, a transverse section reveals, similar to 1), but cell wall of cork cells not thickened, phloem fibers not observed at the terminal portion of phloem, nor oil canals observed in xylem.
1) Peucedanum Praeruptorum Root—To 1 g of pulverized Peucedanum Root add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of (±)-praeruptorin A for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether and hexane (3:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the blue-white to blue-purple fluorescent spot from the standard solution. 2) Angelica Decursiva Root—To 1 g of pulverized Peu-cedanum Root add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 1 mg of nodakenin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl ace-tate, methanol and water (12:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the blue-white to blue-purple fluorescent spot from the standard solution.
Heavy metals <1.07>—Proceed with 1.0 g of pulver-ized Peucedanum Root according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Loss on drying <5.01> Not more than 13.0z. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 20.0z.
Longitudinally quartered or sexpartite globe, 4 – 6 mm in length, 3 – 5 mm in width; externally black to grayish red-brown or grayish white, smooth, but slightly shrunken and coarsely wrinkled. The transverse section almost fan-shaped, light yellow-brown to light grayish brown, and dense in texture. Under a magnifying glass, the surface of the seed coat reveals dense, short hairs; dented hilum at the bottom of the raphe. Seed coat thin, the outer layer dark gray, and the inner layer light gray; two irregu-larly folded cotyledons in the transverse section at one end; two septa from the center of the dorsal side to the ridge separating cotyledons but unrecognizable in the transverse section of the other end having hilum; dark gray secretory pits in the section of the cotyledon. 100 seeds weighing about 3.5 g. When cracked, odor, slight; taste, oily and slightly pun-gent. Total ash <5.01> Not more than 6.0z.
Powdered Phellodendron Bark occurs as a bright yellow to yellow powder. It has a slight odor and an extremely bitter taste, is mucilaginous, and colors the saliva yellow on chewing. Under a microscope <5.01>, Powdered Phellodendron Bark reveals fragments of yellow, thick-walled fiber bundles or fibers, and fibers often accompanied by crystal cell rows; fewer groups of stone cells together with idioblasts; frag-ments of parenchyma cells containing starch grains and oil droplets; fragments of medullary ray and phloem; mucilage cells and mucilage masses. Numerous solitary crystals of cal-cium oxalate, 7 – 20 mm in diameter; starch grains, simple grains and 2- to 4-compound grains, simple grain, 2 – 6 mm in diameter; oil droplets, stained red with sudan III TS.
(1) To 1 g of Powdered Phellodendron Bark add 10 mL of diethyl ether, allow to stand for 10 minutes with occasional shaking, and filter to remove the diethyl ether. Collect the powder on the filter paper, add 10 mL of ethanol (95), allow to stand for 10 minutes with occa-sional shaking, and filter. To 2 to 3 drops of the filtrate add 1 mL of hydrochloric acid, add 1 to 2 drops of hydrogen peroxide TS, and shake: a red-purple color develops. Use the filtrate obtained in (1) as the sample solution. Separately, dissolve 1 mg of Berberine Chloride RS or ber-berine chloride hydrate for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the sever-al spots obtained from the sample solution and a spot with yellow to yellow-green fluorescence from the standard solu-tion show the same color tone and the same Rf value. Stir up Powdered Phellodendron Bark with water: the solution becomes gelatinous owing to mucilage. ‌2098‌ Compound Phellodendron Powder for Cataplasm / Crude Drugs and Related Drugs
Curcuma—Place Powdered Phellodendron Bark on filter paper, drop diethyl ether on it, and allow to stand. Take the powder off the filter paper, and drip 1 drop of po-tassium hydroxide TS: no red-purple color develops. Under a microscope <5.01>, Powdered Phellodendron Bark does not contain gelatinized starch or secretory cells containing yellow-red resin. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 7.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Weigh accurately about 0.5 g of Powdered Phel-lodendron Bark, add 30 mL of a mixture of methanol and dilute hydrochloric acid (100:1), heat under a reflux con-denser for 30 minutes, cool, and filter. Repeat the above procedure twice with the residue, using 30-mL and 20-mL portions of a mixture of methanol and dilute hydrochloric acid (100:1). To obtained residue add 10 mL of methanol, shake well, and filter. Combine the whole filtrates, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine [as berberine chloride (C20H18ClNO4)] ＝ MS × AT/AS MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen-phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. System suitability－ System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in methanol to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, palmatine and berber-ine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Flat or rolled semi-tubular pieces of bark, 2 – 4 mm in thickness; externally grayish yellow-brown to grayish brown, with numerous traces of lenticels; the internal sur-face yellow to dark yellow-brown in color, with fine vertical lines, and smooth; fractured surface fibrous and bright yel-low. Odor, slight; taste, extremely bitter; mucilaginous; it colors the saliva yellow on chewing. Under a microscope <5.01>, a transverse section reveals primary ray expands outward and looks fan shaped in sec-ondary cortex, and sometimes ray differentiated later con-verges outward; groups of stone cells yellow and scattered in primary ray; groups of phloem fibers light yellow to yellow, lined alternately with the other tissue of phloem between rays, and then these tissues show obviously latticework; soli-tary crystals of calcium oxalate, single and compound starch grains observed in parenchyma. JP XVIII Crude Drugs and Related Drugs / Powdered Phellodendron Bark 2097 ‌Identification (1) To 1 g of pulverized Phellodendron Bark add 10 mL of diethyl ether, allow to stand for 10 minutes with occasional shaking, and filter to remove the diethyl ether. Collect the powder on the filter paper, add 10 mL of ethanol (95), allow to stand for 10 minutes with occa-sional shaking, and filter. To 2 to 3 drops of the filtrate add 1 mL of hydrochloric acid, add 1 to 2 drops of hydrogen peroxide TS, and shake: a red-purple color develops. Use the filtrate obtained in (1) as the sample solution. Separately, dissolve 1 mg of Berberine Chloride RS or ber-berine chloride hydrate for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the sever-al spots obtained from the sample solution and a spot with yellow to yellow-green fluorescence from the standard solu-tion show the same color tone and the same Rf value. Stir up pulverized Phellodendron Bark with water: the solution becomes gelatinous owing to mucilage. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 7.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Weigh accurately about 0.5 g of pulverized Phel-lodendron Bark, add 30 mL of a mixture of methanol and dilute hydrochloric acid (100:1), heat under a reflux con-denser for 30 minutes, cool, and filter. Repeat the above procedure twice with the residue, using 30-mL and 20-mL portions of a mixture of methanol and dilute hydrochloric acid (100:1). To the last residue add 10 mL of methanol, shake well, and filter. Combine the whole filtrates, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Berberine Chloride RS (separately determine the water <2.48> in the same manner as Berberine Chloride Hydrate), dissolve in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of berberine in each solution. Amount (mg) of berberine [as berberine chloride (C20H18ClNO4)] ＝ MS × AT/AS MS: Amount (mg) of Berberine Chloride RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 345 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 15 to 25 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 409C Mobile phase: Dissolve 3.4 g of potassium dihydrogen-phosphate and 1.7 g of sodium lauryl sulfate in 1000 mL of a mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of berberine is about 10 minutes. System suitability— System performance: Dissolve 1 mg each of Berberine Chloride RS and palmatine chloride in methanol to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, palmatine and berber-ine are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of berberine is not more than 1.5z.
Phellodendron, Albumin Tannate and Bismuth Subnitrate Powder
オウバク・タンナルビン・ビスマス散
wood of Picrasma quassioides Bennet (Simaroubaceae)
Powdered Picrasma occurs as a grayish white to light yellow powder. It is odorless, and has an extremely bitter and lasting taste. Under a microscope <5.01>, Powdered Picrasma Wood re- veals fragments of vessels of various sizes, xylem fibers and xylem parenchyma cells; fragments of medullary rays con-taining starch grains; all tissues lignified; a few crystals of calcium oxalate observed. Starch grains are 5 to 15 mm in di-ameter. Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Light yellow chips, slices or short pieces of wood; a transverse section reveals distinct annual rings and thin medullary rays; tissue dense in texture. Odorless; taste, extremely bitter and lasting. Under a microscope <5.01>, it reveals medullary rays con-sisting of 1 – 5 cells wide for transverse section, and 5 – 50 cells high for longitudinal cut surface; vessels of spring wood up to about 150 mm in diameter, but those of autumn wood only one-fifth as wide; vessels, single or in groups, scattered in the xylem parenchyma; wall of wood fibers extremely thickened; medullary rays and xylem parenchyma cells con-tain rosette aggregates of calcium oxalate and starch grains. Vivid yellow or red-brown, resinous substance often present in the vessels.
Foreign matter <5.01>—The amount of foreign mat-ter contained in Picrasma Wood does not exceed 1.0z. Total ash <5.01> Not more than 4.0z.
Slightly flattened spherical to irregular-shaped tuber; 0.7 – 2.5 cm in diameter and 0.7 – 1.5 cm in height; externally white to grayish white-yellow; the upper end dent-ed, where the stem has been removed, with root scars dented as numerous small spots on the circumference; dense in tex-ture; cross section white and powdery. Almost odorless; tasteless at first, slightly mucous, but leaving a strong acrid taste. Under a microscope <5.01>, a transverse section reveals mainly tissue of parenchyma filled with starch grains, and scattered with a few mucilage cells containing raphides of calcium oxalate. Starch grains mostly 2- to 3-compound grains, usually 10 – 15 mm in diameter, and simple grains, usually 3 – 7 mm in diameter; raphides of calcium oxalate 25 – 150 mm in length.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Pinellia Tuber according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Pinellia Tuber according to Method 4, and per-form the test (not more than 5 ppm). Rhizome of Arisaema species and others—Under a microscope <5.01>, no mucilage canal is revealed on the out-er layer of cortex. Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 3.5z.
Usually wrinkled and contracted leaf and spike, grayish green to dark yellow-green in color; when soaked in water and smoothed out, the lamina is ovate to orbicular-ovate, 4 – 15 cm in length, 3 – 8 cm in width; apex acute, and base sharply narrowed; margin slightly wavy, with distinct parallel veins; glabrous or nearly glabrous; peti-ole is rather longer than the lamina, and its base is slightly expanded with thin-walled leaf-sheath; scape is 10 – 50 cm in length, one-third to one-half of the upper part forming the spike, with dense florets; the lower part of inflorescence often shows pyxidia; roots usually removed, but, if any, fine roots are closely packed. Odor, slight; practically tasteless.
To 2.0 g of pulverized Plantago Herb add 10 mL of methanol, warm on a water bath for 3 minutes, cool, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride TS on the plate: a dark blue spot appears at an Rf value of about 0.55. Total ash <5.01> Not more than 15.0z. Acid-insoluble ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 14.0z.
Flattened ellipsoidal seed, 2 – 2.5 mm in length, 0.7 – 1 mm in width, 0.3 – 0.5 mm in thickness; ex-ternally brown to yellow-brown and lustrous. Under a mag-nifying glass, the surface of the seed is practically smooth, with the dorsal side protruding like a bow, and with the ven-tral side somewhat dented; micropyle and raphe not observa-ble. 100 seeds weigh about 0.05 g. Almost odorless; taste, slightly bitter and mucous. Under a microscope <5.01>, a transverse section reveals a seed coat consisting of three layers of epidermis composed of cells containing mucilage, a vegetative layer, and a pigment layer of approximately equidiameter cells; in the interior, en-dosperm thicker than seed coat, enclosing two cotyledons.
(1) To 1 g of Plantago Seed add 2 mL of warm water, and allow the mixture to stand for 10 minutes: the seed coat swells to discharge mucilage. (2) To 1.0 g of pulverized Plantago Seed add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, to 0.2 g of powdered plantago seed for thin-layer chromatography add 1 mL of methanol, and warm on a water bath for 3 minutes. After cooling, centrifuge, and use the supernatant liquid as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of acetone, ethyl ace-tate, water and acetic acid (100) (10:10:3:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methox-ybenzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 10 minutes: the spot appeared at an Rf value of about 0.25 obtained from the sample solution has the same color tone with the dark blue spot appeared at an Rf value of about 0.25 from the standard solution.
Foreign matter <5.01>—The amount of foreign mat-ter contained in Plantago Seed does not exceed 2.0z. ‌JP XVIII Crude Drugs and Related Drugs / Powdered Platycodon Root 2101‌ Total ash <5.01> Not more than 5.5z. Acid-insoluble ash <5.01> Not more than 2.0z.
entire plant of Plantago asiatica Linnáe (Plantaginaceae), collected during the flowering season
Powdered Platycodon Root occurs as a light grayish yellow to light grayish brown powder. It has a slight odor, and is tasteless at first, later acrid and bitter. Under a microscope <5.01>, Powdered Platycodon Root reveals numerous fragments of colorless parenchyma cells; fragments of reticulate vessels and scalariform vessels; frag-ments of sieve tubes and lactiferous tubes; fragments of cork layer are sometimes observed. Usually, starch grains are not observed, but very rarely simple grain.
(1) Warm 0.2 g of Powdered Platycodon Root with 2 mL of acetic anhydride on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add carefully 0.5 mL of sulfuric acid to make two layers: a red to red-brown color develops at the zone of contact, and the upper layer ac-quires a blue-green to green color. (2) To 2.0 g of Powdered Platycodon Root add 20 mL of sodium carbonate TS, and shake. Add 5 mL of 1-butanol, shake for 10 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of platycodin D for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (5:3:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution and a spot from the stan-dard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Platycodon Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Platycodon Root according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Platycodon Root does not show fibers, stone cells or other foreign matter. Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 1.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
(1) Warm 0.2 g of pulverized Platycodon Root with 2 mL of acetic anhydride on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add carefully 0.5 mL of sulfuric acid to make two layers: a red to red-brown color develops at the zone of contact, and the upper layer ac-quires a blue-green to green color. (2) To 2.0 g of pulverized Platycodon Root add 20 mL of sodium carbonate TS, and shake. Add 5 mL of 1-butanol, shake for 10 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Separately, dissolve 1 mg of platycodin D for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (5:3:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution and a spot from the stan-dard solution show the same color tone and the same Rf value.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Platycodon Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Platycodon Root according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 25.0z.
terrestrial part of Pogostemon cablin Bentham (Labiatae)
Stems with opposite leaves, leaves wrinkled and shriveled. When smoothed by immersion in water, leaves are obovate to ovate-oblong, 2.5 – 10 cm in length, 2.5 – 7 cm in width, with obtusely serrate margins and peti-oles at the cuneate bases; the upper surface of leaves dark brown, the lower surface grayish brown, both sides covered densely with hairs. Stems are square, solid, grayish green, covered with grayish white to yellow-white hairs; the pith broad, whitish, spongy. Under a magnifying glass, leaf rev-eals hairs, glandular hairs and glandular scales. Odor, distinct; taste, slightly bitter. Under a microscope <5.01>, a transverse section of petiole reveals central portion of the adaxial side protruding remarkably, with collenchyma cells beneath epidermis; vas-cular bundles at the center divided into two groups. Under a microscope <5.01>, a transverse section of the midvein of lamina reveals the adaxial side protruding remarkably, with collenchyma cells beneath epidermis; vascular bundles at the center arranged in fan-shape. Under a microscope <5.01>, a transverse section of stem reveals several-cells-layered collen-chyma beneath epidermis, occasionally with cork layer deve-loped; beneath cortex, collateral vascular bundles arranged in a circle, phloem fibers in groups observed at the outer portion of phloem; oil droplets observed in parenchymat cells of cortex, needle, solitary or columnar crystals of cal-cium oxalate in parenchyma cells of pith.
To 0.5 g of pulverized Pogostemon Herb, add 5 mL of methanol, shake for 3 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of hexane and acetone (9:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-methoxy-benzaldehyde-sulfuric acid TS on the plate, and heat the plate at 1059C for 5 minutes; a blue-purple spot appears at an Rf value of about 0.4. Loss on drying <5.01> Not more than 15.0z (6 hours). Total ash <5.01> Not more than 13.0z. Acid-insoluble ash <5.01> Not more than 3.0z. Essential oil content <5.01> When the test is performed with 50.0 g of pulverized Pogostemon Herb in a flask with 1 mL of silicon resin added, the essential oil content is not less than 0.3 mL.
Powdered Polygala Root occurs as a light yel-low-grayish brown powder. It has a slight odor and a slightly acrid taste. Under a microscope <5.01>, Powdered Polygala Root re- veals fragments of cork layers, pitted vessels, reticulate ves-sels and tracheids; fragments of xylem fibers and xylem parenchyma cells with a small number of simple pits; frag-ments of parenchyma cells containing substances such as oil droplets, rosette aggregates and solitary crystals of calcium oxalate. Oil drop-like contents stained red with sudan III TS.
(1) Shake vigorously 0.5 g of Powdered Polygala Root with 10 mL of water: a lasting fine foam is produced. (2) To 1.0 g of Powdered Polygala Root add 10 mL of a solution of sodium hydroxide (1 in 10), and heat under a re‰ux condenser for 20 minutes. After cooling, add 10 mL of dilute hydrochloric acid, and shake. After cooling, add 10 mL of 1-butanol, shake for 10 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, water and acetic acid (100) (20:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: a red-brown to light brown spot appears at an Rf value of about 0.35.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Polygala Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Polygala Root according to Method 4, and per-form the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Polygala Root does not show stone cells or starch grains. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 6.0z.
Thin, long and bent, cylindrical or tubular root; main root, 10 – 20 cm in length, 0.2 – 1 cm in diameter, sometimes with one to several lateral roots; externally light grayish brown, with coarse longitudinal wrinkles, and with deep lateral furrows cracked to some degree here and there; brittle, and fractured surface not fibrous; under a mag-nifying glass, margin of the transverse section irregularly un-dulate; cortex, comparatively thick, with large cracks here and there; xylem usually round to elliptical, light brown in color, and often tears in a wedge-like shape. ‌JP XVIII Crude Drugs and Related Drugs / Polygonatum Rhizome 2103‌ Odor, slight; taste, slightly acrid.
(1) Shake vigorously 0.5 g of pulverized Polygala Root with 10 mL of water: a lasting fine foam is produced. (2) To 1.0 g of pulverized Polygala Root add 10 mL of a solution of sodium hydroxide (1 in 10), and heat under a re‰ux condenser for 20 minutes. After cooling, add 10 mL of dilute hydrochloric acid, and shake. After cooling, add 10 mL of 1-butanol, shake for 10 minutes, centrifuge, and use the 1-butanol layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, water and acetic acid (100) (20:4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat the plate at 1059C for 10 minutes: a red-brown to light brown spot appears at an Rf value of about 0.35.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Polygala Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Polygala Root according to Method 4, and perform the test (not more than 5 ppm). Stem—When perform the test of foreign matter <5.01>, the amount of the stems contained in Polygala Root does not exceed 10.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than the stems is not more than 1.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 6.0z.
Irregularly cylindrical rhizome, 3 – 10 cm in length, 0.5 – 3 cm in diameter; or irregular massive rhizome, 5 – 10 cm in length, 2 – 6 cm in diameter, occasionally bran-ched; both rhizomes with many cyclic nodes and longitudi-nally striate; externally yellow-brown to black-brown; stem scars, round, concave at their center, and protuberant on the upper surface; root scars on the lower surface; cut surface flat and horny. Odor, slight; taste, slightly sweet. Under a microscope <5.01>, a transverse section of the rhi-zome reveals an epidermis coated with cuticle; inside of epidermis parenchyma lie; numerous vascular bundles and mucilage cells scattered in parenchyma; vascular bundles col-lateral or amphivasal concentric; mucilage cells contain raphides of calcium oxalate.
(1) To 0.5 g of fine cutted Polygonatum Rhizome add 2 mL of acetic anhydride, warm on a water bath for 2 minutes, and filter. To 1 mL of the filtrate add gently 0.5 mL of sulfuric acid: a red-brown color appears at the zone of contact. (2) To 1.0 g of fine cutted Polygonatum Rhizome add 10 mL of dilute hydrochloric acid, boil gently for 2 minutes, and filter. Neutralize the filtrate with sodium hydroxide TS. To 3 mL of this solution add 1 mL of Fehling's TS, and warm: red precipitates appear. ‌2104‌ Polygonum Root / Crude Drugs and Related Drugs
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Polygonutum Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g ‌of pulverized Polygonutum Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
the rhizome of Polygonatum kingianum Collett et Hemsley, Polygonatum
Polygonum Root is nearly fusiform, 10 – 15 cm in length, 2 – 5 cm in diameter; externally red-brown to dark brown; roughly wrinkled; a cross section light red-brown or light grayish brown, with numerous abnormal vas-cular bundles scattering irregularly around the large vascular bundles near center; heavy and hard in texture. Odor, slight and characteristic; taste, astringent and slightly bitter. Under a microscope <5.01>, transverse section reveals the outermost layer to be several cells thick and composed of cork; cork cells contain brown substances; cortex composed of parenchyma; abnormal vascular bundles, exhibiting a ring of cambium; xylem lies inside of the cambium, and phloem outside; fibers lie outside the phloem; central portion of root lignified; parenchymatous cells contain aggregated crystals of calcium oxalate, and both simple and 2- to 8-compound starch grains; navel of starch grain obvious.
To 1 g of pulverized Polygonum Root add 10 mL of methanol, shake for 15 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 2 mL of methanol, and use this as the sample solution. Per-form the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatogra-phy, develop the plate with a mixture of ethyl acetate, water, methanol and acetic acid (100) (200:10:10:3) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): a fluorescent blue-white spot appears at an Rf value of about 0.3.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Polygonum Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Polygonum Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 14.0z (6 hours). Total ash <5.01> Not more than 5.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 17.0z.
Flat or semi-tubular pieces of bark; 3 – 6 mm thick, externally light brown to brown, internal surface smooth, grayish brown to brown, occasionally periderm peeled off; the bark with periderm externally rough and lenticels observed; internal surface with many fine longitudi-nal lines; transversely cut surface grayish brown to brown, fibrous. Odor, slightly characteristics; taste, slightly bitter and astringent. Under a microscope <5.01>, a transverse section reveals cork layer containing solitary crystals and rosette aggregates of calcium oxalate in the bark with periderm; in secondary cortex many stone cells and idioblasts arranged irregularly and parenchyma cells containing solitary crystals and rosette aggregates of calcium oxalate dotted; groups of phloem fibers lined alternately with the other tissue of phloem be-tween rays.
Shake 1 g of pulverized Cherry Bark with 10 mL of dilute hydrochloric acid, and heat in a boiling water bath for 10 minutes. After cooling, add 5 mL of diethyl ether, shake for 10 minutes, centrifuge, and use the diethyl ether layer as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (20:20:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: a crimson spot appears at an Rf value of about 0.5. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Usually cut into small pieces of irregular hexa-gons of about 0.5 cm cube, or cut into longitudinally plate-like pieces 20 – 30 cm in length, 5 – 10 cm in width, and about 1 cm in thickness; externally light grayish yellow to grayish white; transverse section showing concentric annu-late ring or part of it formed by abnormal growth of cambi-um. Under a magnifying glass, phloem light grayish yellow in color; in xylem, numerous vessels appearing as small dots; medullary rays slightly dented; vertical section showing lon-gitudinal patterns formed alternately by fibrous xylem and parenchyma; easily breakable lengthwise, and its section ex-tremely fibrous. Almost odorless; taste, at first slightly sweet, followed by a slight bitterness. Under a microscope <5.01>, a transverse section reveals fiber bundles accompanied by crystal cells in phloem; dis-tinct vessels and xylem fibers in xylem; starch grains numerous in parenchyma, mainly composed of polygonal simple grains, rarely 2- to 3-compound grains, 2 – 18 mm, mostly 8 – 12 mm, in size, with hilum or cleft in the center, and also with striae.
To 2 g of pulverized Pueraria Root add 10 mL of methanol, shake for 3 minutes, filter, and use the fil-trate as the sample solution. Separately, dissolve 1 mg of Puerarin RS or puerarin for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (12:2:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the blue-white fluorescent spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Pueraria Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g ‌JP XVIII Crude Drugs and Related Drugs / Rape Seed Oil 2111 ‌of pulverized Pueraria Root according to Method 4, and per-form the test (not more than 5 ppm).‌‌ Loss on drying <5.01> Not less than 13.0z (6 hours). Total ash <5.01> Not more than 6.0z.
Weigh accurately about 0.3 g of pulverized Pueraria Root, add 50 mL of diluted methanol (1 in 2), and heat under a reflex condenser for 30 minutes, cool, and filter. To the residue add 50 mL of diluted methanol (1 in 2), and per-form as the same as above. Combine the filtrates, add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of Puerarin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), add diluted methanol (1 in 2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of puerarin in each solution. verse section brown to light brown, white small spots com-posed of stone cells in groups observed sporadically. Almost odorless, tasteless. Under a microscope <5.01>, a transverse section reveals a cork layer with scattered cork stone cells; in secondary cor-tex fiber bundles lined almost stepwise, large groups of stone cells arranged irregularly; in parenchyma aggregated crystals of calcium oxalate scattered; adjacent to stone cells and fiber cells, cells containing solitary crystals of calcium oxalate ob-served, and these cells form crystal cell rows in a longitudinal section. Identification To 2 g of pulverized Quercus Bark, add 10 mL of ethyl acetate, shake for 10 minutes, and centrifuge to remove the supernatant liquid. Add 10 mL of acetone to the residue, shake for 10 minutes, centrifuge, and use the super-natant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatog-raphy <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, methanol and water (7:2:1) Amount (mg) of puerarin (C 21H20O9) ＝ M S × AT/AS to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): Two con- MS: Amount (mg) of Puerarin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). ‌Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS and acetonitrile (9:1). Flow rate: Adjust so that the retention time of puerarin is about 15 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the number of theoretical plates and the symme-try coefficient of the peak of puerarin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of puerarin is not more than 1.5z.
Plate-like or semi-tubular pieces of bark, 5 – 15 mm in thickness; externally grayish brown to dark brown, with thick periderm and longitudinal coarse splits; internally brown to light brown, with longitudinal ridges, the trans- secutive blue to blue-white fluorescent spots in different color tone are observed at an Rf value of about 0.4. Then, spray evenly diluted sulfuric acid on the plate, heat the plate at 1059C. Examine under ultraviolet light (main wavelength: 365 nm): one of these spots produces blue to blue-white fluorescence. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 8.5z. Acid-insoluble ash <5.01> Not more than 0.5z.
Ginseng
Red Ginseng
ninjin
Ginseng Radix consists of the root of Panax ginseng C.A. Meyer (Panax schinseng Nees) (Araliaceae), usually with the root bark removed, dried and processed.
Collected in autumn from plants 4-6 years old
Fusiform or cylindrical, often branched, 5-20 cm long, 0.5-3 cm in diameter
Fracture surface is flat, yellowish white
Sweet at first, followed by slight bitterness
Cork layer of several rows of cells; cortex of parenchyma cells containing starch grains and occasionally cluster crystals of calcium oxalate; phloem with sieve tubes and parenchyma; xylem with vessels, tracheids, and parenchyma
Under a microscope, powder shows fragments of cork cells, parenchyma cells containing starch grains (simple or compound, 2-20 μm in diameter), vessels with scalariform or reticulate thickenings, and occasional calcium oxalate crystals
Presence of secretory canals in cortex and phloem; starch grains abundant; calcium oxalate cluster crystals 20-70 μm in diameter
To 2.0 g of powdered Ginseng Radix add 20 mL of methanol, heat under reflux on a water bath for 15 minutes, cool, and filter. Evaporate the filtrate to dryness, add 10 mL of water to the residue, and extract with two 20-mL portions of 1-butanol saturated with water. Combine the 1-butanol extracts, evaporate to dryness, and dissolve the residue in 2 mL of methanol. Use this solution as the sample solution. Separately, dissolve 2 mg of Ginsenoside Rb1 RS in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography. Spot 10 μL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and water (65:35:10) (lower layer) to a distance of about 10 cm, and dry the plate. Spray evenly dilute sulfuric acid on the plate, and heat at 110°C for 5 minutes: spots from the sample solution show the same color tone and Rf values with the spots from the standard solution.
Weigh accurately about 0.5 g of powdered Ginseng Radix, add 30 mL of diluted methanol (3 in 5), heat under reflux on a water bath for 15 minutes, cool, filter, and add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 10 mL of this solution, add 10 mL of water, transfer to a column packed with 2 g of polyamide C for column chromatography, and elute with 50 mL of water, then with 50 mL of diluted methanol (3 in 5). Combine the diluted methanol (3 in 5) eluates, evaporate to dryness under reduced pressure, dissolve the residue in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Ginsenoside Rb1 RS (previously determine the water content), dissolve in methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 μL each of the sample solution and standard solution as directed under Liquid Chromatography according to the following conditions. Calculate the amounts of ginsenoside Rb1 in the sample solution.
1) Kan-jio—Massive or fusiform root, nar-row at one or both ends, 5 – 10 cm in length, 0.5 – 3.0 cm in diameter, sometimes broken or markedly deformed in shape; externally yellow-brown, black-brown or black, with deep, longitudinal wrinkles and constrictions; soft in texture; transversely cut surface yellow-brown, black-brown, or black and peripheral portion darker. Odor, characteristic; taste, slightly sweet at first, followed by a slight bitterness. Under a microscope <5.01>, a transverse section reveals 7 – 15 cellular layers of cork layer; cortex composed entirely of parenchyma; cells containing brown secretes scattered in cor-tex; xylem practically filled with parenchyma; vessels radial-ly lined, mainly reticulate vessels. 2) Juku-jio—Irregularly massive root, or massive or fusiform root, narrow at one or both ends, 5 – 10 cm in length, 0.5 – 3.0 cm in diameter; externally black, usually lustrous, with deep, longitudinal wrinkles and constrictions; soft in texture and mucous; transversely cut surface black. Odor, characteristic; taste, sweet at first, followed by a slight bitterness. Under a microscope <5.01>, a transverse section reveals 7 – 15 cellular layers of cork layer; cortex composed entirely of parenchyma; cells containing brown secretes scattered in cor-tex; xylem practically filled with parenchyma, often paren-chyma partially broken and gaps observed; vessels radially lined, mainly reticulate vessels.
1) Kan-jio—Sake 0.5 g of the fine cutting of Rehmannia Root with 5 mL of water, add 20 mL of meth-anol, shake for 10 minutes, centrifuge, and use the superna-tant liquid as the sample solution. Separately, dissolve 2 mg of stachyose for thin-layer chromatography in 1 mL of a mixture of water and methanol (1:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-propanol, water and methanol (3:2:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1,3-naphthalenediol TS on the plate, heat the plate at 1059C for 5 minutes: one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the spot from the standard solution. When further heat for more than 5 minutes, a blue spot is not ob-served at just lower than the spot mentioned above, or even appears it is only few. 2) Juku-jio—Sake 0.5 g of the fine cutting of Rehmannia Root with 5 mL of water, add 20 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 2 mg of fructose for thin-layer chromatography in 1 mL of a mixture of water and methanol (1:1), and use this solution as the standard so-lution (1). Separately, dissolve 3 mg of manninotriose for thin-layer chromatography in 1 mL of a mixture of water and methanol (1:1), and use this solution as the standard so-lution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and the standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-propanol, water and methanol (3:2:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1,3-naphthalenediol TS on the plate, heat the plate at 1059C for 10 minutes: the principal spot obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution (1). Further-more, one of the several spots from the sample solution has the same color tone and Rf value with the blue spot from the standard solution (2).
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Rehmannia Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Rehmannia Root according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 2.5z.
Rosin occurs as a light yellow to light brown, glassily transparent, brittle mass, the surfaces of which are often covered with a yellow powder. The fractured surface is shell-like and lustrous. It has a slight odor. It melts easily, and burns with a yellow-brown flame. It is freely soluble in ethanol (95), in acetic acid (100) and in diethyl ether. ‌JP XVIII Crude Drugs and Related Drugs / Ryokeijutsukanto Extract 2119‌ A solution of Rosin in ethanol (95) is acidic. Acid value <1.13> 150 – 177 Total ash <5.01> Not more than 0.1z.
Powdered Rhubarb occurs as a brown powder. It has a characteristic odor and a slightly astringent and bit-ter taste; is gritty between the teeth and colors the saliva yel-low on chewing. JP XVIII Crude Drugs and Related Drugs / Compound Rhubarb and Senna Powder 2115 ‌Under a microscope <5.01>, Powdered Rhubarb reveals starch grains, dark brown substances or druses of calcium oxalate, fragments of parenchyma cells containing them, and reticulate vessels. The starch grains are spherical, sim- side A in each solution. Amount (mg) of sennoside A (C ＝ MS × AT/AS × 1/4 42H 38O 20) ple, or 2- to 4-compound grains. Simple grain, 3 – 18 mm in diameter, rarely 30 mm; crystal druses of calcium oxalate, 30 – 60 mm in diameter, sometimes exceeding 100 mm.
To 1.0 g of Powdered Rhubarb add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, cen-trifuge, and use the diethyl ether layer as the sample solu-tion. Separately, dissolve 1 mg of rhein for thin-layer chro-matography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution, and the spot develops a red color on spraying sodium carbonate TS.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Rhubarb according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Rhubarb according to Method 4, and perform the test (not more than 5 ppm). Rhaponticin—To 0.1 g of Powdered Rhubarb add exactly 10 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of rhaponticin for purity in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl for-mate, 2-butanon, water and formic acid (10:7:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the chromato-gram obtained from the sample solution shows no spot hav-ing the same color tone and Rf value with the blue fluores-cent spot from the standard solution. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 13.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Weigh accurately about 0.5 g of Powdered Rhubarb, add exactly 50 mL of a solution of sodium hydro-gen carbonate (1 in 1000), shake for 30 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Sennoside A RS, (separately deter-mine the water <2.48> by coulometric titration, using 10 mg), dissolve in a solution of sodium hydrogen carbonate (1 in 1000) to make exactly 50 mL. Pipet 5 mL of this solution, add a solution of sodium hydrogen carbonate (1 in 1000) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of senno- MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column about 4 – 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 80) and acetonitrile (4:1). Flow rate: Adjust so that the retention time of sennoside A is about 15 minutes. System suitability— System performance: Dissolve 1 mg each of Sennoside A RS and naringin for thin-layer chromatography in a solution of sodium hydrogen carbonate (1 in 1000) to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, sennoside A and naringin are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z.
Ovoid, oblong-ovoid or cylindrical rhizome, often cut crosswise or longitudinally, 4 – 10 cm in diameter, 5 – 15 cm in length. In the case of Rhubarb without most part of cortex, the outer surface is flat and smooth, yellow- Powdered Rhubarb / Crude Drugs and Related Drugs ‌brown to light brown in color, and sometimes exhibiting white, fine reticulations; thick and hard in texture. In the case of Rhubarb with cork layer, externally dark brown or red-black, and with coarse wrinkles; rough and brittle in tex-ture. The fractured surface of Rhubarb is not fibrous; trans-verse section grayish brown, light grayish brown or brown in color, having patterns of black-brown tissue complicated with white and light brown tissues; near the cambium, the patterns often radiate, and in pith, consist of whirls of tis-sues radiated from the center of a small brown circle 1 – 3 mm in diameter and arranged in a ring or scattered irregu-larly. Odor, characteristic; taste, slightly astringent and bitter; when chewed, gritty between the teeth, and coloring the saliva yellow. Under a microscope <5.01>, the transverse section reveals mostly parenchyma cells; small abnormal cambium-rings scattered here and there in the pith; the cambium-rings produce phloem inside and xylem outside, accompanied with 2 to 4 cell rows of medullary rays containing brown-colored substances, and the rays run radiately from the center of the ring towards the outside forming whirls of tissues; paren-chyma cells contain starch grains, brown-colored substances or crystal druses of calcium oxalate.
To 1.0 g of pulverized Rhubarb add 10 mL of water, shake, then add 10 mL of diethyl ether, shake, cen-trifuge, and use the diethyl ether layer as the sample solu-tion. Separately, dissolve 1 mg of rhein for thin-layer chro-matography in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography<2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and water (20:3:2) to a distance of about 7 cm, and air-dry the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution, and the spot develops a red color on spraying sodium carbonate TS.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Rhubarb according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Rhubarb according to Method 4, and perform the test (not more than 5 ppm). Rhaponticin—To 0.1 g of pulverized Rhubarb add exactly 10 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of rhaponticin for purity in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of ethyl for-mate, 2-butanon, water and formic acid (10:7:1:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the chromato-gram obtained from the sample solution shows no spot hav-ing the same color tone and Rf value with the blue fluores-cent spot from the standard solution. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 13.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Weigh accurately about 0.5 g of pulverized Rhubarb, add exactly 50 mL of a solution of sodium hydro-gen carbonate (1 in 1000), shake for 30 minutes, filter, and use the filtrate as the sample solution. Separately, weigh ac-curately about 10 mg of Sennoside A RS, (separately deter-mine the water <2.48> by coulometric titration, using 10 mg) dissolve in a solution of sodium hydrogen carbonate (1 in 1000) to make exactly 50 mL. Pipet 5 mL of this solution, add a solution of sodium hydrogen carbonate (1 in 1000) to make exactly 20 mL and use this solution as the standard so-lution. Perform the test with exactly 10 mL of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of sennoside A in each solution. Amount (mg) of sennoside A (C42H38O20) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4 – 6 mm in inside diam-eter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 80) and acetonitrile (4:1). Flow rate: Adjust so that the retention time of sennoside A is about 15 minutes. System suitability— System performance: Dissolve 1 mg each of Sennoside A RS and naringin for thin-layer chromatography in a solution of sodium hydrogen carbonate (1 in 1000) to make 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, sennoside A and naringin are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z.
Powdered Rose Fruit occurs as a grayish yel-low-brown powder. It has a slight odor, and has a slightly mucilaginous, astringent, bitter, and slightly acid taste. Under a microscope <5.01>, Powdered Rose Fruit reveals fragments of extremely thick-walled hairs 35 – 70 mm in di-ameter, fragments of epidermis and hypodermis containing brown tannin masses, fragments of thin-walled fundamental tissue containing grayish brown substances, fragments of fine vessels, and solitary or twin crystals or rosette agregates of calcium oxalate (components of fruit receptacle); frag-ments of sclerenchyma, fiber groups, fine vessels, and frag-ments of epidermis containing brown tannin and mucilage (components of pericarp); fragments of endosperm com-posed of polygonal cells containing aleuron grains and fatty oil, fragments of outer epidermis composed of polygonal cells containing tannin, and fragments of inner epidermis composed of elongated cells having wavy lateral walls (com-ponents of seed).
Boil gently 1 g of Powdered Rose Fruit with 20 mL of methanol for 2 minutes, and filter. To 5 mL of the filtrate add 0.1 g of magnesium in ribbon form and 0.5 mL of hydrochloric acid, and allow the mixture of stand: a light red to red color develops. Total ash <5.01> Not more than 6.0z.
The pseudocarp, spherical, ellipsoidal or spheroidal, 5 – 9.5 mm in length, 3.5 – 8 mm in diameter; the external surface red to dark brown in color, smooth and lus-trous; often with peduncle about 10 mm in length at one end, and with pentagonal remains of calyx without sepal at the other end; internal wall of receptacle covered densely with silvery hairs; the interior containing 5 – 10 mature nuts; the nut, irregularly angular ovoid, about 4 mm in length, about 2 mm in diameter; external surface, light yellow-brown; obtuse at one end, and slightly acute at the other. Odor, slight; taste of fruit receptacle, sweet and acid, and of nut, mucilaginous at first, later astringent, bitter and ir-ritative.
Boil gently 1 g of pulverized Rose Fruit with 20 mL of methanol for 2 minutes, and filter. To 5 mL of the filtrate add 0.1 g of magnesium in ribbon form and 0.5 mL of hydrochloric acid, and allow the mixture to stand: a light red to red color develops.
Foreign matter <5.01>—The amount of the peduncle and other foreign matter contained in Rose Fruit is not more than 1.0z. Total ash <5.01> Not more than 6.0z.
the tubulous flower of Carthamus tinctorius Linnáe (Compositae) without any treatment or
larger than that of the standard solution
Nearly cylindrical root, 5 – 25 cm in length, 0.3 – 1.5 cm in diameter; slightly curved, often with lateral roots; outer surface red-brown, dark red-brown or black-brown; with irregular rough wrinkles; hard in texture, and easily broken; fracture surface fine or rough with clefts; cor-tex grayish yellow-white or red-brown, xylem light yellow-white or black-brown. Odor, slight; taste, sweet at first and followed by slight bitterness and astringency. Under a microscope <5.01>, a transverse section reveals usually cork layer in the outermost part, or rarely paren-chyma or endodermis at the outside of the cork layer; several sclerenchyma cells observed or not in secondary cortex; cam-bium obvious; vessels radially arranged in secondary xylem, sometimes radial lines of vessels unite in the center of root; xylem fibers surrounding vessels; primary xylem divided into 2 – 3; vessels of secondary xylem mainly pitted vessels and reticulate vessels in a longitudinal section.
To 1.0 g of pulverized Salvia Miltiorrhiza Root add 10 mL of diethyl ether, allow to stand for 10 minutes with occasional shaking, and filter. Evaporate the filtrate on a water bath to dryness, dissolve the residue in 1 mL of ethyl acetate, and use this solution as the sample solu-tion. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chroma-tography. Develop the plate with a mixture of hexane and ethyl acetate (3:1) to a distance of about 10 cm, and air-dry the plate: a red-brown spot at an Rf value of about 0.4 is observed.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Salvia Miltiorrhiza Root according to Method 3, and perform the test. Prepare the control solution with 3.0 Amount (mg) of glycyrrhizic acid (C ＝ MS × AT/AS × 1/2 42H 62O 16) mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Salvia Miltiorrhiza Root according to Method MS: Amount (mg) of Glycyrrhizic Acid RS taken, calcu-lated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in i). System suitability— System repeatability: Proceed as directed in the system suitability in i). System performance: Dissolve 5 mg of monoammonium glycyrrhizinate for resolution check in 20 mL of dilute ethanol. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, the resolution be-tween the peak having the relative retention time of about 0.9 to glycyrrhizic acid and the peak of glycyrrhizic acid is not less than 1.5.
Long and narrow, conical rhizome and root, 15 – 20 cm in length, 0.7 – 1.5 cm in diameter; externally light brown; rhizome reveals dense crosswise wrinkles like ring nodes, and sometimes reveals brown and hair-like remains of leaf sheath; the root reveals many longitudinal wrinkles and scars of rootlets; in a transverse section, cortex is grayish brown in color and reveals many lacunae, and xylem is yellow in color. Odor, slight; taste, slightly sweet.
To 1.0 g of pulverized Saposhnikovia Root and Rhizome, add 5 mL of methanol, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of 4?-O-glucosyl-5-O-methylvisamminol for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 4 mL of the sample solution and 1 mL of the standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of ethyl formate, formic acid, 2-butanone, and water (20:5:5:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 2.0 g of pulverized Saposhnikovia Root and Rhizome according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Saposhnikovia Root and Rhizome according to Method 4, and perform the test (not more than 5 ppm). Foreign matter <5.01>—The amount of stems and other foreign matter is not more than 2.0z. Peucedanum ledebourielloides—Place 1.0 g of pul-verized Saposhnikovia Root and Rhizome in a glass-stop-pered centrifuge tube, add 5 mL of hexane, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, place 1.0 g of peucedanum・ ledebourielloides for purity in a glass-stoppered centrifuge tube, add 5 mL of hexane, shake for 10 minutes, centrifuge, and use the supernatant liquid as the standard solution. Per-form the test with these solutions as directed under Thin-lay-er Chromatography <2.03>. Spot 5 mL each of the sample so-lution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (20:10:1) to a dis-tance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the sample solu-tion shows no spot corresponding to the blue ‰uorescent spot at an Rf value of about 0.4 obtained from the standard solution. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 20.0z.
Chips, slices or short pieces of wood; yellow-red to grayish yellow-brown, sometimes with light brown to grayish white splint woods; hard in texture; a transverse sec-tion shows a pattern like annual ring. Almost odorless; almost tasteless. Under a microscope <5.01>, a transverse section reveals ray composed of 1 – 2 cell rows of slender and long cells; the area between rays filled with fiber cells, and large and oblong vessels scattered there; solitary crystals of calcium oxalate in parenchymatous cells of the innermost of xylem.
To 1 g of pulverized Sappan Wood add 10 mL of methanol, shake for 5 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, water, formic acid and 2-propanol (20:1:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly sodium carbonate TS on the plate, and air-dry the plate: a red-purple spot appears at an Rf value of about 0.7.
Put a small piece of Sappan Wood in calcium hy-droxide TS: no purple-blue color develops. Loss on drying <5.01> Not more than 11.5z (6 hours). Total ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 7.0z.
Nearly cylindrical roots, 5 – 20 cm in length, 1 – 6 cm in diameter; some of them slightly bent, and some-times longitudinally cut; scar of stem dented on the top of the root with crown; externally yellow-brown to grayish brown, with coarse longitudinal wrinkles and fine reticulate furrows, and also with remains of lateral roots; sometimes root from which periderm has been removed; hard and dense ‌JP XVIII Crude Drugs and Related Drugs / Scopolia Rhizome 2133 ‌in texture, and difficult to break. A transverse section is yellow-brown to dark brown, and cambium part has a dark color. Under a magnifying glass, a transverse section reveal obvious medullary rays, large clefts here and there, and brown oil sacs scattered; in old root, pith existing in the cen-ter, and often forming a hollow.‌ Odor, characteristic; taste, bitter.
To 1.0 g of pulverized Saussurea Root add 10 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane and acetone (7:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and cool: a red-purple spot at an Rf value of about 0.5 and a grayish blue to grayish brown spot just below it are observed.
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Saussurea Root according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Saussurea Root according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Add iodine TS dropwise to a trans-verse section: no blue-purple color develops. Total ash <5.01> Not more than 4.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 17.0z.
Sap fruit of irregular sphere or spheroid, about 6 mm in diameter; externally dark red to black-brown in color, with wrinkles, and occasionally with white powder; seeds, kidney-shaped, externally yellow-brown to dark red-brown, lustrous, with distinct raphe on the dorsal side; exter-nal seed coat easily peeled but internal seed coat adhering closely to the albumen. Odor, slight; taste, acid, later astringent and bitter.
To 1.0 g of pulverized Schisandra Fruit add 10 mL of methanol, warm on a water bath for 3 minutes with shaking, cool, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of schisandrin for thin-layer chromatography in 1 mL of methanol, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (10:10:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the sever-al spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard so-lution.
Foreign matter <5.01>—The amount of fruit recep-tacle, peduncle and other foreign matter contained in Schisandra Fruit is not more than 1.0z. Total ash <5.01> Not more than 5.0z.
a blue fluorescent spot with an Rf value of about 0
Chie‰y irregularly branched, slightly curved rhizome, about 15 cm in length, about 3 cm in diameter, oc-casionally longitudinally cut; externally grayish brown, with wrinkles; rhizome has constrictions and nodes; rarely, remains of stem at the apex; stem scars at upper side of each node; roots or root scars on both sides and lower surface of rhizome; fractured surface granular, grayish white to light brown in color; cortex a little lighter in color. Odor characteristic. Under a microscope <5.01>, xylem reveals groups of ves-sels arranged stepwise between medullary rays; xylem sieve tubes accompanied to the groups of vessels; parenchyma cells contain starch grains, and sometimes sand crystals of calcium oxalate.
(1) To 1 g of pulverized Scopolia Rhizome add 10 mL of diethyl ether and 0.5 mL of ammonia TS, shake for 30 minutes, and filter. Wash the residue with 10 mL of diethyl ether, transfer the filtrate and the washing to a separator, add 20 mL of diluted sulfuric acid (1 in 50), shake well, and drain off the acid extract into another separator. Render the solution slightly alkaline with ammonia TS, add 10 mL of diethyl ether, shake well, transfer the diethyl ether layer to a porcelain dish, and evaporate the solvent on a water bath. To the residue add 5 drops of fuming nitric acid, and evaporate the mixture on a water bath to dryness. Cool, dissolve the residue in 1 mL of N,N-dimethylformamide, and add 5 to 6 drops of tetraethylammonium hydroxide TS: a red-purple to purple color develops. (2) Place 2.0 g of pulverized Scopolia Rhizome in a glass-stoppered centrifuge tube, add 30 mL of ammonia TS, sonicate for 5 minutes, and centrifuge. Transfer the superna-tant liquid to a separator, add 40 mL of ethyl acetate, and shake. Drain off the ethyl acetate layer, add 3 g of anhy-
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Scopolia Rhizome according to Method 3, and perform the test. Prepare the control solution with 4.5 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Scopolia Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. ＝ MSA × QTA/QSA × 1/5 × 0.855 Amount (mg) of scopolamine (C17H21NO4) ＝ MSS × QTS/QSS × 1/25 × 0.789 MSA: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis MSS: amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption spectrometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadesilcylanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen-phosphate in 900 mL of water, add 10 mL of triethylamine, adjust with phosphoric acid to pH 3.5, and add water to make 1000 mL. To 9 parts of this solution add 1 part of acetonitrile. Flow rate: Adjust so that the retention time of scopola-mine is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, scopolamine, atropine and the internal standard are ‌JP XVIII Crude Drugs and Related Drugs / Scopolia Extract Powder 2135‌ eluted in this order with the resolution between the peaks of scopolamine and atropine being not less than 11, and with the resolution between the peaks of atropine and the internal standard being not less than 4.
Weigh accurately about 0.7 g of pulverized Scopolia Rhizome, previously dried at 609C for 8 hours, in a glass-stoppered centrifuge tube, and moisten with 15 mL of am-monia TS. To this add 25 mL of diethyl ether, stopper the centrifuge tube tightly, shake for 15 minutes, centrifuge, and separate the diethyl ether layer. To the residue add 25 mL of diethyl ether, proceed in the same manner, and repeat this procedure twice. Combine all the extracts, and evaporate the solvent on a water bath. Dissolve the residue in 5 mL of the mobile phase, add exactly 3 mL of the internal standard so-lution, and add the mobile phase to make 25 mL. Filter this solution through a filter of a porosity of not more than 0.8 mm, discard the first 2 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately about 25 mg of Atropine Sulfate RS (separately deter-mine the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solution as standard stock solution A. Weigh accurately about 25 mg of Scopolamine Hydrobromide RS (separately determine the loss on drying <2.41> under the same conditions as Scopolamine Hydrobromide Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solution as standard stock solution B. Pipet 5 mL of standard stock solution A and 1 mL of standard stock solution B, add exactly 3 mL of the internal standard solution, then add 25 mL of the mobile phase, and use this solution as the standard solution. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QTA and QSA, of the peak area of hyoscyamine (atro-pine), and the ratios, QTS and QSS, of the peak area of scopolamine to that of the internal standard in each solu-tion, calculate the amounts of hyoscyamine and scopolamine by the following equation, and designate the total as the amount of total alkaloids. drous sodium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent Amount (mg) of hyoscyamine (C 17H 23NO3) of the filtrate under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. Separately, dissolve 2 mg of Atropine Sul-fate RS or atropine sulfate hydrate for thin-layer chromatog-raphy and 1 mg of Scopolamine Hydrobromide RS or scopolamine hydrobromide hydrate for thin-layer chroma-tography in 1 mL each of ethanol (95), and use these solu-tions as standard solution (1) and standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution, standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, water and ammonia water (28) (90:7:3) to a distance of about 10 cm, and dry the plate at 809C for 10 minutes. After cooling, spray evenly Dragen-dorff's TS for spraying on the plate: two principal spots ob-tained from the sample solution and each spot from the standard solutions show the same color tone and the same Rf value. Purity (1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Scopolia Rhizome according to Method 3, and perform the test. Prepare the control solution with 4.5 mL of Standard Lead Solution (not more than 15 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Scopolia Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 7.0z. ＝ MSA × QTA/QSA × 1/5 × 0.855 Amount (mg) of scopolamine (C17H21NO4) ＝ MSS × QTS/QSS × 1/25 × 0.789 MSA: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis MSS: amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption spectrometer (wave-length: 210 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadesilcylanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen-phosphate in 900 mL of water, add 10 mL of triethylamine, adjust with phosphoric acid to pH 3.5, and add water to make 1000 mL. To 9 parts of this solution add 1 part of acetonitrile. Flow rate: Adjust so that the retention time of scopola-mine is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, scopolamine, atropine and the internal standard are ‌JP XVIII Crude Drugs and Related Drugs / Scopolia Extract Powder 2135‌ eluted in this order with the resolution between the peaks of scopolamine and atropine being not less than 11, and with the resolution between the peaks of atropine and the internal standard being not less than 4.
Powdered Scutellaria Root occurs as a yellow-brown powder. It is almost odorless, and has a slight, bitter taste. Under a microscope <5.01>, Powdered Scutellaria Root reveals fragments of parenchyma cells containing small amount of simple and compound starch grains, fragments of short reticulate vessel elements and fusiform, stick-like and ellipsoidal to spherical sclerenchyma cells; also a few frag-ments of spiral vessels and xylem fibers are observed.
(1) Heat gently 0.5 g of Powdered Scutel-laria Root with 20 mL of diethyl ether under a reflux con-denser for 5 minutes, cool, and filter. Evaporate the filtrate, dissolve the residue in 10 mL of ethanol (95), and to 3 mL of the solution add 1 to 2 drops of dilute iron (III) chloride TS: a grayish green color develops, and it changes to purple-brown later. (2) To 1 g of Powdered Scutellaria Root add 25 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Baicalin RS or baicalin for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Scutellaria Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Scutellaria Root according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Scutellaria Root does not show crystals of calcium oxalate. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.0z.
Weigh accurately about 0.5 g of Powdered Scutellar-ia Root, add 30 mL of diluted methanol (7 in 10), heat under a reflux condenser for 30 minutes, and cool. Transfer the mixture to a glass-stoppered centrifuge tube, centrifuge, and separate the supernatant liquid. Wash the vessel for the reflux extraction with 30 mL of diluted methanol (7 in 10), transfer the washings to the glass-stoppered centrifuge tube, centrifuge after shaking for 5 minutes, and separate the su-pernatant liquid. To the residue add 30 mL of diluted metha-nol (7 in 10), shake for 5 minutes, centrifuge, and separate the supernatant liquid. Combine all the extracts, add diluted methanol (7 in 10) to make exactly 100 mL, then pipet 2 mL of this solution, add diluted methanol (7 in 10) to make ex-actly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titra-tion, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of the solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 5 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted phosphoric acid (1 in 146) and acetonitrile (18:7). Flow rate: Adjust so that the retention time of baicalin is about 6 minutes. System suitability— System performance: Dissolve 1 mg of Baicalin RS and 2 mg of methyl parahydroxybenzoate for resolution check in methanol to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, baicalin and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌2140‌‌ Senega / Crude Drugs and Related Drugs ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z.
Cone-shaped, cylindrical, semitubular or flat-tened root, 5 – 20 cm in length, 0.5 – 3 cm in diameter; exter-nally yellow-brown, with coarse and marked longitudinal wrinkles, and with scattered scars of lateral root and remains of brown periderm; scars of stem or remains of stem at the crown; sometimes central portion of xylem rotted, often forming a hollow; hard in texture and easily broken; frac-tured surface fibrous and yellow in color. Almost odorless; taste, slightly bitter. Under a microscope <5.01>, a transverse section reveals a remaining cork layer 6 – 20 cells thick, coritical layer com-posed of parenchyma, sclerenchyma cells scattered in cortex; xylem composed of parenchyma, vessels and small amount of xylem fibers observed in xylem; vessels usually in groups and arranged in tangential direction, radial direction or in ir-regular form; in case where central portion of xylem rotted, cork layer observed around hollow; parenchyma cells of co-ritical layer and xylem contain simple and compound starch grains.
(1) Heat gently 0.5 g of pulverized Scutel-laria Root with 20 mL of diethyl ether under a reflux con-denser for 5 minutes, cool, and filter. Evaporate the solvent of the filtrate, dissolve the residue in 10 mL of ethanol (95), and to 3 mL of the solution add 1 to 2 drops of dilute iron (III) chloride TS: a grayish green color develops, and it changes to purple-brown. (2) To 1 g of pulverized Scutellaria Root add 25 mL of methanol, shake for 15 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Baicalin RS or baicalin for thin-layer chromatography in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly iron (III) chloride-methanol TS on the plate: one of the several spots obtained from the sample solution has the same color tone and Rf value with the spot from the standard solution.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Scutellaria Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Scutellaria Root according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 6.0z.
Weigh accurately about 0.5 g of pulverized Scutel-laria Root, add 30 mL of diluted methanol (7 in 10), heat under a reflux condenser for 30 minutes, and cool. Transfer the mixture to a glass-stoppered centrifuge tube, centrifuge, and separate the supernatant liquid. Wash the vessel for the reflux extraction with 30 mL of diluted methanol (7 in 10), transfer the washings to the glass-stoppered centrifuge tube, centrifuge after shaking for 5 minutes, and separate the su-pernatant liquid. To the residue add 30 mL of diluted metha-nol (7 in 10), shake for 5 minutes, centrifuge, and separate the supernatant liquid. Combine all the extracts, add diluted methanol (7 in 10) to make exactly 100 mL, then pipet 2 mL of this solution, add diluted methanol (7 in 10) to make ex-actly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Baicalin RS (separately determine the water <2.48> by coulometric titra-tion, using 10 mg), and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of the solution, add diluted methanol (7 in 10) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of baicalin in each solution. Amount (mg) of baicalin (C21H18O11) ＝ MS × AT/AS × 5 MS: Amount (mg) of Baicalin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 277 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ‌JP XVIII Crude Drugs and Related Drugs / Powdered Scutellaria Root 2139 gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted phosphoric acid (1 in 146) and acetonitrile (18:7). Flow rate: Adjust so that the retention time of baicalin is about 6 minutes. System suitability— System performance: Dissolve 1 mg of Baicalin RS and 2 mg of methyl parahydroxybenzoate for resolution check in methanol to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, baicalin and methyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of baicalin is not more than 1.5z.
Powdered Senega occurs as a light brown pow-der, and has a characteristic odor resembling the aroma of methyl salicylate; taste, sweet at first, but later acrid. Under a microscope <5.01>, Powdered Senega reveals frag- ments of pitted vessels, reticulate vessels and tracheids; frag-ments of xylem fibers with oblique pits; fragments of xylem parenchyma cells with simple pits; fragments of phloem parenchyma containing oily droplets; fragments of exoder-mis often composed of cells suberized and divided into daughter cells; oily droplets stained red by sudan III TS. The parenchyma cells of Powdered Senega do not contain starch grains and crystals of calcium oxalate.
(1) Shake vigorously 0.5 g of Powdered Senega with 10 mL of water: a lasting fine foam is produced. (2) Shake 0.5 g of Powdered Senega with 30 mL of water for 15 minutes, and filter. Take 1 mL of the filtrate, mix with 50 mL of water, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum at about 317 nm.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Senega according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Senega according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, stone cells, starch grains or crystals of calcium oxalate are not ob-served. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Slender, conical root often branched, 3 – 10 cm in length; main root 0.5 – 1.5 cm in diameter; externally light grayish brown to grayish brown; with many longitudi-nal wrinkles and sometimes with twisted protruding lines; tuberously enlarged crown, with remains of stems and red buds; branched rootlets twisted; a transverse section reveals grayish brown cortex and yellowish white xylem; usually round, and sometimes cuneate to semicircular; cortex on the opposite side is thickened. Odor, characteristic, resembling the aroma of methyl salicylate; taste, sweet at first but leaving an acrid taste. Under a microscope <5.01>, a transverse section of the main root reveals a cork layer consisting of several cell rows of light brown cork cells; secondary cortex composed of parenchyma cells and phloen, traversed by medullary rays, 1 to 3 cells wide; medullary rays on xylem not distinct. Its parenchyma cells contain oil droplets, but starch grains and calcium oxalate crystals are absent.
(1) Shake vigorously 0.5 g of pulverized Senega with 10 mL of water: a lasting fine foam is produced. (2) Shake 0.5 g of pulverized Senega with 30 mL of water for 15 minutes, and filter. Take 1 mL of the filtrate, mix with 50 mL of water, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum at about 317 nm.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Senega according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Senega according to Method 4, and perform the test (not more than 5 ppm). Stem—When perform the test of foreign matter <5.01>, the amount of the stems contained in Senega does not exceed 2.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than the stems is not more than 1.0z. Loss on drying <5.01> Not more than 13.0z (6 hours). Total ash <5.01> Not more than 5.0z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 30.0z.
Powdered Senna Leaf occurs as a light yellow to light grayish yellow-green powder. It has a slight odor and a bitter taste. Under a microscope <5.01>, Powdered Senna Leaf reveals fragments of vessels and vein tissue accompanied with crys-tal cell rows; fragments of thick-walled, bent, unicellular hairs; fragments of palisade and spongy tissue; clustered and solitary crystals of calcium oxalate, 10 to 20 mm in diameter.
(1) Macerate 0.5 g of Powdered Senna Leaf in 10 mL of diethyl ether for 2 minutes, and filter. Add 5 mL of ammonia TS to the filtrate: a yellow-red color is produced in the aqueous layer. To the residue of maceration add 10 mL of water, and macerate for 2 minutes. Filter, and use this solution as standard stock solution (2). Pipet 5 mL of the standard stock solution (1) and 10 mL of the standard stock solution (2), add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATa and ASa, of sennoside A, and the peak areas, ATb and ASb, of sennoside B in each solution, calculate the amounts of sennoside A and sennoside B by the following equations, and designate the total as the amount of total sen-noside. Amount (mg) of sennoside A (C42H38O20) ＝ MSa × ATa/ASa × 1/4 Amount (mg) of sennoside B (C H O ) add 5 mL of ammonia TS: a yellow-red color is produced in the aqueous layer. ＝ MSb × ATb/ASb × 1/2 42 38 20 (2) To 2 g of Powdered Senna Leaf add 40 mL of a mixture of tetrahydrofuran and water (7:3), shake for 30 minutes, and centrifuge. Transfer the supernatant liquid to a separator, add 13 g of sodium chloride, and shake for 30 minutes. Separate the aqueous layer with undissolved so-dium chloride, and adjust to pH 1.5 with 1 mol/L hydro-chloric acid TS. Transfer this solution to another separator, shake with 30 mL of tetrahydrofuran for 10 minutes, sepa-rate the tetrahydrofuran layer, and use the tetrahydrofuran layer as the sample solution. Separately, dissolve 1 mg of Sennoside A RS or sennoside A for thin-layer chromatogra-phy in 1 mL of a mixture of tetrahydrofuran and water (7:3), and use this solution as the standard solution. Perform the MSa: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis MSb: Amount (mg) of Sennoside B RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. ‌JP XVIII Crude Drugs and Related Drugs / Shakuyakukanzoto Extract 2143‌‌ Mobile phase: Dissolve 2.45 g of tetra-n-heptylammonium bromide in 1000 mL of a mixture of diluted 1 mol/L acetic acid-sodium acetate buffer solution (pH 5.0) (1 in 10) and acetonitrile (17:8). Flow rate: Adjust so that the retention time of sennoside A is about 26 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sennoside B and sennoside A are eluted in this order with the resolution between these peaks being not less than 15, and the number of theoretical plates of the peak of sen-noside A being not less than 8000. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z.
(1) Foreign matter <5.01>—Under a microscope, stone cells and thick fibers are not observable. (2) Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of Powdered Senna Leaf in a glass-stoppered centrifuge tube, add 25 mL of diluted methanol (7 in 10), shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 10 mL of diluted methanol (7 in 10), shake for 10 minutes, cen-trifuge, and separate the supernatant liquid. Repeat this procedure once more, combine all the extracts, add diluted methanol (7 in 10) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in a solution of sodium hydrogen carbonate (1 in 100) to make exactly 20 mL, and use this solution as standard stock solution (1). Weigh accurately about 10 mg of Sennoside B RS (separately determine the water <2.48> by coulometric [sennoside A (C42 H38 O20 : 862.74) and sennoside B titration, using 10 mg), dissolve in a solution of sodium hydrogen carbonate (1 in 100) to make exactly 20 mL, and (C42H38O20: 862.74)], calculated on the basis of dried material. Description Powdered Senna Leaf occurs as a light yellow to light grayish yellow-green powder. It has a slight odor and a bitter taste. Under a microscope <5.01>, Powdered Senna Leaf reveals fragments of vessels and vein tissue accompanied with crys-tal cell rows; fragments of thick-walled, bent, unicellular hairs; fragments of palisade and spongy tissue; clustered and solitary crystals of calcium oxalate, 10 to 20 mm in diameter. Identification (1) Macerate 0.5 g of Powdered Senna Leaf in 10 mL of diethyl ether for 2 minutes, and filter. Add 5 mL of ammonia TS to the filtrate: a yellow-red color is produced in the aqueous layer. To the residue of maceration add 10 mL of water, and macerate for 2 minutes. Filter, and use this solution as standard stock solution (2). Pipet 5 mL of the standard stock solution (1) and 10 mL of the standard stock solution (2), add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATa and ASa, of sennoside A, and the peak areas, ATb and ASb, of sennoside B in each solution, calculate the amounts of sennoside A and sennoside B by the following equations, and designate the total as the amount of total sen-noside. Amount (mg) of sennoside A (C42H38O20) ＝ MSa × ATa/ASa × 1/4 Amount (mg) of sennoside B (C H O ) add 5 mL of ammonia TS: a yellow-red color is produced in the aqueous layer. ＝ MSb × ATb/ASb × 1/2 42 38 20 (2) To 2 g of Powdered Senna Leaf add 40 mL of a mixture of tetrahydrofuran and water (7:3), shake for 30 minutes, and centrifuge. Transfer the supernatant liquid to a separator, add 13 g of sodium chloride, and shake for 30 minutes. Separate the aqueous layer with undissolved so-dium chloride, and adjust to pH 1.5 with 1 mol/L hydro-chloric acid TS. Transfer this solution to another separator, shake with 30 mL of tetrahydrofuran for 10 minutes, sepa-rate the tetrahydrofuran layer, and use the tetrahydrofuran layer as the sample solution. Separately, dissolve 1 mg of Sennoside A RS or sennoside A for thin-layer chromatogra-phy in 1 mL of a mixture of tetrahydrofuran and water (7:3), and use this solution as the standard solution. Perform the MSa: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis MSb: Amount (mg) of Sennoside B RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. ‌JP XVIII Crude Drugs and Related Drugs / Shakuyakukanzoto Extract 2143‌‌ Mobile phase: Dissolve 2.45 g of tetra-n-heptylammonium bromide in 1000 mL of a mixture of diluted 1 mol/L acetic acid-sodium acetate buffer solution (pH 5.0) (1 in 10) and acetonitrile (17:8). Flow rate: Adjust so that the retention time of sennoside A is about 26 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sennoside B and sennoside A are eluted in this order with the resolution between these peaks being not less than 15, and the number of theoretical plates of the peak of sen-noside A being not less than 8000. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z.
Lanceolate to narrow lanceolate leaflets, 1.5 – 5 cm in length, 0.5 – 1.5 cm in width, light grayish yellow to light grayish yellow-green in color; margin entire, apex acute, base asymmetric, petiole short; under a magnifying glass, vein marked, primary lateral veins running toward the apex along the margin and joining the lateral vein above; lower surface having slight hairs. Odor slight; taste, bitter. Under a microscope <5.01>, a transverse section of Senna Leaf reveals epidermis with thick cuticle, with numerous stomata, and with thick-walled, warty unicellular hairs; epidermal cells are often separated into two loculi by a septum which is in parallel with the surface of the leaf, and contain mucilage in the inner loculus; palisade of a single cel-lular layer under each epidermis; spongy tissue, consisting of 3 to 4 cellular layers, and containing clustered or solitary crystals of calcium oxalate; cells adjacent to vascular bundle, forming crystal cell rows.
(1) Macerate 0.5 g of pulverized Senna Leaf in 10 mL of diethyl ether for 2 minutes, and filter. Add 5 mL of ammonia TS to the filtrate: a yellow-red color is produced in the aqueous layer. To the residue of maceration add 10 mL of water, and macerate for 2 minutes. Filter, and add 5 mL of ammonia TS: a yellow-red color is produced in the aqueous layer. (2) To 2 g of pulverized Senna Leaf add 40 mL of a mixture of tetrahydrofuran and water (7:3), shake for 30 and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol, ethyl acetate, water and acetic acid (100) (40:40:30:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots obtained from the sample solu-tion has the same color tone and Rf value with the red fluo-rescent spot from the standard solution.
(1) Rachis and fruit—When perform the test of foreign matter <5.01>, the amount of rachis and fruits con-tained in Senna Leaf does not exceed 5.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than rachis and fruits contained in Senna Leaf does not exceed 1.0z. Total BHC's and total DDT's <5.01>—Not more than 0.2 ppm, respectively. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 12.0z. Acid-insoluble ash <5.01> Not more than 2.0z.
Weigh accurately about 0.5 g of pulverized Senna Leaf in a glass-stoppered centrifuge tube, add 25 mL of diluted methanol (7 in 10), shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 10 mL of diluted methanol (7 in 10), shake for 10 minutes, centrifuge, and separate the supernatant liquid. Repeat this procedure once more, combine all the extracts, add diluted methanol (7 in 10) to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Sennoside A RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in a solution of sodium hydrogen carbonate (1 in 100) to make exactly 20 mL, and use this solution as standard stock solution (1). Weigh accurately about 10 mg of Sennoside B RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in a solution of sodium hydrogen carbonate (1 in 100) to make exactly 20 mL, and use this solution as standard stock solution (2). Pipet 5 mL of the standard stock solution (1) and 10 mL of the standard stock solution (2), add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATa and ASa, of sennoside A, and the peak areas, ATb and ASb, of sennoside B in each solution, calculate the amounts of sennoside A and sennoside B by the following equations, and designate the total as the amount of total sen-nosides. Amount (mg) of sennoside A (C42H38O20) ＝ MSa × ATa/ASa × 1/4 Amount (mg) of sennoside B (C H O ) minutes, and centrifuge. Transfer the supernatant liquid to a separator, add 13 g of sodium chloride, and shake for 30 ＝ MSb × ATb/ASb × 1/2 42 38 20 minutes. Separate the aqueous layer with undissolved so-dium chloride, and adjust to pH 1.5 with 1 mol/L hydro-chloric acid TS. Transfer this solution to another separator, shake with 30 mL of tetrahydrofuran for 10 minutes, sepa-rate the tetrahydrofuran layer, and use the tetrahydrofuran layer as the sample solution. Separately, dissolve 1 mg of Sennoside A RS or sennoside A for thin-layer chromatogra-phy in 1 mL of a mixture of tetrahydrofuran and water (7:3), MSa: Amount (mg) of Sennoside A RS taken, calculated on the anhydrous basis MSb: Amount (mg) of Sennoside B RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). ‌2142 Powdered Senna Leaf / Crude Drugs and Related Drugs Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 2.45 g of tetra-n-heptylammonium bromide in 1000 mL of a mixture of diluted 1 mol/L acetic acid-sodium acetate buffer solution (pH 5.0) (1 in 10) and acetonitrile (17:8). Flow rate: Adjust so that the retention time of sennoside A is about 26 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sennoside B and sennoside A are eluted in this order with the resolution between these peaks being not less than 15, and the number of theoretical plates of the peak of sen-noside A being not less than 8000. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sennoside A is not more than 1.5z.
Ovate to spatulate seed, 3 – 4 mm in length, about 2 mm in width, about 1 mm in thickness; externally dark brown to black, rarely light brown to brown. Under a magnifying glass, thin ridges are observed on edges. 100 seeds weigh about 0.2 – 0.3 g. Odorless; taste, slightly sweet and oily. Under a microscope <5.01>, transverse section reveals a seed coat consisting of palisade epidermis and flattened Sesame Oil Oleum Sesami
Round or elliptic sections, 0.2 – 0.4 cm in thickness, 1 – 4.5 cm in diameter; cortex on both fractured surfaces, light brown to dark brown; in xylem, grayish brown vessel portions and dark brown medullary rays lined alternately and radially; flank, dark gray, with longitudinal wrinkles and warty protrusions. Almost odorless; taste, bitter. Under a microscope <5.01>, a transverse section reveals extremely thick-walled stone cells in primary cortex and pericycle; irregular-sized vessels lined nearly stepwise in the vessel portion; cells of medullary ray mostly not lignified, and extremely thick-walled and large stone cells scattered here and there; primary cortex containing needle crystals of calcium oxalate; medullary rays containing starch gains, mainly simple grain, 3 – 20 mm in diameter, and small needle crystals of calcium oxalate. Identiˆcation To 1.0 g of pulverized Sinomenium Stem and Rhizome add 5 mL of methanol, shake for 10 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, dis-solve 1 mg of sinomenine for thin-layer chromatography in 1 mL of methanol, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl formate, 1-propanol, water and acet-ic acid (100) (3:3:2:2) to a distance of about 7 cm, and air-d-ry the plate. Spray evenly DragendorŠ's TS for spraying on the plate, air-dry the plate, and spray evenly sodium nitrite TS on the plate: one of the several spots obtained from the sample solution and the spot from the standard solution show the same color tone and the same Rf value. Further, a spot with the same color tone appears immediately below the spot. Total ash <5.01> Not more than 7.0z. Acid-insoluble ash <5.01> Not more than 0.5z.
Powdered Smilax Rhizome occurs as a light yellow-brown powder, and has a slight odor, and is practi-cally tasteless. Under a microscope <5.01>, Powdered Smilax Rhizome reveals starch grains and fragments of parenchyma cells containing them; fragments of raphides of calcium oxalate contained in mucilage masses; fragments of lignified paren-chyma cells of cortical layer; fragments of cork cells and scalariform vessels; starch grains composed mostly of simple grains, and mixed with a few 2- to 4-compound grains 12 – 36 mm in diameter.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Smilax Rhizome according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Smilax Rhizome according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Smilax Rhizome does not show a large quantity of stone cells or thick-walled fibers. Total ash <5.01> Not more than 5.0z.
Flattened and irregular cylindrical tuber, often with node-like branches; usually 5 – 15 cm in length, 2 – 5 cm in diameter; the outer surface grayish yellow-brown to yellow-brown, and the upper surface scattered with knotty remains of stem; transverse section irregular elliptical to obtuse triangular, whitish to reddish white, consisting of extremely thin cortex and mostly of stele. Odor, slight; almost tasteless. Under a microscope <5.01>, a transverse section reveals a 2- to 3-cell-wide cork layer, with extremely narrow cortical layer, usually consisting of a 2- to 4-cell-wide, thick-walled parenchyma cells, showing large mucilage cells here and there; mucilage cell containing raphides of calcium oxalate; stele consisting chiefly of parenchyma cells, and scattered with vascular bundles; parenchyma cells containing starch grains composed mostly of simple grains, 12 – 36 mm in di-ameter, and sometimes mixed with 2- to 4-compound grains.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Smilax Rhizome according to Method 3, and per- ‌2154‌‌ Powdered Smilax Rhizome / Crude Drugs and Related Drugs form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Smilax Rhizome according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 5.0z.
Powdered Sophora Root occurs as a light brown powder. It has a slight odor, and an extremely bitter and lasting taste. Under a microscope <5.01>, Powdered Sophora Root re- veals mainly starch grains and fragments of parenchyma ‌2156‌‌ Soybean Oil / Crude Drugs and Related Drugs cells containing them, fibers, bordered pitted vessels, reticu-late vessels; a few fragments of corky tissue and solitary crystals of calcium oxalate. Starch grains usually composed of 2- to 4-compound grains 15 – 20 mm in diameter, and sim-ple grains 2 – 5 mm in diameter.
To 1.0 g of Powdered Sophora Root add 10 mL of methanol, shake for 10 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 1 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethanol (99.5), ethyl acetate, and ammonia solu-tion (28) (3:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: two brown spots appear at an Rf value of about 0.5.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Sophora Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Sophora Root according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
Cylindrical root, 5 – 20 cm in length, 2 – 3 cm in diameter; externally dark brown to yellow-brown, with distinct longitudinal wrinkles, and with laterally extended lenticels; root without periderm, externally yellow-white, with somewhat fibrous surface; under a magnifying glass, the transverse section, light yellow-brown; cortex, 0.1 – 0.2 cm in thickness, slightly tinged with dark color near cambi-um, forming a crack between xylem. Odor, slight; taste, extremely bitter and lasting.
To 1.0 g of pulverized Sophora Root add 10 mL of methanol, shake for 10 minutes, filter, and use the fil-trate as the sample solution. Perform the test with the sam-ple solution as directed under Thin-layer Chromatography <2.03>. Spot 1 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethanol (99.5), ethyl acetate and ammonia solu-tion (28) (3:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate, and air-dry the plate. Then spray evenly sodium nitrite TS on the plate: two brown spots appear at an Rf value of about 0.5.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Sophora Root according to Method 3, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Sophora Root according to Method 4, and per-form the test (not more than 5 ppm). Stem—When perform the test of foreign matter <5.01>, the amount of its stems contained in Sophora Root does not exceed 10.0z. Foreign matter <5.01>—The amount of foreign mat-ter other than stems is not more than 1.0z. Total ash <5.01> Not more than 6.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
Disk, often slightly bent, 1 – 3 cm in diameter, 0.3 – 0.5 cm in thickness; externally light grayish yellow-green to light grayish brown, covered densely with lustrous appressed hairs radiating from the center to the circumfer-ence; on both sides, the margin and the central part bulged a little; the dot-like micropyle situated at one point on the margin, and from which usually a raised line runs to the cen-ter on one side; extremely hard in texture; when cracked upon soaking in water, the seed coat thin, the interior con-sisting of two horny, light grayish yellow endosperms, and leaving a central narrow cavity at the center; a white embryo, about 0.7 cm in length, situated at one end between the inner surfaces of the endosperms. Odorless.
(1) To 3 g of pulverized Nux Vomica add 3 mL of ammonia TS and 20 mL of chloroform, macerate for 30 minutes with occasional shaking, and filter. Remove most of the chloroform from the filtrate by warming on a water bath, add 5 mL of diluted sulfuric acid (1 in 10), and warm on a water bath while shaking well until the odor of chlo-roform is no longer perceptible. After cooling, filter through a pledget of absorbent cotton, and add 2 mL of nitric acid to 1 mL of the filtrate: a red color develops. (2) To the remaining filtrate obtained in (1) add 1 mL of potassium dichromate TS, and allow to stand for 1 hour: a yellow-red precipitate is produced. Collect the precipitate by ‌JP XVIII Crude Drugs and Related Drugs / Nux Vomica Extract 2079 filtration, and wash with 1 mL of water. Transfer a part of the precipitate to a small test tube, add 1 mL of water, dis-solve by warming, cool, and add 5 drops of sulfuric acid dropwise carefully along the wall of the test tube: the layer of sulfuric acid shows a purple color which turns immedi-ately red to red-brown. Total ash <5.01> Not more than 3.0z. ‌Assay Weigh accurately about 1 g of pulverized Nux Vomica, previously dried at 609C for 8 hours, place in a glass-stoppered centrifuge tube, and moisten with 1 mL of ammonia solution (28). To this solution add 20 mL of diethyl ether, stopper the centrifuge tube tightly, shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 20 mL of diethyl ether, proceed in the same manner, and repeat this procedure three times. Com-bine all the extracts, and evaporate the solvent on a water bath. Dissolve the residue in 10 mL of the mobile phase, add exactly 10 mL of the internal standard solution, and add the mobile phase to make 100 mL. Filter this solution through a membrane filter with a porosity not more than 0.8 mm, dis-card the first 2 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 75 mg of strychnine nitrate for assay (separately deter-mine the loss on drying), and dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratio, QT and QS, of the peak area of strychnine to that of the internal standard. Nux Vomica Extract
Powdered Swertia Herb occurs as a grayish yellow-green to yellow-brown powder. It has a slight odor, and extremely bitter, persistent taste. Under a microscope <5.01>, Powdered Swertia Herb re- veals xylem tissues with fibers (components of stems and roots); assimilation tissues (components of leaves and calyces); striated epidermis (components of stems and peduncles); tissues of corollas and filaments with spiral ves-sels; cells of anthers and their inner walls; spherical pollen grains with granular patterns (components of flowers), about 30 mm in diameter; starch grains are simple grain, about 6 mm in diameter, and very few. Identiˆcation To 0.5 g of Powdered Swertia Herb add 10 mL of methanol, shake for 5 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, dissolve 1 mg of Swertiamarin RS or swertiamarin for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution and the spot from the standard solution show the same color tone and Rf value.
Foreign matter—Under a microscope <5.01>, crys-tals of calcium oxalate, a large quantity of starch grains and groups of stone cells are not observable. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 2.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 20.0z.
Weigh accurately about 1 g of Powdered Swertia Herb in a glass-stoppered centrifuge tube, add 40 mL of methanol, shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 40 mL of methanol, and proceed in the same manner. Combine the extracts, and add methanol to make exactly 100 mL. Pipet 5 mL of the so-lution, add the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of Swertiamarin RS (separately determine the water <2.48> by coulometric titration, using 10 mg), dis-solve in methanol to make exactly 20 mL. Pipet 5 mL of the solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of swertiamarin in each solution. Amount (mg) of swertiamarin (C16H22O10) ＝ MS × AT/AS × 5 MS: Amount (mg) of Swertiamarin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water and acetonitrile (91:9). Flow rate: Adjust so that the retention time of swertiama- rin is about 12 minutes. System suitability— System performance: Dissolve 1 mg each of Sweriamarin RS and theophylline in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, theophylline and swertiama-rin are eluted in this order with the resolution of these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of swertiamarin is not more than 1.5z.
Herb, 10 – 50 cm in length, having flowers, opposite leaves, stems, and, usually, with short, lignified roots; stems square, about 2 mm in diameter, often with branches; the leaves and stems dark green to dark purple or yellow-brown in color; the flowers white to whitish, and the roots yellow-brown. When smoothed by immersing in water, leaves, linear or narrow lanceolate, 1 – 4 cm in length, 0.1 – 0.5 cm in width, entire, and sessile; corolla split deeply as five lobes; the lobes narrow, elongated ellipse shape, and under a magnifying glass, with two elliptical nectaries jux-taposed at the base of the inner surface; the margin of lobe resembles eyelashes; the five stamens grow on the tube of the corolla and stand alternately in a row with corolla-lobes; peduncle distinct. Odor, slight; taste, extremely bitter and persisting. Identiˆcation To 0.5 g of pulverized Swertia Herb add 10 mL of methanol, shake for 5 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, dissolve 1 mg of Swertiamarin RS or swertiamarin for thin-layer chro-matography in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly vanillin-sulfuric acid-ethanol TS for spraying on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution and the spot from the standard solution show the same color tone and Rf value.
Foreign matter <5.01>—The amount of straw and other foreign matters contained in Swertia Herb is not more than 1.0z. Loss on drying <5.01> Not more than 12.0z (6 hours). Total ash <5.01> Not more than 6.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 20.0z.
Weigh accurately about 1 g of moderately fine pow-der of Swertia Herb in a glass-stoppered centrifuge tube, add 40 mL of methanol, shake for 15 minutes, centrifuge, and separate the supernatant liquid. To the residue add 40 mL of methanol, and proceed in the same manner. Combine the extracts, and add methanol to make exactly 100 mL. Pipet 5 mL of the solution, add the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Swertiamarin RS (sepa-rately determine the water <2.48> by coulometric titration, using 10 mg), dissolve in methanol to make exactly 20 mL. Pipet 5 mL of the solution, add the mobile phase to make ex-actly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of swertiamarin in each solution. Amount (mg) of swertiamarin (C16H22O10) ＝ MS × AT/AS × 5 MS: Amount (mg) of Swertiamarin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water and acetonitrile (91:9). Flow rate : Adjust so that the retention time of swertiama- rin is about 12 minutes. System suitability— System performance: Dissolve 1 mg each of Swertiamarin RS and theophylline in the mobile phase to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, theophylline and swertiama-rin are eluted in this order with the resolution of these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of swertiamarin is not more than 1.5z. ‌2158‌ Powdered Swertia Herb / Crude Drugs and Related Drugs
Pentagonal star shaped fruit, composed of five mericarps, 7 – 12 mm in diameter, often each mericarp sepa-rated; externally grayish green to grayish brown; a pair of longer and shorter spines on surface of each mericarp, the longer spine 3 – 7 mm in length, the shorter one 2 – 5 mm in length, numerous small processes on ridge; pericarp hard in texture, cut surface light yellow; each mericarp contains 1 – 3 seeds. Almost odorless; taste, mild at first, followed by bitter-ness. Under a microscope <5.01>, a transverse section reveals epicarp composed of an epidermis; mesocarp composed of parenchyma and sclerenchyma layer; endocarp composed of several-cellular-layered fiber cells; a single-layer of cell be-tween mesocarp and endocarp contain solitary crystals of calcium oxalate; cotyledons of seed contain oil drops and aleurone grains, and occasionally starch grains.
To 2 g of pulverized Tribulus Fruit add 5 mL of methanol, shake for 10 minutes, filter, and use the filtrate as the sample solution. Perform the test with the sample so-lution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate and water (40:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 5 minutes, and examine under ultraviolet light (main wavelength: 365 nm): a bluish white fluorescent spot appears at an Rf value of about 0.4.
(1) Peduncle—When perform the test of foreign matter <5.01>, the amount of peduncle contained in Tribulus Fruit does not exceed 4.0z. (2) Foreign matters <5.01>—Not more than 1.0z of for-eign matters other than peduncle. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 13.0z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.5z.
Irregular cylindrical root, 5 – 10 cm in length, 3 – 5 cm in diameter, often cut lengthwise; externally light yellow-white, and with irregular pattern of vascular bundles appearing as brownish yellow lines; fractured surface some-what fibrous and light yellow in color; under a magnifying glass, the transverse section reveals wide medullary rays and brownish yellow spots or small holes formed by vessels. Almost odorless; taste, slightly bitter.
To 2.0 g of pulverized Trichosanthes Root add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Perform the test with the sample solution as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (20:10:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid on the plate, heat the plate at 1059C for 10 minutes, and examine under ultraviolet light (main wavelength: 365 nm): a light yellow to light yellow-green fluorescent spot appears at an Rf value of about 0.4.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Trichosanthes Root according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Trichosanthes Root according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 4.0z.
Obovate to spatulate leaves, 1 – 3 cm in length, 0.5 – 1.5 cm in width; upper surface yellow-green to dark green; lower surface light yellow-green; margin entire; apex obtuse or round, sometimes retuse; base cuneate; petiole very short; lamina thick with characteristic reticulate vena- tion, and easily broken. Odor, slight; taste, slightly bitter and astringent. Under a microscope <5.01>, the transverse section reveals thick cuticule; parenchyma cells of palisade tissue and sponge tissue being similar in form; in the vascular bundle, medullary ray consisting of 2 to 7 rows of one-cell line, ap-pearing as bones of Japanese fan; polygonal solitary crystals and clustered crystals of calcium oxalate present sparsely in cells on both outer and inner sides of the vascular bundle, but no crystals in mesophyll.
(1) Macerate 0.5 g of pulverized Bearberry Leaf with 10 mL of boiling water, shake the mixture for a few minutes, allow to cool, and filter. Place 1 drop of the fil-trate on filter paper, and add 1 drop of iron (III) chloride TS: a dark purple color appears. (2) To 0.2 g of pulverized Bearberry Leaf add 10 mL of a mixture of ethanol (95) and water (7:3), shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of arbutin for thin-layer chromatography in 1 mL of a mixture of ethanol (95) and water (7:3), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl formate, water and formic acid (8:1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly dilute sulfuric acid upon the plate, and heat the plate at 1059C for 10 minutes: one of the several spots obtained from the sample solution and the spot from the standard solution show the same color tone and the Rf value.
(1) Twig—When perform the test of foreign mat-ter <5.01>, the amount of twigs contained in Bearberry Leaf does not exceed 4.5z. (2) Foreign matter <5.01>—The amount of foreign mat-ter other than twigs contained in Bearberry Leaf does not exceed 2.0z. Total ash <5.01> Not more than 4.0z. Acid-insoluble ash <5.01> Not more than 1.5z.
Weigh accurately about 0.5 g of pulverized Bear-berry Leaf in a glass-stoppered centrifuge tube, add 40 mL of water, shake for 30 minutes, centrifuge, and separate the supernatant liquid. To the residue add 40 mL of water, and proceed in the same manner. To the combined extracts add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of arbutin for assay, previously dried for 12 hours (in vacu-um, silica gel), dissolve in water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of arbutin in each solution. Amount (mg) of arbutin ＝ MS × AT/AS MS: Amount (mg) of arbutin for assay taken Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 280 nm). Column: A stainless steel column 4 – 6 mm in inside diam-eter and 15 – 25 cm in length, packed with octadecylsilanized silica gel (5 – 10 mm in particle diameter). Column temperature: A constant temperature of about 209C. ‌1956‌‌ Beef Tallow / Crude Drugs and Related Drugs Mobile phase: A mixture of water, methanol and 0.1 mol/L hydrochloric acid TS (94:5:1). Flow rate: Adjust so that the retention time of arbutin is about 6 minutes. System suitability— System performance: Dissolve 50 mg each of arbutin for assay, hydroquinone and gallic acid in water to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, arbutin, hydroqui-none and gallic acid are eluted in this order with the resolu-tions among these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of arbutin is not more than 1.5z.
Powdered Japanese Valerian occurs as a dark grayish brown powder. It is somewhat moist to the touch. It has a strong, characteristic odor and a slightly bitter taste. Under a microscope <5.01>, Powdered Japanese Valerian reveals starch grains and fragments of parenchyma cells con-taining them; fragments of pitted vessels, reticulate vessels, ring vessels, and spiral vessels; fragments of exodermis con-taining oil droplets and composed of cells suberized and divided into daughter cells; fragments of yellow stone cells from the rhizome and the stolon; and very rarely, some frag-ments of epidermis and phloem fibers. Starch grains, simple grains 10 – 20 mm in diameter and 2- to 4-compound grains; oil droplets stained red with sudan III TS.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Japanese Valerian according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Japanese Valerian according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 5.0z. Essential oil content <5.01> Perform the test with 50.0 g of Powdered Japanese Valerian provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.2 mL.
Obovoid, short rhizome with numerous, fine and long roots; externally dark brown to grayish brown. The ‌JP XVIII Crude Drugs and Related Drugs / Japanese Zanthoxylum Peel 2041‌ root, 10 – 15 cm in length, 0.1 – 0.3 cm in diameter; exter-nally, with fine longitudinal wrinkles; brittle. The rhizome, 1 – 2 cm in length, 1 – 2 cm in diameter, with buds and remains of stem at the crown; hard in texture and difficult to break; flank of rhizome sometimes accompanied with stol-ons having thick and short or thin, long and extremely small, scaly leaves. Under a magnifying glass, the transverse section reveals a thick, light grayish brown cortex, and a grayish brown stele. Odor, strong and characteristic; taste, slightly bitter.
(1) Heavy metals <1.07>—Proceed with 3.0 g of pulverized Japanese Valerian according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Japanese Valerian according to Method 4, and perform the test (not more than 5 ppm). Total ash <5.01> Not more than 10.0z. Acid-insoluble ash <5.01> Not more than 5.0z. Essential oil content <5.01> Perform the test with 50.0 g of pulverized Japanese Valerian provided that 1 mL of silicon resin is previously added to the sample in the flask: the volume of essential oil is not less than 0.3 mL.
slightly soluble in diethyl ether, and practically insoluble in water and in ethanol (99
Irregularly compressed and often branched massive rhizome; branched parts slightly curved ovoid or ob-long- ovoid, 2 – 4 cm in length, and 1 – 2 cm in diameter; ex-ternal surface grayish yellow to grayish yellow-brown, with wrinkles and ring node; fractured surface brown to dark brown, transparent and horny; under a magnifying glass, a transverse cut surface reveals cortex and stele distinctly divided; vascular bundles scattered throughout the surface. Odor, characteristic; taste, extremely pungent. Under a microscope <5.01>, a transverse section reveals cork layer, cortex and stele in this order from the outside; cortex and stele, divided by an endodermis, composed of parenchyma; vascular bundles surrounded by fibers scat-tered in cortex and stele; oil cells contain yellow oily sub-stances, scattered in parenchyma; parenchyma cells contain solitary crystals of calcium oxalate, and gelatinized starch.
To 2 g of pulverized Processed Ginger add 5 mL of diethyl ether, shake for 10 minutes, filter, and use the filtrate as the sample solution (1). To the residue add 5 mL of methanol, proceed in the same manner as above, and use so obtained solution as the sample solution (2). Separately, dissolve 1 mg of [6]-shogaol for thin-layer chromatography in 2 mL of methanol, and use this solution as the standard solution (1). Separately, dissolve 1 mg of sucrose in 2 mL of methanol, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution (1) and standard solution (1) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (1:1) to a distance of about 7 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS for spraying on the plate, heat the plate at 1059C for 5 minutes, and allow to cool: one of the several spots obtained from the sample solution (1) has the same color tone and Rf value with the spot from the standard solution (1). Spot 10 mL each of the sample solution (2) and standard solution (2) on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of 1-butanol, water and acetic acid (100) (8:5:3) to a distance of about 7 cm, and air-dry the plate. Spray evenly 1,3-naphthalenediol TS on the plate, and heat the plate at 1059C for 5 minutes: one of the several spots obtained from the sample solution (2) has the same color tone and Rf value with the spot from the standard solution (2).
(1) Heavy metals <1.07>—Proceed with 1.0 g of pulverized Processed Ginger according to Method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Arsenic <1.11>—Prepare the test solution with 0.40 g of pulverized Processed Ginger according to Method 4, and perform the test (not more than 5 ppm). Loss on drying <5.01> Not more than 15.0z (6 hours). ‌2110 Prunella Spike / Crude Drugs and Related Drugs ‌Total ash <5.01> Not more than 6.5z. Acid-insoluble ash <5.01> Not more than 1.5z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.0z.
Weigh accurately about 1 g of pulverized Processed Ginger, place in a centrifuge tube, add 30 mL of the mobile phase, shake for 20 minutes, centrifuge, and separate the supernatant liquid. To the residue add 30 mL of the mobile phase, and repeat the extraction twice more. To the com-bined all extracts add the mobile phase to make exactly 100 mL, use this solution as the sample solution. Separately, weigh accurately about 5 mg of [6]-shogaol for assay, dis-solve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of [6]-shogaol in each solution. Amount (mg) of [6]-shogaol ＝ MS × AT/AS MS: Amount (mg) of [6]-shogaol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and water (3:2). Flow rate: Adjust so that the retention time of [6]-shogaol is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-shogaol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-shogaol is not more than 1.5z.
Powdered Ginger occurs as a light grayish brown to light grayish yellow powder. It has a characteristic odor and an extremely pungent taste. Under a microscope <5.01>, Powdered Ginger reveals mainly starch grains and parenchyma cells containing them; also, parenchyma cells containing yellow-brown to dark brown oily substances or single crystals of calcium oxalate; fragments of fibers with distinct pits; fragments of spiral, ring and reticulate vessels, and rarely fragments of cork tissue; starch grains composed of simple, compound or half-compound grains, ovoid, triangular ovoid, ellipsoidal or spherical, with abaxial hilum, usually 10 – 30 mm in long axis.
To 2 g of Powdered Ginger add 5 mL of diethyl ether, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of [6]-gingerol for thin-layer chromatography in 2 mL of metha-nol, and use this solution as the standard solution. Perform Amount (mg) of [6]-gingerol ＝ MS × AT/AS the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu- MS: Amount (mg) of [6]-gingerol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile and phosphoric acid (3800:2200:1). Flow rate: Adjust so that the retention time of [6]-gingerol is about 19 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-gingerol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-gingerol is not more than 1.5z.
(1) Heavy metals <1.07>—Proceed with 3.0 g of Powdered Ginger according to Method 3, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Powdered Ginger according to Method 4, and perform the test (not more than 5 ppm). Foreign matter—Under a microscope <5.01>, Pow- dered Ginger does not show stone cells, lignified parenchyma cells and other foreign matter. Total ash <5.01> Not more than 8.0z.
Weigh accurately about 1 g of Powdered Ginger (separately determine the loss on drying <5.01>, at 1059C for 5 hours), place in a centrifuge tube, add 30 mL of a mixture of methanol and water (3:1), shake for 20 minutes, centri-fuge, and separate the supernatant liquid. To the residue add 30 mL of a mixture of methanol and water (3:1), and repeat the extraction twice more. To the combined all extracts add a mixture of methanol and water (3:1) to make exactly 100 mL, use this solution as the sample solution. Separately, weigh accurately about 5 mg of [6]-gingerol for assay, dis-solve in a mixture of methanol and water (3:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and ‌JP XVIII Crude Drugs and Related Drugs / Ginseng 2011 determine the peak areas, AT and AS, of [6]-gingerol in each solution. 5 mL of the sample solution and 2 mL of the standard solu-tion on a plate of silica gel for thin-layer chromatography. Amount (mg) of [6]-gingerol ＝ MS × AT/AS Develop the plate with a mixture of ethyl acetate, methanol and water (14:5:4) to a distance of about 7 cm, and air-dry MS: Amount (mg) of [6]-gingerol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile and phosphoric acid (3800:2200:1). Flow rate: Adjust so that the retention time of [6]-gingerol is about 19 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of [6]-gingerol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of [6]-gingerol is not more than 1.5z.
Ginger
Fresh Ginger
生姜
shokyo
Zingiberis Rhizoma consists of the rhizome of Zingiber officinale Roscoe (Zingiberaceae), from which the cork layer is usually removed.
Pale yellowish white to light yellowish brown
Irregularly branched, compressed or knotty, 4-10 cm in length, 1-3 cm in diameter
Fracture surface is fibrous, pale yellow
Cork layer usually absent; cortex composed of parenchyma cells containing starch grains, scattered with fiber bundles and oil cells; endodermis distinct; vascular bundles collateral, scattered throughout the cortex and stele; fibers thick-walled; vessels with spiral or reticulate thickenings
Powder shows numerous starch grains (simple or compound, 5-40 μm in diameter), fragments of fibers, vessels with spiral or reticulate thickenings, and oil cells containing yellowish brown contents
Presence of oil cells containing yellowish brown volatile oil; parenchyma cells with calcium oxalate crystals; spiral vessels abundant
To 2.0 g of pulverized Zingiberis Rhizoma add 10 mL of methanol, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of [6]-gingerol RS in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography. Spot 5 μL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (2:3) to a distance of about 10 cm, and dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate, and heat at 105°C for 5 minutes: a spot among the spots from the sample solution and a spot from the standard solution show the same color tone and Rf value.
Weigh accurately about 0.5 g of pulverized Zingiberis Rhizoma, add 20 mL of methanol, heat under reflux on a water bath for 30 minutes, cool, and filter. To the residue add 20 mL of methanol, and repeat the extraction in the same manner. Combine the filtrates, add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of [6]-gingerol RS, dissolve in methanol to make exactly 20 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 μL each of the sample solution and standard solution as directed under Liquid Chromatography.
Ellipsoidal or broad ovoid fruit, 2 – 3 cm in length, 1 – 2 cm in diameter; externally red-brown with coarse wrinkles, or dark grayish red with fine wrinkles, and both lustrous; both ends slightly dented, with a scar of style on one end and a scar of peduncle on the other; epicarp thin and leather; mesocarp thick, dark grayish brown in color, spongy, soft and adhesive; endocarp extremely hard, fusiform, and divided into two loculi; seeds flat and ovoid. Odor, slight and characteristic; taste, sweet.
(1) Rancidity—Jujube has no unpleasant, rancid odor and taste. (2) Total BHC's and total DDT's <5.01> Not more than 0.2 ppm, respectively. Total ash <5.01> Not more than 3.0z.
Jujube Seed is a compressed ovate to orbicu-lar, lenticular seed, 5 – 9 mm in length, 4 – 6 mm in width, 2 – 3 mm in thickness, externally brown to dark red-brown, glossy; hilum at one end, chalaza at the other end; seed coat sightly flexible, covering, milky white endosperm and light yellow embryo. 100 seeds weigh 3.0 – 4.5 g. Odor, slightly oily; taste, mild and slightly oily. Under a microscope <5.01>, transverse section reveals seed coat composed of an upper epidermis, parenchyma and lower epidermis; upper epidermal cells sclerified and elon-gated in radial direction; lower epidermis covered with cuticle; endosperm composed of parenchyma, containing aggregated crystals of calcium oxalate, aleurone grains and starch grains; cotyledons composed of parenchyma that con-tains aleurone grains, starch grains and oil drops.
To 2 g of pulverized Jujube Seed add 10 mL of methanol, and heat under a reflux condenser for 10 minutes. After cooling, filter, and use the filtrate as the sam-ple solution. Perform the test with the sample solution as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL of the sample solution on a plate of silica gel with fluores-cent indicator for thin-layer chromatography, develop the plate with a mixture of acetone, ethyl acetate, water and acetic acid (100) (10:10:3:1) to a distance of about 7 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots appear at the Rf value of about 0.3 and about 0.4, and these spots exhibit a blue-white fluorescence when examined under ultraviolet light (main wavelength: 365 nm) after spraying evenly dilute sulfuric acid on the plate and heating at 1059C for 5 minutes.
Foreign matter <5.01>—Jujube Seed contains not more than 1.0z of the endocarp and other foreign matters. Loss on drying <5.01> Not more than 11.0z (6 hours). Total ash <5.01> Not more than 5.0z. Extract content <5.01> Dilute ethanol-soluble extract: not less than 8.5z.
solution add 7 mL of ethanol (95): the solution remains clear
Compound Scopolia Extract and Diastase Powder
複方ロートエキス・ジアスターゼ散
Crude Glycyrrhiza Extract contains not less than
Glycyrrhiza or the root and stolon of Glycyrrhiza glabra
Keishibukuryogan Extract contains not less than
Nux Vomica Extract Powder contains not less than
Scopolia Extract and Ethyl Aminobenzoate Powder
Scopolia Extract Powder contains not less than
Method of preparation Extract the coarse powder of
Tokishakuyakusan Extract contains not less than
slightly soluble in ethanol (95), and practically insoluble in water
Method of preparation Take moderately fine powder of
miscible with water and with ethanol (95)
Determine the absorption spectrum of a so-lution of 10z Codeine Phosphate Powder (1 in 1000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of 10z Codeine Phosphate Powder is not less than 85z. Start the test with about 0.2 g of 10z Codeine Phosphate Powder, accurately weighed, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 28 mg of codeine phosphate hydrate for assay (separately determine the water <2.48> in the same manner as Codeine Phosphate Hydrate), and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of codeine in each solution. Dissolution rate (z) with respect to the labeled amount of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2H2O) ＝ MS/MT × AT/AS × 18/25 × 1.023 MS: Amount (mg) of codeine phosphate hydrate for assay taken, calculated on the anhydrous basis MT: Amount (g) of 10z Codeine Phosphate Powder Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of codeine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of codeine is not more than 2.0z.
Weigh accurately about 2.5 g of 10z Codeine Phos-phate Powder, dissolve in water to make exactly 100 mL, then pipet 2 mL of this solution, add exactly 10 mL of the in- under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of codeine to that of the internal standard: Amount (mg) of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2H2O) ＝ MS × QT/QS × 5 × 1.023 MS: Amount (mg) of codeine phosphate hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of codeine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, codeine and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of codeine to that of the internal standard is not more than 1.0z.
‌10 dl-Methylephedrine Hydrochloride Powder
To 0.5 g of 10z dl-Methylephedrine Hydro-chloride Powder add 100 mL of water, shake vigorously for 20 minutes, if necessary, filter the solution. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 250 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 264 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of 10z dl-Methylephedrine Hydrochloride Powder is not less than 85z. ‌Start the test with about 0.5 g of 10z dl-Methylephedrine Hydrochloride Powder, accurately weighed, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add exactly 2 mL of the mobile phase, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of dl-methylephedrine hydrochloride for assay, previ-ously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 25 mL of this solution, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of the mobile phase, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of methylephedrine in each solution. Dissolution rate (z) with respect to the labeled amount of dl-methylephedrine hydrochloride (C11H17NO.HCl) ＝ MS/MT × AT/AS × 9/4 MS: Amount (mg) of dl-methylephedrine hydrochloride for assay taken MT: Amount (g) of 10z dl-Methylephedrine Hydrochlo-ride Powder taken Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of methylephedrine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methylephedrine is not more than 2.0z.
Weigh accurately about 0.5 g of 10z dl-Methyl-ephedrine Hydrochloride Powder, add exactly 4 mL of the internal standard solution and 25 mL of water, shake vigor-ously for 20 minutes to dissolve, add water to make 50 mL, filter through a membrane filter with a pore size not exceed-ing 0.45 mm, if necessary, discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of dl-methylephe-drine hydrochloride for assay, previously dried at 1059C for 3 hours, add exactly 4 mL of the internal standard solution and water to make 50 mL, and use this solution as the stand-ard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios of the peak area, QT and QS, of methylephedrine to that of the internal standard. Amount (mg) of dl-methylephedrine hydrochloride (C11H17NO.HCl) ＝ MS × QT/QS MS: Amount (mg) of dl-methylephedrine hydrochloride for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 257 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 13.6 g of potassium dihydrogen phosphate and 3 g of sodium 1-heptane sulfonate in 1000 mL of water, and adjust the pH to 2.5 with phosphoric acid. To 900 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of methylephedrine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, methylephedrine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of methylephedrine to that of the internal standard is not more than 1.0z.
<2.03>. Spot 2 mL each of the sample solution and standard
Pyridoxine Hydrochloride Injection is a color-less or pale yellow, clear liquid. It is gradually affected by light. pH: 3.0 – 6.0
(1) To a volume of Pyridoxine Hydrochlo-ride Injection, equivalent to 0.05 g of Pyridoxine Hydrochlo-ride, add 0.1 mol/L hydrochloric acid TS to make 100 mL. To 2 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible spectro-photometry <2.24>: it exhibits a maximum between 288 nm and 292 nm. (2) To a volume of Pyridoxine Hydrochloride Injection, equivalent to 0.01 g of Pyridoxine Hydrochloride, add water to make 10 mL, and use this solution as the sample solution. Separately, dissolve 0.01 g of Pyridoxine Hydrochloride RS in 10 mL of water, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, and air-dry the plate. Devel-op the plate with a mixture of acetone, tetrahydrofuran, hexane and ammonia solution (28) (65:13:13:9) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solu-tion of sodium carbonate in diluted ethanol (3 in 10) (1 in 20) ‌on the plate. After air-drying, spray evenly a solution of 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol (99.5) (1 in 1000) on the plate: the spots obtained from the sample solution and the standard solution are blue in color and have the same Rf value.‌ Bacterial endotoxins <4.01> Less than 3.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Pyridoxine Hydrochlo-ride Injection, equivalent to about 20 mg of pyridoxine hy-drochloride (C8H11NO3.HCl), dilute with water, if necessary, and add water to make exactly 100 mL. Pipet 25 mL of this solution, add water to make exactly 200 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 0.1 g of Pyridoxine Hydrochloride RS, previously dried in a desiccator (in vacuum, silica gel) for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL, and use this so-lution as the standard solution. Pipet 1 mL each of the sam-ple solution and standard solution, add 2.0 mL of barbital buffer solution, 9.0 mL of 2-propanol and 2.0 mL of a freshly prepared solution of 2,6-dibromo-N-chloro-1,4-ben-zoquinone monoimine in ethanol (95) (1 in 4000), shake well, add 2-propanol to make exactly 25 mL, and allow to stand for 90 minutes. Determine the absorbances, AT and AS, of the subsequent sample solution and subsequent standard so-lution, respectively, at 650 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared in the same manner with 1 mL of water, as the blank. Amount (mg) of pyridoxine hydrochloride (C8H11NO3.HCl) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of Pyridoxine Hydrochloride RS taken
Containers—Hermetic containers, and colored containers may be used.
Absorptive Cream is white in color and is lustrous. It has a slightly characteristic odor.
Acemetacin Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of acemetacin (C21H18ClNO6: 415.82). Method of preparation Prepare as directed under Cap-sules, with Acemetacin. Identification To an amount of powdered contents of Acemetacin Capsules, equivalent to 0.1 g of Acemetacin, add 100 mL of methanol, shake well, and filter. Take 10 mL of the filtrate, and distil the methanol under reduced pres-sure. To the residue add 1 mL of methanol, shake well, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of acemetacin in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of hexane, 4-methyl-2-penta-none and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 319 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of acemetacin (C21H18ClNO6) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of acemetacin for assay taken C: Labeled amount (mg) of acemetacin (C21H18ClNO6) in 1 capsule Assay Take out the contents of not less than 20 Acemeta-cin Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equiva-lent to about 30 mg of acemetacin (C21H18ClNO6), add 40 mL of methanol, shake well, and add methanol to make exactly 50 mL. Filter this solution, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 2 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of acemetacin for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add exactly 2 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the stand-ard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- JP XVIII Official Monographs / Acemetacin Tablets 401 tions, and calculate the ratios, QT and QS, of the peak area of acemetacin to that of the internal standard. Amount (mg) of acemetacin (C21H18ClNO6) ＝ MS × QT/QS ‌MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. under ultraviolet light (main wavelength: 254 nm): the spots obtained from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Acemetacin Tablets add 3 mL of water, and shake until the tablet is disintegrated. Add 15 mL of metha-nol, shake for 20 minutes, and add methanol to make exactly V mL so that each mL contains about 1.
To an amount of powdered contents of Acemetacin Capsules, equivalent to 0.1 g of Acemetacin, add 100 mL of methanol, shake well, and filter. Take 10 mL of the filtrate, and distil the methanol under reduced pres-sure. To the residue add 1 mL of methanol, shake well, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of acemetacin in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of hexane, 4-methyl-2-penta-none and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 319 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of acemetacin (C21H18ClNO6) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of acemetacin for assay taken C: Labeled amount (mg) of acemetacin (C21H18ClNO6) in 1 capsule
Take out the contents of not less than 20 Acemeta-cin Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equiva-lent to about 30 mg of acemetacin (C21H18ClNO6), add 40 mL of methanol, shake well, and add methanol to make exactly 50 mL. Filter this solution, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 2 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of acemetacin for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add exactly 2 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the stand-ard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi- JP XVIII Official Monographs / Acemetacin Tablets 401 tions, and calculate the ratios, QT and QS, of the peak area of acemetacin to that of the internal standard. Amount (mg) of acemetacin (C21H18ClNO6) ＝ MS × QT/QS ‌MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. under ultraviolet light (main wavelength: 254 nm): the spots obtained from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Acemetacin Tablets add 3 mL of water, and shake until the tablet is disintegrated. Add 15 mL of metha-nol, shake for 20 minutes, and add methanol to make exactly V mL so that each mL contains about 1.2 mg of acemetacin (C21H18ClNO6). Centrifuge this solution, filter the superna-tant liquid, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 1 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Mobile phase: To 6 g of acetic acid (100) add water to Amount (mg) of acemetacin (C H ClNO ) make 1000 mL, and adjust the pH to 3.2 with a solution of 1.36 g of sodium acetate trihydrate in 100 mL of water. To ＝ MS × QT/QS × V/25 21 18 6 200 mL of this solution add 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of acemetacin is about 7 minutes. System suitability— System performance: Dissolve 75 mg of acemetacin and 75 mg of indometacin in 50 mL of methanol. To 2 mL of this solution add 2 mL of the internal standard solution, and add methanol to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, acemetacin, indometacin and the internal standard are eluted in this order with the resolutions between the peaks of acemetacin and indometacin and between the peaks of indometacin and the internal standard being not less than 3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of acemetacin to that of the internal standard is not more than 1.0z.
Acemetacin Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of acemetacin (C21H18ClNO6: 415.82). Method of preparation Prepare as directed under Tablets, with Acemetacin. Identification To a quantity of powdered Acemetacin Tablets, equivalent to 0.1 g of Acemetacin, add 100 mL of methanol, shake well, and filter. Take 10 mL of the filtrate, and distil the methanol under reduced pressure. Dissolve the residue in 1 mL of methanol, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 10 mg of acemetacin in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Examine MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Acemetacin Tablets is not less than 80z. Start the test with 1 tablet of Acemetacin Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 33 mg of acemetacin (C21H18ClNO6), and use this solution as the sample solution.
To a quantity of powdered Acemetacin Tablets, equivalent to 0.1 g of Acemetacin, add 100 mL of methanol, shake well, and filter. Take 10 mL of the filtrate, and distil the methanol under reduced pressure. Dissolve the residue in 1 mL of methanol, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 10 mg of acemetacin in 1 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane, 4-methyl-2-pentanone and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Examine MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Acemetacin Tablets is not less than 80z. Start the test with 1 tablet of Acemetacin Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 33 mg of acemetacin (C21H18ClNO6), and use this solution as the sample solution. Separately, weigh accurately about 17 mg of acemetacin for assay, previously dried at 1059C for 2 hours, dissolve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 319 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of acemetacin (C21H18ClNO6) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of acemetacin for assay taken C: Labeled amount (mg) of acemetacin (C21H18ClNO6) in 1 tablet
Weigh accurately the mass of not less than 20 Acemetacin Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 0.6 g of acemetacin (C21H18ClNO6), add 120 mL of methanol, shake for 20 minutes, and add methanol to make exactly 200 mL. Centri-fuge this solution, filter the supernatant liquid, discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add exactly 1 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of acemetacin for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 1 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the standard solution. Perform the test with ‌10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of acemetacin to that of the internal standard. Amount (mg) of acemetacin (C21H18ClNO6) ＝ MS × QT/QS × 20 MS: Amount (mg) of acemetacin for assay taken Internal standard solution—A solution of hexyl parahy-droxybenzoate in methanol (1 in 250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 6 g of acetic acid (100) add water to make 1000 mL, and adjust the pH to 3.2 with a solution of 1.36 g of sodium acetate trihydrate in 100 mL of water. To 200 mL of this solution add 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of acemetacin is about 7 minutes. System suitability— System performance: Dissolve 75 mg of acemetacin and 75 mg of indometacin in 50 mL of methanol. To 4 mL of this solution add 1 mL of the internal standard solution, and add methanol to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, acemetacin, indometacin and the internal standard are eluted in this order with the resolutions between the peaks of acemetacin and indometacin and between the peaks of indometacin and the internal standard being not less than 3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of acemetacin to that of the internal standard is not more than 1.0z.
Paracetamol Tablets 500 mg
Acetaminophen 500 mg Tablet
Acetaminophen Tablets contain not less than 95% and not more than 105% of the labeled amount of Acetaminophen (C8H9NO2).
To one tablet add 50 mL of diluted methanol (3 in 5), shake vigorously, and filter. Use the filtrate as the sample solution. Separately, dissolve 10 mg of Acetaminophen RS in diluted methanol (3 in 5) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 μL each of the sample solution and standard solution as directed under Liquid Chromatography according to the following conditions: the retention time of the principal peak from the sample solution agrees with that from the standard solution.
900 mL of phosphate buffer solution, pH 5.8
Not less than 85% of the labeled amount of acetaminophen dissolves in 30 minutes
Use one tablet as the sample. Perform the test at 50 revolutions per minute. Withdraw not less than 20 mL of the dissolution medium at the specified time, and filter through a membrane filter with a pore size not exceeding 0.45 μm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V' mL so that each mL contains about 7 μg of acetaminophen. Determine the absorbance of this solution at 243 nm as directed under Ultraviolet-visible Spectrophotometry, using a solution prepared by adding the dissolution medium to 5 mL of Acetaminophen RS solution in methanol (1 in 10,000) to make exactly 100 mL as the standard solution. Calculate the dissolution rate of Acetaminophen Tablets.
Take 1 tablet of Acetaminophen Tablets, add 50 mL of diluted methanol (3 in 5), shake vigorously for 10 minutes, add diluted methanol (3 in 5) to make exactly V mL so that each mL contains about 10 mg of acetaminophen, and filter through a membrane filter with a pore size not exceeding 0.45 μm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 5 mL of this solution, add diluted methanol (3 in 5) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Acetaminophen RS, add diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, add diluted methanol (3 in 5) to make exactly 50 mL. Pipet 5 mL of this solution, add diluted methanol (3 in 5) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbance of the sample solution and standard solution at 243 nm as directed under Ultraviolet-visible Spectrophotometry. Calculate the content uniformity of 10 tablets.
Weigh accurately and powder not less than 20 Acetaminophen Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of acetaminophen, add 20 mL of diluted methanol (3 in 5), shake for 10 minutes, add diluted methanol (3 in 5) to make exactly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 μm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add diluted methanol (3 in 5) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Acetaminophen RS, add diluted methanol (3 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 μL each of the sample solution and standard solution as directed under Liquid Chromatography according to the following conditions. Calculate the amount of acetaminophen.
Store at room temperature, protect from moisture
Acetylcholine Chloride for Injection is a prepara-tion for injection which is dissolved before use. It contains not less than 98.
Acetylcholine Chloride for Injection occurs as white, crystals or crystalline powder. It is very soluble in water, and freely soluble in ethanol (95). It is extremely hygroscopic.
(1) Determine the infrared absorption spec-trum of Acetylcholine Chloride for Injection, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of Acetylcholine Chloride for Injection (1 in 10) responds to Qualitative Tests <1.09> (2) for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Acetylcholine Chloride for Injection in 10 mL of water: the solution is clear and colorless. Acidity—Dissolve 0.10 g of Acetylcholine Chloride for Injection in 10 mL of freshly boiled and cooled water, and add 1 drop of bromothymol blue TS, and 0.30 mL of 0.01 mol/L sodium hydroxide VS: the solution is blue in color. Heavy metals <1.07>—Proceed with 2.0 g of Acetyl- choline Chloride for Injection according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Uniformity of dosage units <6.02> It meets the requirement Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Acetylcholine chloride—Weigh accurately the contents of not less than 10 Acetylcholine Chloride for Injec-tions. Weigh accurately about 0.5 g of the contents, dissolve in 15 mL of water, then add exactly 40 mL of 0.1 mol/L so-dium hydroxide VS, stopper loosely, and heat on a water bath for 30 minutes. Cool quickly, and titrate <2.50> the excess sodium hydroxide with 0.05 mol/L sulfuric acid VS (indicator: 3 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 18.17 mg of C7H16ClNO2 (2) Chlorine—Titrate <2.50> the solution, which has been titrated in (1), with 0.1 mol/L silver nitrate VS (indicator: 3 drops of fluorescein sodium TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 3.545 mg of Cl
(1) Weigh a quantity of powdered Aspirin Tablets, equivalent to 0.1 g of Aspirin, add 10 mL of water, and boil for 5 to 6 minutes. After cooling, filter, and add 1 to 2 drops of iron (III) chloride TS to the filtrate: a red-violet color develops. (2) Weigh a portion of powdered Aspirin Tablets, equivalent to 0.5 g of Aspirin, extract with two 10-mL por-tions of warm ethanol (95), and filter the combined extracts. Evaporate the filtrate to dryness, and boil the residue with 10 mL of sodium carbonate TS for 5 minutes. Proceed as di-rected in the Identification (2) under Aspirin.
Salicylic acid—Take a portion of the powdered Aspirin Tablets, equivalent to 1.0 g of Aspirin, shake with 15 mL of ethanol (95) for 5 minutes, filter, discard the first 5 mL of the filtrate, and add 1.0 mL of the subsequent filtrate to a solution which is prepared by transferring 1 mL of freshly prepared dilute ammonium iron (III) sulfate TS to a Nessler tube and diluting with water to make 50 mL. Pro-ceed as directed in the Purity (2) under Aspirin.
Weigh accurately and powder not less than 20 Aspirin Tablets. Weigh accurately a portion of the powder, equivalent to about 1.5 g of aspirin (C9H8O4), add exactly 50 mL of 0.5 mol/L sodium hydroxide VS, and proceed as directed in the Assay under Aspirin. Each mL of 0.5 mol/L sodium hydroxide VS ＝ 45.04 mg of C9H8O4
Aciclovir for Injection is a preparation for injection which is dissolved before use. It contains not less than 95.
Aciclovir for Injection occurs as white to pale yellow-white, light masses or powder.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. pH Being specified separately when the drug is granted approval based on the Law. droxide TS to make exactly 100 mL. Pipet 15 mL of this solution, add 70 mL of water and 5 mL of 2 mol/L hy-drochloric acid TS, then add water to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in dilute sodium hydroxide TS to make exactly 20 mL. Pipet 15 mL of this solution, add 70 mL of water and 5 mL of 2 mol/L hydro-chloric acid TS, then add water to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 255 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using 0.1 mol/L hydrochloric acid TS as the blank. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × AT/AS × 5 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis
Aciclovir for Syrup is a preparation for syrup, which is suspended before use. It contains not less than 95.
Dissolve an amount of Aciclovir for Syrup, equivalent to 12 mg of Aciclovir, in 0.1 mol/L hydrochloric acid TS to make 50 mL. To 2 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Aciclovir for Syrup in single-dose packages meets the requirement of the Content unifor-mity test. the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 11 mg of Aciclovir RS (separately de-termine the water <2.48> in the same manner as Aciclovir), and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 254 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of aciclovir (C8H11N5O3) ＝ MS/MT × AT/AS × 1/C × 1800 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis MT: Amount (g) of Aciclovir for Syrup taken C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 g
Weigh accurately an amount of Aciclovir for Syrup, previously powdered if necessary, equivalent to about 0.2 g of aciclovir (C8H11N5O3), add 20 mL of diluted sodium hy-droxide TS (1 in 10), sonicate to disintegrate, then add water to make exactly 200 mL, and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, pipet 5 mL of the sub-sequent filtrate, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Aciclovir RS (separately deter-mine the water <2.48> in the same manner as Aciclovir), add exactly 10 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of aciclovir to that of the internal standard. To the total content of 1 package of Aciclovir for Syrup Amount (mg) of aciclovir (C H N O ) add 2V/25 mL of diluted sodium hydroxide TS (1 in 10), and sonicate to disintegrate, add water to make exactly ＝ MS × QT/QS × 20 8 11 5 3 V mL so that each mL contains about 0.8 mg of aciclovir (C8H11N5O3), and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 50 mL, and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × QT/QS × V/10 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis Internal standard solution—A solution of parahydroxyben-zoic acid in the mobile phase (1 in 1250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Aciclovir for Syrup is not less than 85z. Start the test with an accurately weighed amount of Aciclovir for Syrup, equivalent to about 0.2 g of aciclovir (C8H11N5O3), withdraw not less than 5 mL of the medium at MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis Internal standard solution—A solution of parahydroxyben-zoic acid in the mobile phase (1 in 1250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate dihydrate and 0.85 g of sodium 1-octanesulfonate in 900 mL of water, adjust to pH 3.0 with phosphoric acid, add water to make 950 mL, and add 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of aciclovir is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, aciclovir and the internal standard are eluted in this order with the resolution between these peaks being not less ‌JP XVIII Official Monographs / Aclarubicin Hydrochloride 413‌ than 20. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of aciclovir to that of the internal standard is not more than 1.0z.
Aciclovir Granules contain not less than 93.0z and not more than 107.0z of the labeled amount of aciclovir (C8H11N5O3: 225.20). Method of preparation Prepare as directed under Gran-ules, with Aciclovir. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> Perform the test ac-cording to the following method: Aciclovir Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Aciclovir Granules, add 100 mL of dilute sodium hydroxide TS, sonicate with occasional shaking, and add dilute sodium hydroxide TS to make exactly 200 mL. Filter this solution, discard the first 20 mL of the filtrate, pipet V mL of the sub-sequent filtrate, add dilute sodium hydroxide TS to make ex-actly V? mL so that each mL contains about 1 mg of aciclovir (C8H11N5O3).
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> Perform the test ac-cording to the following method: Aciclovir Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Aciclovir Granules, add 100 mL of dilute sodium hydroxide TS, sonicate with occasional shaking, and add dilute sodium hydroxide TS to make exactly 200 mL. Filter this solution, discard the first 20 mL of the filtrate, pipet V mL of the sub-sequent filtrate, add dilute sodium hydroxide TS to make ex-actly V? mL so that each mL contains about 1 mg of aciclovir (C8H11N5O3). Pipet 15 mL of this solution, add 50 mL of water and 5.8 mL of 2 mol/L hydrochloric acid TS, add water to make exactly 100 mL. Pipet 5 mL of this solu-tion, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Then, ‌JP XVIII Official Monographs / Aciclovir Injection 409 proceed as directed in the Assay. Amount (mg) of acyclovir (C8H11N5O3) ＝ MS × AT/AS × V?/V × 8 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Aciclovir Granules is not less than 85z. Start the test with an accurately weighed amount of Aciclovir Granules, equivalent to about 0.4 g of aciclovir (C8H11N5O3), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 252 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of aciclovir (C8H11N5O3) ＝ MS/MT × AT/AS × 1/C × 1800 MS: Amount (mg) of aciclovir RS taken, calculated on the anhydrous basis MT: Amount (g) of Aciclovir Granules taken C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 g Aciclovir Injection
Aciclovir Injection is an aqueous injection. It contains not less than 95.
Aciclovir Injection occurs as a colorless or pale yellow, clear liquid.
To a volume of Aciclovir Injection, equiva-lent to 25 mg of Aciclovir, add 0.5 mol/L hydrochloric acid TS to make 100 mL. To 2 mL of this solution add 0.5 mol/L hydrochloric acid TS to make 50 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 254 nm and 258 nm. Bacterial endotoxins <4.01> Less than 0.5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Aciclovir Injection, equiva-lent to about 25 mg of aciclovir (C8H11N5O3), add 0.1 mol/L Assay Powder Aciclovir Granules, and weigh accurately a portion of the powder, equivalent to about 0.1 g of aciclovir (C8H11N5O3), add 60 mL of dilute sodium hydroxide TS, sonicate for 15 minutes, then add dilute sodium hydroxide TS to make exactly 100 mL, and filter. Discard the first 20 mL of filtrate, pipet 15 mL of the subsequent filtrate, add 50 mL of water and 5.8 mL of 2 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in dilute sodium hydroxide TS to make exactly 25 mL. Pipet 15 mL of this solution, add 50 mL of water and 5.8 mL of 2 mol/L hydrochloric acid TS, hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 5 mL of the internal standard solu-tion, then add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 5 mL of the internal standard solution, then add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of aciclovir to that of the internal standard. add water to make exactly 100 mL. Pipet 5 mL of this solu- Amount (mg) of aciclovir (C H N O ) tion, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. De- ＝ MS × QT/QS 8 11 5 3 termine the absorbances, AT and AS, at 255 nm of the sam-ple solution and standard solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>, using 0.1 mol/L hy-drochloric acid TS as the blank. Amount (mg) of acyclovir (C8H11N5O3) ＝ MS × AT/AS × 4 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis
Aciclovir Ointment contains not less than 95.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm.
Clarity and color of solution—Dissolve an amount of Aciclovir for Injection, equivalent to 0.25 g of Aciclovir, in 10 mL of water: the solution is clear and is not more colored than the following control solution. Control solution: To 2.5 mL of Matching Fluid F add diluted dilute hydrochloric acid (1 in 10) to make 100 mL. Water <2.48> Not more than 7.5z (0.1 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.25 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately an amount of Aciclovir Ointment, equivalent to about 10 mg of aciclovir (C8H11N5O3), add Purity Clarity and color of solution—Dissolve an amount of Aciclovir for Injection, equivalent to 0.25 g of Aciclovir, in 10 mL of water: the solution is clear and is not more colored than the following control solution. Control solution: To 2.5 mL of Matching Fluid F add diluted dilute hydrochloric acid (1 in 10) to make 100 mL. Water <2.48> Not more than 7.5z (0.1 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.25 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Weigh accurately the mass of the contents of not less than 10 Aciclovir for Injection. Weigh accurately an amount of the contents, equivalent to about 0.1 g of aciclovir (C8H11N5O3), and dissolve in dilute sodium hy- mL of dilute sodium hydroxide TS, warm if necessary, and dissolve by shaking. After cooling, add water to make exactly 100 mL. Pipet 15 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Aciclovir RS (separately, determine the water <2.48> in the same manner as Aciclovir), and dissolve in dilute sodium hydroxide TS to make exactly 20 mL. Pipet 10 mL of this solution, and add 15 mL of dilute sodium hydroxide TS and water to make exactly 100 mL. Pipet 15 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 255 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using 0.1 mol/L hydrochloric acid TS as the blank. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Metal Particles <6.01> It meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Particle diameter Being specified separately when the drug is granted approval based on the Law.
Weigh accurately a portion of Aciclovir Ophthalmic Ointment, equivalent to about 15 mg of aciclovir (C8H11N5O3), add exactly 20 mL of hexane and exactly 20 mL of dilute sodium hydroxide TS, and shake vigorously. Centrifuge this mixture, discard the upper layer, pipet 1 mL of the lower layer, add 70 mL of water and 5 mL of 2 mol/L hydrochloric acid TS, then add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 15 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in dilute sodium hydroxide TS to make exactly 20 mL. Pipet 1 mL of this solution, add 70 mL of water and 5 mL of 2 mol/L hydrochloric acid TS, then add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 255 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Amount (mg) of acyclovir (C8H11N5O3) ＝ MS × AT/AS MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis
Aciclovir Syrup is a suspension syrup. It contains not less than 93.
To a volume of thoroughly shaken Aciclovir Syrup, equivalent to 80 mg of Aciclovir, add 0.1 mol/L hydrochloric acid TS to make 100 mL. Centrifuge this solution, to 1 mL of the supernatant liquid add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Aciclovir Syrup is not less than 85z. Start the test with an exact volume of thoroughly shaken Aciclovir Syrup, equivalent to about 0.4 g of aciclovir (C8H11N5O3), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and centrifuge. Pipet 2 mL of the supernatant liquid, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 252 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of aciclovir (C8H11N5O3) ＝ MS/VT × AT/AS × 1/C × 1800 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis VT: Amount (mL) of Aciclovir Syrup taken C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 mL
Shake thoroughly Aciclovir Syrup. To an exact volume of the syrup, equivalent to about 80 mg of aciclovir (C8H11N5O3), add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, then add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 5 mL of the internal standard solution, then add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this so-lution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of aciclovir to that of the internal standard. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × QT/QS × 2 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis Internal standard solution—A solution of nicotinic acid in 0.1 mol/L hydrochloric acid TS (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1.45 g of phosphoric acid and 25 mL of dilute acetic acid add water to make 900 mL. Adjust this so-lution to pH 2.5 with 1 mol/L sodium hydroxide TS, and add water to make 1000 mL. To 950 mL of this solution add 50 mL of methanol. ‌412 Flow rate: Adjust so that the retention time of aciclovir is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and aciclovir are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of aciclovir to that of the internal standard is not more than 1.0z.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Aciclovir Tablets is not less than 80z. Start the test with 1 tablet of Aciclovir Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 8.9 mg of aciclovir (C8H11N5O3), and use this so-lution as the sample solution. Separately, weigh accurately about 22 mg of aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 252 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of aciclovir (C8H11N5O3) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of Acyclovir RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of aciclovir (C8H11N5O3) in 1 tablet
Weigh accurately the mass of not less than 20 Aciclovir Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of aciclovir (C8H11N5O3), add 60 mL of dilute sodium hydroxide TS, and sonicate for 15 minutes, then add dilute sodium hydroxide TS to make exactly 100 mL, and filter. Discard the first 20 mL of filtrate, pipet 15 mL of the subsequent filtrate, add 50 mL of water and 5.8 mL of 2 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Aciclovir RS (separately determine the water <2.48> in the same manner as Aciclovir), and dissolve in dilute sodium hydroxide TS to make exactly 25 mL. Pipet 15 mL of this solution, add 50 mL of water and 5.8 mL of 2 mol/L hydrochloric acid TS, add water to make exactly 100 mL. Pipet 5 mL of this solu-tion, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, at 255 nm of the sam-ple solution and standard solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>, using 0.1 mol/L hy-drochloric acid TS as the blank. Amount (mg) of aciclovir (C8H11N5O3) ＝ MS × AT/AS × 4 MS: Amount (mg) of Aciclovir RS taken, calculated on the anhydrous basis
Acrinol and Zinc oxide Ointment is yellow in color.
(1) Shake 0.5 g of Acrinol and Zinc Oxide Ointment with 5 mL of diethyl ether, 5 mL of dilute hydro-chloric acid and 2 to 3 drops of sodium nitrite TS, and allow to stand: a dark red color develops in the water layer (acrinol). Ignite 0.5 g of Acrinol and Zinc Oxide Ointment to char, and dissolve the residue in 5 mL of dilute hydrochloric acid: the solution responds to Qualitative Tests <1.09> for zinc salt. Shake 0.5 g of Acrinol and Zinc Oxide Ointment with 5 mL of diethyl ether, 1 mL of acetic acid (100) and 5 mL of water, separate the water layer, and use the water layer as the sample solution. Dissolve 5 mg of acrinol in 1 mL of acetic acid (100) and 5 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of diethyl ether, ethanol (95) and acetic acid (100) (40:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the spots from the sample solution and the standard solution exhibit a blue fluorescence and show the same Rf value. ‌JP XVIII Official Monographs / Adrenaline 417‌
Ajmaline Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of ajmaline (C20H26N2O2: 326.43). Method of preparation Prepare as directed under Tablets, with Ajmaline. Identification (1) Shake a quantity of powdered Ajmaline Tablets, equivalent to 0.1 g of Ajmaline, with 30 mL of chlo-roform, and filter. Evaporate the filtrate on a water bath to dryness. With the residue, proceed as directed in the Identifi-cation under Ajmaline. (2) Dissolve 0.01 g of the residue of (1) in 100 mL of ethanol (95). To 10 mL of this solution add ethanol (95) to make 50 mL, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 247 nm and 251 nm and between 291 nm and 294 nm, and a minimum between 269 nm and 273 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ajmaline Tablets add 150 mL of 2nd fluid for dissolution test, shake to disintegrate the tablet, then add 1 cm Absorbance <2.24> E 1z (249 nm): 257 – 271 (after drying, 2nd fluid for dissolution test to make exactly 200 mL, and 2 mg, ethanol (95), 100 mL). filter this solution through a membrane filter with a pore size E 1z 1 cm (292 nm): 85 – 95 (after drying, 2 mg, ethanol (95), not exceeding 0.8 mm. Discard the first 10 mL of the filtrate, 100 mL). D Optical rotation <2.49> [a]20: ＋136 – ＋1519(after drying, 0.5 g, chloroform, 50 mL, 100 mm). Purity Related substances—Dissolve 0.10 g of Ajmaline in 10 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each pipet V mL of the subsequent filtrate equivalent to about 0.5 mg of ajmaline (C20H26N2O2), add 2nd fluid for dissolution test to make exactly 10 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of ajmaline for assay, previously dried in vacuum at 809C for 3 hours, dissolve in 2nd fluid for dissolution test to make ex-actly 500 mL, and use this solution as the standard solution. Determine the absorbances at 288 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. of the sample solution and standard solution on a plate of Amount (mg) of ajmaline (C H N O ) silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of chloroform, ace- ＝ MS × AT/AS × 1/V × 4 2 2 tone and diethylamine (5:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.6 g, in vacu-um, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g). Assay Weigh accurately about 0.3 g of Ajmaline, previ-ously dried, dissolve in 50 mL of acetic anhydride and 50 mL of acetone for nonaqueous titration, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 16.32 mg of C H N O MS: Amount (mg) of ajmaline for assay taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Ajmaline Tablets is not less than 75z. Start the test with 1 tablet of Ajmaline Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of ajmaline (C20H26N2O2), and use this solution as the sample solution.
(1) Shake a quantity of powdered Ajmaline Tablets, equivalent to 0.1 g of Ajmaline, with 30 mL of chlo-roform, and filter. Evaporate the filtrate on a water bath to dryness. With the residue, proceed as directed in the Identifi-cation under Ajmaline. (2) Dissolve 0.01 g of the residue of (1) in 100 mL of ethanol (95). To 10 mL of this solution add ethanol (95) to make 50 mL, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 247 nm and 251 nm and between 291 nm and 294 nm, and a minimum between 269 nm and 273 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ajmaline Tablets add 150 mL of 2nd fluid for dissolution test, shake to disintegrate the tablet, then add 1 cm Absorbance <2.24> E 1z (249 nm): 257 – 271 (after drying, 2nd fluid for dissolution test to make exactly 200 mL, and 2 mg, ethanol (95), 100 mL). filter this solution through a membrane filter with a pore size E 1z 1 cm (292 nm): 85 – 95 (after drying, 2 mg, ethanol (95), not exceeding 0.8 mm. Discard the first 10 mL of the filtrate, 100 mL). D Optical rotation <2.49> [a]20: ＋136 – ＋1519(after drying, 0.5 g, chloroform, 50 mL, 100 mm).
Related substances—Dissolve 0.10 g of Ajmaline in 10 mL of chloroform, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each pipet V mL of the subsequent filtrate equivalent to about 0.5 mg of ajmaline (C20H26N2O2), add 2nd fluid for dissolution test to make exactly 10 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of ajmaline for assay, previously dried in vacuum at 809C for 3 hours, dissolve in 2nd fluid for dissolution test to make ex-actly 500 mL, and use this solution as the standard solution. Determine the absorbances at 288 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. of the sample solution and standard solution on a plate of Amount (mg) of ajmaline (C H N O ) silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of chloroform, ace- ＝ MS × AT/AS × 1/V × 4 2 2 tone and diethylamine (5:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (0.6 g, in vacu-um, 809C, 3 hours). Residue on ignition <2.44> Not more than 0.2z (0.5 g).
Weigh accurately about 0.3 g of Ajmaline, previ-ously dried, dissolve in 50 mL of acetic anhydride and 50 mL of acetone for nonaqueous titration, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 16.32 mg of C H N O MS: Amount (mg) of ajmaline for assay taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Ajmaline Tablets is not less than 75z. Start the test with 1 tablet of Ajmaline Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of ajmaline (C20H26N2O2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of ajmaline for assay, previously dried in vacuum at 809C for 3 hours, dis-solve in the dissolution medium to make exactly 500 mL, and 20 26 2 2 ‌JP XVIII Official Monographs / Alacepril 421 use this solution as the standard solution. Determine the ab-sorbances, AT and AS, of the sample solution and standard solution at 288 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of ajmaline (C20H26N2O2) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of ajmaline for assay taken C: Labeled amount (mg) of ajmaline (C20H26N2O2) in 1 tablet Assay Weigh accurately and powder not less than 20 Ajmaline Tablets. Weigh accurately a portion of the powder, equivalent to about 0.3 g of ajmaline (C20H26N2O2), add 15 mL of ammonia solution (28), and extract with four 25-mL portions of chloroform. Combine the chloroform extracts, wash with 10 mL of water, add 5 g of anhydrous sodium sulfate, shake well, and filter. Wash the container and the residue with two 10-mL portions of chloroform, and filter. Evaporate the combined filtrate on a water bath to dryness, dissolve the residue in 50 mL of acetic anhydride and 50 mL of acetone for nonaqueous titration, and titrate <2.50> with 0.05 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 16.32 mg of C20H26N2O2
Alacepril Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of alacepril (C20H26N2O5S: 406.50). Method of preparation Prepare as directed under Tablets, with Alacepril. Identification Shake well a quantity of powdered Alacepril Tablets, equivalent to 0.1 g of Alacepril, with 10 mL of ethanol (95), filter, and use the filtrate as the sample solu-tion. Separately, dissolve 10 mg of alacepril in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, de-velop the plate with a mixture of ethanol (99.5) and hexane (2:1) to a distance of about 10 cm, and air-dry the plate. Ex-amine under ultraviolet light (main wavelength: 254 nm): the principal spot from the sample solution and the spot from the standard solution show the same color tone and Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Alacepril Tablets add 2 mL of water, soni-cate to disperse the particle, and add exactly 2 mL of the internal standard solution for every 10 mg of alacepril (C20H26N2O5S). To this solution add a suitable amount of methanol, extract for 15 minutes with the aid of ultrasonic wave while occasional shaking, and shake more 15 minutes. Add methanol to make V mL so that each mL of the solu-tion contains about 0.
Shake well a quantity of powdered Alacepril Tablets, equivalent to 0.1 g of Alacepril, with 10 mL of ethanol (95), filter, and use the filtrate as the sample solu-tion. Separately, dissolve 10 mg of alacepril in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, de-velop the plate with a mixture of ethanol (99.5) and hexane (2:1) to a distance of about 10 cm, and air-dry the plate. Ex-amine under ultraviolet light (main wavelength: 254 nm): the principal spot from the sample solution and the spot from the standard solution show the same color tone and Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Alacepril Tablets add 2 mL of water, soni-cate to disperse the particle, and add exactly 2 mL of the internal standard solution for every 10 mg of alacepril (C20H26N2O5S). To this solution add a suitable amount of methanol, extract for 15 minutes with the aid of ultrasonic wave while occasional shaking, and shake more 15 minutes. Add methanol to make V mL so that each mL of the solu-tion contains about 0.5 mg of alacepril (C20H26N2O5S), cen-trifuge, and use the supernatant liquid as the sample solu- <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of alacepril to that of the internal standard. Amount (mg) of alacepril (C20H26N2O5S) ＝ MS × QT/QS × V/50 MS: Amount (mg) of alacepril for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in methanol (3 in 20,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate of a 12.5-mg tablet and a 25-mg tablet in 30 minutes is not less than 75z, and that of a 50-mg tablet in 30 minutes is not less than 70z. Start the test with 1 tablet of Alacepril Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 14 mg of alacepril (C20H26N2O5S), and use this so-lution as the sample solution. Separately, weigh accurately about 14 mg of alacepril for assay, previously dried at 1059C for 3 hours, dissolve in 2 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT1 and AS1, at 230 nm, and AT2 and AS2, at 300 nm, of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of alacepril (C20H26N2O5S) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 90 MS: Amount (mg) of alacepril for assay taken C: Labeled amount (mg) of alacepril (C20H26N2O5S) in 1 tablet
Weigh accurately, and powder not less than 20 Alacepril Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of alacepril (C20H26N2O5S), moisten with 2 mL of water, add exactly 3 mL of the internal standard solution and 40 mL of methanol, sonicate for 15 minutes, cool, and add methanol to make 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of alacepril for assay, previously dried at 1059C for 3 hours, add exactly 3 mL of the internal standard solution, dissolve with methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of alacepril to that of the internal standard. tion. Separately, weigh accurately about 25 mg of alacepril for assay, previously dried at 1059C for 3 hours, add exactly Amount (mg) of alacepril (C 20H26N2O5S) ＝ M S × QT/QS 5 mL of the internal standard solution and methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography MS: Amount (mg) of alacepril for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in methanol (1 in 2000). ‌JP XVIII Official Monographs / L-Alanine 423 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100), acetonitrile, methanol and tetrahydrofuran (13:5:1:1). Flow rate: Adjust so that the retention time of alacepril is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, alacepril and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of alacepril to that of the internal standard is not more than 1.0z.
Aldioxa Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of aldioxa (C4H7AlN4O5: 218.10). Method of preparation Prepare as directed under Gran-ules, with Aldioxa. Identification (1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. (2) To a quantity of powdered Aldioxa Granules, equivalent to 0.2 g of Aldioxa, add 10 mL of dilute hydro-chloric acid, boil for 5 minutes, and filter: the cooled filtrate responds to Qualitative Tests <1.09> for aluminum salt. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Aldioxa Granules in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Aldioxa Granules add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add sodium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet V mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly V? mL so that each mL contains about 20 mg of aldioxa (C4H7AlN4O5), and use this solution as the sample solution.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. (2) To a quantity of powdered Aldioxa Granules, equivalent to 0.2 g of Aldioxa, add 10 mL of dilute hydro-chloric acid, boil for 5 minutes, and filter: the cooled filtrate responds to Qualitative Tests <1.09> for aluminum salt. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Aldioxa Granules in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Aldioxa Granules add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add sodium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet V mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly V? mL so that each mL contains about 20 mg of aldioxa (C4H7AlN4O5), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of aldioxa (C4H7AlN4O5) ＝ MS × AT/AS × V?/V × 1/25 MS: Amount (mg) of aldioxa for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Aldioxa Granules is not less than 85z. Start the test with an accurately weighed amount of Aldioxa Granules, equivalent to about 0.1 g of aldioxa (C4H7AlN4O5), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 10 mL of the subsequent filtrate, add diluted ammo-nia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of al-dioxa for assay, previously dried at 1059C for 2 hours, and dissolve in sodium fluoride-hydrochloric acid TS to make ex-actly 25 mL. Pipet 1 mL of this solution, add diluted ammo-nia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 223 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. ‌426‌ Dissolution rate (z) with respect to the labeled amount of aldioxa (C4H7AlN4O5) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount (mg) of aldioxa for assay taken MT: Amount (g) of Aldioxa Granules taken C: Labeled amount (mg) of aldioxa (C4H7AlN4O5) in 1 g
Weigh accurately an amount of powdered Aldioxa Granules, equivalent to about 0.1 g of aldioxa (C4H7AlN4O5), add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add sodium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet 2 mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of aldioxa for assay, previously dried at 1059C for 2 hours, and dissolve in so-dium fluoride-hydrochloric acid TS to make exactly 100 mL. Pipet 4 mL of this solution, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 223 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of aldioxa (C H AlN O ) tively. Start the test with 1 tablet of Aldioxa Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add diluted ammonia-ammonium chloride buffer solu-tion (pH 10.0) (1 in 10) to make exactly V? mL so that each mL contains about 22 mg of aldioxa (C4H7AlN4O5), and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of aldioxa for assay, previously dried at 1059C for 2 hours, and dissolve in sodium fluoride-hydrochloric acid TS to make exactly 25 mL. Pipet 1 mL of this solution, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 223 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of aldioxa (C4H7AlN4O5) ＝ MS × AT/AS × V?/V × 1/C × 72 MS: Amount (mg) of aldioxa for assay taken C: Labeled amount (mg) of aldioxa (C4H7AlN4O5) in 1 ＝ MS × AT/AS × 2 4 7 4 5 tablet MS: Amount (mg) of aldioxa for assay taken
Aldioxa Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of aldioxa (C4H7AlN4O5: 218.10). Method of preparation Prepare as directed under Tablets, with Aldioxa. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Aldioxa Tablets add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add so-dium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet V mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly V? mL so that each mL contains about 20 mg of aldioxa (C4H7AlN4O5), and use this solution as the sample solution.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Aldioxa Tablets add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add so-dium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet V mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly V? mL so that each mL contains about 20 mg of aldioxa (C4H7AlN4O5), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of aldioxa (C4H7AlN4O5) ＝ MS × AT/AS × V?/V × 1/25 MS: Amount (mg) of aldioxa for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 50-mg tablet and in 30 minutes of 100-mg tablet are not less than 80z and not less than 70z, respec-
Weigh accurately, and powder not less than 20 Aldioxa Tablets. Weigh accurately a portion of the powder, equivalent to about 0.1 g of aldioxa (C4H7AlN4O5), add 80 mL of sodium fluoride-hydrochloric acid TS, shake for 20 minutes, add sodium fluoride-hydrochloric acid TS to make exactly 100 mL, and filter. Pipet 2 mL of the filtrate, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of aldioxa for assay, previously dried at 1059C for 2 hours, and dissolve in sodium fluoride-hydrochloric acid TS to make exactly 100 mL. Pipet 4 mL of this solution, add diluted ammonia-ammonium chloride buffer solution (pH 10.0) (1 in 10) to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 223 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of aldioxa (C4H7AlN4O5) ＝ MS × AT/AS × 2 MS: Amount (mg) of aldioxa for assay taken
アレンドロン酸ナトリウム注射液
Alendronate Sodium Injection is an aqueous injec-tion. It contains not less than 95.
Alendronate Sodium Injection is a clear, color-less liquid.
Use Alendronate Sodium Injection as the sample solution. Separately, dissolve 33 mg of alendronate sodium hydrate in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of water, pyridine, acetic acid (100) and ethyl acetate (1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ace-tone (1 in 50) on the plate, and heat at 1009C for 10 minutes: the principal spots from the sample solution and standard solution show a blue-purple color and the same Rf value. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 119 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to Membrane-filter method: it meets the requirement.
To an exactly measured volume of Alendronate So-dium Injection, equivalent to about 5 mg of alendronic acid (C4H13NO7P2), add a solution of disodium dihydrogen ethyl-enediamine tetraacetate dihydrate (1 in 500) to make exactly 100 mL, and use this solution as the sample stock solution. Separately, weigh accurately about 33 mg of Alendronate Sodium RS (separately determine the loss on drying <2.41> under the same conditions as Alendronate Sodium Hydrate), and dissolve in a solution of disodium dihydrogen ethylene-diamine tetraacetate dihydrate (1 in 500) to make exactly 100 mL. Pipet 10 mL of this solution, add a solution of diso-dium dihydrogen ethylenediamine tetraacetate dihydrate (1 in 500) to make exactly 50 mL, and use this solution as the standard stock solution. Pipet 5 mL each of the sample stock solution and standard stock solution, add exactly 5 mL of a solution of sodium tetraborate decahydrate (19 in 500) and exactly 4 mL of a solution of 9-fluorenylmethyl chlorofor-mate in acetonitrile (1 in 1000), shake for 30 seconds, and allow to stand at room temperature for 25 minutes. Then, add 25 mL of dichloromethane to them, shake for 45 seconds, centrifuge, and use the supernatant liquid so ob-tained as the sample solution and the standard solution, respectively. Perform the test with 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of alen-dronic acid in each solution. Amount (mg) of alendronic acid (C4H13NO7P2) ＝ MS × AT/AS × 1/5 × 0.919 MS: Amount (mg) of Alendronate Sodium RS taken, cal-culated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 14.7 g of trisodium citrate dihy-drate and 8.7 g of dipotassium hydrogen phosphate in 900 ‌JP XVIII Official Monographs / Alendronate Sodium Tablets 429 mL of water, adjust to pH 8.0 with phosphoric acid, and add water to make 1000 mL. To 750 mL of this solution add 200 mL of acetonitrile for liquid chromatography and 50 mL of methanol. Flow rate: Adjust so that the retention time of alendronic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of alendronic acid are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above opera-tions conditions, the relative standard deviation of the peak of alendronic acid is not more than 1.0z.
Alendronate Sodium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of alendronic acid (C4H13NO7P2: 249.10). 85z. Start the test with 1 tablet of Alendronate Sodium Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and centrifuge. Pipet V mL of the supernatant liquid, add water to make exactly V? mL so that each mL contains about 6 mg of alendronic acid (C4H13NO7P2), and use this solution as the sample stock so-lution.
To a quantity of powdered Alendronate Sodium Tablets, equivalent to 25 mg of alendronic acid (C4H13NO7P2), add 25 mL of water, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh 33 mg of alendronate sodium hydrate, and dissolve in 25 mL of water, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of water, pyridine, acetic acid (100) and ethyl ace-tate (1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat the plate at 1009C for 10 minutes: the principal spots from the sample solution and standard solution show a blue-purple color and the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Alendronate Sodium Tablets add 0.1 mol/L trisodium citrate TS to make exactly 100 mL, and stir until the tablet is completely disintegrated. Centrifuge this solu-tion, pipet V mL of the supernatant liquid, and add 0.1 mol/L trisodium citrate TS to make exactly V? mL so that each mL contains about 25 mg of alendronic acid (C4H13NO7P2), and use this solution as the sample stock solution. Then, proceed as directed in the Assay. Amount (mg) of alendronic acid (C4H13NO7P2) ＝ MS × AT/AS × V?/V × 2/25 × 0.919 MS: Amount (mg) of Alendronate Sodium RS taken, cal-culated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Alendronate Sodium Tablets is not less than ＝ MS × AT/AS × V?/V × 1/C × 108/5 × 0.919 MS: Amount (mg) of Alendronate Sodium RS taken, cal-culated on the dried basis C: Labeled amount (mg) of alendronic acid (C4H13NO7P2) in 1 tablet
Weigh accurately and powder not less than 20 Alendronate Sodium Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of alendronic acid (C4H13NO7P2), add 0.1 mol/L trisodium citrate TS to make exactly 1000 mL, stir for 30 minutes, and centrifuge. Pipet 5 mL of the supernatant liquid, add 0.1 mol/L trisodium citrate TS to make exactly 10 mL, and use this solution as the sample stock solution. Separately, weigh accurately about 39 mg of Alendronate Sodium RS (separately deter-mine the loss on drying <2.41> under the same conditions as Alendronate Sodium Hydrate), dissolve in 0.1 mol/L triso-dium citrate TS to make exactly 50 mL. Pipet 2 mL of this solution, add 0.1 mol/L trisodium citrate TS to make exactly 50 mL, and use this solution as the standard stock solution. Pipet 5 mL each of the sample stock solution and standard stock solution, add exactly 5 mL of a solution of sodium tetraborate decahydrate (19 in 500) and exactly 4 mL of a so-lution of 9-fluprenylmethyl chloroformate in acetonitrile (1 in 1000), shake for 30 seconds, and allow to stand at room temperature for 25 minutes. Then, add 25 mL of dichloro-methane to them, shake for 45 seconds, centrifuge, and use the supernatant liquid so obtained as the sample solution and the standard solution, respectively. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of alendronic acid in each solution. Amount (mg) of alendronic acid (C4H13NO7P2) ＝ MS × AT/AS × 8/5 × 0.919 MS: Amount (mg) of Alendronate Sodium RS taken, cal-culated on the dried basis ‌430‌ Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 14.7 g of trisodium citrate dihy-drate and 7.1 g of anhydrous disodium hydrogen phosphate in 900 mL of water, adjust to pH 8.0 with phosphoric acid, and add water to make 1000 mL. To 750 mL of this solution add 200 mL of acetonitrile for liquid chromatography and 50 mL of methanol. Flow rate: Adjust so that the retention time of alendronic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of alendronic acid are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above opera-tions conditions, the relative standard deviation of the peak of alendronic acid is not more than 1.0z.
Allopurinol Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of allopurinol (C5H4N4O: 136.11). Method of preparation Prepare as directed under Tablets, with Allopurinol. Identification (1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 248 nm and 252 nm. (2) To a quantity of powdered Allopurinol Tablets, equivalent to 0.1 g of Allopurinol, add 5 mL of a solution of diethylamine (1 in 10), shake well, add 5 mL of methanol, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 0.1 g of allopurinol in 5 mL of a solution of diethylamine (1 in 10), add 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 2.5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 2-butanone, ammonia solution (28) and 2-methoxyethanol (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spots obtained from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Allopurinol Tablets add V/10 mL of 0.05 mol/L sodium hydroxide TS, shake well, and sonicate for 10 minutes. After cooling, add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 0.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 248 nm and 252 nm. (2) To a quantity of powdered Allopurinol Tablets, equivalent to 0.1 g of Allopurinol, add 5 mL of a solution of diethylamine (1 in 10), shake well, add 5 mL of methanol, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 0.1 g of allopurinol in 5 mL of a solution of diethylamine (1 in 10), add 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 2.5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 2-butanone, ammonia solution (28) and 2-methoxyethanol (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spots obtained from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Allopurinol Tablets add V/10 mL of 0.05 mol/L sodium hydroxide TS, shake well, and sonicate for 10 minutes. After cooling, add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 0.5 mg of allopurinol (C5H4N4O), and filter through a mem-brane filter with a pore size not exceeding 0.8 mm. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 0.1 mol/L of hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of allopurinol for assay, previously dried at 1059C for 4 hours, dissolve in 10 mL of 0.05 mol/L sodium hydroxide TS, and add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 250 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of allopurinol (C5H4N4O) ＝ MS × AT/AS × V/100 MS: Amount (mg) of allopurinol for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Allopurinol Tablets is not less than 80z. Start the test with 1 tablet of Allopurinol Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of allopurinol (C5H4N4O), and use this solution as the sample solution. Separately, weigh accurately about 11 mg of allopurinol for assay, previously dried at 105 9C for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 250 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of allopurinol (C5H4N4O) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of allopurinol for assay taken C: Labeled amount (mg) of allopurinol (C5H4N4O) in 1 tablet ‌432
(1) Heavy metals <1.07>—Proceed with 2.0 g of Alminoprofen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Alminoprofen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Alminoprofen in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of these solu-tions by the automatic integration method: the area of the peak other than alminoprofen obtained from the sample so-lution is not larger than 1/5 times the peak area of almino-profen from the standard solution. Furthermore, the total Amount (mg) of allopurinol (C5H4N4O) ＝ M S × AT/AS area of the peaks other than alminoprofen from the sample solution is not larger than the peak area of alminoprofen MS: Amount (mg) of allopurinol for assay taken
Weigh accurately the mass of not less than 20 Allopurinol Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 0.1 g of allopurinol (C5H4N4O), add 20 mL of 0.05 mol/L sodium hydroxide TS, shake well, and treat with ultrasonic waves for 10 minutes. After cooling, add 0.1 mol/L hydrochloric acid TS to make exactly 200 mL, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of allopurinol for assay, previously dried at 1059C for 4 hours, dissolve in 20 mL of 0.05 mol/L sodium hydroxide TS, and add 0.1 mol/L hydrochloric acid TS to make exactly 200 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand-ard solution at 250 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Alminoprofen according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Alminoprofen according to Method 3, and perform the test (not more than 2 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Alminoprofen in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of these solu-tions by the automatic integration method: the area of the peak other than alminoprofen obtained from the sample so-lution is not larger than 1/5 times the peak area of almino-profen from the standard solution. Furthermore, the total Amount (mg) of allopurinol (C5H4N4O) ＝ M S × AT/AS area of the peaks other than alminoprofen from the sample solution is not larger than the peak area of alminoprofen MS: Amount (mg) of allopurinol for assay taken
Alminoprofen Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of alminoprofen (C13H17NO2: 219.28). Method of preparation Prepare as directed under Tablets, with Alminoprofen. Identification Take an amount of powdered Alminoprofen Tablets, equivalent to 30 mg of Alminoprofen, add ethanol (99.5) to make 100 mL, shake thoroughly, and centrifuge. To 2 mL of the supernatant liquid add ethanol (99.5) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 253 nm and 257 nm, and between 298 nm and 302 nm. Purity Related substances—Conduct this procedure using light-resistant vessels. Powder 10 tablets of Alminoprofen Tablets, weigh a portion of the powder equivalent to 50 mg of Alminoprofen, add 50 mL of the mobile phase, shake for 15 minutes, add the mobile phase to make exactly 100 mL, centrifuge, and use the supernatant liquid as the sample solu-tion. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the area of the peak other than alminoprofen obtained from the sample solution is not larger than 1/2 times the peak area of alminoprofen from the standard solution. Furthermore, the total area of the peaks other than alminoprofen from the sample solution is not larger than 2 times the peak area of alminoprofen from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Alminoprofen. System suitability— Proceed as directed in the system suitability in the Purity in Assay under Alminoprofen. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Alminoprofen Tablets add 5 mL of water, shake until the tablet is disintegrated, add 50 mL of ethanol (99.5), shake for 20 minutes, then add ethanol (99.5) to make exactly 100 mL, and centrifuge. Pipet 3 mL of the su-pernatant liquid, add ethanol (99.5) to make exactly 50 mL. Pipet V mL of this solution, add ethanol (99.5) to make exactly V? mL so that each mL contains about 6 mg of alminoprofen (C13H17NO2), and use this solution as the sam-ple solution.
Take an amount of powdered Alminoprofen Tablets, equivalent to 30 mg of Alminoprofen, add ethanol (99.5) to make 100 mL, shake thoroughly, and centrifuge. To 2 mL of the supernatant liquid add ethanol (99.5) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 253 nm and 257 nm, and between 298 nm and 302 nm.
Related substances—Conduct this procedure using light-resistant vessels. Powder 10 tablets of Alminoprofen Tablets, weigh a portion of the powder equivalent to 50 mg of Alminoprofen, add 50 mL of the mobile phase, shake for 15 minutes, add the mobile phase to make exactly 100 mL, centrifuge, and use the supernatant liquid as the sample solu-tion. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of each solution by the automatic integration method: the area of the peak other than alminoprofen obtained from the sample solution is not larger than 1/2 times the peak area of alminoprofen from the standard solution. Furthermore, the total area of the peaks other than alminoprofen from the sample solution is not larger than 2 times the peak area of alminoprofen from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Alminoprofen. System suitability— Proceed as directed in the system suitability in the Purity in Assay under Alminoprofen. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Alminoprofen Tablets add 5 mL of water, shake until the tablet is disintegrated, add 50 mL of ethanol (99.5), shake for 20 minutes, then add ethanol (99.5) to make exactly 100 mL, and centrifuge. Pipet 3 mL of the su-pernatant liquid, add ethanol (99.5) to make exactly 50 mL. Pipet V mL of this solution, add ethanol (99.5) to make exactly V? mL so that each mL contains about 6 mg of alminoprofen (C13H17NO2), and use this solution as the sam-ple solution. Then, proceed as directed in the Assay. Amount (mg) of alminoprofen (C13H17NO2) ＝ MS × AT/AS × V?/V × 1/3 MS: Amount (mg) of alminoprofen for assay taken Dissolution <6.10> When the test is performed at 50 revolu- tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of Alminoprofen Tablets is not less than 80z. Start the test with 1 tablet of Alminoprofen Tablets, with-draw not less than 20 mL of the medium at specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add 0.05 mol/L sodium hydroxide TS to make exactly V? mL so that each mL contains about 8.9 mg of almino-profen (C13H17NO2), and use this solution as the sample so-lution. Separately, weigh accurately about 30 mg of almino-profen for assay, previously dried in vacuum for 1 hour using phosphorus (V) oxide as the dessicant, and dissolve in 0.05 mol/L sodium hydroxide TS to make exactly 100 mL. Pipet 3 mL of this solution, add 0.05 mol/L sodium hydrox-ide TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 245 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of alminoprofen (C13H17NO2) ＝ MS × AT/AS × V?/V × 1/C × 27 MS: Amount (mg) of alminoprofen for assay taken C: Labeled amount (mg) of alminoprofen (C13H17NO2) in 1 tablet
Weigh accurately the mass of not less than 20 tablets of Alminoprofen Tablets, and powder. Weigh accurately an amount equivalent to about 60 mg of alminoprofen (C13H17NO2), add ethanol (99.5) and shake well, add ethanol (99.5) to make exactly 200 mL, and centrifuge. Pipet 2 mL of the supernatant liquid, add ethanol (99.5) to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 30 mg of alminoprofen for assay, previously dried in vacuum for 1 hour using phospho-rus (V) oxide as the dessicant, dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 2 mL of this solution, add ethanol (99.5) to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at the wavelength of maximum absorption at about 255 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of alminoprofen (C13H17NO2) ＝ MS × AT/AS × 2 MS: Amount (mg) of alminoprofen for assay taken
Alprostadil Injection is an emulsion-type injection. It contains not less than 80.
Alprostadil Injection occurs as a white emul-sion and is slightly viscous. It has a distinctive odor.
To a quantity of Alprostadil Injection, cor-responding to 10 mg of Alprostadil, add 2 mL of acetonitrile, shake well, and centrifuge. To 3.5 mL of the supernatant Amount (mg) of alprostadil (C 20H34O5) ＝ M S × QT/QS liquid add 7 mL of diluted phosphoric acid (1 in 1000), and then run this solution on a column (prepared by filling a 10 MS: Amount (mg) of Alprostadil RS taken Internal standard solution—A solution of dimethyl phtha-late in the mixture of acetonitrile for liquid chromatography and water (9:1) (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 196 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 9.07 g of potassium dihydrogen phosphate in water to make 1000 mL. Adjust the pH to 6.3 with a solution prepared by dissolving 9.46 g of disodium hydrogen phosphate in water to make 1000 mL, and dilute to 10 times its volume with water. To 360 mL of this solution add 110 mL of acetonitrile for liquid chromatography and 30 mL of methanol for liquid chromatography. Flow rate: Adjust so that the retention time of alprostadil is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, alprostadil and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of alprostadil to that of the internal standard is not more than 1.0z.
(1) Heavy metals <1.07>—Proceed with 4.0 mL of Alprostadil Injection according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 5 ppm). Prostaglandin A1—Use the sample solution obtained in the Assay as the sample solution. Separately, weigh accu-rately about 10 mg of prostaglandin A1, previously dried for 4 hours in a desiccator (in vacuum, phosphorus (V) oxide), and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 2.5 mL of this solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add exactly 1 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 40 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, calculate the ratios, QT and QS, of the peak area of prostaglandin A1 to that of the internal standard, and calcu-late the amount of prostaglandin A1 converted to alprostadil using the following equation: not more than 3.0 mg per a volume, equivalent to 5 mg of alprostadil (C20H34O5). JP XVIII Official Monographs / Alprostadil Injection 437 Amount ( mg) of prostaglandin A1 (C20H32O4), converted to alprostadil ＝ MS × QT/QS × 1/2 × 1.054 Bacterial endotoxins <4.01> Less than 10 EU/mL. Extractable volume <6.05> It meets the requirement. MS: Amount (mg) of prostaglandin A1 taken Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Internal standard solution—Dissolve 50 mg of 1-naphthol in 20 mL of ethanol (99.5). To 3 mL of this solution add the mobile phase to make 100 mL. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 5 mL. Confirm that the peak area of prostaglandin A1 ob-tained with 40 mL of this solution is equivalent to 14 to 26z of that with 40 mL of the standard solution. Peroxide—Pipet 4 mL of Alprostadil Injection, place in a glass-stoppered flask, add 15 mL of a mixture of acetic acid (100) and isooctane (3:2), previously having undergone a 30 minute nitrogen substitution, and dissolve with gentle shaking. To this solution add 0.5 mL of saturated potassium iodide TS, replace the inside of the vessel with nitrogen, and shake for exactly 5 minutes. Then, add 0.5 mL of starch TS, shake vigorously, add 15 mL of water, and shake vigorously. Under a stream of nitrogen, titrate <2.50> with 0.01 mol/L sodium thiosulfate VS until the color of the solution disap-pears. Separately, perform a blank determination using 4 mL of water, and make any necessary correction. Calculate the amount of peroxides using the following equation: not more than 0.5 meq/L. Amount (meq/L) of peroxides ＝ V × 2.5 V: Amount (mL) of 0.01 mol/L sodium thiosulfate VS consumed Free fatty acids—Pipet 3 mL of Alprostadil Injec-tion, add exactly 15 mL of a mixture of 2-propanol, heptane and 0.5 mol/L sulfuric acid TS (40:10:1), and shake for 1 minute. After leaving for 10 minutes, add exactly 9 mL of heptane and exactly 9 mL of water, shake the test tube by in-verting 10 times, leave for 15 minutes, and pipet 9 mL of the supernatant liquid. To this solution, add 3 mL of a solution prepared by combining 1 volume of Nile blue solution (1 in 5000) washed 5 times with heptane and 9 volumes of ethanol (99.5), and use this solution as the sample solution. Titrate <2.50> this solution with 0.02 mol/L sodium hydroxide VS under a stream of nitrogen. Separately, dissolve 5.65 g of oleic acid in heptane to make exactly 200 mL, and use this solution as the standard solution. Pipet 25 mL of the stand-ard solution, add 2 drops of phenolphthalein TS, titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS until a light red color develops, and determine the correction fac-tor f. Pipet 30 mL of the standard solution and add heptane to make exactly 200 mL. Pipet 3 mL of this solution, add exactly 15 mL of a mixture of 2-propanol, heptane and 0.5 mol/L sulfuric acid TS (40:10:1), and shake for 1 minute. After leaving for 10 minutes, add exactly 6 mL of heptane and exactly 12 mL of water, shake the test tube by inverting 10 times, and then titrate <2.50> in the same manner as for the sample solution. Determine the volume (mL), VT and VS, of 0.02 mol/L sodium hydroxide VS consumed by the sam-ple and standard solutions: the amount of free fatty acid is not more than 12.0 meq/L. Amount (meq/L) of free fatty acids ＝ VT/VS × f × 15 Sterility <4.06> Perform the test according to the Mem-brane filter method: it meets the requirement. However, use the sample solution consisting of equal volume of Alprostadil Injection and a solution prepared by adding water to 0.1 g of polysorbate 80 to make 100 mL. Particle diameter Being specified separately when the drug is granted approval based on the Law.
Measure exactly a volume of Alprostadil Injection corresponding to 5 mg of alprostadil (C20H34O5), add exactly 1 mL of the internal standard solution, shake, and use this solution as the sample solution. Separately, weigh accurately about 5 mg of Alprostadil RS, previously dried in a desicca-tor (in vacuum, phosphorus (V) oxide) for 4 hours, dissolve in ethanol (99.5) to make exactly 50 mL, and use this solu-tion as standard stock solution. Pipet 2.5 mL of the standard stock solution, add the mobile phase to make exactly 50 mL, pipet 1 mL, add exactly 1 mL of the internal standard solu-tion, and use this solution as the standard solution. Perform the test with 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions using an apparatus equipped with an automatic pretreatment device (using a postcolumn reaction), and calculate the ratios, QT and QS, of the peak area of alprostadil to that of the internal standard. Amount ( mg) of alprostadil (C20H34O5) ＝ MS × QT/QS MS: Amount (mg) of Alprostadil RS taken Internal standard solution—Dissolve 50 mg of 1-naphthol in 20 mL of ethanol (99.5). To 3 mL of this solution add the mobile phase to make 100 mL. Operating conditions— Equipment: Liquid chromatograph consisting of 2 pumps for pumping the mobile phase and the reaction reagent, an automatic pretreatment device, column, reaction coil, detec-tor, and recording apparatus. Use a reaction coil that is maintained at a constant temperature. Detector: An ultraviolet absorption photometer (wave-length: 278 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 609C. Reaction coil: Polytetrafluoroethylene tube 0.5 mm in inside diameter and 10 m in length. Mobile phase: Dissolve 9.07 g of potassium dihydrogen phosphate in water to make 1000 mL and adjust the pH to 6.3 by adding a solution prepared by dissolving 9.46 g of disodium hydrogen phosphate in water to make 1000 mL. To 1 volume of this solution add 9 volumes of water. To 3 volumes of this solution add 1 volume of acetonitrile for liquid chromatography. Reaction reagent: Potassium hydroxide TS. Reaction temperature: A constant temperature of about 609C. Mobile phase flow rate: Adjust so that the retention time of alprostadil is about 7 minutes. Reaction reagent flow rate: 0.5 mL per minute. Automatic pretreatment device: Composed of a pretreat- ment column, pump for pumping pretreatment column wash ‌438 solution, and routing valve for 2 high pressure flow paths. Pretreatment column: A stainless steel column 4 mm in inside diameter and 2.5 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Pretreatment column wash solution: Ethanol (99.5). Flow rate of wash solution: A constant flow rate of about 2.0 mL per minute. Flow path operating conditions: Change the flow path operating conditions at the times shown in the table below using the valves shown in the figure. Time of switchover (minutes) Valve 9.0 9.1 *1) *2) RVA RVB *1) After the internal standard has completely eluted *2) 0.1 minutes after *1) System suitability— System performance: Dissolve 10 mg of prostaglandin A1, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, in ethanol (99.5) to make 100 mL. To 2.5 mL of this solution add 2.5 mL of the standard stock solu-tion, and add the mobile phase to make 50 mL. To 1 mL of this solution add 1 mL of the internal standard solution, shake, and perform the test under the above conditions with 40 mL of the solution. Alprostadil, prostaglandin A1 and the internal standard are eluted in this order, and the resolution between the peaks of alprostadil and prostaglandin A1 is not less than 10, and that between prostaglandin A1 and the in-ternal standard is not less than 2.0. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of alprostadil to that of the internal standard is not more than 2.0z.
Alum Solution is a clear, colorless liquid. It has the odor of the mentha oil and an astringent taste.
(1) To 5 mL of Alum Solution add 3 mL of ammonium chloride TS and 1 mL of ammonia TS: a white, gelatinous precipitate is produced, which changes to red upon the addition of 5 drops of alizarin red S TS (aluminum sulfate). Place 100 mL of Alum Solution in an evaporating dish, evaporate on a water bath to dryness, and dissolve the residue in 5 mL of water: the solution responds to Qualita-tive Tests <1.09> for potassium salt. Alum Solution responds to the Qualitative Tests <1.09> (1) and (2) for sulfate.
Pipet 50 mL of Alum Solution, add exactly 30 mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetra-acetate VS, and further add 20 mL of acetic acid-ammonium acetate buffer solution (pH 4.8). Boil for 5 minutes, cool, add 55 mL of ethanol (95), and titrate <2.50> with 0.02 mol/L zinc acetate VS (indicator: 2 mL of dithizone TS), until the color of the solution changes from light dark green to light red. Perform a blank determination in the same manner. Each mL of 0.02 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS ＝ 9.488 mg of AlK(SO4)2.12H2O
Amikacin Sulfate for Injection is a preparation for injection, which is dissolved before use. It contains not less than 90.
Amikacin Sulfate for Injection occurs as white to yellow-white masses or powder.
Dissolve an amount of Amikacin Sulfate for Injection, equivalent to 25 mg (potency) of Amikacin Sul-fate, in 1 mL of water, and use this solution as the sample solution. Separately, dissolve 25 mg (potency) of Amikacin Sulfate RS in 1 mL of water, and use this solution as the standard solution. Then, proceed as directed in the Identifi-cation (2) under Amikacin Sulfate. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> Dissolve an amount of Amikacin Sulfate for In-jection, equivalent to 0.1 g (potency) of Amikacin Sulfate, in 10 mL of water: the pH of this solution is 6.0 to 7.5.
Clarity and color of solution—Dissolve an amount of Amikacin Sulfate for Injection, equivalent to 0.5 g (po-tency) of Amikacin Sulfate, in 5 mL of water: the solution is clear, and the absorbance at 405 nm of the solution deter-mined as directed under Ultraviolet-visible Spectrophotome-try <2.24> is not more than 0.15. Loss on drying <2.41> Not more than 4.0z (1 g, in vacu-um, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the content of not less than 10 Amikacin Sulfate for Injection. Weigh accurately a portion of the content, equivalent to about 50 mg (potency) of Amikacin Sulfate, dissolve in water to make exactly 50 mL. Separately, weigh accurately an amount of Amikacin Sulfate RS, equivalent to about 50 mg (potency), and dis-solve in water to make exactly 50 mL. Transfer exactly 200 mL each of these solutions to separate glass stoppered tubes, and proceed as directed in the Assay under Amikacin Sul-fate.
Amikacin Sulfate Injection is an aqueous injection. It contains not less than 90.
Amikacin Sulfate Injection occurs as a color-less or pale yellow clear liquid.
To a volume of Amikacin Sulfate Injection, equivalent to 0.1 g (potency) of Amikacin Sulfate, add water to make 4 mL, and use this solution as the sample solution. Separately, dissolve 25 mg (potency) of Amikacin Sulfate RS in 1 mL of water, and use this solution as the standard solu-tion. Then, proceed as directed in the Identifiction (2) under Amikacin Sulfate. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> 6.0 – 7.5 Amount [ mg (potency)] of amikacin (C ＝ MS × HT/HS × 1000 22H43N5O13) Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po- tency). MS: Amount [mg (potency)] of Amikacin Sulfate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 2.72 g of potassium dihydrogen-phosphate in 800 mL of water, adjust to pH 6.5 with a solu-tion of potassium hydroxide (1 in 40), and add water to make 1000 mL. To 280 mL of this solution add 720 mL of methanol, and mix. Flow rate: Adjust so that the retention time of amikacin Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take exactly a volume of Amikacin Sulfate Injec-tion, equivalent to about 0.1 g (potency) of Amikacin Sul-fate, and add water to make exactly 100 mL. Separately, weigh accurately an amount of Amikacin Sulfate RS, equivalent to about 50 mg (potency), and add water to make exactly 50 mL. Take exactly 200 mL each of these solutions into stoppered test tubes, then proceed as directed in the Assay under Amikacin Sulfate. derivative is about 9 minutes. System suitability— System performance: Dissolve about 5 mg (potency) of Amount [mg (potency)] of amikacin (C ＝ MS × HT/HS × 2 22H43N5O13) Amikacin Sulfate and about 5 mg (potency) of Kanamycin Sulfate in 5 mL of water. Transfer 200 mL of this solution in a glass-stoppered test tube, add 3 mL of pyridine and 2 mL of a solution of 2,4,6-trinitrobenzenesulfonic acid (1 in 100), stopper tightly, heat in a water bath at 709C for 30 minutes. After cooling, add 2 mL of acetic acid (100). When the procedure is run with 20 mL of this solution under the above operating conditions, amikacin derivative and kanamycin derivative are eluted in this order with the resolution between MS: Amount [mg (potency)] of Amikacin Sulfate RS taken
Aminophylline Injection is an aqueous injection. It contains not less than 75.
Aminophylline Injection is a clear and color-less liquid. It has a slightly bitter taste. It gradually changes in color by light. pH: 8.0 – 10.0
To a volume of Aminophylline Injection, equivalent to 0.75 g of Aminophylline Hydrate, add water to make 30 mL. Proceed with this solution as directed in the Identification under Aminophylline Hydrate. Bacterial endotoxins <4.01> Less than 0.6 EU/mg.
(1) Theophylline—Pipet a volume of Aminophyl-line Injection, equivalent to about 39.4 mg of theophylline (C7H8N4O2) (about 50 mg of Aminophylline Hydrate), add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of theophylline for assay, previously dried at 1059C for 4 hours, dissolve in water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of theophylline in each solution. Amount (mg) of theophylline (C7H8N4O2) ＝ MS × AT/AS MS: Amount (mg) of theophylline for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and methanol (4:1). Flow rate: Adjust so that the retention time of theophyl-line is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of theophylline are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of theophylline is not more than 1.0z. (2) Ethylenediamine—To an accurately measured volume of Aminophylline Injection, equivalent to about 30 mg of ethylenediamine (C2H8N2) (about 0.2 g of Aminophyl-line Hydrate), add water to make 30 mL, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS (indicator: 2 to 3 drops of bromophenol blue TS). Each mL of 0.1 mol/L hydrochloric acid VS ＝ 3.005 mg of C H N 2 8 2 Description Aminophylline Injection is a clear and color-less liquid. It has a slightly bitter taste. It gradually changes in color by light. pH: 8.0 – 10.0 Identification To a volume of Aminophylline Injection, equivalent to 0.75 g of Aminophylline Hydrate, add water to make 30 mL. Proceed with this solution as directed in the Identification under Aminophylline Hydrate. Bacterial endotoxins <4.01> Less than 0.6 EU/mg.
To 1 mL of the sample stock solution ob-tained in the Assay add the mobile phase to make 50 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 239 nm and 243 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Amiodarone Hydrochloride Tablets add 160 mL of the mobile phase, sonicate for 10 minutes, add the mobile phase to make exactly 200 mL, and centrifuge. Pipet V mL of the supernatant liquid, equivalent to about 1 mg of amiodarone hydrochloride (C25H29I2NO3.HCl), add the mobile phase to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 25 mg of amiodarone hydrochloride for assay, previ-ously dried at 509C for 4 hours under reduced pressure not exceeding 0.3 kPa, and dissolve in the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of amiodarone in each solution. Amount (mg) of amiodarone hydrochloride (C25H29I2NO3.HCl) ＝ MS × AT/AS × 8/V MS: Amount (mg) of amiodarone for assay taken Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amiodarone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amiodarone is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 30 minutes of Amiodarone Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Amiodarone Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add exactly V mL of methanol, then add a mixture of the dissolution medium and methanol (1:1) to make exactly V? mL so that each mL contains about 11 mg of amiodarone hydrochloride (C25H29I2NO3.HCl), and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of amiodarone hydrochloride for assay, previously dried at 509C for 4 hours under reduced pressure not exceeding 0.3 kPa, and dissolve in methanol to make ex-actly 50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the dissolution medium, then add a mixture of the disso-lution medium and methanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand-ard solution at 241 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a mixture of the dissolution medium and methanol (1:1) as the blank. Dissolution rate (z) with respect to the labeled amount of amiodarone hydrochloride (C25H29I2NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of amiodarone hydrochloride for assay taken C: Labeled amount (mg) of amiodarone hydrochloride (C25H29I2NO3.HCl) in 1 tablet
Weigh accurately the mass of not less than 20 Amio-darone Hydrochloride Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 50 mg of amiodarone hydrochloride (C25H29I2NO3.HCl), add 80 mL of the mobile phase, sonicate for 10 minutes, and add the ‌452 mobile phase to make exactly 100 mL. Centrifuge this solu-tion, and use the supernatant liquid as the sample stock solu-tion. Pipet 2 mL of the stock solution, add exactly 2 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately 25 mg of amiodarone hydrochloride for assay, previously dried at 509C for 4 hours under reduced pressure not exceeding 0.3 kPa, and dissolve in the mobile phase to make exactly 50 mL. Pipet 2 mL of this so-lution, add exactly 2 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of amiodarone to that of the internal standard. Amount (mg) of amiodarone hydrochloride (C25H29I2NO3.HCl) ＝ MS × QT/QS × 2 MS: Amount (mg) of amiodarone hydrochloride for assay taken Internal standard solution—A solution of chlorhexidine hy-drochloride in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of acetonitrile for liquid chroma-tography, a solution of sodium laurylsulfate (1 in 50) and phosphoric acid (750:250:1). Flow rate: Adjust so that the retention time of amioda-rone is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and amiodarone are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of amiodarone to that of the internal standard is not more than 1.0z.
Amitriptyline Hydrochloride Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of amitriptyline hydrochloride (C20H23N.HCl: 313.86). Method of preparation Prepare as directed under Tablets, with Amitriptyline Hydrochloride. Identification (1) Weigh a quantity of powdered Ami-triptyline Hydrochloride Tablets, equivalent to 0.1 g of Ami-triptyline Hydrochloride. Add 10 mL of chloroform, shake thoroughly, and filter. Evaporate the filtrate on a water bath to about 2 mL, add diethyl ether until turbidity is produced, and allow to stand. Filter the crystals formed through a glass filter (G4), and proceed as directed in the Identification (1) and (2) under Amitriptyline Hydrochloride. (2) Determine the absorption spectrum of a solution of the crystals obtained in (1) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 238 nm and 240 nm, and a minimum be-tween 228 nm and 230 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Amitriptyline Hydrochloride Tablets add 50 mL of diluted methanol (1 in 2), shake to disintegrate the tablet, then add diluted methanol (1 in 2) to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 10 mg of amitriptyline hydrochloride (C20H23N.
(1) Weigh a quantity of powdered Ami-triptyline Hydrochloride Tablets, equivalent to 0.1 g of Ami-triptyline Hydrochloride. Add 10 mL of chloroform, shake thoroughly, and filter. Evaporate the filtrate on a water bath to about 2 mL, add diethyl ether until turbidity is produced, and allow to stand. Filter the crystals formed through a glass filter (G4), and proceed as directed in the Identification (1) and (2) under Amitriptyline Hydrochloride. (2) Determine the absorption spectrum of a solution of the crystals obtained in (1) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 238 nm and 240 nm, and a minimum be-tween 228 nm and 230 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Amitriptyline Hydrochloride Tablets add 50 mL of diluted methanol (1 in 2), shake to disintegrate the tablet, then add diluted methanol (1 in 2) to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 10 mg of amitriptyline hydrochloride (C20H23N.HCl), and use this so-lution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of amitriptyline hydrochloride (C20H23N.HCl) ＝ MS × AT/AS × V?/V × 1/20 MS: Amount (mg) of Amitriptyline Hydrochloride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Amitriptyline Hydrochloride Tablets is not less than 70z. Start the test with 1 tablet of Amitriptyline Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL of the filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of amitriptyline hydrochloride (C20H23N.HCl), and use this solution as the sample solution. Separately, weigh accurately about 55 mg of Amitriptyline Hydrochloride RS, previously dried at 1059C for 2 hours, and dissolve in the dissolution medium to make exactly 250 mL. Pipet 5 mL of this solu-tion, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand- ard solution at 239 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of amitriptyline hydrochloride (C20H23N.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Amitriptyline Hydrochloride RS taken C: Labeled amount (mg) of amitriptyline hydrochloride (C20H23N.HCl) in 1 tablet
Weigh accurately and powder not less than 20 Amitriptyline Hydrochloride Tablets. Weigh accurately a portion of the powder, equivalent to about 20 mg of ami-triptyline hydrochloride (C20H23N.HCl), and add 75 mL of diluted methanol (1 in 2). After shaking for 30 minutes, add diluted methanol (1 in 2) to make exactly 100 mL, and filter. Discard the first 20-mL portion of the filtrate, measure exactly the subsequent 5-mL portion, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Amitriptyline Hydrochloride RS, previously dried at 1059C for 2 hours, and dissolve in diluted methanol (1 in 2) to make exactly 100 mL. Measure exactly 5 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 239 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, respec-tively. Amount (mg) of amitriptyline hydrochloride (C20H23N.HCl) ＝ MS × AT/AS MS: Amount (mg) of Amitriptyline Hydrochloride RS taken
Amlexanox Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of amlexanox (C16H14N2O4: 298.29). Method of preparation Prepare as directed under Tablets, with Amlexanox. Identification (1) Take an amount of powdered Amlexa-nox Tablets, equivalent to 10 mg of Amlexanox, add 100 mL of ethanol (99.5), shake vigorously, and filter. Pipet 1 mL of the filtrate, add 25 mL of ethanol (99.5), and use this solu-tion as the sample solution. Determine the absorption spec-trum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits absorption maxima between 240 nm and 244 nm, between 285 nm and 289 nm, and between 341 nm and 352 nm. (2) Observe the sample solution obtained in (1) under ultraviolet light (main wavelength: 365 nm): the solution shows a blue-white fluorescence. MS: Amount (mg) of Amlexanox RS taken Internal standard solution—A solution of 3-nitroaniline in the mobile phase (1 in 500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Amlexanox Tablets is not less than 80z. Start the test with 1 tablet of Amlexanox Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.
(1) Take an amount of powdered Amlexa-nox Tablets, equivalent to 10 mg of Amlexanox, add 100 mL of ethanol (99.5), shake vigorously, and filter. Pipet 1 mL of the filtrate, add 25 mL of ethanol (99.5), and use this solu-tion as the sample solution. Determine the absorption spec-trum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits absorption maxima between 240 nm and 244 nm, between 285 nm and 289 nm, and between 341 nm and 352 nm. (2) Observe the sample solution obtained in (1) under ultraviolet light (main wavelength: 365 nm): the solution shows a blue-white fluorescence. MS: Amount (mg) of Amlexanox RS taken Internal standard solution—A solution of 3-nitroaniline in the mobile phase (1 in 500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Amlexanox Tablets is not less than 80z. Start the test with 1 tablet of Amlexanox Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of amlexanox (C16H14N2O4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Amlexanox RS, previously dried at 1059C for 2 hours, and dissolve in 2 mL of dilute sodium hydroxide TS, add the dissolution medium to make exactly 50 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, at 350 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Dissolution rate (z) with respect to the labeled amount of amlexanox (C16H14N2O4) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Amlexanox RS taken C: Labeled amount (mg) of amlexanox (C16H14N2O4) in 1 tablet
Weigh accurately not less than 20 Amlexanox Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 15 mg of amlexanox (C16H14N2O4), add exactly 10 mL of the internal standard so-lution, add 80 mL of the mobile phase, shake vigorously for 5 minutes, and then add the mobile phase to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 30 mg of Amlexanox RS, previously dried at 1059C for 2 hours, and dissolve in the mobile phase to make exactly 50 mL. Pipet 25 mL of this solution, add exactly 10 mL of the inter-nal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Amlexanox. Uniformity of dosage units <6.02> Perform the Mass varia- tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Amount (mg) of amlexanox (C ＝ MS × QT/QS × 1/2 16H14N2O4) Take 1 tablet of Amlexanox Tablets, add exactly 0.6 mL of the internal standard solution per 1 mg of amlexanox (C16H14N2O4), add the mobile phase to make exactly V mL so there is about 167 mg of amlexanox (C16H14N2O4) per mL, disintegrate the tablet, and then shake vigorously for 5 minutes. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 30 mg of Amlexanox RS, previously dried at 1059C for 2 hours, and dissolve in the mobile phase to make exactly MS: Amount (mg) of Amlexanox RS taken Internal standard solution—A solution of 3-nitroaniline in the mobile phase (1 in 500).
Amlodipine Besilate Orally Disintegrating Tablets
アムロジピンベシル酸塩口腔内崩壊錠
To an amount of powdered Amlodipine Besilate Orally Disintegrating Tablets, equivalent to 7 mg of Amlodipine Besilate, add 200 mL of 0.01 mol/L hydrochlo-ric acid-methanol TS, treat with ultrasonic waves, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 358 nm and 362 nm.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and the mobile phase A (3:2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- having the relative retention time of about 4.5 to amlodipine from the sample solution is not larger than 1.8 times the peak area of amlodipine from the standard solution, and the area of the peak having the relative retention time of about 0.16 to amlodipine and the peaks other than mentioned above from the sample solution is not larger than 2/5 times the peak area of amlodipine from the standard solution. Furthermore, the total area of the peaks other than amlodi-pine and the peak having the relative retention time of about 0.16 to amlodipine from the sample solution is not larger than 2.8 times the peak area of amlodipine from the stand-ard solution. For the areas of the peaks, having the relative retention time of about 0.45 and about 4.5 to amlodipine, multiply their correction factors, 2.0 and 1.9, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 4.1 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 6.0 with a solu-tion of 5.4 g of disodium hydrogen phosphate dodecahy-drate in 500 mL of water. To 500 mL of this solution add 500 mL of methanol. Mobile phase B: Dissolve 4.1 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 6.0 with a solu-tion of 5.4 g of disodium hydrogen phosphate dodecahy-drate in 500 mL of water. To 50 mL of this solution add 950 mL of methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: Adjust so that the retention time of amlodipine is about 10 minutes. Time span of measurement: About 5 times as long as the retention time of amlodipine. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and the mobile phase A (3:2) to make exactly 50 mL. Confirm that the peak area of amlodipine obtained with 30 mL of this solution is equivalent to 14 to 26z of that with 30 mL of the standard solution. System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amlodipine are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlodipine is not more than 2.0z. ‌458 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Amlodipine Besilate Orally Disintegrating Tablets add 4V/5 mL of a mixture of the mobile phase and methanol (1:1), disperse the particles by sonicating, add a mixture of the mobile phase and methanol (1:1) to make exactly V mL so that each mL of the solution contains about 0.14 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S). Centrifuge this solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × AT/AS × V × 1/250 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Disintegration Being specified separately when the drug is granted approval based on the Law. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Am-lodipine Besilate Orally Disintegrating Tablets, and powder them. Weigh accurately a portion of this powder, equivalent to about 7 mg of amlodipine besilate (C20H25ClN2O5. C6H6O3S), add 40 mL of a mixture of the mobile phase and methanol (1:1), disperse the particles with the aid of ultra-sonic waves, and add a mixture of the mobile phase and methanol (1:1) to make exactly 50 mL. Centrifuge this solu-tion, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 35 mg of Amlodipine Besilate RS (separately, determine the water <2.48> in the same manner as Amlodipine Besilate), add 150 mL of a mix-ture of the mobile phase and methanol (1:1), dissolve with the aid of ultrasonic waves, then add a mixture of the mobile phase and methanol (1:1) to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 30 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of amlodipine in each solution. Amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.1 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 6.0 with a solu-tion of 5.4 g of disodium hydrogen phosphate dodecahy-drate in 500 mL of water. To 400 mL of this solution add 600 mL of methanol. Flow rate: Adjust so that the retention time of amlodipine is about 10 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con- ditions, the number of theoretical plates and the symmetry factor of the peak of amlodipine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlodipine is not more than 1.0z.
Amlodipine Besilate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of amlodipine besilate (C20H25ClN2O5. C6H6O3S: 567.05). Method of preparation Prepare as directed under Tablets, with Amlodipine Besilate. Identification To a quantity of powdered Amlodipine Besi-late Tablets, equivalent to 2.5 mg of Amlodipine Besilate, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake vigorously, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, and between 358 nm and 362 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Amlodipine Besilate Tablets add 10 mL of water to disintegrate, disperse by sonicating with occasional shaking, add the mobile phase to make exactly V mL so that each mL contains about 69 mg of amlodipine besilate (C20H25ClN2O5.
To a quantity of powdered Amlodipine Besi-late Tablets, equivalent to 2.5 mg of Amlodipine Besilate, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake vigorously, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, and between 358 nm and 362 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Amlodipine Besilate Tablets add 10 mL of water to disintegrate, disperse by sonicating with occasional shaking, add the mobile phase to make exactly V mL so that each mL contains about 69 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S), and shake for 60 minutes. Centri-fuge this solution, pipet 10 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × QT/QS × V/500 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of isobutyl para-hydroxybenzoate in the mobile phase (3 in 20,000). Dissolution Being specified separately when the drug is granted approval based on the Law.
To 20 Amlodipine Besilate Tablets add 100 mL of water to disintegrate, disperse with the aid of ultrasonic waves with occasional shaking, add the mobile phase to make exactly 1000 mL, and shake for 60 minutes. Centrifuge this solution, pipet a volume of the supernatant liquid, equivalent to about 0.7 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S), add exactly 5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 35 mg of Amlodipine Besilate RS (sepa-rately, determine the water <2.48> in the same manner as Amlodipine Besilate), and dissolve in the mobile phase to make exactly 250 mL. Pipet 5 mL of this solution, add ex-actly 5 mL of the internal standard solution, add the mobile ‌JP XVIII Official Monographs / Amobarbital 459‌ phase to make 25 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of am-lodipine to that of the internal standard. Amount (mg) of amlodipine besilate (C H ClN O .C H O S) to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 2.5 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 5 ppm). Potassium permanganate-reducing substances—To 10.0 mL of Ammonia Water add 40 mL of dilute sulfuric acid while cooling, and add 0.10 mL of 0.02 mol/L potas-sium permanganate VS: the red color of the potassium per- 20 25 2 5 6 6 3 manganate does not disappear within 10 minutes. ＝ MS × QT/QS × 1/50 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of isobutyl para-hydroxybenzoate in the mobile phase (3 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and potassium dihy-drogen phosphate (41 in 10,000) (13:7). Flow rate: Adjust so that the retention time of amlodipine is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, amlodipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of amlodipine to that of the internal stand-ard is not more than 1.0z.
Amosulalol Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of amosulalol hydrochloride (C18H24N2O5S.HCl: 416.92). Method of preparation Prepare as directed under Tablets, with Amosulalol Hydrochloride. Identification To a quantity of powdered Amosulalol Hydrochloride Tablets, equivalent to 50 mg of Amosulalol Hydrochloride, add 25 mL of 0.1 mol/L hydrochloric acid TS, shake well, and then centrifuge. To 2.5 mL of the super-natant liquid add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 270 nm and 274 nm, and a shoulder between 275 nm and 281 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Amosulalol Hydrochloride Tablets, disin-tegrate by adding 2 mL of 0.1 mol/L hydrochloric acid TS, add 15 mL of methanol, and shake well. Add methanol to make exactly V mL so that each mL contains about 0.
To a quantity of powdered Amosulalol Hydrochloride Tablets, equivalent to 50 mg of Amosulalol Hydrochloride, add 25 mL of 0.1 mol/L hydrochloric acid TS, shake well, and then centrifuge. To 2.5 mL of the super-natant liquid add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 270 nm and 274 nm, and a shoulder between 275 nm and 281 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Amosulalol Hydrochloride Tablets, disin-tegrate by adding 2 mL of 0.1 mol/L hydrochloric acid TS, add 15 mL of methanol, and shake well. Add methanol to make exactly V mL so that each mL contains about 0.4 mg of amosulalol hydrochloride (C18H24N2O5S.HCl), and centri-fuge. Pipet 5 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution and the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of amosulalol hydrochloride for assay (separately determine the water <2.48> in the same manner as Amosulalol Hydrochloride), and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of amosulalol hydrochloride (C18H24N2O5S.HCl) ＝ MS × QT/QS × V/50 MS: Amount (mg) of amosulalol hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl para-hydroxybenzoate in methanol (1 in 6250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Amosulalol Hydrochloride Tablets is not less than 75z. Start the test with 1 tablet of Amosulalol Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.5 mg of amosulalol hydro-chloride (C18H24N2O5S.HCl), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of amosulalol hydrochloride for assay (separately determine the water <2.48> in the same manner as Amosulalol Hydro-chloride), and dissolve in water to make exactly 200 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the amosulalol peak areas, AT and AS, in each solution. Dissolution rate (z) with respect to the labeled amount of amosulalol hydrochloride (C18H24N2O5S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 45/2 MS: Amount (mg) of amosulalol hydrochloride for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of amosulalol hydrochloride (C18H24N2O5S.HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.7 by adding a soluion prepared by dissolving 3.58 g of disodi-um hydrogen phosphate dodecahydrate in water to make 1000 mL. To 670 mL of this solution add 330 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of amosulalol is about 5 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amosulalol are not less than 4000 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times ‌462 with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amosulalol is not more than 1.0z.
Take 10 Amosulalol Hydrochloride Tablets, add 20 mL of 0.1 mol/L hydrochloric acid TS, and shake well to disintegrate. Add 120 mL of methanol, again shake well, add methanol to make exactly 200 mL, and then centrifuge. Pipet a volume of supernatant liquid corresponding to about 5 mg of amosulalol hydrochloride (C18H24N2O5S.HCl), add exactly 5 mL of the internal standard solution, add the mo-bile phase to make 50 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of amosulalol hydrochloride for assay (separately determine the water <2.48> in the same manner as Amosulalol Hydrochlo-ride), and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand- Amoxapine
Amoxicillin Capsules contain not less than 92.0z and not more than 105.0z of the labeled potency of Amoxicillin (C16H19N3O5S: 365.40). Method of preparation Prepare as directed under Cap-sules, with Amoxicillin Hydrate. Identification Take out the contents of Amoxicillin Cap-sules, to a quantity of the contents, equivalent to 8 mg (potency) of Amoxicillin Hydrate, add 2 mL of 0.01 mol/L hydrochloric acid TS, shake for 30 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve an amount equivalent to 8 mg (potency) of Amoxicillin RS in 2 mL of 0.01 mol/L hydrochloric acid TS, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of tetrahydrofuran, water and formic acid (50:5:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (95) (1 in 20) on the plate, and heat the plate at 1109C for 15 minutes: the principal spot obtained from the sample solu-tion and the spot from the standard solution show a red-purple color and the same Rf value. Purity Related substances—Take out the contents of Amoxicillin Capsules, to a quantity of the contents, equiva-lent to 0.1 g (potency) of Amoxicillin Hydrate, add 30 mL of a solution of boric acid (1 in 200), shake for 15 minutes, and add a solution of boric acid (1 in 200) to make 50 mL. Cen-trifuge this solution, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add a solution of boric acid (1 in 200) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than amoxicillin obtained from the sample solution is not larger than the peak area of amoxicillin from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Amoxicillin Hydrate. System suitability— Test for required detectability and system repeatability: Proceed as directed in the system suitability in the Purity (3) under Amoxicillin Hydrate. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amoxicillin is not less than 2500 and not more than 1.5, respectively. Water <2.48> Not more than 15.0z (0.1 g, volumetric titration, direct titration). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Amoxicillin Capsules is not less than 75z. Start the test with 1 capsule of Amoxicillin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg (potency) of Amoxicillin Hydrate, and use this solution as the sample solution.
Amphotericin B occurs as a yellow to orange powder. It is freely soluble in dimethylsulfoxide and practically insoluble in water and in ethanol (95).
Take out the contents of Amoxicillin Cap-sules, to a quantity of the contents, equivalent to 8 mg (potency) of Amoxicillin Hydrate, add 2 mL of 0.01 mol/L hydrochloric acid TS, shake for 30 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve an amount equivalent to 8 mg (potency) of Amoxicillin RS in 2 mL of 0.01 mol/L hydrochloric acid TS, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of tetrahydrofuran, water and formic acid (50:5:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (95) (1 in 20) on the plate, and heat the plate at 1109C for 15 minutes: the principal spot obtained from the sample solu-tion and the spot from the standard solution show a red-purple color and the same Rf value.
Related substances—Take out the contents of Amoxicillin Capsules, to a quantity of the contents, equiva-lent to 0.1 g (potency) of Amoxicillin Hydrate, add 30 mL of a solution of boric acid (1 in 200), shake for 15 minutes, and add a solution of boric acid (1 in 200) to make 50 mL. Cen-trifuge this solution, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add a solution of boric acid (1 in 200) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than amoxicillin obtained from the sample solution is not larger than the peak area of amoxicillin from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Amoxicillin Hydrate. System suitability— Test for required detectability and system repeatability: Proceed as directed in the system suitability in the Purity (3) under Amoxicillin Hydrate. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amoxicillin is not less than 2500 and not more than 1.5, respectively. Water <2.48> Not more than 15.0z (0.1 g, volumetric titration, direct titration). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Amoxicillin Capsules is not less than 75z. Start the test with 1 capsule of Amoxicillin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg (potency) of Amoxicillin Hydrate, and use this solution as the sample solution. Separately, weigh accurately an amount equivalent to about 28 mg (potency) of Amoxicillin RS, dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amoxicillin in each solution. Dissolution rate (z) with respect to the labeled amount of amoxicillin (C16H19N3O5S) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount [mg (potency)] of Amoxicillin RS taken C: Labeled amount [mg (potency)] of amoxicillin (C16H19N3O5S) in 1 capsule Operating conditions— Proceed as directed in the operating conditions in the Assay under Amoxicillin Hydrate. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amoxicillin are not less than 2500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amoxicillin is not more than 1.5z.
Weigh accurately the mass of not less than 10 Amoxicillin Capsules, take out the contents, and weigh accu- JP XVIII Official Monographs / Amphotericin B 465 ‌rately the mass of the emptied shells. Weigh accurately an amount equivalent to about 0.1 g (potency) of Amoxicillin Hydrate, add 70 mL of water, shake for 15 minutes, and add water to make exactly 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately an amount equivalent to about 20 mg (po-tency) of Amoxicillin RS, dissolve in water to make exactly 20 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amoxicillin in each solution. It contains not less than 840 mg (potency) per mg, calculated on the dried basis. The potency of Am-photericin B is expressed as mass (potency) of am-photericin B (C47H73NO17). Description Amphotericin B occurs as a yellow to orange powder. It is freely soluble in dimethylsulfoxide and practically insoluble in water and in ethanol (95). Identification (1) Dissolve 5 mg of Amphotericin B in 10 mL of dimethylsulfoxide. To 1 mL of this solution add 5 mL of phosphoric acid: a blue color develops between the two layers, and the solution becomes blue by shaking. After ad- Amount [mg (potency)] of amoxicillin (C ＝ MS × AT/AS × 5 16H19N3O5S) dition of 15 mL of water it becomes yellow to light yellow-brown by shaking. (2) Dissolve 25 mg of Amphotericin B in 5 mL of MS: Amount [mg (potency)] of Amoxicillin RS taken Operating conditions— Column temperature, mobile phase, and flow rate: Pro-ceed as directed in the operating conditions in the Assay under Amoxicillin Hydrate. Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amoxicillin are not less than 2500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of peak areas of amoxicillin is not more than 1.0z.
Amphotericin B for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Amphotericin B for Injection occurs as yellow to orange, powder or masses.
To an amount of Amphotericin B for Injec-tion, equivalent to 25 mg (potency) of Amphotericin B, add 5 mL of dimethylsulfoxide and 45 mL of methanol, and shake. To 1 mL of this solution add methanol to make 50 mL, and filter if necessary. Determine the absorption spec-trum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 361 nm and 365 nm, between 380 nm and 384 nm and between 403 nm and 407 nm. pH <2.54> Dissolve an amount of Amphotericin B for In-jection, equivalent to 50 mg (potency) of Amphotericin B, in 10 mL of water. To 1 mL of this solution add water to make 50 mL: 7.2 – 8.0.
Clarity and color of solution—Dissolve an amount of Amphotericin B for Injection, equivalent to 50 mg (po-tency) of Amphotericin B, in 10 mL of water: the solution is clear and yellow to orange. Loss on drying <2.41> Not more than 8.0z (0.3 g, in vacu- um, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 3.0 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: 105.0z). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, preparation of cylin-der-agar plate and standard solutions—Proceed as directed in the Assay under Amphotericin B. Sample solutions—Prepare using light-resistant con-tainers. Weigh accurately an amount of Amphotericin B for Injection, equivalent to about 50 mg (potency), dissolve in dimethylsulfoxide to make exactly 50 mL, and use this solu-tion as the sample stock solution. Measure exactly a suitable quantity of the sample stock solution, add dimethylsulfoxide to make solutions so that each mL contains about 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these so-lutions, add 0.2 mol/L phosphate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Containers—Hermetic containers. Storage—Light-resistant, at a cold place.
Amphotericin B Syrup contain not less than 90.0z and not more than 115.0z of the labeled potency of amphotericin B (C47H73NO17: 924.08). Method of preparation Prepare as directed under Syrup, with Amphotericin B. Identification To an amount of Amphotericin B Syrup, equivalent to 25 mg (potency) of Amphotericin B, add 5 mL of dimethylsulfoxide and 45 mL of methanol, and shake. To 1 mL of this solution add methanol to make 50 mL, and filter, if necessary. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 361 nm and 365 nm, between 380 nm and 384 nm and between 403 nm and 407 nm. pH <2.54> 5.0 – 7.0 Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/mL and 5 × 101 CFU/mL, respec-tively. Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, preparation of cylin-der-agar plate and standard solutions—Proceed as directed ‌JP XVIII Official Monographs / Anhydrous Ampicillin 467‌ in the Assay under Amphotericin B. Sample solutions—Prepare using light-resistant con-tainers. Weigh accurately an amount of Amphotericin B Syrup, equivalent to about 0.1 g (potency), add about 70 mL of dimethylsulfoxide, shake, then add dimethylsulfoxide to make exactly 100 mL, and use this solution as the sample stock solution. Measure exactly a suitable amount of the sample stock solution, add dimethylsulfoxide to make solu-tions so that each mL contains about 200 mg (potency) and 50 mg (potency).
To an amount of Amphotericin B Syrup, equivalent to 25 mg (potency) of Amphotericin B, add 5 mL of dimethylsulfoxide and 45 mL of methanol, and shake. To 1 mL of this solution add methanol to make 50 mL, and filter, if necessary. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 361 nm and 365 nm, between 380 nm and 384 nm and between 403 nm and 407 nm. pH <2.54> 5.0 – 7.0 Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/mL and 5 × 101 CFU/mL, respec-tively.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, preparation of cylin-der-agar plate and standard solutions—Proceed as directed ‌JP XVIII Official Monographs / Anhydrous Ampicillin 467‌ in the Assay under Amphotericin B. Sample solutions—Prepare using light-resistant con-tainers. Weigh accurately an amount of Amphotericin B Syrup, equivalent to about 0.1 g (potency), add about 70 mL of dimethylsulfoxide, shake, then add dimethylsulfoxide to make exactly 100 mL, and use this solution as the sample stock solution. Measure exactly a suitable amount of the sample stock solution, add dimethylsulfoxide to make solu-tions so that each mL contains about 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these solutions, add 0.2 mol/L phosphate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration sample solution and low concentration sample solution, re-spectively.
Amphotericin B Tablets contain not less than 90.0z and not more than 120.0z of the labeled potency of amphotericin B (C47H73NO17: 924.08). Method of preparation Prepare as directed under Tablets, with Amphotericin B. Identification To an amount of pulverized Amphotericin B Tablets, equivalent to 25 mg (potency) of Amphotericin B, add 5 mL of dimethylsulfoxide and 45 mL of methanol, and shake. To 1 mL of this solution add methanol to make 50 mL, and filter, if necessary. Determine the absorption spec-trum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 361 nm and 365 nm, between 380 nm and 384 nm and between 403 nm and 407 nm. Loss on drying <2.41> Not more than 5.0z (0.3 g, in vacu-um, 609C, 3 hours). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: 105.0z). Disintegration <6.09> Perform the test using the disk: it meets the requirement. Assay Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, preparation of cylin-der-agar plate and standard solutions—Proceed as directed in the Assay under Amphotericin B. Sample solutions—Prepare using light-resistant con-tainers. Weigh accurately and powder not less than 20 tablets of Amphotericin B Tablets. Weigh accurately a part of the powder, equivalent to about 0.1 g (potency), add about 70 mL of dimethylsulfoxide, shake, then add dimethylsulfoxide to make exactly 100 mL, centrifuge, and use the supernatant liquid as the sample stock solution. Measure exactly a suita-ble amount of the sample stock solution, add dimethylsul-foxide to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency).
To an amount of pulverized Amphotericin B Tablets, equivalent to 25 mg (potency) of Amphotericin B, add 5 mL of dimethylsulfoxide and 45 mL of methanol, and shake. To 1 mL of this solution add methanol to make 50 mL, and filter, if necessary. Determine the absorption spec-trum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 361 nm and 365 nm, between 380 nm and 384 nm and between 403 nm and 407 nm. Loss on drying <2.41> Not more than 5.0z (0.3 g, in vacu-um, 609C, 3 hours). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: 105.0z). Disintegration <6.09> Perform the test using the disk: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, preparation of cylin-der-agar plate and standard solutions—Proceed as directed in the Assay under Amphotericin B. Sample solutions—Prepare using light-resistant con-tainers. Weigh accurately and powder not less than 20 tablets of Amphotericin B Tablets. Weigh accurately a part of the powder, equivalent to about 0.1 g (potency), add about 70 mL of dimethylsulfoxide, shake, then add dimethylsulfoxide to make exactly 100 mL, centrifuge, and use the supernatant liquid as the sample stock solution. Measure exactly a suita-ble amount of the sample stock solution, add dimethylsul-foxide to make solutions so that each mL contains 200 mg (potency) and 50 mg (potency). Pipet 1 mL each of these so-lutions, add 0.2 mol/L phosphate buffer solution (pH 10.5) to make exactly 20 mL, and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Ampicillin Sodium and Sulbactam Sodium for Injection
注射用アンピシリンナトリウム･スルバクタムナトリウム
Ampicillin Sodium and Sulbactam Sodium for Injection occurs as a white to yellowish white powder.
(1) The retention times of ampicillin ob-tained from the sample solution and the standard solution observed in the Assay are the same, and the peak area of am-picillin observed in the Assay obtained from the sample solu-tion is 2.8 to 3.6 times the peak area of ampicillin observed in the test performed with 10 mL of the sample solution ob-tained in the Assay as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. (2) The retention times of sulbactam obtained from the sample solution and the standard solution observed in the Assay are the same, and the peak area of sulbactam observed in the Assay obtained from the sample solution is 2.0 to 2.6 times the peak area of sulbactam observed in the test per-formed with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. pH <2.54> The pH of a solution prepared by dissolving an amount of Ampicillin Sodium and Sulbactam Sodium for Injection, equivalent to 1.0 g (potency) of ampicillin (C16H19N3O4S), in 10 mL of water is between 8.0 and 10.0.
(1) Clarity and color of solution—Dissolve an amount of Ampicillin Sodium and Sulbactam Sodium for Injection, equivalent to 1.0 g (potency) of ampicillin (C16H19N3O4S), in 10 mL of water: the solution is clear. Determine the absorption of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absor-bance at 425 nm is not more than 0.10. (2) Total penicilloic acid—Weigh accurately about 25 mg Amount [mg (potency)] of sulbactam (C8H11NO5S) ＝ MS2 × QTb/QSb × V/10 MS1: Amount [mg (potency)] of Ampicillin RS taken MS2: Amount [mg (potency)] of Sulbactam RS taken Internal standard solution—A solution of parahydroxyben-zoic acid in the mobile phase (1 in 1000). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Ampicillin Sodium and Sulbactam Sodium for Injection. Weigh accurately an amount of a por-tion of the contents, equivalent to about 0.25 g (potency) of ampicillin (C16H19N3O4S), and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add ex-actly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Ampicil-lin RS, equivalent to about 50 mg (potency), and an amount of Sulbactam RS, equivalent to about 25 mg (potency), dis-solve in the mobile phase, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QTa and QTb, of the peak areas of ampicillin and sulbactam to that of the internal standard obtained from the sample solution, and the ratios, QSa and QSb, of the peak areas of ampicillin and sulbactam to that of the internal standard from the standard solution. of Ampicillin Sodium and Sulbactam Sodium for Injection, Amount [mg (potency)] of ampicillin (C H N O S) place in a glass-stoppered flask, dissolve in 25 mL of 0.02 mol/L phosphate buffer solution (pH 3.0), add exactly 5 mL ＝ MS1 × QTa/QSa × 5 16 19 3 4 of 0.005 mol/L iodine VS, stopper the flask, allow to stand for 5 minutes, and titrate <2.50> with 0.005 mol/L sodium thiosulfate VS (indicator: 1.0 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction: the amount of total penicilloic acid (as C16H21N3O5S: 367.42) is not more than 3.0z. Each mL of 0.005 mol/L sodium thiosulfate VS ＝ 0.2064 mg of C16H21N3O5S Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test (T: 105.0z). Dissolve 1 Ampicillin Sodium and Sulbactam Sodium for Injection in the mobile phase to make exactly V mL so that each mL contains 5 mg (potency) of ampicillin (C16H19N3O4S). Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 50 mL, and use this solution as the sample so-lution. Then, proceed as directed in the Assay. Amount [mg (potency)] of sulbactam (C8H11NO5S) ＝ MS2 × QTb/QSb × 5 MS1: Amount [mg (potency)] of Ampicillin RS taken MS2: Amount [mg (potency)] of Sulbactam RS taken Internal standard solution—A solution of parahydroxyben-zoic acid in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of 0.02 mol/L phosphate buffer (pH 3.0) and acetonitrile for liquid chromatography (23:2). Flow rate: Adjust so that the retention time of the internal standard is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sulbactam, the internal standard and ampicillin are eluted in this order, and either resolution between these Amount [mg (potency)] of ampicillin (C ＝ MS1 × QTa/QSa × V/10 16H19N3O4S) peaks is not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌JP XVIII Official Monographs / Ampiroxicam Capsules 473‌ ing conditions, the relative standard deviation of the peak area of sulbactam is not more than 1.0z. Containers and storage—Hermetic containers. Plastic con-tainers for aqueous injections may be used. Ampiroxicam
Ampicillin Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Ampicillin Sodium for Injection occurs as white to light yellow-white, crystals or crystalline powder.
Proceed as directed in the Identification (1) under Ampicillin Sodium. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> The pH of a solution prepared by dissolving an amount of Ampicillin Sodium for Injection, equivalent to 1.0 g (potency) of Ampicillin Sodium, in 10 mL of water is 8.0 to 10.0.
Clarity and color of solution—Dissolve an amount of Ampicillin Sodium for Injection, equivalent to 0.25 g (potency) of Ampicillin Sodium, in 0.75 mL of water: the solution is clear. Perform the test with the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance at 400 nm is not more than 0.40. Water <2.48> Not more than 3.0z (0.2 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.075 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Ampicillin Sodium for Injection. Weigh accurately an amount of a portion of the contents, equivalent to about 50 mg (potency) of Ampicillin Sodium, add exactly 5 mL of the internal standard solution and dis-solve. Then add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Ampicillin RS, equivalent to about 50 mg (potency), add exactly 5 mL of the internal standard so-lution and dissolve. Then add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ampicillin to that of the internal standard. Amount [mg (potency)] of ampicillin (C16H19N3O4S) ＝ MS × QT/QS MS: Amount [mg (potency)] of Ampicillin RS taken Internal standard solution—A solution of guaifenesin in the mobile phase (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.94 mg of diammonium hydrogen phosphate in 850 mL of water, add 100 mL of acetonitrile, add phosphoric acid to adjust the pH to 5.0, then add water to make exactly 1000 mL. Flow rate: Adjust so that the retention time of ampicillin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ampicillin and the internal standard are eluted in this order with the resolution between these peaks being not less than 26. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ampicillin to that of the internal standard is not more than1.0z.
Ampiroxicam Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of ampiroxicam (C20H21N3O7S: 447.46). Method of Preparation Prepare as directed under Cap-sules, with Ampiroxicam. Identification Take out the contents of Ampiroxicam Cap-sules, to a quantity of the contents, equivalent to 10 mg of Ampiroxicam, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake well, and centrifuge. To 5 mL of the su-pernatant liquid add 0.01 mol/L hydrochloric acid-methanol TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 318 nm and 322 nm. ‌Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Ampiroxicam Cap-sules, add acetonitrile to make exactly V mL so that each mL contains about 0.
Take out the contents of Ampiroxicam Cap-sules, to a quantity of the contents, equivalent to 10 mg of Ampiroxicam, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake well, and centrifuge. To 5 mL of the su-pernatant liquid add 0.01 mol/L hydrochloric acid-methanol TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 318 nm and 322 nm. ‌Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Ampiroxicam Cap-sules, add acetonitrile to make exactly V mL so that each mL contains about 0.27 mg of ampiroxicam (C20H21N3O7S). Stir for 30 minutes, then centrifuge, and use the supernatant liq-uid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of ampiroxicam (C20H21N3O7S) ＝ MS × AT/AS × V/100 MS: Amount (mg) of ampiroxicam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Ampiroxicam Capsules is not less than 70z. Start the test with 1 capsule of Ampiroxicam Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 15 mg of ampiroxicam (C20H21N3O7S), and use this solution as the sample solution. Separately, weigh accurately about 30 mg of ampiroxicam for assay, previously dried at 1059C for 3 hours, dissolve in 5 mL of acetonitrile, and add the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, at 320 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of ampiroxicam (C20H21N3O7S) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of ampiroxicam for assay taken C: Labeled amount (mg) of ampiroxicam (C20H21N3O7S) in 1 capsule
Take out the contents of not less than 20 Ampiroxic-am Capsules, weigh accurately the mass of the contents, and powder if necessary. Weigh accurately a portion of the powder, equivalent to about 13.5 mg of ampiroxicam (C20H21N3O7S), and add acetonitrile to make exactly 50 mL. Stir for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 27 mg of ampiroxicam for assay, previously dried at 1059C for 3 hours, dissolve in acetonitrile to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following condition, and de-termine the peak areas, AT and AS, of ampiroxicam in each solution. Amount (mg) of ampiroxicam (C20H21N3O7S) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of ampiroxicam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (3 in 500), methanol, and acetonitrile (5:3:2). Flow rate: Adjust so that the retention time of ampiroxic-am is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ampiroxicam are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ampiroxicam is not more than 1.0z.
Aprindine Hydrochloride Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of aprindine hydrochloride (C22H30N2.HCl: 358.95). Method of preparation Prepare as directed under Cap-sules, with Aprindine Hydrochloride. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, it exhibits maxima between 264 nm and 268 nm, and between 271 nm and 275 nm. ‌JP XVIII Official Monographs / Arbekacin Sulfate 477 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Aprindine Hydro-chloride Capsules, add 30 mL of a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125), shake vigorously for 20 minutes, add a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly V mL so that each mL contains about 0.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, it exhibits maxima between 264 nm and 268 nm, and between 271 nm and 275 nm. ‌JP XVIII Official Monographs / Arbekacin Sulfate 477 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Aprindine Hydro-chloride Capsules, add 30 mL of a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125), shake vigorously for 20 minutes, add a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly V mL so that each mL contains about 0.2 mg of aprindine hydrochloride (C22H30N2.HCl), and filter. Discard the first 5 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Proceed as directed in the Assay. Amount (mg) of aprindine hydrochloride (C22H30N2.HCl) ＝ MS × AT/AS × V/250 MS: Amount (mg) of aprindine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Aprindine Hydrochloride Capsules is not less than 80z. Start the test with 1 capsule of Aprindine Hydrochloride Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL factor of the peak of aprindine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of aprindine is not more than 1.5z.
Take out the contents of not less than 20 Aprindine Hydrochloride Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of aprindine hydrochlo-ride (C22H30N2.HCl), add 60 mL of a solution of hydrochlo-ric acid in diluted ethanol (1 in 2) (1 in 125), shake vigorously for 20 minutes, and add a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly 100 mL. Pipet 10 mL of this solution, add a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly 50 mL, and filter, Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of aprindine hydrochloride for assay, previously dried in vacuum at 609C for 4 hours, and dissolve in a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly 50 mL. Pipet 10 mL of this solution, add a solution of hydrochloric acid in diluted ethanol (1 in 2) (1 in 125) to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 265 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. so that each mL contains about 11 mg of aprindine hydro-chloride (C22H30N2.HCl), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of aprin- Amount (mg) of aprindine hydrochloride (C ＝ MS × AT/AS × 2 22H 30N2.HCl) dine hydrochloride for assay, previously dried in vacuum at 609C for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and determine the peak areas, AT and AS, of aprindine in each solution. Dissolution rate (z) with respect to the labeled amount of aprindine hydrochloride (C22H30N2.HCl) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of aprindine hydrochloride for assay taken C: Labeled amount (mg) of aprindine hydrochloride (C22H30N2.HCl) in 1 capsule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.40 g of potassium dihydrogen phosphate in 500 mL of water, and adjust the pH to 3.0 with hydrochloric acid. To 500 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of aprindine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry MS: Amount (mg) of aprindine hydrochloride for assay taken
Arbekacin Sulfate Injection is an aqueous injection. It contains not less than 90.
Arbekacin Sulfate Injection occurs as a clear and colorless liquid.
To 0.2 mL of Arbekacin Sulfate Injection add 1 mL of water, and use this solution as the sample solu-tion. Separately, dissolve 10 mg of Arbekacin Sulfate RS in 1 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop with a mixture of am-monia solution (28), methanol, chloroform and ethanol (95) (7:6:4:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 809C for 10 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show a purple-brown color and the same Rf value. Osmotic pressure ratio <2.47> 0.8 – 1.2 (for the preparation intended for intramuscular use). pH <2.54> 6.0 – 8.0 Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, Culture medium and Standard solu-tions: Proceed as directed in the Assay under Arbekacin Sul-fate. Sample solutions—Take exactly a volume of Arbeka-cin Sulfate Injection, equivalent to about 20 mg (potency), and add water to make exactly 50 mL. Take exactly a suita-ble amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and low concen-tration sample solution, respectively.
Ascorbic Acid and Calcium Pantothenate Tablets
アスコルビン酸･パントテン酸カルシウム錠
Ascorbic Acid and Calcium Pantothenate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of L-ascorbic acid (C6H8O6: 176.12) and not less than 93.0z and not more than 107.0z of the labeled amount of calcium pantothenate (C18H32CaN2O10: 476.53). Method of preparation Prepare as directed under Tablets, with Ascorbic Acid and Calcium Pantothenate. Identification (1) To a quantity of powdered Ascorbic Acid and Calcium Pantothenate Tablets, equivalent to 0.5 g of Ascorbic Acid, add 30 mL of water, shake for 1 minute, and filter. Proceed as directed in the Identification (1) under Ascorbic Acid using 5 mL each of the filtrate. (2) To a quantity of powdered Ascorbic Acid and Cal-cium Pantothenate Tablets, equivalent to 3 mg of Calcium Pantothenate, add 20 mL of ethanol (95), shake vigorously for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 3 mg of calcium pantothenate in 20 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and dilute acetic acid (5:3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (95) (1 in 200) on the plate, and heat the plate at 1209C for 20 minutes: one of the spots obtained from the sample solution and the spot from the standard solution are purple in color and their Rf value are the same. Uniformity of dosage units <6.02> (1) L-Ascorbic acid—Perform the Mass variation test, or the Content uniformity test according to the following method: it meets the require-ment. To 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets add 100 mL of a solution of metaphosphoric acid (1 in 50), stir thoroughly, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6 (2) Calcium pantothenate—Perform the test according to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets add exactly V mL of the internal standard solution so that each mL contains about 0.
(1) To a quantity of powdered Ascorbic Acid and Calcium Pantothenate Tablets, equivalent to 0.5 g of Ascorbic Acid, add 30 mL of water, shake for 1 minute, and filter. Proceed as directed in the Identification (1) under Ascorbic Acid using 5 mL each of the filtrate. (2) To a quantity of powdered Ascorbic Acid and Cal-cium Pantothenate Tablets, equivalent to 3 mg of Calcium Pantothenate, add 20 mL of ethanol (95), shake vigorously for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 3 mg of calcium pantothenate in 20 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and dilute acetic acid (5:3:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in ethanol (95) (1 in 200) on the plate, and heat the plate at 1209C for 20 minutes: one of the spots obtained from the sample solution and the spot from the standard solution are purple in color and their Rf value are the same. Uniformity of dosage units <6.02> (1) L-Ascorbic acid—Perform the Mass variation test, or the Content uniformity test according to the following method: it meets the require-ment. To 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets add 100 mL of a solution of metaphosphoric acid (1 in 50), stir thoroughly, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6 (2) Calcium pantothenate—Perform the test according to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets add exactly V mL of the internal standard solution so that each mL contains about 0.15 mg of calcium panto-thenate (C18H32CaN2O10), shake vigorously for 15 minutes, filter through a membrane filter with a pore size not exceed- 1st fluid for dissolution test to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 243 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24> within 1 hour after withdrawing the medium, using 1st fluid for dissolution test as the blank. Dissolution rate (z) with respect to the labeled amount of L-ascorbic acid (C6H8O6) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Ascorbic Acid RS taken C: Labeled amount (mg) of L-ascorbic acid (C6H8O6) in 1 tablet (2) Calcium pantothenate—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Ascorbic Acid and Cal-cium Pantothenate Tablets is not less than 75z. Start the test with 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets, withdraw 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 1st fluid for dissolution test to make exactly V? mL so that each mL contains about 3.3 mg of calcium pantothenate (C18H32CaN2O10), and use this solu-tion as the sample solution. Separately, weigh accurately about 16.5 mg of Calcium Pantothenate RS (separately de-termine the loss on drying <2.41> under the same conditions as Calcium Pantothenate), and dissolve in water to make ex-actly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of pan-tothenic acid in each solution. Dissolution rate (z) with respect to the labeled amount of calcium pantothenate (C H CaN O ) ing 0.45 mm, and use the filtrate as the sample solution. 18 32 2 10 Then, proceed as directed in the Assay (2). Amount (mg) of calcium pantothenate (C H CaN O ) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Calcium Pantothenate RS taken, cal- ＝ MS × QT/QS × V/200 18 32 2 10 culated on the dried basis C: Labeled amount (mg) of calcium pantothenate MS: Amount (mg) of Calcium Pantothenate RS taken, cal-culated on the dried basis Internal Standard Solution—A solution of acetaminophen (1 in 50,000). Dissolution <6.10> (1) L-Ascorbic acid—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Ascorbic Acid and Calcium Pantothenate Tablets is not less than 85z. Start the test with 1 tablet of Ascorbic Acid and Calcium Pantothenate Tablets, withdraw 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 1st fluid for dissolution test to make exactly V? mL so that each mL contains about 11 mg of L-ascorbic acid (C6H8O6), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Ascor-bic Acid RS, previously dried in a desiccator (silica gel) for 24 hours, dissolve in water to make exactly 100 mL, and warm at 379C for 1 hour. Pipet 5 mL of this solution, add (C18H32CaN2O10) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized sili-cone polymer coated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 7.80 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust to pH 2.6 with phosphoric acid, and add water to make 1000 mL. To 970 mL of this solution add 30 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of pantothenic acid is about 10 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pantothenic acid are not less than 5000 ‌JP XVIII Official Monographs / L-Aspartic Acid 487 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pantothenic acid is not more than 2.0z.
(1) L-Ascorbic acid—Weigh accurately the mass of not less than 20 tablets of Ascorbic Acid and Calcium Pan-tothenate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of L-ascorbic acid (C6H8O6), add 50 mL of a solution of metaphosphoric acid (1 in 50), stir thoroughly, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6 (2) Calcium pantothenate—Weigh accurately the mass of not less than 20 tablets of Ascorbic Acid and Calcium Pantothenate Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 3 mg of calcium pan-tothenate (C18H32CaN2O10), add exactly 20 mL of the inter-nal standard solution, shake for 15 minutes, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh ac-curately about 30 mg of Calcium Pantothenate RS (separate-ly determine the loss on drying <2.41> under the same condi-tions as Calcium Pantothenate), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 20 mL of the internal standard solution, and use this solu-tion as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of pantothenic acid to that of the internal standard. L-Aspartic Acid
Atorvastatin Calcium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O: 1209.39). Method of preparation Prepare as directed under Tablets, with Atorvastatin Calcium Hydrate. Identification To a quantity of powdered Atorvastatin Cal-cium Tablets, equivalent to 10 mg of Atorvastatin Calcium Hydrate, add 50 mL of methanol, shake thoroughly, and centrifuge. To 2.5 mL of the supernatant liquid add metha-nol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro- ＝ MS × QT/QS × V/200 × 1.047 MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of 1,3-dinitroben-zene in methanol (1 in 2500). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and atorvastatin are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of atorvastatin to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Atorvastatin Calcium Tablets is not less than 80z. Start the test with 1 tablet of Atorvastatin Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 6 mg of atorvastatin calcium hy-drate (C66H68CaF2N4O10.
To a quantity of powdered Atorvastatin Cal-cium Tablets, equivalent to 10 mg of Atorvastatin Calcium Hydrate, add 50 mL of methanol, shake thoroughly, and centrifuge. To 2.5 mL of the supernatant liquid add metha-nol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro- ＝ MS × QT/QS × V/200 × 1.047 MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of 1,3-dinitroben-zene in methanol (1 in 2500). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and atorvastatin are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of atorvastatin to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Atorvastatin Calcium Tablets is not less than 80z. Start the test with 1 tablet of Atorvastatin Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 6 mg of atorvastatin calcium hy-drate (C66H68CaF2N4O10.3H2O), and use this solution as the sample solution. Separately, weigh accurately about 60 mg of Atorvastatin Calcium RS (separately determine the water <2.48> in the same manner as Atorvastatin Calcium Hy- drate), and dissolve in a mixture of water and methanol (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add ‌494 water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of atorvastatin in each solution. Dissolution rate (z) with respect to the labeled amount of atorvastatin calcium hydrate (C H CaF N O .3H O) Mobile phase: Dissolve 10.5 g of citric acid monohydrate in 900 mL of water, adjust to pH 4.0 with ammonia solution (28), and add water to make 1000 mL. To 530 mL of this solution add 270 mL of acetonitrile and 200 mL of tetra-hydrofuran. Flow rate: Adjust so that the retention time of atorvasta-tin is about 9 minutes. System suitability— 66 68 2 4 10 2 System performance: When the procedure is run with 10 ＝ MS × AT/AS × V?/V × 1/C × 9 × 1.047 MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of atorvastatin are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of atorvastatin is not more than 2.0z.
To 20 Atorvastatin Calcium Tablets add 3V/5 mL of a mixture of water and methanol (1:1), and disintegrate the tablet by shaking. Add exactly V/10 mL of the internal mL of the standard solution under the above operating con-ditions, the internal standard and atorvastatin are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of atorvastatin to that of the internal standard is not more than 1.0z.
Atropine Sulfate Injection is an aqueous injection. It contains not less than 93.
Atropine Sulfate Injection is a clear, colorless liquid. pH: 4.0 – 6.0
(1) Evaporate a volume of Atropine Sul-fate Injection, equivalent to 1 mg of Atropine Sulfate Hy-drate, on a water bath to dryness. Proceed with the residue as directed in the Identification (1) under Atropine Sulfate Hydrate. Evaporate an exactly measured volume of Atropine Sulfate Injection, equivalent to 5 mg of Atropine Sulfate Hydrate, on a water bath to dryness. After cooling, dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solution. If insoluble substance remains, crush it, allow to stand, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of Atropine Sulfate RS in 2 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetone, water and ammonia water (28) (90:7:3) to a distance of about 10 cm, and dry the plate at 809C for 10 minutes. After cooling, spray evenly Dragendorff's TS for spraying on the plate: the spots obtained from the sample solution and the standard solution show an orange color and the same Rf value. Atropine Sulfate Injection responds to Qualitative Tests <1.09> for sulfate. Bacterial endotoxins <4.01> Less than 75 EU/mg. Extractable volume <6.05> It meets the requirements. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test: it meets the requirement.
To an exactly measured volume of Atropine Sulfate Injection, equivalent to about 5 mg of atropine sulfate hy-drate [(C17H23NO3)2.H2SO4.H2O], add exactly 3 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Atropine Sulfate RS (separately deter-mine the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), and dissolve in water to make ex-actly 50 mL. Pipet 10 mL of this solution, add exactly 3 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of atropine to that of the inter-nal standard. Amount (mg) of atropine sulfate hydrate [(C17H23NO3)2.H2SO4.H2O] ＝ MS × QT/QS × 1/5 × 1.027 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated based on the dried basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 0.4 g of sodium lauryl sulfate add 500 mL of diluted phosphoric acid (1 in 1000) to dissolve, and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL ‌496 of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of atropine is about 16 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and atropine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of atropine to that of the internal standard is not more than 1.5z.
Auranofin Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of auranofin (C20H34AuO9PS: 678.48). Method of preparation Prepare as directed under Tablets, with Auranofin. Identification Put an amount of powdered Auranofin Tablets, equivalent to 11 mg of Auranofin, in a porcelain crucible, and heat weakly to carbonize. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid, heat cau-tiously at first then incinerate by ignition. After cooling, add 4 mL of aqua regia to the residue, dissolve by warming, and add 16 mL of water. To 5 mL of this solution add 0.5 mL of tin (II) chloride TS: a purple to red-brown color is deve-loped. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Auranofin Tablets add 2 mL of water, dis-integrate the tablet by sonicating, add exactly 2 mL of the internal standard solution for every 3 mg of auranofin (C20H34AuO9PS), and add 2 mL of a mixture of water and acetonitrile (1:1). Shake for 15 minutes, then add a mixture of water and acetonitrile (1:1) to make V mL so that each mL contains 0.
Put an amount of powdered Auranofin Tablets, equivalent to 11 mg of Auranofin, in a porcelain crucible, and heat weakly to carbonize. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid, heat cau-tiously at first then incinerate by ignition. After cooling, add 4 mL of aqua regia to the residue, dissolve by warming, and add 16 mL of water. To 5 mL of this solution add 0.5 mL of tin (II) chloride TS: a purple to red-brown color is deve-loped. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Auranofin Tablets add 2 mL of water, dis-integrate the tablet by sonicating, add exactly 2 mL of the internal standard solution for every 3 mg of auranofin (C20H34AuO9PS), and add 2 mL of a mixture of water and acetonitrile (1:1). Shake for 15 minutes, then add a mixture of water and acetonitrile (1:1) to make V mL so that each mL contains 0.3 mg of auranofin (C20H34AuO9PS), centri-fuge, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of auranofin (C20H34AuO9PS) ＝ MS × QT/QS × V/100 MS: Amount (mg) of Auranofin RS taken Internal standard solution—A solution of butyl para-hydroxybenzoate in acetonitrile (9 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Auranofin Tablets is not less than 85z. Start the test with 1 tablet of Auranofin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 3.3 mg of auranofin (C20H34AuO9PS), and use this solution as the sample solution. Separately, weigh accu-rately about 30 mg of Auranofin RS, previously dried at 1059C for 3 hours, and dissolve in acetonitrile to make ex-actly 50 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> under the following conditions, and determine the peak areas, AT and AS, of auranofin in each solution. Dissolution rate (z) with respect to the labeled amount of auranofin (C20H34AuO9PS) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of Auranofin RS taken C: Labeled amount (mg) of auranofin (C20H34AuO9PS) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay under Auranofin. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of auranofin are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of auranofin is not more than 1.0z.
Accurately weigh the mass of not less than 20 Auranofin Tablets, and powder them. Weigh accurately a portion of the powder, equivalent to about 60 mg of auranofin (C20H34AuO9PS), add 40 mL of water, disperse the particles with the aid of ultrasonic waves, then add ex-actly 40 mL of the internal standard solution, add 40 mL of a mixture of water and acetonitrile (1:1), and shake for 15 minutes. To this solution add a mixture of water and aceto-nitrile (1:1) to make 200 mL, centrifuge, and use the super-natant liquid as the sample solution. Separately, weigh accu-rately about 30 mg of Auranofin RS, previously dried at 1059C for 3 hours, dissolve in 60 mL of a mixture of water and acetonitrile (1:1), add exactly 20 mL of the internal standard solution, then add water to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as ‌498 directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of auranofin to that of the internal standard. Method <1.06>, using 20 mL of water as the absorbing liquid: the test solution responds to Qualitative Tests <1.09> for sulfate. (4) Dissolve 0.01 g of Azathioprine in 2 mol/L hydro- Amount (mg) of auranofin (C H AuO PS) chloric acid TS to make 100 mL. Dilute 5 mL of the solution ＝ MS × QT/QS × 2 20 34 9 with water to make 50 mL. Determine the absorption spec-trum of the solution as directed under Ultraviolet-visible MS: Amount (mg) of Auranofin RS taken Internal standard solution—A solution of butyl para-hydroxybenzoate in acetonitrile (9 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Auranofin. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, auranofin and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of auranofin to that of the internal standard is not more than 1.0z.
Azathioprine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of azathioprine (C9H7N7O2S: 277.26). Method of preparation Prepare as directed under Tablets, with Azathioprine. Identification (1) Weigh a quantity of powdered Azathioprine Tablets, equivalent to 0.01 g of Azathioprine. Add 50 mL of water, shake well while warming, and filter. Proceed with 5 mL of the filtrate as directed in the Identifi-cation (1) under Azathioprine. Proceed with 1 mL of the filtrate obtained in (1) as directed in the Identification (2) under Azathioprine. Determine the absorption spectrum of the sample so-lution in the Assay as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 278 nm and 282 nm. Weigh a quantity of powdered Azathioprine Tablets, equivalent to 0.1 g of Azathioprine to the labeled amount. Add 10 mL of a solution of ammonia solution (28) in ethanol (95) (1 in 10), shake well, filter, and use the filtrate MS: Amount (mg) of Azathioprine RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Azathioprine Tablets is not less than 80z. Start the test with 1 tablet of Azathioprine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of azathioprine (C9H7N7O2S), and use this solution as the sample solution.
(1) Weigh a quantity of powdered Azathioprine Tablets, equivalent to 0.01 g of Azathioprine. Add 50 mL of water, shake well while warming, and filter. Proceed with 5 mL of the filtrate as directed in the Identifi-cation (1) under Azathioprine. Proceed with 1 mL of the filtrate obtained in (1) as directed in the Identification (2) under Azathioprine. Determine the absorption spectrum of the sample so-lution in the Assay as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 278 nm and 282 nm. Weigh a quantity of powdered Azathioprine Tablets, equivalent to 0.1 g of Azathioprine to the labeled amount. Add 10 mL of a solution of ammonia solution (28) in ethanol (95) (1 in 10), shake well, filter, and use the filtrate MS: Amount (mg) of Azathioprine RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Azathioprine Tablets is not less than 80z. Start the test with 1 tablet of Azathioprine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of azathioprine (C9H7N7O2S), and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of Azathioprine RS, previously dried at 1059C for 5 hours, and dissolve in water to make exactly 100 mL. Pipet 6 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 280 nm of the sample solution and standard solution as directed under Ultraviolet- visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of azathioprine (C9H7N7O2S) ＝ MS × AT/AS × V?/V × 1/C × 108 MS: Amount (mg) of Azathioprine RS taken C: Labeled amount (mg) of azathioprine (C9H7N7O2S) in 1 tablet
Weigh accurately and powder not less than 20 Azathioprine Tablets. Weigh accurately a portion of the powder, equivalent to about 0.1 g of azathioprine (C9H7N7O2S), add 20 mL of dimethylsulfoxide for ultravio-let-visible spectrophotometry, shake well, add 0.1 mol/L hydrochloric acid TS to make exactly 500 mL, and filter. Discard the first 20 mL of the filtrate, measure exactly 3 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 0.1 g of ‌500 Azathioprine RS, previously dried at 1059C for 5 hours, dis-solve in 20 mL of dimethylsulfoxide for ultraviolet-visible spectrophotometry, and add 0.1 mol/L hydrochloric acid TS to make exactly 500 mL. Measure exactly 3 mL of this solu-tion, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 280 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of azathioprine (C H N O S) Arsenic <1.11>—Prepare the test solution with 1.0 g of Azelastine Hydrochloride according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Azelastine Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> ＝ MS × AT/AS 9 7 7 2 according to the following conditions. Determine each peak area of these solutions by the automatic integration method: MS: Amount (mg) of Azathioprine RS taken
To a quantity of Azelastine Hydrochloride Granules, equivalent to 2 mg of Azelastine Hydrochloride, add 30 mL of 0.1 mol/L hydrochloric acid TS, and sonicate for 30 minutes. After cooling, add 0.1 mol/L hydrochloric acid TS to make 50 mL, and centrifuge. Determine the ab-sorption spectrum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 283 nm and 287 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 45 minutes of Azelastine Hydrochloride Granules is not less than 80z. Start the test with accurately weighed amount of Azelastine Hydrochloride Granules, equivalent to about 1 mg of azelastine hydrochloride (C22H24ClN3O.HCl), with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 50 mg of azelastine hydrochloride for assay, previ-ously dried at 1059C for 2 hours, and dissolve in the dissolu-tion medium to make exactly 250 mL. Pipet 1 mL of this so-lution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of azelastine in each solution. Dissolution rate (z) with respect to the labeled amount of azelastine hydrochloride (C22H24ClN3O.HCl) ＝ MS/MT × AT/AS × 1/C × 9/5 MS: Amount (mg) of azelastine hydrochloride for assay taken MT: Amount (g) of Azelastine Hydrochloride Granules taken C: Labeled amount (mg) of azelastine hydrochloride (C22H24ClN3O.HCl) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of azelastine are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of azelastine is not more than 2.0z.
Weigh accurately an amount of Azelastine Hydro-chloride Granules, equivalent to about 2 mg of azelastine hy-drochloride (C22H24ClN3O.HCl), add 50 mL of 0.1 mol/L hydrochloric acid TS, sonicate for 20 minutes, add 40 mL of ethanol (99.5), add exactly 5 mL of the internal standard so-lution, and add ethanol (99.5) to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of azelastine hydrochloride for assay, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 50 mL. Pipet 10 mL of this solution, and add 0.1 mol/L hydrochlo-ric acid TS to make exactly 50 mL. Pipet 10 mL of this solu-tion, add 40 mL of 0.1 mol/L hydrochloric acid TS and 40 mL of ethanol (99.5), add exactly 5 mL of the internal stand-ard solution, add ethanol (99.5) to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of azelastine to that of the internal standard. Amount (mg) of azelastine hydrochloride (C22H24ClN3O.HCl) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of azelastine hydrochloride for assay taken Internal standard solution—Dissolve 0.2 g of 2-ethylhexyl parahydroxybenzoate in ethanol (99.5) to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and a solution of sodium lauryl sulfate in diluted acetic acid (100) (1 in 250) (1 in 500) (11:9). Flow rate: Adjust so that the retention time of azelastine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, azelastine and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of azelastine to that of the internal standard is not more than 1.0z.
Azelnidipine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of azelnidipine (C33H34N4O6: 582.65). Method of preparation Prepare as directed under Tablets, with Azelnidipine. Identification Powder Azelnidipine Tablets. Weigh a por-tion of the powder, equivalent to 4 mg of Azelnidipine, add 150 mL of ethanol (99.5), sonicate for 15 minutes, then add ethanol (99.5) to make 200 mL. Centrifuge this solution, filter the supernatant liquid through a glass wool filter with a pore size not exceeding 0.7 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 253 nm and 257 JP XVIII Official Monographs / Azelnidipine Tablets 503 nm and between 339 nm and 346 nm. Purity Related substances—Conduct this procedure using light-resistant vessels. Powder Azelnidipine Tablets. Weigh a portion of the powder, equivalent to 10 mg of Azelnidipine, add 10 mL of a mixture of acetonitrile and water (4:1), agitate gently, then disperse to fine particles by sonicating for 15 minutes. Centrifuge this solution, and use the super-natant liquid as the sample solution. Pipet 2 mL of the sam-ple solution, add a mixture of acetonitrile and water (4:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the areas of the peaks, having the relative retention times of about 0.10, about 0.13, about 0.50, and about 1.42 to azelnidipine, obtained from the sample solu-tion, are not larger than 9/20 times, 1/5 times, 2/5 times, and 2/5 times the peak area of azelnidipine obtained from the standard solution, respectively, the area of the peak, other than azelnidipine and the peaks mentioned above, is not larger than 1/10 times the peak area of azelnidipine from the standard solution. Furthermore, the total area of these peaks other than azelnidipine is not larger than 1.75 times the peak area of azelnidipine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Azelnidipine. Time span of measurement: About 2 times as long as the retention time of azelnidipine. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (4:1) to make exactly 20 mL. Confirm that the peak area of azelnidipine obtained with 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 10 mL of the standard solu-tion. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of azelnidipine are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of azelnidipine is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Azelnidipine Tablets add exactly 1 mL of ether in a mixture of acetonitrile and water (4:1) (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Azelnidipine Tablets is not less than 75z. Start the test with 1 tablet of Azelnidipine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 8.
Powder Azelnidipine Tablets. Weigh a por-tion of the powder, equivalent to 4 mg of Azelnidipine, add 150 mL of ethanol (99.5), sonicate for 15 minutes, then add ethanol (99.5) to make 200 mL. Centrifuge this solution, filter the supernatant liquid through a glass wool filter with a pore size not exceeding 0.7 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 253 nm and 257 JP XVIII Official Monographs / Azelnidipine Tablets 503 nm and between 339 nm and 346 nm.
Related substances—Conduct this procedure using light-resistant vessels. Powder Azelnidipine Tablets. Weigh a portion of the powder, equivalent to 10 mg of Azelnidipine, add 10 mL of a mixture of acetonitrile and water (4:1), agitate gently, then disperse to fine particles by sonicating for 15 minutes. Centrifuge this solution, and use the super-natant liquid as the sample solution. Pipet 2 mL of the sam-ple solution, add a mixture of acetonitrile and water (4:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the areas of the peaks, having the relative retention times of about 0.10, about 0.13, about 0.50, and about 1.42 to azelnidipine, obtained from the sample solu-tion, are not larger than 9/20 times, 1/5 times, 2/5 times, and 2/5 times the peak area of azelnidipine obtained from the standard solution, respectively, the area of the peak, other than azelnidipine and the peaks mentioned above, is not larger than 1/10 times the peak area of azelnidipine from the standard solution. Furthermore, the total area of these peaks other than azelnidipine is not larger than 1.75 times the peak area of azelnidipine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Azelnidipine. Time span of measurement: About 2 times as long as the retention time of azelnidipine. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (4:1) to make exactly 20 mL. Confirm that the peak area of azelnidipine obtained with 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 10 mL of the standard solu-tion. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of azelnidipine are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of azelnidipine is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Azelnidipine Tablets add exactly 1 mL of ether in a mixture of acetonitrile and water (4:1) (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Azelnidipine Tablets is not less than 75z. Start the test with 1 tablet of Azelnidipine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 8.9 mg of azelnidipine (C33H34N4O6), and use this solution as the sample solution. Separately, weigh accurately about 45 mg of azelnidipine for assay, previously dried in vacuum at 709C for 5 hours, dis-solve in ethanol (99.5) to make exactly 25 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, at 270 nm of the sam-ple solution and standard solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of azelnidipine (C33H34N4O6) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of azelnidipine for assay taken C: Labeled amount (mg) of azelnidipine (C33H34N4O6) in 1 tablet
Weigh accurately the mass of not less than 20 Azel-nidipine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of azelnidipine (C33H34N4O6), add exactly 25 mL of the internal standard so-lution, add 50 mL of a mixture of acetonitrile and water (4:1). After sonicating for 10 minutes, add a mixture of acetonitrile and water (4:1) to make 100 mL. Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (4:1) to make 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 50 mg of azelnidipine for assay, previously dried in vacuum at 709C for 5 hours, dissolve in exactly 25 mL of the internal standard solution, and add a mixture of acetonitrile and water (4:1) to make 100 mL. To 5 mL of this solution add a mixture of acetonitrile and water (4:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of azelnidipine to that of the in-ternal standard. the internal standard solution per 2 mg of azelnidipine (C33H34N4O6), and add a mixture of acetonitrile and water Amount (mg) of azelnidipine (C33H34N4O6) ＝ M S × QT/QS (4:1) to make 32 mL. Disintegrate the tablet with occasional shaking, and sonicate for 10 minutes. Centrifuge this solu-tion, pipet V mL of the supernatant liquid, equivalent to 2.5 mg of azelnidipine (C33H34N4O6), add a mixture of aceto-nitrile and water (4:1) to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of azelnidipine (C33H34N4O6) ＝ MS × QT/QS × 8/5V MS: Amount (mg) of azelnidipine for assay taken Internal standard solution—A solution of 2,2?-dinaphthyl- MS: Amount (mg) of azelnidipine for assay taken Internal standard solution—2,2?-dinaphthylether in a mix-ture of acetonitrile and water (4:1) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.9 g of potassium dihydrogen ‌504 phosphate in 300 mL of water, add 700 mL of acetonitrile, then adjust to pH 6.0 with dilute sodium hydroxide TS. Flow rate: Adjust so that the retention time of azelnidi-pine is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, azelnidipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of azelnidipine to that of the internal standard is not more than 1.0z.
Azosemide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of azosemide (C H ClN O S : 370.84). Melting point: about 2269C (with decomposition). 12 11 6 2 2 Identification (1) Determine the absorption spectrum of a solution of Azosemide in dilute sodium hydroxide TS (3 in 500,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Azosemide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Purity (1) Chloride <1.03>—To 1.0 g of Azosemide add 60 mL of dilute sodium hydroxide TS, dissolve by warming. After cooling, add 0.5 mL of nitric acid and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.032z). Heavy metal <1.07>—Proceed with 1.0 g of Azo- semide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Primary aromatic amines—Dissolve 20 mg of Azo-semide in 5 mL of N,N-dimethylformamide, add 12 mL of water, 1.0 mL of a solution of sodium nitrite (1 in 200) and 2.0 mL of diluted hydrochloric acid (1 in 10) under ice-cool-ing, shake, and allow to stand for 3 minutes. Add 1.0 mL of ammonium amidosulfate TS, shake thoroughly, allow to stand for 3 minutes, and add 1.0 mL of a solution of N-1-naphthylethylenediamine dihydrochloride (1 in 200). Shake this solution, and add N,N-dimethylformamide to make ex-actly 50 mL. Determine the absorbance of this solution at 540 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared in the same man-ner with 5 mL of N,N-dimethylformamide as the blank: the absorbance is not more than 0.15. Method of preparation Prepare as directed under Tablets, with Azosemide. Identification To a quantity of powdered Azosemide Tablets, equivalent to 60 mg of Azosemide, add dilute so-dium hydroxide TS to make 100 mL, shake, and filter. To 1 mL of the filtrate add dilute sodium hydroxide TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 234 nm and 238 nm, be- tween 272 nm and 276 nm and between 324 nm and 330 nm. Purity Primary aromatic amines—To a quantity of pow-dered Azosemide Tablets, equivalent to 20 mg of Azo-semide, add 5 mL of N,N-dimethylformamide, and allow to stand with occasional shaking. Add 12 mL of water, 1.0 mL of a solution of sodium nitrite (1 in 200) and 2.0 mL of diluted hydrochloric acid (1 in 10) under ice-cooling, shake, and allow to stand for 3 minutes. Add 1.0 mL of ammonium amidosulfate TS, shake thoroughly, and allow to stand for 3 minutes. Add 1.0 mL of a solution of N-1-naphthylethylenediamine dihydrochloride (1 in 200), and shake. Add N,N-dimethylformamide to make exactly 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Determine the absorbance of the sample solution at 540 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared in the same man-ner with 5 mL of N,N-dimethylformamide as the blank: the absorbance is not more than 0.15. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Azosemide Tablets add dilute sodium hy-droxide TS to make exactly V mL so that each mL contains about 0.
(1) Determine the absorption spectrum of a solution of Azosemide in dilute sodium hydroxide TS (3 in 500,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Azosemide, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.
(1) Chloride <1.03>—To 1.0 g of Azosemide add 60 mL of dilute sodium hydroxide TS, dissolve by warming. After cooling, add 0.5 mL of nitric acid and filter. To 30 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.032z). Heavy metal <1.07>—Proceed with 1.0 g of Azo- semide according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Primary aromatic amines—Dissolve 20 mg of Azo-semide in 5 mL of N,N-dimethylformamide, add 12 mL of water, 1.0 mL of a solution of sodium nitrite (1 in 200) and 2.0 mL of diluted hydrochloric acid (1 in 10) under ice-cool-ing, shake, and allow to stand for 3 minutes. Add 1.0 mL of ammonium amidosulfate TS, shake thoroughly, allow to stand for 3 minutes, and add 1.0 mL of a solution of N-1-naphthylethylenediamine dihydrochloride (1 in 200). Shake this solution, and add N,N-dimethylformamide to make ex-actly 50 mL. Determine the absorbance of this solution at 540 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared in the same man-ner with 5 mL of N,N-dimethylformamide as the blank: the absorbance is not more than 0.15. Method of preparation Prepare as directed under Tablets, with Azosemide. Identification To a quantity of powdered Azosemide Tablets, equivalent to 60 mg of Azosemide, add dilute so-dium hydroxide TS to make 100 mL, shake, and filter. To 1 mL of the filtrate add dilute sodium hydroxide TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 234 nm and 238 nm, be- tween 272 nm and 276 nm and between 324 nm and 330 nm. Purity Primary aromatic amines—To a quantity of pow-dered Azosemide Tablets, equivalent to 20 mg of Azo-semide, add 5 mL of N,N-dimethylformamide, and allow to stand with occasional shaking. Add 12 mL of water, 1.0 mL of a solution of sodium nitrite (1 in 200) and 2.0 mL of diluted hydrochloric acid (1 in 10) under ice-cooling, shake, and allow to stand for 3 minutes. Add 1.0 mL of ammonium amidosulfate TS, shake thoroughly, and allow to stand for 3 minutes. Add 1.0 mL of a solution of N-1-naphthylethylenediamine dihydrochloride (1 in 200), and shake. Add N,N-dimethylformamide to make exactly 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Determine the absorbance of the sample solution at 540 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared in the same man-ner with 5 mL of N,N-dimethylformamide as the blank: the absorbance is not more than 0.15. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Azosemide Tablets add dilute sodium hy-droxide TS to make exactly V mL so that each mL contains about 0.6 mg of azosemide (C12H11ClN6O2S2), shake thor-oughly, and centrifuge. Pipet 10 mL of the supernatant liq-uid, and add dilute sodium hydroxide TS to make exactly 100 mL. Pipet 10 mL of this solution, add dilute sodium hy-droxide TS to make exactly 50 mL, and use this solution as ‌506 the sample solution. Separately, weigh accurately about 60 mg of azosemide for assay, previously dried at 1059C for 3 culate the ratios, QT and QS, of the peak area of azosemide to that of the internal standard. hours, and dissolve in dilute sodium hydroxide TS to make Amount (mg) of azosemide (C H ClN O S ) exactly 100 mL. Pipet 10 mL of this solution, and add dilute sodium hydroxide TS to make exactly 100 mL. Pipet 10 mL ＝ MS × QT/QS 12 11 6 2 2 of this solution, add dilute sodium hydroxide TS to make ex-actly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 274 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>. Amount (mg) of azosemide (C12H11ClN6O2S2) ＝ MS × AT/AS × V/100 MS: Amount (mg) of azosemide for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 60 minutes of a 30-mg tablet and in 90 minutes of a 60-mg tablet are not less than 70z, respectively. Start the test with 1 tablet of Azosemide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add the dissolution medium to make exactly V? mL so that each mL contains about 33 mg of azosemide (C12H11ClN6O2S2). Pipet 8 mL of this solution, add 0.2 mol/L sodium hydroxide TS to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 22 mg of azosemide for assay, previously dried at 1059C for 3 hours, and dissolve in 0.2 mol/L sodium hy-droxide TS to make exactly 100 mL. Pipet 5 mL of this solu-tion, and add 0.2 mol/L sodium hydroxide TS to make ex-actly 50 mL. Pipet 15 mL of this solution, add the dissolu-tion medium to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 274 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared by adding 0.2 mol/L so-dium hydroxide TS to 8 mL of the dissolution medium to make 20 mL as the blank. Dissolution rate (z) with respect to the labeled amount of azosemide (C12H11ClN6O2S2) ＝ MS × AT/AS × V?/V × 1/C × 135 MS: Amount (mg) of azosemide for assay taken C: Labeled amount (mg) of azosemide (C12H11ClN6O2S2) in 1 tablet
Weigh accurately the mass of not less than 20 tablets of Azosemide Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 60 mg of azosemide (C12H11ClN6O2S2), add dilute sodium hydroxide TS to make exactly 100 mL, shake thoroughly, and filter. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent fil- MS: Amount (mg) of azosemide for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.03 mol/L potassium dihy-drogen phosphate solution, acetonitrile and methanol (55:27:18). Flow rate: Adjust so that the retention time of azosemide is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, azosemide and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of azosemide to that of the internal standard is not more than 1.0z.
Aztreonam for Injection is a preparation for injec-tion which is dissolved before use. It contains not less than 93.
Aztreonam for Injection is white to yellow- ‌508‌ white masses or powder.
(1) Dissolve an amount of Aztreonam for Injection, equivalent to 6 mg (potency) of Aztreonam, in 1 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 3 minutes, add 1 mL of acidic ammonium iron (III) sulfate TS, and mix: a red-brown color develops. (2) Dissolve an amount of Aztreonam for Injection, equivalent to 3 mg (potency) of Aztreonam, in 100 mL of water, and determine the absorption spectrum of the solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 289 nm and 293 nm. pH <2.54> The pH of a solution prepared by dissolving an amount of Aztreonam for Injection, equivalent to 1.0 g (potency) of Aztreonam, in 10 mL of water is 4.5 to 7.0.
Clarity and color of solution—Dissolve an amount of Aztreonam for Injection, equivalent to 1.0 g (potency) of Aztreonam, in 10 mL of water: the solution is clear, and its absorbance <2.24> at 450 nm is not more than 0.06. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take an amount of Aztreonam for Injection, equivalent to about 5 g (potency) of Aztreonam, dissolve the contents with a suitable amount of water, and transfer to a 100-mL volumetric flask. Wash each container with water, combine the washings and the solution, and add water to make exactly 100 mL. Pipet 10 mL of this solution, and add water to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Aztreo-nam RS, equivalent to about 20 mg (potency), dissolve in a suitable amount of water, add exactly 10 mL of the internal standard solution and water to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Aztreonam. Bacampicillin Hydrochloride
Baclofen Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of baclofen (C10H12ClNO2: 213.66). Method of preparation Prepare as directed under Tablets, with Baclofen. Identification (1) To a portion of powdered Baclofen Tablets, equivalent to 0.01 g of Baclofen, add 10 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and proceed as directed in the Identifi-cation (1) under Baclofen. To a portion of powdered Baclofen Tablets, equiva-lent to 25 mg of Baclofen, add 50 mL of 0.1 mol/L hydro-chloric acid TS, shake for 15 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 257 nm and 261 nm, between 264 nm and 268 nm, and between 272 nm and 276 nm. To a portion of powdered Baclofen Tablets, equiva-lent to 0.01 g of Baclofen, add 2 mL of a mixture of metha-nol and acetic acid (100) (4:1), shake well, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 0.01 g of Baclofen RS in 2 mL of a mixture of meth-anol and acetic acid (100) (4:1), and use this solution as the standard solution. Perform the test with these solutions as fuge, pipet 5 mL of the supernatant liquid, add 2 drops of phenolphthalein TS, neutralize with dilute sodium hydroxide TS, then add water to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 25 mg of Baclofen RS (separately determine the water <2.48> in the same manner as Baclofen), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add 2 drops of phenolphthalein TS, neutralize with dilute sodium hydroxide TS, then add water to make exactly 50 mL, and use this solution as the standard solution. To exactly 2 mL each of the sample solution and standard solution add 4 mL of ninhydrin-tin (II) chloride TS, mix, heat on a water bath for 20 minutes, then immedi-ately shake vigorously for 2 minutes. After cooling, add a mixture of water and 1-propanol (1:1) to make them exactly 25 mL, and determine the absorbances, AT and AS, of them at 570 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution obtained with 2 mL of water by the same procedure as above as the blank. Amount (mg) of baclofen (C10H12ClNO2) ＝ MS × AT/AS × V/50 MS: Amount (mg) of Baclofen RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 500 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Baclofen Tablets is not less than 70z. Start the test with 1 tablet of Baclofen Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent, add water to make exactly V? mL so that each mL contains about 10 mg of baclofen (C10H12ClNO2), and use this solution as the sample solution.
(1) To a portion of powdered Baclofen Tablets, equivalent to 0.01 g of Baclofen, add 10 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and proceed as directed in the Identifi-cation (1) under Baclofen. To a portion of powdered Baclofen Tablets, equiva-lent to 25 mg of Baclofen, add 50 mL of 0.1 mol/L hydro-chloric acid TS, shake for 15 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 257 nm and 261 nm, between 264 nm and 268 nm, and between 272 nm and 276 nm. To a portion of powdered Baclofen Tablets, equiva-lent to 0.01 g of Baclofen, add 2 mL of a mixture of metha-nol and acetic acid (100) (4:1), shake well, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 0.01 g of Baclofen RS in 2 mL of a mixture of meth-anol and acetic acid (100) (4:1), and use this solution as the standard solution. Perform the test with these solutions as fuge, pipet 5 mL of the supernatant liquid, add 2 drops of phenolphthalein TS, neutralize with dilute sodium hydroxide TS, then add water to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 25 mg of Baclofen RS (separately determine the water <2.48> in the same manner as Baclofen), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add 2 drops of phenolphthalein TS, neutralize with dilute sodium hydroxide TS, then add water to make exactly 50 mL, and use this solution as the standard solution. To exactly 2 mL each of the sample solution and standard solution add 4 mL of ninhydrin-tin (II) chloride TS, mix, heat on a water bath for 20 minutes, then immedi-ately shake vigorously for 2 minutes. After cooling, add a mixture of water and 1-propanol (1:1) to make them exactly 25 mL, and determine the absorbances, AT and AS, of them at 570 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution obtained with 2 mL of water by the same procedure as above as the blank. Amount (mg) of baclofen (C10H12ClNO2) ＝ MS × AT/AS × V/50 MS: Amount (mg) of Baclofen RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 500 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Baclofen Tablets is not less than 70z. Start the test with 1 tablet of Baclofen Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent, add water to make exactly V? mL so that each mL contains about 10 mg of baclofen (C10H12ClNO2), and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Baclofen RS (separately determine the water <2.48> in the same manner as Baclofen), dissolve in water to make ex-actly 100 mL, then pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and the standard solution at 220 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of baclofen (C10H12ClNO2) ＝ MS × AT/AS × V?/V × 1/C × 50 MS: Amount (mg) of Baclofen RS taken, calculated on the directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer anhydrous basis C: Labeled amount (mg) of baclofen (C10 tablet H12 ClNO2) in 1 chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spot from the sam-ple solution and that from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Baclofen Tablets add 5 mL of 0.1 mol/L hy-drochloric acid TS, disperse the tablet into small particles by sonicating, then shake for 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL
Weigh accurately and powder not less than 20 Baclofen Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of baclofen (C10H12ClNO2), add 130 mL of 0.1 mol/L hydrochloric acid TS, shake for 10 minutes, add 0.1 mol/L hydrochloric acid TS to make ex-actly 200 mL, and centrifuge. Pipet 10 mL of the superna-tant liquid, add 2 drops of phenolphthalein TS, neutralize with dilute sodium hydroxide TS, add water to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.25 g of Baclofen RS (separately determine the water content <2.48> in the same manner as Baclofen), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 10 mL of this solu- contains about 0.5 mg of baclofen (C 10H 12ClNO2). Centri- tion, and add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 10 mL of this solution, add 2 drops of ‌512 ‌phenolphthalein TS, neutralize with dilute sodium hydroxide TS, add water to make exactly 50 mL, and use this solution as the standard solution. Pipet 2 mL each of the sample solu-tion and the standard solution, to each add 4 mL of nin-hydrin-stannous chloride TS, shake, heat on a water bath for 20 minutes, and shake at once vigorously for 2 minutes. After cooling, to each solution add a mixture of water and 1-propanol (1:1) to make exactly 25 mL. Determine the absor-bances, AT and AS, of these solutions at 570 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a blank prepared with 2 mL of water in the same manner. of Bamethan Sulfate in 20 mL of water: the solution is clear, and has no more color than the following control solution. Control solution: To 1.5 mL of Matching Fluid O add diluted hydrochloric acid (1 in 40) to make 200 mL. Chloride <1.03>—Perform the test with 3.5 g of Bamethan Sulfate. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.002z). Heavy metals <1.07>—Proceed with 2.0 g of Bame- than Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Amount (mg) of baclofen (C ＝ MS × AT/AS × 1/5 10H12ClNO2) Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Bamethan Sulfate according to Method 3, and perform MS: Amount (mg) of Baclofen RS taken, calculated on the anhydrous basis
Shake well a quantity of powdered Benidi-pine Hydrochloride Tablets, equivalent to 10 mg of Benidi-pine Hydrochloride, with 100 mL of methanol, and centri-fuge. To 10 mL of the supernatant liquid add methanol to make 100 mL, and use this solution as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, and between 350 nm and 360 nm.
Dehydro derivative—Powder Benidipine Hydro-chloride Tablets in an agate mortar. To an amount of the powder, equivalent to 20 mg of Benidipine Hydrochloride, add about 80 mL of a mixture of diluted phosphoric acid (1 in 500) and methanol (1:1), shake well, and add the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly 100 mL. Filter through a membrane filter with pore size of 0.45 mm, and use the filtrate as the sample solu-tion. Separately, dissolve 20 mg of benidipine hydrochloride for assay in the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly 100 mL. Pipet 1 mL of this solution, add the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of dehydro derivative having the relative retention time of about 0.75 to benidipine obtained from the sample solu-tion is not larger than 1/2 times the peak area of benidipine from the standard solution. For the area of the peak of de-hydro derivative, multiply the correction factor 1.6. Operating conditions— Perform as directed in the operating conditions in the Assay. System suitability— Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mixture of diluted phos-phoric acid (1 in 500) and methanol (1:1) to make exactly 20 mL. Confirm that the peak area of benidipine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 6 mg of benidipine hydro-chloride and 5 mg of benzoin in 200 mL of a mixture of water and methanol (1:1). When the procedure is run with 10 mL of this solution under the above operating conditions, benzoin and benidipine are eluted in this order with the reso-lution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of benidipine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Benidipine Hydrochloride Tablets add 40 mL of a mixture of diluted phosphoric acid (1 in 500) and methanol (1:1), shake to disintegrate, and add a suitable amount of the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly V mL of a solution, con-taining 40 mg of benidipine hydrochloride (C28H31N3O6.HCl) per mL. Centrifuge the solution, pipet 20 mL of the superna-tant liquid, add exactly 10 mL of the internal standard solu- tion and the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of benidipine hydrochloride (C28H31N3O6.HCl) ＝ MS × QT/QS × V/1000 MS: Amount (mg) of benidipine hydrochloride for assay taken Internal standard solution—A solution of benzoin in a mix-ture of water and methanol (1:1) (13 in 200,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate of a 2-mg tablet and a 4-mg tablet in 30 minutes is not less than 80z, and that of a 8-mg tablet in 45 minutes is not less than 85z. Start the test with 1 tablet of Benidipine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL, and add the dissolution medium to make exactly V? mL so that each mL contains about 2.2 mg of benidipine hydrochloride (C28H31N3O6.HCl). Pipet 5 mL of this solution, add exactly 5 mL of the mobile phase, and use this solution as the sample solution. Separately, weigh accu-rately about 22 mg of benidipine hydrochloride for assay, previously dried at 1059C for 2 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this so-lution, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, and add the mobile phase to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the dissolution medium, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of benidipine in each solution. Dissolution rate (z) of benidipine hydrochloride (C28H31N3O6.HCl) with respect to the labeled amount ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of benidipine hydrochloride for assay taken C: Labeled amount (mg) of benidipine hydrochloride (C28H31N3O6.HCl) in 1 tablet. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0) and acetonitrile (11:9). Flow rate: Adjust so that the retention time of benidipine is about 5 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of benidipine are not less than 3000 and not more than 2.0, respectively. ‌518 System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of benidipine is not more than 1.5z.
Weigh accurately the mass of not less than 20 Benidipine Hydrochloride Tablets, and powder using an agate mortar. Weigh accurately a part of the powder, equivalent to about 8 mg of benidipine hydrochloride (C28H31N3O6.HCl), add about 150 mL of a mixture of diluted phosphoric acid (1 in 500) and methanol (1:1), shake, then add the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly 200 mL, and centrifuge. Pipet 20 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution and the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of benidipine hydrochloride for assay, previously dried at 1059C for 2 hours, and dis-solve in the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and the mixture of diluted phosphoric acid (1 in 500) and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of benidipine to that of the internal standard. Amount (mg) of benidipine hydrochloride (C28H31N3O6.HCl) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of benidipine hydrochloride for assay taken Internal standard solution—A solution of benzoin in a mix-ture of water and methanol (1:1) (13 in 200,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogen phosphate TS (pH 3.0), methanol and tetrahydrofu-ran (65:27:8). Flow rate: Adjust so that the retention time of benidipine is about 20 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and benidipine are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of benidipine to that of the internal standard is not more than 1.0z.
Benzalkonium Chloride Concentrated Solution 50
Benzalkonium Chloride Solution is an aqueous solution containing not more than 50.0 w/vz of ben-zalkonium chloride. It contains not less than 93.
Benzalkonium Chloride Solution is a clear, colorless to light yellow liquid, having a characteristic odor. Description Benzalkonium Chloride Concentrated Solu-tion 50 is a colorless to light yellow liquid or jelly-like fluid, and has a characteristic odor. It is very soluble in water and in ethanol (95), and practi-cally insoluble in diethyl ether. A solution prepared by adding water to it vigorously foams when shaken.
(1) Dissolve 0.4 g of Benzalkonium Chlo-ride Concentrated Solution 50 in 1 mL of sulfuric acid, add 0.1 g of sodium nitrate, and heat for 5 minutes on a water bath. After cooling, add 10 mL of water and 0.5 g of zinc powder, heat for 5 minutes, cool, and filter: the filtrate re-sponds to Qualitative Tests <1.09> for primary aromatic amines. The color of the solution is red. To 2 mL of a solution of Benzalkonium Chloride Concentrated Solution 50 (1 in 500) add a mixture of 0.2 mL of a solution of bromophenol blue (1 in 2000) and 0.5 mL of sodium hydroxide TS: a blue color develops. Add 4 mL of chloroform to this solution, and shake vigorously: the blue color shifts to the chloroform layer. Collect the chloroform layer, and add dropwise, with stirring, a solution of sodium ‌JP XVIII Official Monographs / Benzbromarone 521 lauryl sulfate (1 in 1000): the chloroform layer turns color-less. Determine the absorption spectrum of a solution of Benzalkonium Chloride Concentrated Solution 50 in 0.1 mol/L hydrochloric acid TS (1 in 1000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum of Benzalkonium Chloride: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. To 1 mL of a solution of Benzalkonium Chloride Concentrated Solution 50 (1 in 50) add 2 mL of ethanol (95), 0.5 mL of dilute nitric acid and 1 mL of silver nitrate TS: a white precipitate is produced. This precipitate does not dis-solve on the addition of dilute nitric acid, but dissolves on the addition of ammonia TS.
(1) Clarity and color of solution—Dissolve 2.0 g of Benzalkonium Chloride Concentrated Solution 50 in 10 mL of water: the solution is clear and colorless to light yel-low. (2) Petroleum ether-soluble substances—To 6.0 g of Benzalkonium Chloride Concentrated Solution 50 add water to make 50 mL, then add 50 mL of ethanol (99.5) and 5 mL of 0.5 mol/L sodium hydroxide TS, and extract with three 50-mL portions of petroleum ether. Combine the petroleum ether extracts, and wash with three 50-mL portions of dilute ethanol. After shaking well with 10 g of anhydrous sodium sulfate, filter through a dry filter paper, and wash the filter paper with two 10-mL portions of petroleum ether. Evaporate the petroleum ether on a water bath by heating, and dry the residue at 1059C for 1 hour: the residue is not more than 1.0z. Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.3 g of Benzalkonium Chloride Concentrated Solution 50, and dissolve in 75 mL of water. Adjust the pH to between 2.6 and 3.4 by adding dropwise diluted dilute hydrochloric acid (1 in 2), add 1 drop of methyl orange TS, and titrate <2.50> with 0.02 mol/L sodium tetraphenylboron VS until the color of the solution becomes red. Each mL of 0.02 mol/L sodium tetraphenylborate VS ＝ 7.080 mg of C22H40ClN
Benzethonium Chloride Solution contains not less than 93.
Benzethonium Chloride Solution is a clear, colorless liquid. It is odorless. It foams strongly when shaken.
(1) Evaporate a volume of Benzethonium Chloride Solution, equivalent to 0.2 g of Benzethonium Chloride, on a water bath to dryness, and proceed with the residue as directed in the Identification (1) under Benzetho-nium Chloride. To a volume of Benzethonium Chloride Solution, equivalent to 0.01 g of Benzethonium Chloride, add water to make 10 mL, proceed with 2 mL of this solution as directed in the Identification (2) under Benzethonium Chloride. To a volume of Benzethonium Chloride Solution, equivalent to 1 g of Benzethonium Chloride, and add water or concentrate on a water bath to make 10 mL. To 1 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 500 mL, and determine the abosorption spectrum as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 262 nm and 264 nm, between 268 nm and 270 nm, and between 274 nm and 276 nm. To a volume of Benzethonium Chloride Solution, equivalent to 0.1 g of Benzethonium Chloride, add water, or concentrate on a water bath, if necessary, to make 10 mL, and proceed with 1 mL of this solution as directed in the Identification (4) under Benzethonium Chloride.
(1) Nitrite—Add 1.0 mL of Benzethonium Chlo-ride Solution to a mixture of 1 mL of a solution of glycine (1 in 10) and 0.5 mL of acetic acid (31): no gas is evolved. (2) Oxidizing substances—To 5 mL of Benzethonium Chloride Solution add 0.5 mL of potassium iodide TS and 2 to 3 drops of dilute hydrochloric acid: no yellow color is produced.
Pipet a volume of Benzethonium Chloride Solution, equivalent to about 0.2 g of benzethonium chloride (C27H42ClNO2), dilute with water to make 75 mL, if neces-sary, and proceed as directed in the Assay under Benzetho-nium Chloride. ‌JP XVIII Official Monographs / Benzyl Alcohol 523‌ Each mL of 0.02 mol/L sodium tetraphenylboron VS ＝ 8.962 mg of C27H42ClNO2
Bepotastine Besilate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of bepotastine besilate (C21H25ClN2O3.C6H6O3S: 547.06). Method of preparation Prepare as directed under Tablets, with Bepotastine Besilate. Identification To an amount of powdered Bepotastine Besilate Tablets, equivalent to 2 mg of Bepotastine Besilate, add 40 mL of water, shake thoroughly, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Bepotastine Besilate Tablets add exactly V/5 mL of the internal standard solution, then add the mo-bile phase to make V mL so that each mL contains about 0.
To an amount of powdered Bepotastine Besilate Tablets, equivalent to 2 mg of Bepotastine Besilate, add 40 mL of water, shake thoroughly, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Bepotastine Besilate Tablets add exactly V/5 mL of the internal standard solution, then add the mo-bile phase to make V mL so that each mL contains about 0.4 mg of bepotastine besilate (C21H25ClN2O3.C6H6O3S), shake ple solution. Separately, weigh accurately about 0.11 g of bepotastine besilate for assay (separately determine the water <2.48> and the residual solvent in the same manner as Bepotastine Besilate), and dissolve in water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of bepotastine in each solution. Dissolution rate (z) with respect to the labeled amount of bepotastine besilate (C21H25ClN2O3.C6H6O3S) ＝ MS × AT/AS × V?/V × 1/C × 9/5 MS: Amount (mg) of bepotastine besilate for assay taken, calculated on the anhydrous and residual solvent-free basis C: Labeled amount (mg) of bepotastine besilate (C21H25ClN2O3.C6H6O3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bepotastine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bepotastine is not more than 1.5z.
Weigh accurately the mass of not less than 20 tablets of Bepotastine Besilate Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 10 mg of vigorously for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the bepotastine besilate (C21H25 ClN2O3.C6H6O3S), add exactly 5 first 5 mL of the filtrate, to 2 mL of the subsequent filtrate add the mobile phase to make 10 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of bepotastine besilate (C21H25ClN2O3.C6H6O3S) ＝ MS × QT/QS × V/50 MS: Amount (mg) of bepotastine besilate for assay taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in acetonitrile (1 in 4500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate mL of the internal standard solution, then add 20 mL of the mobile phase, shake thoroughly for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, to 2 mL of the subsequent filtrate add the mobile phase to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of bepotastine besilate for assay (separately determine the water <2.48> and the residual sol-vent in the same manner as Bepotastine Besilate), add ex-actly 10 mL of the internal standard solution, and dissolve in the mobile phase to make 50 mL. To 2 mL of this solution add the mobile phase to make 10 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of bepotastine to that of the internal standard. ‌JP XVIII Official Monographs / Beraprost Sodium 531 Amount (mg) of bepotastine besilate (C21H25ClN2O3.C6H6O3S) ＝ MS × QT/QS × 1/2 MS: Amount (mg) of bepotastine besilate for assay taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in acetonitrile (1 in 4500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A solution of sodium 1-pentanesulfonate in a mixture of 0.05 mol/L potassium dihydrogen phosphate TS (pH 3.0) and acetonitrile (7:3) (1 in 1000). Flow rate: Adjust so that the retention time of bepotastine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, bepotastine and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of bepotastine to that of the internal standard is not more than 1.0z.
Beraprost Sodium Tablets contain not less than 95.0z and not more than 105.0z of the labeled System performance: When the procedure is run with 15 mL of the solution for system suitability test under the above amount of beraprost sodium (C 24H29 NaO5: 420.47). operating conditions, the resolution between the two peaks of beraprost is not less than 1.5. System repeatability: When the test is repeated 6 times with 15 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the total area of the two peaks of beraprost is not more than 2.0z. Loss on drying <2.41> Not more than 3.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, silica gel, 609C, 5 hours). Isomer ratio Dissolve 10 mg of Beraprost Sodium in 5 mL of methanol, and use this solution as the sample solution. Perform the test with 15 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine the areas, Aa of the peak which appears at the retention time about 25 minutes, and Ab of the peak which appears at about 27 minutes: Ab/ Aa is between 0.90 and 1.10. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water and acetic acid (100) (600:400:1). Flow rate: Adjust so that the retention time of the second eluting peak of beraprost is about 27 minutes. System suitability— System performance: When the procedure is run with 15 mL of the sample solution under the above operating condi-tions, the resolution between the two peaks of beraprost is not less than 1.2. Method of preparation Prepare as directed under Tablets, with Beraprost Sodium. Identification Powder Beraprost Sodium Tablets. To a portion of the powder, equivalent to 0.2 mg of Beraprost Sodium, add 10 mL of water, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To the filtrate add 1 mL of 0.1 mol/L hydrochloric acid TS, ex-tract with two 50-mL portions of ethyl acetate, combine the extracts, and evaporate in reduced pressure at 409C. Dis-solve the residue in 1 mL of methanol, use this solution as the sample solution. Separately, dissolve 1 mg of beraprost sodium in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with the upper layer of a mixture of 11 volumes of ethyl acetate, 10 volumes of water, 4 volumes of isooctane and 2 volumes of acetic acid (100) to a distance of about 10 cm, air-dry the plate, and heat at 1209C for 30 minutes. After cooling, spray evenly a mixture of ethanol (99.5), water, sulfuric acid and 4-methoxybenzaldehide (17:2:1:1) on the plate, and heat the plate at 1209C for 3 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Beraprost Sodium Tablets add exactly V mL of the internal standard solution so that each mL contains about 2 mg of beraprost sodium (C24H29NaO5), shake at 309C for 30 minutes, filter through a membrane filter with a pore size not exceeding 0.
Powder Beraprost Sodium Tablets. To a portion of the powder, equivalent to 0.2 mg of Beraprost Sodium, add 10 mL of water, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To the filtrate add 1 mL of 0.1 mol/L hydrochloric acid TS, ex-tract with two 50-mL portions of ethyl acetate, combine the extracts, and evaporate in reduced pressure at 409C. Dis-solve the residue in 1 mL of methanol, use this solution as the sample solution. Separately, dissolve 1 mg of beraprost sodium in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with the upper layer of a mixture of 11 volumes of ethyl acetate, 10 volumes of water, 4 volumes of isooctane and 2 volumes of acetic acid (100) to a distance of about 10 cm, air-dry the plate, and heat at 1209C for 30 minutes. After cooling, spray evenly a mixture of ethanol (99.5), water, sulfuric acid and 4-methoxybenzaldehide (17:2:1:1) on the plate, and heat the plate at 1209C for 3 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Beraprost Sodium Tablets add exactly V mL of the internal standard solution so that each mL contains about 2 mg of beraprost sodium (C24H29NaO5), shake at 309C for 30 minutes, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Then, proceed as directed in the Assay. System repeatability: When the test is repeated 6 times with 15 mL of the sample solution under the above operating conditions, the relative standard deviation of the total area Amount (mg) of beraprost sodium (C ＝ MS × QT/QS × V/10,000 24H 29NaO5) of the two peak of beraprost is not more than 2.0z.
Weigh accurately about 0.1 g of Beraprost Sodium, previously dried, dissolve in 30 mL of diluted ethanol with freshly boiled and cooled water (7 in 10), add exactly 2 mL of 0.2 mol/L hydrochloric acid TS, and titrate <2.50> with 0.025 mol/L sodium hydroxide-ethanol VS from the first equivalence point to the second equivalence point (potentio-metric titration). Each mL of 0.025 mol/L sodium hydroxide-ethanol VS ＝ 10.51 mg of C24H29NaO5
Betahistine Mesilate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of betahistine mesilate (C H N .2CH O S: directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by 328.41). 8 12 2 4 3 the automatic integration method: the area of the peak other than betahistine obtained from the sample solution is not larger than 1/10 times the peak area of betahistine from the standard solution, and the total area of the peaks other than the peak of betahistine from the sample solution is not larger than 1/2 times the peak area of betahistine from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 261 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 5 mL of diethylamine and 20 mL of acetic acid (100) add water to make 1000 mL. Dissolve 2.3 g of sodium lauryl sulfate in 630 mL of this solution, and add 370 mL of acetonitrile. Flow rate: Adjust so that the retention time of betahistine is about 5 minutes. Time span of measurement: About 3 times as long as the retention time of betahistine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mixture of water and acetonitrile (63:37) to make exactly 50 mL. Confirm that the peak area of betahistine obtained with 20 mL of this solution is equiva-lent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg of betahistine mesi-late and 10 mg of 2-vinylpyridine in 50 mL of the mixture of water and acetonitrile (63:37). To 2 mL of this solution add the mixture of water and acetonitrile (63:37) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 2-vinylpyridine and beta-histine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betahistine is not more than 1.0z Method of preparation Prepare as directed under Tablets, with Betahistine Mesilate. Identification To 5 mL of the sample solution obtained in the Assay add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 259 nm and 263 nm. Purity Related substances—Powder not less than 20 Beta-histine Mesilate Tablets. To a portion of the powder, equiva-lent to about 50 mg of Betahistine Mesilate, add 10 mL of a mixture of water and acetonitrile (63:37), agitate for 10 minutes by sonicating, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solu-tion, add the mixture of water and acetonitrile (63:37) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having the relative retention time of about 1.9 to betahistine obtained from the sample solution, is not larger than 3/5 times the peak area of betahistine from the standard solution, and the total area of the peaks other than betahistine from the sample solution is not larger than the peak area of betahistine from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 8 times as long as the retention time of betahistine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mixture of water and acetonitrile (63:37) to make exactly 50 mL. Confirm that the peak area of betahistine obtained with 20 mL of this solution is equiva-lent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg of betahistine mesi-late and 10 mg of 2-vinylpyridine in 50 mL of the mixture of JP XVIII Official Monographs / Betahistine Mesilate Tablets 537 water and acetonitrile (63:37). To 2 mL of this solution add the mixture of water and acetonitrile (63:37) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 2-vinylpyridine and beta-histine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betahistine is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Betahistine Mesilate Tablets add exactly V mL of 0.1 mol/L hydrochloric acid TS so that each mL contains about 0.
To 5 mL of the sample solution obtained in the Assay add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 259 nm and 263 nm.
Related substances—Powder not less than 20 Beta-histine Mesilate Tablets. To a portion of the powder, equiva-lent to about 50 mg of Betahistine Mesilate, add 10 mL of a mixture of water and acetonitrile (63:37), agitate for 10 minutes by sonicating, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solu-tion, add the mixture of water and acetonitrile (63:37) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having the relative retention time of about 1.9 to betahistine obtained from the sample solution, is not larger than 3/5 times the peak area of betahistine from the standard solution, and the total area of the peaks other than betahistine from the sample solution is not larger than the peak area of betahistine from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 8 times as long as the retention time of betahistine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mixture of water and acetonitrile (63:37) to make exactly 50 mL. Confirm that the peak area of betahistine obtained with 20 mL of this solution is equiva-lent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg of betahistine mesi-late and 10 mg of 2-vinylpyridine in 50 mL of the mixture of JP XVIII Official Monographs / Betahistine Mesilate Tablets 537 water and acetonitrile (63:37). To 2 mL of this solution add the mixture of water and acetonitrile (63:37) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, 2-vinylpyridine and beta-histine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betahistine is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Betahistine Mesilate Tablets add exactly V mL of 0.1 mol/L hydrochloric acid TS so that each mL contains about 0.4 mg of betahistine mesilate (C8H12N2.2CH4O3S), agitate for about 10 minutes by sonicating to disintegrate the tablet, then centrifuge, and use the supernatant liquid as the sample solution. Proceed as directed in the Assay. Amount (mg) of betahistine mesilate (C8H12N2.2CH4O3S) ＝ MS × AT/AS × V/250 MS: Amount (mg) of betahistine mesilate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Betahistine Mesilate Tablets is not less than 85z. Start the test with 1 tablet of Betahistine Mesilate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 6.7 mg of betahistine mesilate (C8H12N2. 2CH4O3S), and use this solution as the sample solution. Separately, weigh accurately about 17 mg of betahistine mesilate for assay, previously dried under reduced pressure with phosphorous (V) oxide at 709C for 24 hours, and dis-solve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of betahistine in each solution. Dissolution rate (z) with respect to the labeled amount of betahistine mesilate (C8H12N2.2CH4O3S) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of betahistine mesilate for assay taken C: Labeled amount (mg) of betahistine mesilate (C8H12N2.2CH4O3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of betahistine are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak area of betahistine is not more than 2.0z.
Weigh accurately the mass of not less than 20 Beta-histine Mesilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of beta-histine mesilate (C8H12N2.2CH4O3S), add 40 mL of 0.1 mol/L hydrochloric acid TS, agitate for 10 minutes by sonicating, and add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 0.1 g of betahistine mesilate for assay, previously dried under reduced pressure with phosphorous (V) oxide at 709C for 24 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 10 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01>, ac-cording to the following conditions, and determine the peak areas, AT and AS, of betahistine in each solution. Amount (mg) of betahistine mesilate (C8H12N2.2CH4O3S) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of betahistine mesilate for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 261 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 5 mL of diethylamine and 20 mL of acetic acid (100) add water to make 1000 mL. In 630 mL of this solution dissolve 2.3 g of sodium lauryl sulfate, and add 370 mL of acetonitrile. Flow rate: Adjust so that the retention time of betahistine is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of betahistine are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of betahistine is not more than 1.0z.
Betamethasone Tablets contain not less than 90.0z and not more than 107.0z of the labeled amount of betamethasone (C22H29FO5: 392.46). Method of preparation Prepare as directed under Tablets, with Betamethasone. Identification Pulverize Betamethasone Tablets. To a por-tion of the powder, equivalent to 2 mg of Betamethasone, add 20 mL of methanol, shake for 5 minutes, and filter. Evaporate the filtrate on a water bath to dryness, dissolve the residue after cooling in 2 mL of methanol, filter if neces-sary, and use this as the sample solution. Separately, dissolve 2 mg of Betamethasone RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop with a mixture of 1-butanol, water and acetic anhydride (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the principal spot ob-tained from the sample solution and the spot from the stand- the peak area of betamethasone to that of the internal stand-ard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Betamethasone Tablets is not less than 85z. Start the test with 1 tablet of Betamethasone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL of the filtrate, add water to make exactly V? mL so that each mL contains about 0.
Pulverize Betamethasone Tablets. To a por-tion of the powder, equivalent to 2 mg of Betamethasone, add 20 mL of methanol, shake for 5 minutes, and filter. Evaporate the filtrate on a water bath to dryness, dissolve the residue after cooling in 2 mL of methanol, filter if neces-sary, and use this as the sample solution. Separately, dissolve 2 mg of Betamethasone RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop with a mixture of 1-butanol, water and acetic anhydride (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the principal spot ob-tained from the sample solution and the spot from the stand- the peak area of betamethasone to that of the internal stand-ard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Betamethasone Tablets is not less than 85z. Start the test with 1 tablet of Betamethasone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL of the filtrate, add water to make exactly V? mL so that each mL contains about 0.56 mg of betamethasone (C22H29FO5), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Betamethasone RS, previ-ously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make ex-actly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of be-tamethasone in each solution. Dissolution rate (z) with respect to the labeled amount ard solution show the same Rf value. of betamethasone (C 22H 29FO5) Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Betamethasone Tablets add V mL of water so that each mL contains about 50 mg of betamethasone (C22H29FO5). Add exactly 2V mL of the internal standard so-lution, shake vigorously for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Betamethasone RS, previ-ously dried for 4 hours in a desiccator (in vacuum, phospho-rus (V) oxide), dissolve in acetonitrile to make exactly 200 mL. Pipet 5 mL of this solution, add exactly 20 mL of the internal standard solution, add 5 mL of water, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of betamethasone to that of the internal standard. ＝ MS × AT/AS × V?/V × 1/C × 9/5 MS: Amount (mg) of Betamethasone RS taken C: Labeled amount (mg) of betamethasone (C22H29FO5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 241 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (3:2). Flow rate: Adjust so that the retention time of betametha- sone is about 7 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con- Amount (mg) of betamethasone (C ＝ MS × QT/QS × V/400 22H 29FO5) ditions, the number of theoretical plates and the symmetry factor of the peak of betamethasone are not less than 3000 and not more than 2.0, respectively. MS: Amount (mg) of Betamethasone RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in acetonitrile (1 in 40,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, betamethasone and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of betamethasone is not more than 2.0z.
Weigh accurately the mass of not less than 20 Beta-methasone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 5 mg of betamethasone (C22H29FO5), add 25 mL of water, then add exactly 50 mL of the internal standard solution, and shake vigorously for 10 minutes. Filter through a membrane filter with pore size not exceeding 0.5 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of Betamethasone RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in acetonitrile to make ex- ‌540 actly 50 mL. Pipet 5 mL of this solution, add exactly 20 mL of the internal standard solution and 5 mL of water, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of betamethasone to that of the internal standard. Amount (mg) of betamethasone (C22H29FO5) ＝ MS × QT/QS × 1/4 MS: Amount (mg) of Betamethasone RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in acetonitrile (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (3:2). Flow rate: Adjust so that the retention time of betametha- sone is about 4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, betamethasone and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of betamethasone to that of the internal stand-ard is not more than 1.0z.
Betamethasone Valerate and Gentamicin Sulfate Cream
ベタメタゾン吉草酸エステル･ゲンタマイシン硫酸塩クリーム
(1) To a quantity of Betamethasone Valer-ate and Gentamicin Sulfate Cream, equivalent to about 1.2 mg of Betamethasone Valerate, add 20 mL of methanol and 20 mL of hexane, shake vigorously for 10 minutes, and allow to stand. Take 15 mL of the lower layer, evaporate the layer to dryness on a water bath under a current of nitrogen. To the residue add 1 mL of ethyl acetate, mix, and use as the sample solution. Separately, dissolve about 18 mg of Be-tamethasone Valerate RS in 20 mL of ethyl acetate, and use this solution as the standard solution. Perform the test with mL of water, shake vigorously for 10 minutes, and centri-fuge. To 3 mL of the lower layer add 1 mL of dilute sodium hydroxide TS and 2 mL of ninhydrin TS, and heat in a water bath at 90 – 959C for 10 minutes: a purple to dark purple color develops. pH <2.54> To a quantity of Betamethasone Valerate and Gentamicin Sulfate Cream, equivalent to 6 mg of Beta-methasone Valerate, add 15 mL of water, and mix while warming on a water bath to make a milky liquid: the pH of the cooled liquid is between 4.0 and 6.0.
Related substances—Weigh accurately an amount of Betamethasone Valerate and Gentamicin Sulfate Cream, equivalent to about 1 mg of Betamethasone Valerate, and add 10 mL of a mixture of methanol and water (7:3). Warm in a water bath at 609C for 5 minutes, and shake vigorously for 20 minutes. Repeat this procedure 2 times. After cooling for 15 minutes with ice, centrifuge for 5 minutes, take away the bubbles from the upper surface, and filter the remaining liquid. Discard first 2 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Perform the test with 150 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, determine each peak area by the automatic integration method, and calculate these amounts by the area percentage method: the amount of the substance other than betametha-sone valerate is not more than 3.5z, and the total amount of them is not more than 7.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of water, acetonitrile and metha-nol (12:7:1). Flow rate: Adjust so that the retention time of betametha-sone valerate is about 16 minutes. Time span of measurement: About 2.5 times as long as the retention time of betamethasone valerate beginning after the solvent peak. The peaks of the compounding ingredients are not determined. System suitability— Test for required detectability: Dissolve 20 mg of Beta-methasone Valerate in 100 mL of a mixture of methanol and water (7:3). To exactly 1 mL of this solution add the mixture of methanol and water (7:3) to make exactly 100 mL, and use this solution as the solution for system suitability test. To exactly 2.5 mL of the solution for system suitability test add the mixture of methanol and water (7:3) to make exactly 50 mL. Confirm that the peak area of betamethasone valerate obtained with 150 mL of this solution is equivalent to 3.5 to 6.5z of that with 150 mL of the solution for system suita- ‌544 bility test. System performance: When the procedure is run with 150 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of betamethasone valerate are not less than 4000 and 0.8 to 1.3, respectively. System repeatability: When the test is repeated 6 times with 150 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of betamethasone valerate is not more than 2.0z.
(1) Betamethasone valerate—Weigh accurately an amount of Betamethasone Valerate and Gentamicin Sulfate Cream, equivalent to about 1 mg of betamethasone valerate (C27H37FO6), add 10 mL of a mixture of methanol and water (7:3), and add exactly 10 mL of the internal standard solu-tion. After warming in a water bath at 609C for 5 minutes, shake vigorously for 20 minutes. Repeat this procedure twice, cool with ice for 15 minutes, centrifuge for 5 minutes, then filter the supernatant liquid, discard the first 5 mL of filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 25 mg of Beta-methasone Valerate RS, previously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, and add the mixture of metha-nol and water (7:3) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard so-lution, mix, and use this solution as the standard solution. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of betamethasone valerate to that of the internal standard. Amount (mg) of betamethasone valerate (C27H37FO6) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of Betamethasone Valerate RS taken Internal standard solution—Dissolve 20 mg of beclometa-sone dipropionate in 10 mL of methanol, and add the mix-ture of methanol and water (7:3) to make 200 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 2.1 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (13:7). Flow rate: Adjust so that the retention time of betametha- sone valerate is about 16 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, betamethasone valerate and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of betamethasone valerate to that of the internal standard is not more than 1.0z. Gentamicin sulfate—Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, agar media for base layer and seed layer, agar medium for transferring test organisms, and standard solutions—Proceed as directed in the Assay under Gentamicin Sulfate. Sample solutions—Weigh accurately an amount of Betamethasone Valerate and Gentamicin Sulfate Cream, equivalent to about 1 mg (potency) of Gentamicin Sulfate, add 100 mL of 0.1 mol/L phosphate buffer solution (pH 8.0) previously warmed to about 859C, and shake well to dissolve. After cooling, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 250 mL to make the high concentration sample solution, which contains 4 mg (potency) per mL. Pipet a suitable amount of the high concentration sample solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) so that each mL contains 1 mg (potency), and use this solution as the low concentration sample solution.
Betamethasone Valerate and Gentamicin Sulfate Ointment
ベタメタゾン吉草酸エステル･ゲンタマイシン硫酸塩軟膏
Betamethasone Valerate and Gentamicin Sulfate Ointment contains not less than 95.
(1) To a quantity of Betamethasone Valer-ate and Gentamicin Sulfate Ointment, equivalent to 1.2 mg of Betamethasone Valerate, add 20 mL of methanol and 20 mL of hexane, and disperse the ointment by sonicating. Shake vigorously for 5 minutes, centrifuge for 5 minutes, cool for 15 minutes with ice, and take 15 mL of the lower layer. Evaporate the layer to dryness on a water bath under a current of nitrogen. To the residue add 1 mL of ethyl ace-tate, sonicate, filter, if necessary, and use the filtrate as the sample solution. Separately, dissolve 18 mg of Betametha-sone Valerate RS in 20 mL of ethyl acetate, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, de-velop the plate with ethyl acetate to a distance of about 10 cm, and air-dry the plate. Spray evenly alkaline blue tetrazo-lium TS on the plate, and heat the plate at 1009C: the princi-pal spot obtained from the sample solution and the spot from the standard solution are purple in color, and their Rf values are the same. (2) To a quantity of Betamethasone Valerate and Gen-tamicin Sulfate Ointment, equivalent to 2 mg (potency) of Gentamicin Sulfate, add 20 mL of hexane and 10 mL of water, shake vigorously for 10 minutes, and centrifuge. To 3 mL of the lower layer add 1 mL of dilute sodium hydrox-ide TS and 2 mL of ninhydrin TS, and heat in a water bath at 90 – 959C for 10 minutes: a red-brown color develops. pH <2.54> To a quantity of Betamethasone Valerate and Gentamicin Sulfate Ointment, equivalent to 6 mg of Beta-methasone Valerate, add 15 mL of water, and warm on a ‌JP XVIII Official Monographs / Betamipron 545 water bath to dissolve. After cooling, separate the water layer: the pH of the layer is between 4.0 and 7.0.
(1) Betamethasone valerate—Weigh accurately an amount of Betamethasone Valerate and Gentamicin Sulfate Ointment, equivalent to about 1 mg of betamethasone valer-ate (C27H37FO6), add 10 mL of a mixture of methanol and water (7:3), and add exactly 10 mL of the internal standard solution. After warming in a water bath at 759C for 5 minutes, shake vigorously for 10 minutes. Repeat this proce-dure once more, cool with ice for 15 minutes, filter, discard the first 5 mL of filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of Betamethasone Valerate RS, previously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, and add the mixture of methanol and water (7:3) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal stand-ard solution, mix, and use this solution as the standard solu-tion. Perform the test with 3 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of betametha-sone valerate to that of the internal standard. Amount (mg) of betamethasone valerate (C27H37FO6) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of Betamethasone Valerate RS taken Internal standard solution—Dissolve 20 mg of beclometa-sone dipropionate in 10 mL of methanol, and add the mix-ture of methanol and water (7:3) to make 200 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 2.1 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (13:7). Flow rate: Adjust so that the retention time of betametha- sone valerate is about 16 minutes. System suitability— System performance: When the procedure is run with 3 mL of the standard solution under the above operating condi-tions, betamethasone valerate and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 3 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of betamethasone valerate to that of the internal standard is not more than 1.0z. Gentamicin sulfate—Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, agar media for base layer and seed layer, agar medium for transferring test organisms, and standard solutions—Proceed as directed in the Assay under Gentamicin Sulfate. Sample solutions—Weigh accurately an amount of Betamethasone Valerate and Gentamicin Sulfate Ointment, equivalent to about 1 mg (potency) of Gentamicin Sulfate, transfer to a separator, add 50 mL of petroleum ether and exactly 100 mL of 0.1 mol/L phosphate buffer solution (pH 8.0), shake for 10 minutes, and allow to stand. Pipet a suita-ble amount of the water layer, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and low concentration sample solution, respectively.
Mix well an amount of powdered Bezafibrate Extended-release Tablets, equivalent to 0.1 g of Bezafibrate, with 100 mL of methanol, and filter. To 1 mL of the filtrate add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 227 nm and 231 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citric acid buffer solu-tion (pH 7.2) as the dissolution medium, the dissolution rates of a 100-mg tablet in 1.5 hours, in 2.5 hours and in 8 hours are 15 – 45z, 35 – 65z and not less than 80z, respec-tively, and those of a 200-mg tablet in 1.5 hours, in 2.5 hours and in 8 hours are 15 – 45z, 30 – 60z and not less than 75z, respectively. Start the test with 1 tablet of Bezafibrate Extended-release Tablets, withdraw exactly 20 mL of the medium at the speci-fied minutes after starting the test, and immediately fill up the dissolution medium each time with exactly 20 mL of fresh dissolution medium, previously warmed to 37 ± 0.59C. Filter these media through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL, add the dis-solution medium to make exactly V? mL so that each mL contains about 13 mg of bezafibrate (C19H20ClNO4), and use these solutions as the sample solutions. Separately, weigh ac-curately about 66 mg of bezafibrate for assay, previously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this so-lution as the standard solution. Determine the absorbances, AT(n) (n ＝ 1,2,3) and AS, of the sample solutions and stand-ard solution at 228 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Ø A » Dissolution rate (z) in each case of n with respect to the labeled amount of bezafibrate (C19H20ClNO4) for 20 minutes. Add diluted 0.5 mol/L ammonium acetate TS (1 in 50) to make 100 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 20 mg of bezafibrate for assay, previously dried at 1059C for 3 hours, dissolve in 60 mL of methanol, add exactly 10 mL of the internal standard solution and diluted 0.5 mol/L ammo-nium acetate TS (1 in 50) to make 100 mL, and use this solu-tion as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of bezafibrate to that of the internal standard. Amount (mg) of bezafibrate (C19H20ClNO4) ＝ MS × QT/QS MS: Amount (mg) of bezafibrate for assay taken Internal standard solution—A solution of 4-nitrophenol in methanol (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and diluted acetic acid (100) (1 in 100) (9:4). Flow rate: Adjust so that the retention time of bezafibrate is about 6 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the internal standard and bezafibrate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of bezafibrate to that of the internal standard is not more than 1.0z.
Bisoprolol Fumarate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of bisoprolol fumarate [(C H NO ) .C H O : 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to 766.96]. 18 31 4 2 4 4 4 the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peaks other than bisoprolol obtained from the sample so-lution is not larger than 1/2 times the peak area of bisoprolol from the standard solution. Furthermore, the total of the areas of peaks other than bisoprolol from the sample solu-tion is not larger than the peak area of bisoprolol from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 800 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of bisoprolol is about 8 minutes. Time span of measurement: About 2 times as long as the retention time of bisoprolol, beginning after the fumaric acid peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of water and acetonitrile (4:1) to make exactly 20 mL. Confirm that the peak area of bi-soprolol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bisoprolol is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 809C, 5 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.6 g of Bisoprolol Method of preparation Prepare as directed under Tablets, with Bisoprolol Fumarate. Identification To a quantity of powdered Bisoprolol Fumarate Tablets, equivalent to 10 mg of Bisoprolol Fumarate, add 60 mL of methanol, shake vigorously for 10 minutes, add methanol to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. De-termine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 271 nm and 275 nm. Purity Related substances—This is applied to 0.625-mg tablets. Shake vigorously for 10 minutes a portion of pow-dered Bisoprolol Fumarate Tablets, equivalent to 5 mg of Bi-soprolol Fumarate, with exactly 20 mL of a mixture of water and acetonitrile (3:1), filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of the peak other than bisoprolol and the peak having the rela-tive retention time of about 0.8 to bisoprolol by the area per-centage method: the amount of the two peaks, having rela-tive retention time of about 1.2 and about 3.8 to bisoprolol, are not more than 1.0z, respectively, the amount of the peak other than the peaks mentioned above is not more than 0.2z, and the total amount of the peaks other than bi-soprolol is not more than 2.5z. For the area of the peak, having the relative retention time of about 1.2 to bisoprolol, multiply the correction factor 5. Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. Time span of measurement: About 5 times as long as the retention time of bisoprolol, beginning after the peak of fumaric acid. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water and acetonitrile (3:1) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suita-bility test, and add a mixture of water and acetonitrile (3:1) to make exactly 20 mL. Confirm that the peak area of bi-soprolol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suita-bility test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of bisoprolol is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Bisoprolol Fumarate Tablets, disintegrate by adding 8 mL of water, and add water to make exactly 10 mL, and then filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the fil-trate, pipet V mL of the subsequent filtrate, add water to MS: Amount (mg) of bisoprolol fumarate for assay taken C: Labeled amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4] in 1 tablet Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bisoprolol is not more than 2.0z. Assay Weigh accurately not less than 20 Bisoprolol Fumarate Tablets and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of bisoprolol make exactly V? mL so that each mL contains about 62.
To a quantity of powdered Bisoprolol Fumarate Tablets, equivalent to 10 mg of Bisoprolol Fumarate, add 60 mL of methanol, shake vigorously for 10 minutes, add methanol to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. De-termine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 271 nm and 275 nm.
Related substances—This is applied to 0.625-mg tablets. Shake vigorously for 10 minutes a portion of pow-dered Bisoprolol Fumarate Tablets, equivalent to 5 mg of Bi-soprolol Fumarate, with exactly 20 mL of a mixture of water and acetonitrile (3:1), filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of the peak other than bisoprolol and the peak having the rela-tive retention time of about 0.8 to bisoprolol by the area per-centage method: the amount of the two peaks, having rela-tive retention time of about 1.2 and about 3.8 to bisoprolol, are not more than 1.0z, respectively, the amount of the peak other than the peaks mentioned above is not more than 0.2z, and the total amount of the peaks other than bi-soprolol is not more than 2.5z. For the area of the peak, having the relative retention time of about 1.2 to bisoprolol, multiply the correction factor 5. Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. Time span of measurement: About 5 times as long as the retention time of bisoprolol, beginning after the peak of fumaric acid. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water and acetonitrile (3:1) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suita-bility test, and add a mixture of water and acetonitrile (3:1) to make exactly 20 mL. Confirm that the peak area of bi-soprolol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suita-bility test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of bisoprolol is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Bisoprolol Fumarate Tablets, disintegrate by adding 8 mL of water, and add water to make exactly 10 mL, and then filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the fil-trate, pipet V mL of the subsequent filtrate, add water to MS: Amount (mg) of bisoprolol fumarate for assay taken C: Labeled amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4] in 1 tablet Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bisoprolol is not more than 2.0z.
Weigh accurately about 0.6 g of Bisoprolol Method of preparation Prepare as directed under Tablets, with Bisoprolol Fumarate. Identification To a quantity of powdered Bisoprolol Fumarate Tablets, equivalent to 10 mg of Bisoprolol Fumarate, add 60 mL of methanol, shake vigorously for 10 minutes, add methanol to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. De-termine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 271 nm and 275 nm. Purity Related substances—This is applied to 0.625-mg tablets. Shake vigorously for 10 minutes a portion of pow-dered Bisoprolol Fumarate Tablets, equivalent to 5 mg of Bi-soprolol Fumarate, with exactly 20 mL of a mixture of water and acetonitrile (3:1), filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of the peak other than bisoprolol and the peak having the rela-tive retention time of about 0.8 to bisoprolol by the area per-centage method: the amount of the two peaks, having rela-tive retention time of about 1.2 and about 3.8 to bisoprolol, are not more than 1.0z, respectively, the amount of the peak other than the peaks mentioned above is not more than 0.2z, and the total amount of the peaks other than bi-soprolol is not more than 2.5z. For the area of the peak, having the relative retention time of about 1.2 to bisoprolol, multiply the correction factor 5. Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. Time span of measurement: About 5 times as long as the retention time of bisoprolol, beginning after the peak of fumaric acid. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water and acetonitrile (3:1) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suita-bility test, and add a mixture of water and acetonitrile (3:1) to make exactly 20 mL. Confirm that the peak area of bi-soprolol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suita-bility test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of bisoprolol is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Bisoprolol Fumarate Tablets, disintegrate by adding 8 mL of water, and add water to make exactly 10 mL, and then filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the fil-trate, pipet V mL of the subsequent filtrate, add water to MS: Amount (mg) of bisoprolol fumarate for assay taken C: Labeled amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4] in 1 tablet Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 750 mL of this solution add 250 mL of acetonitrile. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bisoprolol are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bisoprolol is not more than 2.0z. Assay Weigh accurately not less than 20 Bisoprolol Fumarate Tablets and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of bisoprolol make exactly V? mL so that each mL contains about 62.5 mg of bisoprolol fumarate [(C18H31NO4)2.C4H4O4], and use this fumarate [(C18H31 NO4)2.C4H4O4], add 70 mL of a mixture of solution as the sample solution. Separately, weigh accurately about 20 mg of bisoprolol fumarate for assay, previously dried under reduced pressure at 809C for 5 hours, using phosphorus (V) oxide as a dessicant, and dissolve in water to make exactly 200 mL. Pipet 15 mL of this solution, add water to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 271.5 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of bisoprolol fumarate [(C18H31NO4)2.C4H4O4] ＝ MS × AT/AS × V?/V × 3/100 MS: Amount (mg) of bisoprolol fumarate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 water and acetonitrile (3:1) and exactly 10 mL of the internal standard solution, shake vigorously for 10 minutes, and add the mixture of water and acetonitrile (3:1) to make 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 3 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of bisoprolol fumarate for assay, previously dried in vacuum at 809C for 5 hours using phosphorus (V) oxide as the dessicant, add ex-actly 10 mL of the internal standard solution, dissolve in the mixture of water and acetonitrile (3:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of bisoprolol to that of the internal standard. mL of 2nd fluid for dissolution test as the dissolution me- Amount (mg) of bisoprolol fumarate [(C H NO ) .C H O ] dium, the dissolution rate in 30 minutes of Bisoprolol Fumarate Tablets is not less than 85z. ＝ MS × QT/QS 18 31 4 2 4 4 4 Start the test with 1 tablet of Bisoprolol Fumarate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 0.7 mg of bisoprolol fumarate [(C18H31NO4)2.C4H4O4], and use this solution as the sample solution. Separately, weigh accurately about 14 mg of bisoprolol fumarate for assay, previously dried in vacuum at 809C for 5 hours, using phosphorus (V) oxide as a dessicant, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 1 mL of this solution, add the dissolu-tion medium to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of bisoprolol in each solution. Dissolution rate (z) with respect to the labeled amount of bisoprolol fumarate [(C18H31NO4)2.C4H4O4] ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of bisoprolol fumarate for assay taken Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mixture of water and acetonitrile (3:1) (1 in 250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.08 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 800 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of bisoprolol is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- ‌JP XVIII Official Monographs / Bleomycin Hydrochloride 559 ditions, fumaric acid, bisoprolol and the internal standard are eluted in this order with the resolution between the peaks of bisoprolol and the internal standard being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of bisoprolol to that of the internal standard is not more than 1.0z.
ブロムフェナクナトリウム点眼液
Bromfenac Sodium Ophthalmic Solution is an aque-ous ophthalmic preparation. It contains not less than 90.
Bromfenac Sodium Ophthalmic Solution oc-curs as a clear and yellow liquid. Identiˆcation To a volume of Bromfenac Sodium Ophthal-mic Solution, equivalent to 1 mg of Bromfenac Sodium Hy-drate, add a solution of sodium hydrogen carbonate (21 in 2500) to make 50 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 266 nm and 270 nm, and between 377 nm and 381 nm. pH Being speciˆed separately when the drug is granted ap-proval based on the Law.
Related substances—Being speciˆed separately when the drug is granted approval based on the Law. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Pipet a volume of Bromfenac Sodium Ophthalmic Solution, equivalent to about 2 mg of bromfenac sodium hy-drate (C15H11BrNNaO3.11/2 H2O), add the mobile phase to make exactly 20 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 20 mg of Brom-fenac Sodium RS (separately determine the water <2.48> in the same manner as Bromfenac Sodium Hydrate), and dis-solve in the mobile phase to make exactly 20 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of bromfenac in each solution. Amount (mg) of bromfenac sodium hydrate (C15H11BrNNaO3.11/2H2O) ＝ MS × AT/AS × 1/10 × 1.076 MS: Amount (mg) of Bromfenac Sodium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.98 g of diammonium hydrogen phosphate in 750 mL of water, adjust to pH 7.3 with phos-phoric acid, and add 250 mL of acetonitrile. Flow rate: Adjust so that the retention time of bromfenac is about 18 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bromfenac are not less than 13,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bromfenac is not more than 1.0z.
Brotizolam Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of brotizolam (C15H10BrClN4S: 393.69). Method of preparation Prepare as directed under Tablets, with Brotizolam. Identification Shake a quantity of powdered Brotizolam Tablets, equivalent to 0.1 mg of Brotizolam, with 10 mL of methanol, and centrifuge. Determine the absorption spec-trum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 239 nm and 243 nm. JP XVIII Official Monographs / Brotizolam Tablets 569 Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the areas, AT and AS, of brotizolam in each solution. Dissolution rate (z) with respect to the labeled amount sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform of brotizolam (C 15H 10BrClN4S) the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than brotizolam obtained from the sample solution is not larger than 1.5 times the peak area of brotizolam from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of brotizolam, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 10 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of brotizolam obtained with 40 mL of this solution is equivalent to 7 to 13z of that ob-tained with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of brotizolam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of brotizolam is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Brotizolam Tablets add exactly V mL of the mobile phase so that each mL contains about 25 mg of brotizolam (C15H10BrClN4S), shake for 15 minutes, centri-fuge, and use the supernatant liquid as the sample solution.
Shake a quantity of powdered Brotizolam Tablets, equivalent to 0.1 mg of Brotizolam, with 10 mL of methanol, and centrifuge. Determine the absorption spec-trum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 239 nm and 243 nm. JP XVIII Official Monographs / Brotizolam Tablets 569
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the areas, AT and AS, of brotizolam in each solution. Dissolution rate (z) with respect to the labeled amount sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform of brotizolam (C 15H 10BrClN4S) the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks other than brotizolam obtained from the sample solution is not larger than 1.5 times the peak area of brotizolam from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of brotizolam, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 10 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of brotizolam obtained with 40 mL of this solution is equivalent to 7 to 13z of that ob-tained with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of brotizolam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of brotizolam is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Brotizolam Tablets add exactly V mL of the mobile phase so that each mL contains about 25 mg of brotizolam (C15H10BrClN4S), shake for 15 minutes, centri-fuge, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of brotizolam (C15H10BrClN4S) ＝ MS × AT/AS × V/1000 MS: Amount (mg) of brotizolam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Brotizolam Tablets is not less than 85z. Start the test with 1 tablet of Brotizolam Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.14 mg of brotizolam (C15H10BrClN4S), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of brotizolam for assay, previously dried at 1059C for 3 hours, dissolve in 10 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak ＝ MS × AT/AS × V?/V × 1/C × 9/20 MS: Amount (mg) of brotizolam for assay taken C: Labeled amount (mg) of brotizolam (C15H10BrClN4S) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: A mixture of water and acetonitrile (63:37). Flow rate: Adjust so that the retention time of brotizolam is about 7 minutes. System suitability— System performance: When the procedure is run with 200 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of brotizolam are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 200 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of brotizolam is not more than 2.0z.
Weigh accurately the mass of not less than 20 Brotizolam Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.25 mg of brotizolam (C15H10BrClN4S), add exactly 10 mL of the mobile phase, and shake for 15 minutes. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of brotizolam for assay, previ-ously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 40 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of brotizolam in each solution. Amount (mg) of brotizolam (C15H10BrClN4S) ＝ MS × AT/AS × 1/100 MS: Amount (mg) of brotizolam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.1 g of ammonium carbonate in 1000 mL of water. To 600 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of brotizolam is about 3 minutes. System suitability— System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of brotizolam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌570 ‌area of brotizolam is not more than 1.0z.
(1) To a quantity of powdered Bucillamine Tablets, equivalent to 0.1 g of Bucillamine, add 0.1 g of so-dium hydrogen carbonate and 10 mL of water, shake well, filter, and add 1 or 2 drops of ninhydrin TS to the filtrate: it exhibits a red-brown color. (2) To a quantity of powdered Bucillamine Tablets, equivalent to 0.1 g of Bucillamine, add 25 mL of water, shake well, and filter. To 5 mL of the filtrate, add 2 mL of JP XVIII Official Monographs / Bucillamine Tablets 571 dilute sodium hydroxide TS and 1 or 2 drops of sodium pentacyanonitrosylferrate (III) TS: it exhibits a red-purple color. Uniformity of dosage units <6.02>—Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Store the sample solution and standard solution in a cold place until performing the measurements. Take 1 tablet of Bucillamine Tablets, add exactly 1 mL of the internal stand-ard solution per 0.1 g of bucillamine (C7H13NO3S2), then add 3 mL of water and 6 mL of methanol per 0.1 g of bucilla-mine (C7H13NO3S2), and stir well until the tablet completely disintegrated. To 1 mL of this solution add the mobile phase to make 25 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of bucillamine (C7H13NO3S2) ＝ MS × QT/QS × C × 1/200 MS: Amount (mg) of bucillamine for assay taken C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1 tablet Internal standard solution—A solution of 4-fluorobenzoic acid in methanol (1 in 100). Dissolution <6.10>—When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Bucillamine Tablets is not less than 80z. Store the sample solution and standard solution in a cold place until performing the measurements. Start the test with 1 tablet of Bucillamine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately an amount of bucilla-mine for assay equivalent to the labeled amount of the tablet, previously dried in vacuum at 609C for 6 hours using phosphorus (V) oxide as a dessicant, and dissolve in metha-nol to make exactly 10 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of bucillamine in each solution. Dissolution rate (z) with respect to the labeled amount of bucillamine (C7H13NO3S2) ＝ MS × AT/AS × 1/C × 90 MS: Amount (mg) of bucillamine for assay taken C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and methanol (11:9). Flow rate: Adjust so that the retention time of bucillamine is about 4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bucillamine are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bucillamine is not more than 2.0z.
Store the sample solution and standard solution in a cold place until performing the measurements. Take 10 tablets of Bucillamine Tablets, add exactly 1 mL of the inter-nal standard solution per 0.1 g of bucillamine (C7H13NO3S2), add 3 mL of water and 6 mL of methanol, and stir well until the tablets completely disintegrated. To 1 mL of this solution add the mobile phase to make 25 mL, filter through a mem-brane filter with a pore size not exceeding 0.45 mm, and use this solution as the sample solution. Separately, weigh accu-rately about 0.2 g of bucillamine for assay, previously dried in vacuum at 609C for 6 hours using phosphorus (V) oxide as a dessicant, add exactly 2 mL of the internal standard solution, and add 6 mL of water and 12 mL of methanol. To 1 mL of this solution add the mobile phase to make 25 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use this solution as the standard so-lution. Perform the test with 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of bucilla-mine to that of the internal standard. Amount (mg) of bucillamine (C7H13NO3S2) ＝ MS × QT/QS × C × 1/200 MS: Amount (mg) of bucillamine for assay taken C: Labeled amount (mg) of bucillamine (C7H13NO3S2) in 1 tablet Internal standard solution—A solution of 4-fluorobenzoic acid in methanol (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and methanol (3:2). Flow rate: Adjust so that the retention time of bucillamine is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, bucillamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of bucillamine to that of the internal standard is not more than 1.0z.
Buformin Hydrochloride Delayed-release Tablets
To a quantity of powdered Buformin Hy-drochloride Delayed-release Tablets, equivalent to 0.1 g of Buformin Hydrochloride, add 10 mL of water, shake well, and then filter. To 4 mL of the filtrate add 1 mL of a mix-ture of hydrogen peroxide TS, sodium pentacyanonitro-sylferrate (III) TS and a solution of sodium hydroxide (1 in 10) (2:1:1): the solution exhibits a red to red-purple color. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Buformin Hydrochloride Delayed-release Tablets add 5 mL of a mixture of ethanol (99.5) and acetone (1:1), disperse the pellicle to smaller using ultrasonic waves, add exactly 10 mL of the internal standard solution per 50 mg of buformin hydrochloride (C6H15N5.HCl), and then add diluted acetonitrile (1 in 2) to make 13V/20 mL. Disintegrate the tablet using ultrasonic waves, then shake for 20 minutes, and add diluted acetonitrile (1 in 2) to make V mL so that each mL contains about 0.5 mg of buformin hydrochloride (C6H15N5.HCl) per mL. Centrifuge this solution, to 1 mL of the supernatant liquid, add the mobile phase to make 50 mL. If necessary, filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Then, proceed as directed in the Assay. Dissolution <6.10> When the tests are performed at 50 revolutions per minute according to the Paddle method, using 900 mL each of 1st fluid for dissolution test and 2nd fluid for dissolution test as the dissolution medium, the dis-solution rate in 120 minutes of Buformin Hydrochloride Delayed-release Tablets using 1st fluid is not more than 5z, and that in 90 minutes of Buformin Hydrochloride Delayed-release Tablets using 2nd fluid is not less than 80z. Start the test with 1 tablet of Buformin Hydrochloride Delayed-release Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the relevant dissolution medium to make exactly V? mL so that each mL contains about 56 mg of buformin hydrochloride (C6H15N5.HCl), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of buformin hy-drochloride for assay, previously dried at 1059C for 3 hours, and dissolve in the relevant dissolution medium to make ex-actly 100 mL. Pipet 4 mL of this solution, add the relevant dissolution medium to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of buformin in each solution. Dissolution rate (z) with respect to the labeled amount of buformin hydrochloride (C6H15N5.HCl) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of buformin hydrochloride for assay taken C: Labeled amount (mg) of buformin hydrochloride (C6H15N5.HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of a solution of sodium perchlorate in diluted phosphoric acid (1 in 1000) (7 in 500) and acetonitrile (7:1). Flow rate: Adjust so that the retention time of buformin is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of buformin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of buformin is not more than 2.0z. Amount (mg) of buformin hydrochloride (C H N .HCl)
Add 20 mL of a mixture of ethanol (99.5) and ＝ MS × QT/QS × V/50 6 15 5 acetone (1:1) to an amount of Buformin Hydrochloride Delayed-release Tablets equivalent to 0.5 g of buformin hy- MS: Amount (mg) of buformin hydrochloride for assay taken Internal standard solution—A solution of p-acetanisidide in diluted acetonitrile (1 in 2) (1 in 150). drochloride (C6H15N5.HCl), disperse the pellicles to smaller using ultrasonic waves, and then add 100 mL of diluted acetonitrile (1 in 2). Disintegrate the tablets with the aid of ultrasonic waves, shake for 20 minutes, and then add diluted acetonitrile (1 in 2) to make exactly 200 mL. Centrifuge this JP XVIII Official Monographs / Buformin Hydrochloride Tablets 575 ‌solution, pipet 10 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and then add diluted acetonitrile (1 in 2) to make 50 mL. Pipet 1 mL of this solu-tion, and add the mobile phase to make 50 mL. If necessary, filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solu-tion. Separately, weigh accurately about 25 mg of buformin hydrochloride for assay, previously dried at 1059C for 3 hours, dissolve in an adequate amount of diluted acetonitrile (1 in 2), add exactly 5 mL of the internal standard solution, and then add diluted acetonitrile (1 in 2) to make 50 mL. To 1 mL of this solution add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of buformin to that of the inter-nal standard. Amount (mg) of buformin hydrochloride (C6H15N5.HCl) ＝ MS × QT/QS × 20 MS: Amount (mg) of buformin hydrochloride for assay taken Internal standard solution—A solution of p-acetanisidide in diluted acetonitrile (1 in 2) (1 in 150). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 233 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of a solution of sodium perchlorate (7 in 250) and acetonitrile (7:1). Flow rate: Adjust so that the retention time of buformin is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, buformin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of buformin to that of the internal standard is not more than 1.0z.
Buformin Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of buformin hydrochloride (C6H15N5.HCl: 193.68). Method of preparation Prepare as directed under Tablets, with Buformin Hydrochloride. Identification To a quantity of powdered Buformin Hy-drochloride Tablets, equivalent to 1 g of Buformin Hydro- chloride, add 100 mL of water, shake well, and then filter. To 4 mL of the filtrate add 1 mL of dilute sodium penta-cyanonitrosylferrate (III)-potassium hexacyanoferrate (III) TS: the solution exhibits a red-brown color. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take 1 tablet of Buformin Hydrochloride Tablets, add water to make exactly 200 mL, and then treat with ultrasonic waves for 5 minutes. Take 40 mL of this solution and centri-fuge. Pipet V mL of the supernatant liquid equivalent to about 0.5 mg of buformin hydrochloride (C6H15N5.HCl), add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of buformin hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 233 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of buformin hydrochloride (C6H15N5.HCl) ＝ MS × AT/AS × 2/V MS: Amount (mg) of buformin hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Buformin Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Buformin Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 5.
To a quantity of powdered Buformin Hy-drochloride Tablets, equivalent to 1 g of Buformin Hydro- chloride, add 100 mL of water, shake well, and then filter. To 4 mL of the filtrate add 1 mL of dilute sodium penta-cyanonitrosylferrate (III)-potassium hexacyanoferrate (III) TS: the solution exhibits a red-brown color. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take 1 tablet of Buformin Hydrochloride Tablets, add water to make exactly 200 mL, and then treat with ultrasonic waves for 5 minutes. Take 40 mL of this solution and centri-fuge. Pipet V mL of the supernatant liquid equivalent to about 0.5 mg of buformin hydrochloride (C6H15N5.HCl), add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of buformin hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 233 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of buformin hydrochloride (C6H15N5.HCl) ＝ MS × AT/AS × 2/V MS: Amount (mg) of buformin hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Buformin Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Buformin Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 5.6 mg of buformin hy-drochloride (C6H15N5.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 28 mg of buformin hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 233 nm. Dissolution rate (z) with respect to the labeled amount of buformin hydrochloride (C6H15N5.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of buformin hydrochloride for assay taken C: Labeled amount (mg) of buformin hydrochloride (C6H15N5.HCl) in 1 tablet
Weigh accurately not less than 20 Buformin Hydro-chloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 60 mg of buformin hydro-chloride (C6H15N5.HCl), add water to make exactly 200 mL, and treat with ultrasonic waves for 5 minutes. Take 40 mL of this solution, centrifuge, pipet 2 mL of the supernatant liquid, add water to make exactly 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 60 mg of buformin hydrochloride for assay, previ-ously dried at 1059C for 3 hours, and dissolve in water to ‌576 make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and determine the absorbances, AT and AS, at 233 nm. mix them, and add diluted hydrochloric acid (1 in 40) to make exactly 100 mL. (2) Chloride <1.03>—Mix well 0.5 g of Bumetanide with 0.7 g of potassium nitrate and 1.2 g of anhydrous sodium carbonate, transfer, in small portions, to a red-hot platinum crucible, and heat to red-hot until the reaction is complete. Amount (mg) of buformin hydrochloride (C H N .HCl) After cooling, to the residue add 14 mL of dilute sulfuric ＝ MS × AT/AS 6 15 5 acid and 6 mL of water, boil for 5 minutes, filter, wash the residue with 10 mL of water, combine the filtrate and the MS: Amount (mg) of buformin hydrochloride for assay taken
Butenafine Hydrochloride Cream contains not less than 95.
To an amount of Butenafine Hydrochloride Cream, equivalent to 20 mg of Butenafine Hydrochloride, add 20 mL of acetonitrile, and warm on a water bath to melt the bases. Shake thoroughly, add an appropriate amount of sodium chloride, and allow to stand for 30 minutes in an ice cold water keeping not exceeding 09C to separate out the bases. Centrifuge, collect the supernatant liquid, add an ap-propriate amount of sodium chloride to the liquid, allow to stand for 1 hour in an ice cold water keeping not exceeding 09C, and filter while cooling. To 1 mL of the filtrate add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be- ‌582 tween 272 nm and 276 nm, between 281 nm and 285 nm, be- tween 311 nm and 315 nm, and between 316 nm and 320 nm, and a shoulder between 289 nm and 299 nm.
Weigh accurately a quantity of Butenafine Hydro-chloride Cream, equivalent to about 5 mg of butenafine hy-drochloride (C23H27N.HCl), add 20 mL of methanol, and add exactly 10 mL of the internal standard solution. Warm this in a water bath for 5 minutes, and shake vigorously for 20 minutes. Then, cool in an ice bath for 15 minutes, centri-fuge, and filter the supernatant liquid with a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of butenafine hydrochloride for assay, previously dried in vacuum at 609C for 3 hours using phosphorus (V) oxide as desiccant, and dissolve in methanol to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 10 mL of the inter-nal standard solution, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of butena-fine to that of the internal standard. Amount (mg) of butenafine hydrochloride (C23H27N.HCl) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of butenafine hydrochloride for assay taken Internal standard solution—A solution of diphenyl in meth-anol (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 5 cm in length, packed with octylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and diluted 0.5 Butenafine Hydrochloride Solution
Butenafine Hydrochloride Solution is a liquid for external use. It contains not less than 95.
To an amount of Butenafine Hydrochloride Solution, equivalent to 10 mg of Butenafine Hydrochloride, add methanol to make 200 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 272 nm and 276 nm, between 281 nm and 285 nm, be- tween 311 nm and 315 nm, and between 316 nm and 320 nm, and a shoulder between 289 nm and 299 nm.
To an exact volume of Butenafine Hydrochloride Solution, equivalent to about 20 mg of butenafine hydro-chloride (C23H27N.HCl), add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 4 mL of the in-ternal standard solution, then add methanol to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of butenafine hydrochloride for assay, previously dried in vacuum at 609C for 3 hours using phosphorus (V) oxide as desiccant, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add exactly 4 mL of the internal standard solution, then add methanol to make 25 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of butenafine to that of the internal standard. mol/L ammonium acetate TS (1 in 500) (4:1). Flow rate: Adjust so that the retention time of butenafine is about 2.5 minutes. Amount (mg) of butenafine hydrochloride (C ＝ MS × QT/QS 23H 27N.HCl) System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and butenafine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of butenafine to that of the internal standard is not more than 1.0z.
Butenafine Hydrochloride Spray contains not less than 95.
To an amount of Butenafine Hydrochloride Spray, equivalent to 10 mg of Butenafine Hydrochloride, add methanol to make 200 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 272 nm and 276 nm, between 281 nm and 285 nm, between 311 nm and 315 nm, and between 316 nm and 320 nm, and a shoulder between 289 nm and 299 nm.
To an exact volume of Butenafine Hydrochloride Spray, equivalent to about 20 mg of butenafine hydrochlo-ride (C23H27N.HCl), add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 4 mL of the internal standard solution, then add methanol to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of butenafine hydrochloride for assay, previously dried in vacuum at 609C for 3 hours using phosphorus (V) oxide as desiccant, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add ex-actly 4 mL of the internal standard solution, then add meth-anol to make 25 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of butena-fine to that of the internal standard. Amount (mg) of butenafine hydrochloride (C23H27N.HCl) ＝ MS × QT/QS MS: Amount (mg) of butenafine hydrochloride for assay taken Internal standard solution—A solution of diphenyl in meth-anol (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 5 cm in length, packed with octylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and diluted 0.5 mol/L ammonium acetate TS (1 in 500) (4:1). Flow rate: Adjust so that the retention time of butenafine is about 2.5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and butenafine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of butenafine to that of the internal standard is not more than 1.0z.
Cadralazine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of cadralazine (C12H21N5O3: 283.33). Method of preparation Prepare as directed under Tablets, with Cadralazine. Identification To a quantity of powdered Cadralazine Tablets, equivalent to 20 mg of Cadralazine, add 50 mL of 0.05 mol/L sulfuric acid TS, shake well, and centrifuge. To ‌588 1 mL of the supernatant liquid add 0.05 mol/L sulfuric acid TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 247 nm and 251 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cadralazine Tablets add 30 mL of 0.05 mol/L sulfuric acid TS, shake well to disintegrate, and add 0.05 mol/L sulfuric acid TS to make exactly 50 mL. Centri-fuge this solution, pipet 3 mL of the supernatant liquid, add 0.05 mol/L sulfuric acid TS to make exactly V mL so that each mL contains about 6 mg of cadralazine (C12H21N5O3), and use this solution as the sample solution.
To a quantity of powdered Cadralazine Tablets, equivalent to 20 mg of Cadralazine, add 50 mL of 0.05 mol/L sulfuric acid TS, shake well, and centrifuge. To ‌588 1 mL of the supernatant liquid add 0.05 mol/L sulfuric acid TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 247 nm and 251 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cadralazine Tablets add 30 mL of 0.05 mol/L sulfuric acid TS, shake well to disintegrate, and add 0.05 mol/L sulfuric acid TS to make exactly 50 mL. Centri-fuge this solution, pipet 3 mL of the supernatant liquid, add 0.05 mol/L sulfuric acid TS to make exactly V mL so that each mL contains about 6 mg of cadralazine (C12H21N5O3), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of cadralazine for assay, previously dried at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfuric acid TS to make exactly 100 mL. Pipet 3 mL of this solution, add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 249 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of cadralazine (C12H21N5O3) ＝ MS × AT/AS × V/200 MS: Amount (mg) of cadralazine for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Cadralazine Tablets is not less than 80z. Start the test with 1 tablet of Cadralazine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of cadralazine (C12H21N5O3), and use this solution as the sample solution. Separately, weigh accu-rately about 30 mg of cadralazine for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 200 mL. Pipet 4 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 254 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of cadralazine (C12H21N5O3) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of cadralazine for assay taken C: Labeled amount (mg) of cadralazine (C12H21N5O3) in 1 tablet
To 10 Cadralazine Tablets add 70 mL of 0.05 mol/L sulfuric acid TS, shake well to disintegrate, add 0.05 mol/L sulfuric acid TS to make exactly 200 mL. Centrifuge this so-lution, pipet a volume of the supernatant liquid, equivalent to about 2.5 mg of cadralazine (C12H21N5O3), add exactly 5 mL of the internal standard solution, add water to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of cadralazine for assay, previously dried at 1059C for 3 hours, and dissolve in 0.05 mol/L of sulfuric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add water to make 25 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cadralazine to that of the internal standard. Amount (mg) of cadralazine (C12H21N5O3) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of cadralazine for assay taken Internal standard solution—A solution of p-toluenesul-fonamide in acetonitrile (1 in 50). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 13.6 g of sodium acetate trihydrate in 800 mL of water, adjust the pH to 5.8 with dilute acetic acid, and add water to make 1000 mL. To 860 mL of this so-lution add 140 mL of acetonitrile. Flow rate: Adjust so that the retention time of cadralazine is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cadralazine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cadralazine to that of the internal standard is not more than 1.0z.
Calcium Chloride Injection is an aqueous injection. It contains not less than 95.
Calcium Chloride Hydrate occurs as white, granules or masses. It is odorless. It is very soluble in water, and soluble in ethanol (95), and practically insoluble in diethyl ether. It is deliquescent.
A solution of Calcium Chloride Hydrate (1 in 10) responds to Qualitative Tests <1.09> for calcium salt and for chloride. pH <2.54> The pH of a solution of 1.0 g of Calcium Chlo-ride Hydrate in 20 mL of freshly boiled and cooled water is between 4.5 and 9.2.
(1) Clarity and color of solution—A solution of 1.0 g of Calcium Chloride Hydrate in 20 mL of water is clear The concentration of Calcium Chloride Injection is expressed as the quantity of calcium chloride (CaCl2). Method of preparation Prepare as directed under Injec-tion, with Calcium Chloride Hydrate. Description Calcium Chloride Injection is a clear, colorless liquid. Identification Calcium Chloride Injection responds to Qualitative Tests <1.09> for calcium salt and for chloride. pH <2.54> 4.5 – 7.5 Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. ‌JP XVIII Official Monographs / Calcium Folinate Hydrate 597 Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Calcium Chloride Injection, equivalent to about 0.4 g of calcium chloride (CaCl2), and proceed as directed in the Assay under Calcium Chloride Hydrate. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.220 mg of CaCl2
Candesartan Cilexetil and Amlodipine Besylate Tablets
カンデサルタン シレキセチル･アムロジピンベシル酸塩錠
Candesartan Cilexetil and Amlodipine Besylate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of candesartan cilexetil (C33H34N6O6: 610.66) and amlodipine besylate (C20H25ClN2O5.C6H6O3S: 567.05). Method of preparation Prepare as directed under Tablets, with Candesartan Cilexetil and Amlodipine Besylate. Identification (1) Shake thoroughly a quantity of pow-dered Candesartan Cilexetil and Amlodipine Besylate Tablets, equivalent to 8 mg of Candesartan Cilexetil, with 20 mL of 0.01 mol/L hydrochloric acid TS, and centrifuge. Remove the supernatant liquid, to the residue add 20 mL of 0.01 mol/L hydrochloric acid TS, shake thoroughly, and centrifuge. Remove the supernatant liquid, to the residue add 40 mL of methanol, shake thoroughly, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 5 mL of the filtrate add methanol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 252 nm and 256 nm, and between 302 nm and 307 nm. (2) Shake thoroughly a quantity of powdered Candesar-tan Cilexetil and Amlodipine Besylate Tablets, equivalent to 2.5 mg of Amlodipine Besylate, with 20 mL of 0.01 mol/L hydrochloric acid TS, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 5 mL of the filtrate add methanol to make 25 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 236 nm and 240 nm, and between 360 nm and 364 nm. Purity Related substances—Shake vigorously for 20 minutes a quantity of powdered Candesartan Cilexetil and Amlodipine Besylate Tablets, equivalent to 8 mg of Can-desartan Cilexetil, with 20 mL of diluting solution, and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add diluting solution to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.8 to candesartan cilexetil, obtained from the sample solution is not larger than 1.5 times the peak area of can-desartan cilexetil from the standard solution, the area of the peaks, having a relative retention time of about 0.9, about 1.1 and about 1.2 from the sample solution is not larger than 1/2 times the peak area of candesartan cilexetil from the standard solution, the area of the peak, having a relative retention time of about 1.4 from the sample solution, is not larger than the peak area of candesartan cilexetil from the standard solution, and the area of the peak other than can-desartan cilexetil and the peaks mentioned above from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the standard solution. Further-more, the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard so-lution. Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 253 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile and trifluoroacetic acid (4000:1000:1). Mobile phase B: A mixture of acetonitrile, water and trifluoroacetic acid (4000:1000:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 100 → 50 0 → 50 15 – 50 50 → 0 50 → 100 50 – 60 Flow rate: 1.0 mL per minute. Time span of measurement: For 60 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add diluting solution to make exactly 50 mL. Confirm that the peak area of candesartan cilexetil ob-tained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that obtained with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 100,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following methods: it meets the requirements of the Content uniformity test. Candesartan cilexetil—To 1 tablet of Candesartan Cilexetil and Amlodipine Besylate Tablets add exactly 20 mL of diluting solution, shake for 20 minutes to disintegrate the tablet, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/5 mL of the internal standard solution, then add diluting solution to make V? mL so that each mL contains about 0.
(1) Shake thoroughly a quantity of pow-dered Candesartan Cilexetil and Amlodipine Besylate Tablets, equivalent to 8 mg of Candesartan Cilexetil, with 20 mL of 0.01 mol/L hydrochloric acid TS, and centrifuge. Remove the supernatant liquid, to the residue add 20 mL of 0.01 mol/L hydrochloric acid TS, shake thoroughly, and centrifuge. Remove the supernatant liquid, to the residue add 40 mL of methanol, shake thoroughly, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 5 mL of the filtrate add methanol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 252 nm and 256 nm, and between 302 nm and 307 nm. (2) Shake thoroughly a quantity of powdered Candesar-tan Cilexetil and Amlodipine Besylate Tablets, equivalent to 2.5 mg of Amlodipine Besylate, with 20 mL of 0.01 mol/L hydrochloric acid TS, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 5 mL of the filtrate add methanol to make 25 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 236 nm and 240 nm, and between 360 nm and 364 nm.
Related substances—Shake vigorously for 20 minutes a quantity of powdered Candesartan Cilexetil and Amlodipine Besylate Tablets, equivalent to 8 mg of Can-desartan Cilexetil, with 20 mL of diluting solution, and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add diluting solution to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.8 to candesartan cilexetil, obtained from the sample solution is not larger than 1.5 times the peak area of can-desartan cilexetil from the standard solution, the area of the peaks, having a relative retention time of about 0.9, about 1.1 and about 1.2 from the sample solution is not larger than 1/2 times the peak area of candesartan cilexetil from the standard solution, the area of the peak, having a relative retention time of about 1.4 from the sample solution, is not larger than the peak area of candesartan cilexetil from the standard solution, and the area of the peak other than can-desartan cilexetil and the peaks mentioned above from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the standard solution. Further-more, the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard so-lution. Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 253 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile and trifluoroacetic acid (4000:1000:1). Mobile phase B: A mixture of acetonitrile, water and trifluoroacetic acid (4000:1000:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 100 → 50 0 → 50 15 – 50 50 → 0 50 → 100 50 – 60 Flow rate: 1.0 mL per minute. Time span of measurement: For 60 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add diluting solution to make exactly 50 mL. Confirm that the peak area of candesartan cilexetil ob-tained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that obtained with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 100,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following methods: it meets the requirements of the Content uniformity test. Candesartan cilexetil—To 1 tablet of Candesartan Cilexetil and Amlodipine Besylate Tablets add exactly 20 mL of diluting solution, shake for 20 minutes to disintegrate the tablet, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/5 mL of the internal standard solution, then add diluting solution to make V? mL so that each mL contains about 0.16 mg of candesartan cilexetil (C33H34N6O6), and use this solution as the sample solution. Then, proceed as directed in the Assay (1). Amount (mg) of candesartan cilexetil (C33H34N6O6) ＝ MS × QT/QS × V?/V × 2/25 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in diluting solution (1 in 2500). Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Amlodipine besylate—To 1 tablet of Candesartan Cilexetil and Amlodipine Besylate Tablets add exactly 20 mL of diluting solution, shake for 20 minutes to disintegrate the tablet, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/5 mL of the internal standard solution, then add diluting solution to make V? mL so that each mL contains about 70 mg of amlodipine besylate (C H ClN O .C H O S), and use this use this solution as the sample solution. Separately, weigh accurately about 45 mg of candesartan cilexetil for assay (separately, determine the water <2.48> in the same manner as Candesartan Cilexetil), and dissolve in acetonitrile to make exactly 50 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of candesartan cilexetil in each solution. Dissolution rate (z) with respect to the labeled amount of candesartan cilexetil (C33H34N6O6) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of candesartan cilexetil (C33H34N6O6) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Flow rate: Adjust so that the retention time of candesar-tan cilexetil is about 6.5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 2000 and not more than 1.5, respectively. 20 25 2 5 6 6 3 System repeatability: When the test is repeated 6 times solution as the sample solution. Then, proceed as directed in the Assay (2). Amount (mg) of amlodipine besylate (C20H25ClN2O5.C6H6O3S) ＝ MS × QT/QS × V?/V × 1/25 MS: Amount (mg) of Amlodipine Besylate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in diluting solution (1 in 2500). Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Dissolution <6.10> (1) Candesartan cilexetil—When the test is performed at 75 revolutions per minute according to the Paddle method, using 900 mL of a solution, prepared by dissolving 1 g of polysorbate 80 in 2nd fluid for dissolution test to make 1000 mL, as the dissolution medium, the disso-lution rate in 45 minutes of Candesartan Cilexetil and Amlodipine Besylate Tablets is not less than 80z. Start the test with 1 tablet of Candesartan Cilexetil and Amlodipine Besylate Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the disso-lution medium to make exactly V? mL so that each mL con- tains about 8.9 mg of candesartan cilexetil (C H N O ), and with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 1.0z. (2) Amlodipine besylate—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dis-solution rate in 30 minutes of Candesartan Cilexetil and Amlodipine Besylate Tablets is not less than 80z. Start the test with 1 tablet of Candesartan Cilexetil and Amlodipine Besylate Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the disso-lution medium to make exactly V? mL so that each mL con-tains about 3.9 mg of amlodipine besylate (C20H25ClN2O5. C6H6O3S), and use this solution as the sample solution. Separately, weigh accurately about 39 mg of Amlodipine Besylate RS (separately, determine the water <2.48> in the same manner as Amlodipine Besylate), and dissolve in aceto-nitrile to make exactly 50 mL. Pipet 5 mL of this solution, add acetonitrile to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine 33 34 6 6 JP XVIII Official Monographs / Candesartan Cilexetil and Amlodipine Besylate Tablets 617 the peak areas, AT and AS, of amlodipine in each solution. Dissolution rate (z) with respect to the labeled amount of amlodipine besylate (C H ClN O .C H O S) Column temperature: A constant temperature of about 259C. Mobile phase: To 7 mL of triethylamine add water to 20 25 2 5 6 6 3 make 1000 mL, and adjust to pH 6.5 with phosphoric acid. ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of Amlodipine Besylate RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of amlodipine besylate (C20H25ClN2O5.C6H6O3S) in 1 tablet Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay (1). Flow rate: Adjust so that the retention time of amlodipine is about 4 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of amlodipine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlodipine is not more than 1.0z.
(1) Candesartan cilexetil—Weigh accurately the mass of not less than 20 Candesartan Cilexetil and Amlodi-pine Besylate Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 8 mg of candesartan cilexetil (C33H34N6O6), add exactly 20 mL of diluting solu-tion, shake vigorously for 20 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 5 mL of the filtrate, pipet 10 mL of the subse-quent filtrate, add exactly 5 mL of the internal standard solution, then add diluting solution to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of candesartan cilexetil for assay (separately, determine the water <2.48> in the same manner as Candesartan Cilexetil), dissolve in diluting solution to make exactly 100 mL, and use this solution as the candesar-tan cilexetil standard stock solution. Pipet 10 mL of the can-desartan cilexetil standard stock solution, add exactly 5 mL of the internal standard solution, then add diluting solution to make 25 mL, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of candesartan To 800 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of candesar- tan cilexetil is about 31 minutes. System suitability— System performance: Mix 10 mL of the candesartan cilex-etil standard stock solution and 5 mL of the amlodipine be-sylate standard stock solution prepared in the Assay (2), add 5 mL of the internal standard solution, then add diluting so-lution to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, am-lodipine, the internal standard and candesartan cilexetil are eluted in this order and the resolution between the peaks of the internal standard and candesartan cilexetil is not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of candesartan cilexetil to that of the internal standard is not more than 1.0z. (2) Amlodipine besylate—Weigh accurately the mass of not less than 20 Candesartan Cilexetil and Amlodipine Besy-late Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3.5 mg of amlodipine besylate (C20H25ClN2O5.C6H6O3S), add exactly 20 mL of diluting so-lution, shake vigorously for 20 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 5 mL of the filtrate, pipet 10 mL of the subse-quent filtrate, add exactly 5 mL of the internal standard so-lution, then add diluting solution to make 25 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 35 mg of Amlodipine Besylate RS (separately, determine the water <2.48> in the same manner as Amlodi-pine Besylate), dissolve in diluting solution to make exactly 100 mL, and use this solution as the amlodipine besylate standard stock solution. Pipet 5 mL of the amlodipine besy-late standard stock solution, add exactly 5 mL of the internal standard solution, then add diluting solution to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of amlodipine to that of the internal standard. Amount (mg) of amlodipine besylate (C H ClN O .C H O S) cilexetil to that of the internal standard. 20 25 2 5 6 6 3 Amount (mg) of candesartan cilexetil (C33H34N6O6) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in diluting solution (1 in 2500). Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ＝ MS × QT/QS × 1/10 MS: Amount (mg) of Amlodipine Besylate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in diluting solution (1 in 2500). Diluting solution: To 3.5 mL of triethylamine add water to make 500 mL, and adjust to pH 3.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay (1). Flow rate: Adjust so that the retention time of amlodipine is about 2.5 minutes. System suitability— System performance: Mix 10 mL of the candesartan cilex- ‌618 etil standard stock solution prepared in the Assay (1) and 5 mL of the amlodipine besylate standard stock solution, add 5 mL of the internal standard solution, then add diluting so-lution to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, am-lodipine, the internal standard and candesartan cilexetil are eluted in this order and the resolution between the peaks of amlodipine and the internal standard is not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of amlodipine to that of the internal standard is not more than 1.0z.
Candesartan Cilexetil and Hydrochlorothiazide Tablets
カンデサルタン シレキセチル･ヒドロクロロチアジド錠
Candesartan Cilexetil and Hydrochlorothiazide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of candesartan cilexetil (C33H34N6O6: 610.66) and hydrochlorothiazide (C7H8ClN3O4S2: 297.74). Method of preparation Prepare as directed under Tablets, with Candesartan Cilexetil and Hydrochlorothiazide. Identification (1) To an amount of powdered Candesar-tan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 4 mg of Candesartan Cilexetil, add 5 mL of acetone, shake thoroughly, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the filtrate to dryness with the aid of a current of nitrogen. Dissolve the residue in 0.5 mL of ace-tone, and use this solution as the sample solution. Sepa-rately, dissolve 40 mg of candesartan cilexetil in 5 mL of ace-tone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate and acetic acid (100) (10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the spot having a larger Rf value among the spots obtained from the sample solution is the same with that of the spot from the standard solution. (2) To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of Hydrochlorothiazide, add 5 mL of acetone, shake thor-oughly, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the filtrate to dryness with the aid of a current of nitrogen. Dissolve the residue in 0.5 mL of acetone, and use this solution as the sample solution. Separately, dissolve 50 mg of hydrochlorothiazide in 4 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio- let light (main wavelength: 254 nm): the Rf value of the spot having a smaller Rf value among the spots obtained from the sample solution is the same with that of the spot from the standard solution. Purity Related substances—(i) To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 4 mg of Candesartan Cilexetil, add 10 mL of a mixture of acetonitrile and water (3:2), shake vigorously for 10 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having a relative retention time of about 0.5 to candesartan cilexetil, obtained from the sample solution is not larger than 1.5 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak, having a relative retention time of about 0.8, about 1.1 and about 1.5, from the sample solution is not larger than 1/2 times the peak area of can-desartan cilexetil from the standard solution, the area of the peak, having a relative retention time of about 2.0, from the sample solution is not larger than the peak area of candesar-tan cilexetil from the standard solution, and the area of the peak, other than candesartan cilexetil and the peaks men-tioned above, from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the stand-ard solution. Furthermore, the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard solution. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and time span of meas-urement: Proceed as directed in the operating conditions in the Purity (2) under Candesartan Cilexetil. Flow rate: 0.6 mL per minute. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Confirm that the peak area of candesartan cilexetil obtained with 10 mL of this solution is equivalent to 1.4z to 2.6z of that with 10 mL of the stand-ard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 12,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of Hydrochlorothiazide, add 10 mL of a mixture of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) and acetonitrile (3:1), shake vigorously for 10 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the fil-trate, and use the subsequent filtrate as the sample solution. JP XVIII Official Monographs / Candesartan Cilexetil and Hydrochlorothiazide Tablets 619 Pipet 1 mL of the sample solution, add a mixture of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) and aceto-nitrile (3:1) to make exactly 100 mL, and use this solution as <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of candesartan cilex-etil to that of the internal standard. the standard solution. Perform the test with exactly 10 mL Amount (mg) of candesartan cilexetil (C H N O ) each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- ＝ MS × QT/QS × V × 1/1250 33 34 6 6 lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having a relative retention time of about 0.9 and about 3.2 to hydro-chlorothiazide, obtained from the sample solution is not larger than the peak area of hydrochlorothiazide from the standard solution, and the area of the peak, other than hy-drochlorothiazide and the peaks mentioned above, from the sample solution is not larger than 1/5 times the peak area of hydrochlorothiazide from the standard solution. Further-more, the total area of the peaks other than hydrochloro-thiazide from the sample solution is not larger than 2 times the peak area of hydrochlorothiazide from the standard so-lution. For the area of the peak, having a relative retention time of about 0.8 and about 0.9 to hydrochlorothiazide, multiply their correction factors, 1.4 and 0.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (2). Time span of measurement: For 30 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of 0.05 mol/L sodium dihy-drogen phosphate TS (pH 3.0) and acetonitrile (3:1) to make exactly 50 mL. Confirm that the peak area of hydrochloro-thiazide obtained with 10 mL of this solution is equivalent to 1.4z to 2.6z of that obtained with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of hydrochlorothiazide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following methods: it meets the requirements of the Content uniformity test. Candesartan cilexetil—To 1 tablet of Candesartan Cilexetil and Hydrochlorothiazide Tablets add exactly V/10 mL of the internal standard solution, add a mixture of aceto-nitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make V mL so that each mL contains about 40 mg of candesartan cilexetil (C33H34N6O6).
(1) To an amount of powdered Candesar-tan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 4 mg of Candesartan Cilexetil, add 5 mL of acetone, shake thoroughly, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the filtrate to dryness with the aid of a current of nitrogen. Dissolve the residue in 0.5 mL of ace-tone, and use this solution as the sample solution. Sepa-rately, dissolve 40 mg of candesartan cilexetil in 5 mL of ace-tone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate and acetic acid (100) (10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the spot having a larger Rf value among the spots obtained from the sample solution is the same with that of the spot from the standard solution. (2) To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of Hydrochlorothiazide, add 5 mL of acetone, shake thor-oughly, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the filtrate to dryness with the aid of a current of nitrogen. Dissolve the residue in 0.5 mL of acetone, and use this solution as the sample solution. Separately, dissolve 50 mg of hydrochlorothiazide in 4 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio- let light (main wavelength: 254 nm): the Rf value of the spot having a smaller Rf value among the spots obtained from the sample solution is the same with that of the spot from the standard solution.
Related substances—(i) To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 4 mg of Candesartan Cilexetil, add 10 mL of a mixture of acetonitrile and water (3:2), shake vigorously for 10 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having a relative retention time of about 0.5 to candesartan cilexetil, obtained from the sample solution is not larger than 1.5 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak, having a relative retention time of about 0.8, about 1.1 and about 1.5, from the sample solution is not larger than 1/2 times the peak area of can-desartan cilexetil from the standard solution, the area of the peak, having a relative retention time of about 2.0, from the sample solution is not larger than the peak area of candesar-tan cilexetil from the standard solution, and the area of the peak, other than candesartan cilexetil and the peaks men-tioned above, from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the stand-ard solution. Furthermore, the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard solution. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and time span of meas-urement: Proceed as directed in the operating conditions in the Purity (2) under Candesartan Cilexetil. Flow rate: 0.6 mL per minute. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Confirm that the peak area of candesartan cilexetil obtained with 10 mL of this solution is equivalent to 1.4z to 2.6z of that with 10 mL of the stand-ard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 12,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. To an amount of powdered Candesartan Cilexetil and Hydrochlorothiazide Tablets, equivalent to 6.25 mg of Hydrochlorothiazide, add 10 mL of a mixture of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) and acetonitrile (3:1), shake vigorously for 10 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the fil-trate, and use the subsequent filtrate as the sample solution. JP XVIII Official Monographs / Candesartan Cilexetil and Hydrochlorothiazide Tablets 619 Pipet 1 mL of the sample solution, add a mixture of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) and aceto-nitrile (3:1) to make exactly 100 mL, and use this solution as <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of candesartan cilex-etil to that of the internal standard. the standard solution. Perform the test with exactly 10 mL Amount (mg) of candesartan cilexetil (C H N O ) each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- ＝ MS × QT/QS × V × 1/1250 33 34 6 6 lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having a relative retention time of about 0.9 and about 3.2 to hydro-chlorothiazide, obtained from the sample solution is not larger than the peak area of hydrochlorothiazide from the standard solution, and the area of the peak, other than hy-drochlorothiazide and the peaks mentioned above, from the sample solution is not larger than 1/5 times the peak area of hydrochlorothiazide from the standard solution. Further-more, the total area of the peaks other than hydrochloro-thiazide from the sample solution is not larger than 2 times the peak area of hydrochlorothiazide from the standard so-lution. For the area of the peak, having a relative retention time of about 0.8 and about 0.9 to hydrochlorothiazide, multiply their correction factors, 1.4 and 0.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (2). Time span of measurement: For 30 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of 0.05 mol/L sodium dihy-drogen phosphate TS (pH 3.0) and acetonitrile (3:1) to make exactly 50 mL. Confirm that the peak area of hydrochloro-thiazide obtained with 10 mL of this solution is equivalent to 1.4z to 2.6z of that obtained with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of hydrochlorothiazide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following methods: it meets the requirements of the Content uniformity test. Candesartan cilexetil—To 1 tablet of Candesartan Cilexetil and Hydrochlorothiazide Tablets add exactly V/10 mL of the internal standard solution, add a mixture of aceto-nitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make V mL so that each mL contains about 40 mg of candesartan cilexetil (C33H34N6O6). Shake for 20 minutes to disintegrate the tablet, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), dissolve in acetonitrile to make ex-actly 50 mL, and use this solution as the candesartan cilexetil standard stock solution. Pipet 4 mL of the candesartan cilex-etil standard stock solution, add exactly 10 mL of the inter-nal standard solution, add a mixture of acetonitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make 100 mL, and use this solution as the standard solution. MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis. Internal standard solution—A solution of benzophenone in acetonitrile (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 5.5) (11:9). Flow rate: Adjust so that the retention time of candesar-tan cilexetil is about 7 minutes. System suitability— System performance: Mix 4 mL of the candesartan cilex-etil standard stock solution and 10 mL of the hydrochloro-thiazide standard stock solution obtained in (2), add 10 mL of the internal standard solution, and add a mixture of acetonitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, hydrochlorothiazide, candesartan cilexetil and the internal standard are eluted in this order, and the resolu-tion between the peaks of hydrochlorothiazide and candesar-tan cilexetil is not less than 7, and the resolution between the peaks of candesartan cilexetil and the internal standard is not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of candesartan cilexetil to that of the internal standard is not more than 1.0z. Hydrochlorothiazide—To 1 tablet of Candesartan Cilexetil and Hydrochlorothiazide Tablets add exactly V/10 mL of the internal standard solution, add a mixture of aceto-nitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make V mL so that each mL contains about 63 mg of hydrochlorothiazide (C7H8ClN3O4S2). Shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 31 mg of Hydrochlorothiazide RS (separately determine the loss on drying <2.41> under the same conditions as Hydrochloro-thiazide), dissolve in acetonitrile to make exactly 50 mL, and use this solution as the hydrochlorothiazide standard stock solution. Pipet 10 mL of the hydrochlorothiazide standard stock solution, add exactly 10 mL of the internal standard solution, add a mixture of acetonitrile and 0.05 mol/L so-dium dihydrogen phosphate TS (pH 3.0) (3:2) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of hydrochlorothiazide to that of the internal standard. Amount (mg) of hydrochlorothiazide (C H ClN O S ) Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography ＝ MS × QT/QS × V × 1/500 7 8 3 4 2 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis. Internal standard solution—A solution of benzophenone in acetonitrile (1 in 10,000). Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay (2). Mobile phase: A mixture of acetonitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 5.5) (11:9). Flow rate: Adjust so that the retention time of hydrochlo-rothiazide is about 3.5 minutes. System suitability— System performance: Mix 4 mL of the candesartan cilex-etil standard stock solution obtained in (1) and 10 mL of the hydrochlorothiazide standard stock solution, add 10 mL of the internal standard solution, and add a mixture of aceto-nitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) (3:2) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, hydrochlorothiazide, candesartan cilexetil and the in-ternal standard are eluted in this order, and the resolution between the peaks of hydrochlorothiazide and candesartan cilexetil is not less than 7, and the resolution between the peaks of candesartan cilexetil and the internal standard is not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of hydrochlorothiazide to that of the internal standard is not more than 1.0z. Dissolution <6.10> (1) Candesartan cilexetil—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of a solution, prepared by dissolving 1 g of polysorbate 80 in 2nd fluid for dissolution test to make 1000 mL, as the dissolution medium, the disso-lution rate in 45 minutes of Candesartan Cilexetil and Hy-drochlorothiazide Tablets is not less than 75z. Start the test with 1 tablet of Candesartan Cilexetil and Hydrochlorothiazide Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) to make exactly V? mL so that each mL contains about 2.2 mg of can-desartan cilexetil (C33H34N6O6), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), dissolve in acetonitrile to make exactly 100 mL, and use this solution as the candesartan cilexetil standard stock solution. Pipet 2 mL of the candesartan cilexetil standard stock solu-tion, add dissolution medium to make exactly 100 mL. Pipet 10 mL of this solution, add 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with ex-actly 40 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of candesartan cilexetil in each solution. Dissolution rate (z) with respect to the labeled amount of candesartan cilexetil (C33H34N6O6) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of candesartan cilexetil (C33H34N6O6) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units (1). System suitability— System performance: Mix 2 mL each of the candesartan cilexetil standard stock solution and the hydrochlorothiazide standard stock solution obtained in (2), and add the dissolu-tion medium to make 100 mL. To 10 mL of this solution add 10 mL of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0). When the procedure is run with 40 mL of this solution under the above operating conditions, hydrochlorothiazide and candesartan cilexetil are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 1.0z. (2) Hydrochlorothiazide—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of a solution, prepared by dissolving 1 g of polysorbate 80 in 2nd fluid for dissolution test to make 1000 mL, as the dissolution medium, the dissolution rate in 45 minutes of Candesartan Cilexetil and Hydrochlorothiazide Tablets is not less than 80z. Start the test with 1 tablet of Candesartan Cilexetil and Hydrochlorothiazide Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) to make exactly V? mL so that each mL contains about 3.5 mg of hy-drochlorothiazide (C7H8ClN3O4S2), and use this solution as the sample solution. Separately, weigh accurately about 38 mg of Hydrochlorothiazide RS (separately determine the loss on drying <2.41> under the same conditions as Hydrochloro-thiazide), dissolve in acetonitrile to make exactly 100 mL, and use this solution as the hydrochlorothiazide standard stock solution. Pipet 2 mL of the hydrochlorothiazide stand-ard stock solution, add dissolution medium to make exactly 100 mL. Pipet 10 mL of this solution, add 0.05 mol/L so-dium dihydrogen phosphate TS (pH 3.0) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of hydrochlorothiazide in each solution. Dissolution rate (z) with respect to the labeled amount of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis C: Labeled amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay (2). Mobile phase: A mixture of acetonitrile and 0.05 mol/L sodium dihydrogen phosphate TS (pH 5.5) (11:9). Flow rate: Adjust so that the retention time of hydrochlo-rothiazide is about 3.5 minutes. JP XVIII Official Monographs / Candesartan Cilexetil and Hydrochlorothiazide Tablets 621 System suitability— System performance: Mix 2 mL each of the candesartan cilexetil standard stock solution obtained in (1) and the hy-drochlorothiazide standard stock solution, and add the dis-solution medium to make 100 mL. To 10 mL of this solution add 10 mL of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0). When the procedure is run with 40 mL of this solu-tion under the above operating conditions, hydrochloro-thiazide and candesartan cilexetil are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z.
(1) Candesartan cilexetil—Weigh accurately the mass of not less than 20 Candesartan Cilexetil and Hydro-chlorothiazide Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 4 mg of candesartan cilexetil (C33H34N6O6), add exactly 10 mL of the internal standard solution, add a mixture of acetonitrile and water (3:2) to make 100 mL, and shake vigorously for 10 minutes. Allow to stand for 5 minutes, and filter the supernatant liq-uid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), dissolve in acetonitrile to make ex-actly 50 mL. Pipet 4 mL of this solution, add exactly 10 mL of the internal standard solution, add a mixture of aceto-nitrile and water (3:2) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of candesartan cilexetil to that of the internal standard. (2) Hydrochlorothiazide—Weigh accurately the mass of not less than 20 Candesartan Cilexetil and Hydrochloro-thiazide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 6.25 mg of hydrochloro-thiazide (C7H8ClN3O4S2), add exactly 10 mL of the internal standard solution, add a mixture of 0.05 mol/L sodium di-hydrogen phosphate TS (pH 3.0) and acetonitrile (3:1) to make 100 mL, and shake vigorously for 10 minutes. Allow to stand for 5 minutes, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 31 mg of Hydrochlorothiazide RS (separately determine the loss on drying <2.41> under the same conditions as Hydrochlorothiazide), dissolve in aceto-nitrile to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add a mixture of 0.05 mol/L sodium dihydrogen phosphate TS (pH 3.0) and acetonitrile (3:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of hydrochlorothiazide to that of the internal standard. Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis Internal standard solution—A solution of m-hydroxya-cetophenone in acetonitrile (1 in 6500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- Amount (mg) of candesartan cilexetil (C H N O ) ter and 15 cm in length, packed with octadecylsilanized silica ＝ MS × QT/QS × 2/25 33 34 6 6 gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of acenaphthene in acetonitrile (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Flow rate: Adjust so that the retention time of candesar-tan cilexetil is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and candesartan cilexetil are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of candesartan cilexetil to that of the internal standard is not more than 1.0z. 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS (pH 5.5) and acetonitrile (3:1). Flow rate: Adjust so that the retention time of hydrochlo-rothiazide is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, hydrochlorothiazide and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of hydrochlorothiazide to that of the internal standard is not more than 1.0z.
Candesartan Cilexetil Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of candesartan cilexetil (C33H34N6O6: 610.66). Method of preparation Prepare as directed under Tablets, with Candesartan Cilexetil. Identification Powder Candesartan Cilexetil Tablets. To a portion of the powder, equivalent to 1 mg of Candesartan Cilexetil, add 50 mL of methanol, shake vigorously for 10 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits absorption maxima between 252 nm and 256 nm and between 302 nm and 307 nm. Purity Related substances—Powder not less than 10 Can-desartan Cilexetil Tablets. To a portion of the powder, equivalent to 6 mg of Candesartan Cilexetil, add 15 mL of a mixture of acetonitrile and water (3:2), shake vigorously for 10 minutes, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water 0 – 30 100 → 0 0 → 100 Flow rate: 0.8 mL per minute. Time span of measurement: For 30 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of acetonitrile and water (3:2) to make exactly 20 mL. Confirm that the peak area of can-desartan cilexetil obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard so-lution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 12,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Water <2.48> Not more than 0.3z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Candesartan Cilex-etil, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak having the relative retention time of about 0.5 to candesartan cilexetil obtained from the sample solution is not larger than 1.5 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak having the relative reten-tion time of about 0.8, about 1.1 and about 1.5 to candesar-tan cilexetil from the sample solution is not larger than 1/2 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak having the relative reten-tion time of about 2.0 to candesartan cilexetil from the sam-ple solution is not larger than the peak area of candesartan cilexetil from the standard solution, the area of the peak other than candesartan cilexetil, the peak having the relative retention time of about 0.4 to candesartan cilexetil and the peaks mentioned above from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the standard solution, and the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Mobile phase B: A mixture of acetonitrile, water and acetic acid (100) (90:10:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. exactly V? mL so that each mL contains about 2.
Powder Candesartan Cilexetil Tablets. To a portion of the powder, equivalent to 1 mg of Candesartan Cilexetil, add 50 mL of methanol, shake vigorously for 10 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits absorption maxima between 252 nm and 256 nm and between 302 nm and 307 nm.
Related substances—Powder not less than 10 Can-desartan Cilexetil Tablets. To a portion of the powder, equivalent to 6 mg of Candesartan Cilexetil, add 15 mL of a mixture of acetonitrile and water (3:2), shake vigorously for 10 minutes, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water 0 – 30 100 → 0 0 → 100 Flow rate: 0.8 mL per minute. Time span of measurement: For 30 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of acetonitrile and water (3:2) to make exactly 20 mL. Confirm that the peak area of can-desartan cilexetil obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard so-lution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 12,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Water <2.48> Not more than 0.3z (0.5 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Candesartan Cilex-etil, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak having the relative retention time of about 0.5 to candesartan cilexetil obtained from the sample solution is not larger than 1.5 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak having the relative reten-tion time of about 0.8, about 1.1 and about 1.5 to candesar-tan cilexetil from the sample solution is not larger than 1/2 times the peak area of candesartan cilexetil from the stand-ard solution, the area of the peak having the relative reten-tion time of about 2.0 to candesartan cilexetil from the sam-ple solution is not larger than the peak area of candesartan cilexetil from the standard solution, the area of the peak other than candesartan cilexetil, the peak having the relative retention time of about 0.4 to candesartan cilexetil and the peaks mentioned above from the sample solution is smaller than 1/10 times the peak area of candesartan cilexetil from the standard solution, and the total area of the peaks other than candesartan cilexetil from the sample solution is not larger than 4 times the peak area of candesartan cilexetil from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Mobile phase B: A mixture of acetonitrile, water and acetic acid (100) (90:10:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. exactly V? mL so that each mL contains about 2.2 mg of can-desartan cilexetil (C33H34N6O6), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), and dissolve in acetonitrile to make exactly 50 mL. Pipet 5 mL of this solution, add acetonitrile to make exactly 50 mL. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Pipet 1 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the 0 – 30 100 → 0 0 → 100 Flow rate: 0.8 mL per minute. Time span of measurement: For 30 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add a mixture of acetonitrile and water (3:2) to make exactly 20 mL. Confirm that the peak area of can-desartan cilexetil obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard so-lution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 12,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Candesartan Cilexetil Tablets add 30 mL of a mixture of acetonitrile and water (3:2), shake vigorously for 20 minutes, then add a mixture of acetonitrile and water (3:2) to make exactly V mL so that each mL contains about 40 mg of candesartan cilexetil (C33H34N6O6), centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), and dissolve in aceto-nitrile to make exactly 50 mL. Pipet 4 mL of this solution, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 305 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of candesartan cilexetil (C33H34N6O6) ＝ MS × AT/AS × V/1250 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of polysorbate 20 (1 in 100) as the dissolu-tion medium, the dissolution rate in 45 minutes of Candesar-tan Cilexetil Tablets is not less than 75z. Start the test with 1 tablet of Candesartan Cilexetil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make sample solution and standard solution as directed under Liq-uid Chromatography <2.01>, and determine the peak areas, AT and AS, of candesartan cilexetil in each solution. Dissolution rate (z) with respect to the labeled amount of candesartan cilexetil (C33H34N6O6) ＝ MS × AT/AS × V?/V × 1/C × 18/5 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of candesartan cilexetil (C33H34N6O6) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of candesartan cilexetil are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of candesartan cilexetil is not more than 2.0z. Assay Weigh accurately the mass of not less than 20 Can-desartan Cilexetil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 6 mg of can-desartan cilexetil (C33H34N6O6), add exactly 15 mL of the in-ternal standard solution, then add a mixture of acetonitrile and water (3:2) to make 150 mL, shake vigorously for 10 minutes, and allow to stand. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of candesartan cilexetil for assay (separately determine the water <2.48> in the same manner as Candesartan Cilexetil), dissolve in acetonitrile to make ex-actly 50 mL. Pipet 4 mL of this solution, add exactly 10 mL of the internal standard solution, then add a mixture of acetonitrile and water (3:2) to make 100 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of candesartan cilexetil to that of the internal standard. Amount (mg) of candesartan cilexetil (C33H34N6O6) ＝ MS × QT/QS × 3/25 MS: Amount (mg) of candesartan cilexetil for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of acenaphthene in acetonitrile (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌JP XVIII Official Monographs / Candesartan Cilexetil and Amlodipine Besylate Tablets 615 length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (57:43:1). Flow rate: Adjust so that the retention time of candesar-tan cilexetil is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and candesartan cilexetil are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of candesartan cilexetil to that of the internal standard is not more than 1.0z.
Carboplatin Injection is an aqueous injection. It contains not less than 95.
Carboplatin Injection is a clear, colorless to pale yellow liquid.
(1) To an amount of Carboplatin Injec-tion, equivalent to 20 mg of Carboplatin, add 2 to 3 drops of diluted tin (II) chloride TS (1 in 15), and allow to stand for 30 minutes: a yellowish brown precipitate is formed. (2) Evaporate to dryness a volume of Carboplatin Injec-tion, equivalent to 10 mg of Carboplatin, in a water bath at not exceeding 309C under vacuum. Determine the infrared absorption spectrum of the residue as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>: it exhibits absorption at the wave numbers of about 3270 cm－1, 2990 cm－1, 2960 cm－1, 1645 cm－1, 1610 cm－1, 1381 cm－1 and 1348 cm－1. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) 1,1-Cyclobutanedicarboxylic acid—To an ex-act volume of Carboplatin Injection, equivalent to 20 mg of Carboplatin, add the mobile phase to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of 1,1-cyclobutanedicarboxyl-ic acid, and dissolve in the mobile phase to make exactly 100 mL. Pipet 4 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of 1,1-cyclobutanedicarboxylic acid in each solution, and calculate ‌630 the amount of 1,1-cyclobutanedicarboxylic acid by the fol-lowing formula: it is not more than 0.7z. Amount (z) of 1,1-cyclobutanedicarboxylic acid ＝ MS × AT/AS × 1/25 MS: Amount (mg) of 1,1-cyclobutanedicarboxylic acid taken Operating conditions— Proceed as directed in the operating conditions in the Purity (1) under Carboplatin. System suitability— Proceed as directed in the system suitability in the Purity under Carboplatin. Related substances—To a volume of Carboplatin In-jection, equivalent to 10 mg of Carboplatin, add water to make 10 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of these peaks by the area percentage method: the total amount of the peaks other than carboplatin is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phases A and B, and flow rate: Proceed as directed in the operating conditions in the Assay under Carboplatin. Flowing of mobile phase, and time span of measurement: Proceed as directed in the operating conditions in the Purity (2) under Carboplatin. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Carboplatin. Test for required detectability, and system repeatability: Proceed as directed in the system suitability in the Purity (2) under Carboplatin. Bacterial endotoxins <4.01> Less than 0.2 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Carboplatin Injection, equivalent to about 20 mg of carboplatin (C6H12N2O4Pt), add water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Carboplatin RS (separately determine the loss on dry-ing <2.41> under the same conditions as Carboplatin), dis-solve in water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of carboplatin in each solution. Amount (mg) of carboplatin (C6H12N2O4Pt) ＝ MS × AT/AS × 4/5 MS: Amount (mg) of Carboplatin RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 8.5 g of tetrabutylammonium hydrogensulfate in 80 mL of water, add 3.4 mL of phos-phoric acid, and adjust to pH 7.5 with a solution of sodium hydroxide (43 in 100). To 10 mL of this solution add 880 mL of water and 10 mL of acetonitrile. Flow rate: Adjust so that the retention time of carboplatin is about 4 minutes. System suitability— System performance: To a solution of 25 mg of carbopla-tin in 20 mL of water add 2.5 mL of a solution of 65 mg of 1,3-phenylenediamine hydrochloride in 50 mL of water, and add water to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, carboplatin and 1,3-phenylenediamine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carboplatin is not more than 1.0z.
Carvedilol Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of car-vedilol (C24H26N2O4: 406.47). Method of preparation Prepare as directed under Tablets, with Carvedilol. Identification Powder Carvedilol Tablets. To a portion of the powder, equivalent to 20 mg of Carvedilol, add 10 mL of methanol, shake well, and filter. To 0.5 mL of the filtrate add methanol to make 200 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 222 nm and 226 nm, between 241 nm and 245 nm, be- tween 284 nm and 288 nm, between 317 nm and 321 nm and between 330 nm and 334 nm. Purity Related substances—In this procedure the sample solution should be stored not exceeding 59C and used within 24 hours after preparation. Powder Carvedilol Tablets. Dis-solve a portion of the powder, equivalent to 12.5 mg of Car-vedilol, add an adequate amount of the mobile phase and disperse the particles with the aid of ultrasonic waves, if nec-essary, add the mobile phase to make 100 mL, and shake for 30 minutes. Filter through a membrane filter with a pore size not exceeding 0.22 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the two peaks, having the rela-tive retention time between 1.7 and 1.9 and between 2.0 and to carvedilol, obtained from the sample solution of 1.25-mg or 2.5-mg tablet is not larger than 3/10 times and 1.6 times the peak area of carvedilol from the standard solu-tion, respectively, the area of the peak other than carvedilol and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of carvedilol from the standard solution, and the total area of the peaks other than carvedilol from the sample solution is not larger than 2.2 times the peak area of carvedilol from the standard solu-tion. The area of the two peaks, having the relative retention time between 1.7 and 1.9 and between 2.0 and 3.1, from the sample solution of 10-mg or 20-mg tablet is not larger than 1/10 times and 2/5 times the peak area of carvedilol from the standard solution, respectively, the area of the peak other than carvedilol and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of carvedilol from the standard solution, and the total area of the peaks other than carvedilol from the sample solu-tion is not larger than 3/5 times the peak area of carvedilol from the standard solution. For the area of the peak, having the relative retention time between 1.7 and 1.9 to carvedilol, multiply the correction factor 1.25. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 10 times as long as the retention time of carvedilol, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of carvedilol obtained with 50 mL of this solution is equivalent to 3.5 to 6.5z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of carvedilol are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carvedilol is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Carvedilol Tablets add 70 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1), shake until the tablet is completely disintegrated, then add a JP XVIII Official Monographs / Carvedilol Tablets 639 mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly V? mL so that each mL contains about 5 mg of carvedilol (C24H26N2O4), and use this solution as the sample solution.
Powder Carvedilol Tablets. To a portion of the powder, equivalent to 20 mg of Carvedilol, add 10 mL of methanol, shake well, and filter. To 0.5 mL of the filtrate add methanol to make 200 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 222 nm and 226 nm, between 241 nm and 245 nm, be- tween 284 nm and 288 nm, between 317 nm and 321 nm and between 330 nm and 334 nm.
Related substances—In this procedure the sample solution should be stored not exceeding 59C and used within 24 hours after preparation. Powder Carvedilol Tablets. Dis-solve a portion of the powder, equivalent to 12.5 mg of Car-vedilol, add an adequate amount of the mobile phase and disperse the particles with the aid of ultrasonic waves, if nec-essary, add the mobile phase to make 100 mL, and shake for 30 minutes. Filter through a membrane filter with a pore size not exceeding 0.22 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the two peaks, having the rela-tive retention time between 1.7 and 1.9 and between 2.0 and to carvedilol, obtained from the sample solution of 1.25-mg or 2.5-mg tablet is not larger than 3/10 times and 1.6 times the peak area of carvedilol from the standard solu-tion, respectively, the area of the peak other than carvedilol and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of carvedilol from the standard solution, and the total area of the peaks other than carvedilol from the sample solution is not larger than 2.2 times the peak area of carvedilol from the standard solu-tion. The area of the two peaks, having the relative retention time between 1.7 and 1.9 and between 2.0 and 3.1, from the sample solution of 10-mg or 20-mg tablet is not larger than 1/10 times and 2/5 times the peak area of carvedilol from the standard solution, respectively, the area of the peak other than carvedilol and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of carvedilol from the standard solution, and the total area of the peaks other than carvedilol from the sample solu-tion is not larger than 3/5 times the peak area of carvedilol from the standard solution. For the area of the peak, having the relative retention time between 1.7 and 1.9 to carvedilol, multiply the correction factor 1.25. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 10 times as long as the retention time of carvedilol, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of carvedilol obtained with 50 mL of this solution is equivalent to 3.5 to 6.5z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of carvedilol are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carvedilol is not more than 1.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Carvedilol Tablets add 70 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1), shake until the tablet is completely disintegrated, then add a JP XVIII Official Monographs / Carvedilol Tablets 639 mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly V? mL so that each mL contains about 5 mg of carvedilol (C24H26N2O4), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of carvedilol for assay, previously dried at 1059C for 2 hours, and dissolve in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL. Pipet 2 mL of this solution, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 240 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of carvedilol (C24H26N2O4) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of carvedilol for assay taken Dissolution <6.10> (1) 10-mg tablet and 20-mg tablet When the test is performed at 75 revolutions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 30 minutes of Carvedilol Tablets is not less than 80z. Start the test with 1 tablet of Carvedilol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of carvedilol (C24H26N2O4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of carvedilol for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 200 mL. Pipet 2 mL of this solu-tion, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 240 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of carvedilol (C24H26N2O4) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of carvedilol for assay taken C: Labeled amount (mg) of carvedilol (C24H26N2O4) in 1 tablet
Weigh accurately the mass of not less than 20 Car-vedilol Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg of carvedilol (C24H26N2O4), add exactly 5 mL of the internal standard so-lution, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make 250 mL, and shake for 30 minutes. To 2 mL of this solution, add the mobile phase to make 20 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of carvedilol for assay, previously dried at 1059C for 2 hours, add exactly 5 mL of the internal standard solution, and add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make 250 mL. To 2 mL of this solution add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> under the following conditions, and calculate the ratios, QT and QS, of the peak area of carvedilol to that of the internal standard. assay, previously dried at 1059C for 2 hours, and dissolve in Amount (mg) of carvedilol (C H N O ) methanol to make exactly 50 mL. Pipet 2 mL of this solu-tion, add the dissolution medium to make exactly 100 mL, ＝ MS × QT/QS 24 26 2 4 and use this solution as the standard solution. Determine the absorbances, AT and AS, at 285 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of carvedilol (C24H26N2O4) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of carvedilol for assay taken C: Labeled amount (mg) of carvedilol (C24H26N2O4) in 1 tablet (2) 1.25-mg tablet and 2.5-mg tablet When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 20 minutes is not less than 75z. Start the test with 1 tablet of Carvedilol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 1.4 mg of carvedilol (C24H26N2O4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of carvedilol for MS: Amount (mg) of carvedilol for assay taken Internal standard solution—A solution of isoamyl parahy-droxybenzoate in the mobile phase (1 in 70). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.7 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.0 with a solution prepared by dissolving 0.7 g of dipotassium hydrogen phosphate in water to make 200 mL. To 450 mL of this solution add 550 mL of methanol. Flow rate: Adjust so that the retention time of carvedilol is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, carvedilol and the internal standard are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ‌640 the peak area of carvedilol to that of the internal standard is not more than 1.0z.
Cefaclor Capsules contain not less than 90.0z and not more than 110.0z of the labeled potency of cefaclor (C15H14ClN3O4S: 367.81). Method of preparation Prepare as directed under Cap-sules, with Cefaclor. Identification Shake vigorously a quantity of the contents of Cefaclor Capsules, equivalent to 20 mg (potency) of Cefaclor, with 10 mL of water, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, dissolve 20 mg of Cefaclor RS in 10 mL of water, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of acetonitrile, water, ethyl acetate and formic acid (30:10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Purity Related substances—Weigh accurately not less than 5 Cefaclor Capsules, open the capsules and carefully take out the contents, mix well, and powder, if necessary. Wash the empty capsules with a little amount of diethyl ether, if necessary, allow the capsules to stand at room temperature to vaporize adhering diethyl ether, and weigh accurately the capsules to calculate the mass of the contents. Weigh accu-rately a quantity of the contents, equivalent to about 0.25 g (potency) of Cefaclor, shake with 40 mL of 0.1 mol/L phos-phate buffer solution (pH 4.5) for 10 minutes, add the same buffer solution to make exactly 50 mL, and filter through a 0.45-mm pore-size membrane filter. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Pipet 2.5 mL of this solution, add the same buffer solution to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas by the auto-matic integration method. Calculate the amount of each related substance by the following equation: the amount of each related substance is not more than 0.5z, and the total amount of the related substances is not more than 2.5z. If necessary, correct the fluctuation of the base line by per-forming the test in the same manner with 20 mL of 0.1 mol/L phosphate buffer solution (pH 4.5). Amount (z) of each related substance ＝ MS/MT × AT/AS × MM/C × 25/2 Total amount (z) of the related substances ＝ MS/MT × SAT/AS × MM/C × 25/2 MS: Amount [mg (potency)] of Cefaclor RS taken MT: Amount (mg) of the contents of Cefaclor Capsules taken MM: Average mass (mg) of the contents in 1 capsule AT: Area of each peak other than cefaclor and solvent from the sample solution AS: Peak area of cefaclor from the standard solution C: Labeled potency [mg (potency)] of Cefaclor in 1 cap-sule Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefaclor. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Confirm that the peak area of cefaclor obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefaclor are not less than 40,000 and 0.8 to 1.3, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefaclor is not more than 2.0z. Water <2.48> Not more than 8.0z (0.2 g, volumetric titra- ‌642 tion, back titration). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefaclor Capsules is not less than 80z. Start the test with 1 capsule of Cefaclor Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg (potency) of Cefaclor, and use this so-lution as the sample solution.
Shake vigorously a quantity of the contents of Cefaclor Capsules, equivalent to 20 mg (potency) of Cefaclor, with 10 mL of water, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, dissolve 20 mg of Cefaclor RS in 10 mL of water, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of acetonitrile, water, ethyl acetate and formic acid (30:10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value.
Related substances—Weigh accurately not less than 5 Cefaclor Capsules, open the capsules and carefully take out the contents, mix well, and powder, if necessary. Wash the empty capsules with a little amount of diethyl ether, if necessary, allow the capsules to stand at room temperature to vaporize adhering diethyl ether, and weigh accurately the capsules to calculate the mass of the contents. Weigh accu-rately a quantity of the contents, equivalent to about 0.25 g (potency) of Cefaclor, shake with 40 mL of 0.1 mol/L phos-phate buffer solution (pH 4.5) for 10 minutes, add the same buffer solution to make exactly 50 mL, and filter through a 0.45-mm pore-size membrane filter. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Pipet 2.5 mL of this solution, add the same buffer solution to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas by the auto-matic integration method. Calculate the amount of each related substance by the following equation: the amount of each related substance is not more than 0.5z, and the total amount of the related substances is not more than 2.5z. If necessary, correct the fluctuation of the base line by per-forming the test in the same manner with 20 mL of 0.1 mol/L phosphate buffer solution (pH 4.5). Amount (z) of each related substance ＝ MS/MT × AT/AS × MM/C × 25/2 Total amount (z) of the related substances ＝ MS/MT × SAT/AS × MM/C × 25/2 MS: Amount [mg (potency)] of Cefaclor RS taken MT: Amount (mg) of the contents of Cefaclor Capsules taken MM: Average mass (mg) of the contents in 1 capsule AT: Area of each peak other than cefaclor and solvent from the sample solution AS: Peak area of cefaclor from the standard solution C: Labeled potency [mg (potency)] of Cefaclor in 1 cap-sule Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefaclor. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Confirm that the peak area of cefaclor obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefaclor are not less than 40,000 and 0.8 to 1.3, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefaclor is not more than 2.0z. Water <2.48> Not more than 8.0z (0.2 g, volumetric titra- ‌642 tion, back titration). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefaclor Capsules is not less than 80z. Start the test with 1 capsule of Cefaclor Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg (potency) of Cefaclor, and use this so-lution as the sample solution. Separately, weigh accurately about 20 mg (potency) of Cefaclor RS, and dissolve in water to make exactly 20 mL. Pipet 1 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 265 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of cefaclor (C15H14ClN3O4S) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefaclor RS taken C: Labeled amount [mg (potency)] of Cefaclor in 1 cap-sule
Weigh accurately not less than 5 Cefaclor Capsules, open the capsules and carefully take out the contents, mix well, and powder, if necessary. Wash the empty capsules with a little amount of diethyl ether, if necessary, allow the capsules to stand at room temperature to vaporize adhering diethyl ether, and weigh accurately the capsules to calculate the mass of the contents. Weigh accurately a quantity of the contents, equivalent to about 0.1 g (potency) of Cefaclor, shake vigorously with 60 mL of 0.1 mol/L phosphate buffer solution (pH 4.5) for 10 minutes, add the same buffer solu-tion to make exactly 100 mL, and centrifuge. Pipet 10 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 50 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Cefaclor. Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) ＝ MS × QT/QS × 2 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700).
Cefaclor Combination Granules contain gastric-soluble granules and enteric-soluble granules in one package. It contains cefaclor (C15H14ClN3O4S: 367.
Shake vigorously a quantity of Cefaclor Combination Granules, equivalent to 20 mg (potency) of Cefaclor according to the labeled total potency, with 10 mL of water, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 20 mg of Cefaclor RS in 10 mL of water, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of acetonitrile, water, ethyl acetate and formic acid (30:10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value.
Related substances—Take out the total content of not less than 5 packages of Cefaclor Combination Granules, add a small amount of 0.1 mol/L phosphate buffer solution (pH 4.5), grind, add 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigorously for 10 minutes, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly V mL so that each mL contains about 5 mg (potency) of Cefaclor according to the labeled total potency. Pipet 10 mL of this solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 25 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (po-tency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Pipet 2 mL of this solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area in each solution by the automatic integration method. Calculate the amount of each related substance by the following equation: the amount of each related substance is not more than 0.6z, and the total amount of the related substances is not more than 2.8z. If necessary, correct the fluctuation of the base line by per-forming the test in the same manner with 50 mL of 0.1 mol/L phosphate buffer solution (pH 4.5). Amount (z) of each related substance ＝ MS × AT/AS × V/4 ×{1/(C × T )} Total amount (z) of the related substances ＝ MS × SAT/AS × V/4 ×{1/(C × T )} JP XVIII Official Monographs / Cefaclor Combination Granules 643 MS: Amount [mg (potency)] of Cefaclor RS taken AT: Area of each peak other than cefaclor, solvent and excipient from the sample solution SAT: Total area of the peaks other than cefaclor, solvent and excipient from the sample solution AS: Peak area of cefaclor from the standard solution C: Labeled total potency [mg (potency)] of Cefaclor in 1 package T: Number (pack) of Cefaclor Combination Granules Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefaclor. System suitability— Test for required detectability: Pipet 1 mL of standard so-lution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Confirm that the peak area of cefaclor obtained from 50 mL of this solution is equivalent to 3.5 to 6.5z of that obtained from 50 mL of the standard so-lution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefaclor are not less than 40,000 and between 0.8 and 1.3, respectively. System repeatability: When the test is repeated 3 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefaclor is not more than 2.0z. Water <2.48> Not more than 5.5z (0.3 g, volumetric titra-tion, back titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Total potency—Take out the total content of 1 pack-age of Cefaclor Combination Granules, add a little amount of 0.1 mol/L phosphate buffer solution (pH 4.5), grind well, add the same buffer solutions to make exactly V mL so that each mL contains about 3.8 mg (potency) of Cefaclor ac-cording to the labeled total potency after shaking vigorously for 10 minutes, and centrifuge. Pipet 3 mL of the superna-tant liquid, add exactly 10 mL of the internal standard solu-tion and the same buffer solution to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Cefaclor RS, equivalent to about 50 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solu-tion (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this so-lution, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this so-lution as the standard solution. Then, proceed as directed in the Assay under Cefaclor. standard solution and the same buffer solution to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 50 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefaclor. Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) ＝ MS × QT/QS × V/35 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 60 minutes of Cefaclor Combi-nation Granules is between 35z and 45z. Start the test with the total content of 1 package of Cefaclor Combination Granules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 20 mg (potency) of Cefaclor according to the labeled potency of gastric-soluble granule, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (po-tency), and dissolve in the dissolution medium to make ex-actly 20 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 265 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) of cefaclor (C15H14ClN3O4S) with respect to the labeled potency ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefaclor RS taken C: Labeled total potency [mg (potency)] of Cefaclor in 1 pack Separately, when the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Cefaclor Combination Granules is not less than 70z. Amount [mg (potency)] of cefaclor (C ＝ MS × QT/QS × V/15 15H14ClN3O4S) Start the test with the total content of 1 package of Cefaclor Combination Granules, withdraw not less than 20 mL of the medium at the specified minute after starting the MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700). Potency of gastric-soluble granule—Take out the total content of 1 package of Cefaclor Combination Gran-ules stir gently with 60 mL of 0.1 mol/L phosphate buffer solution (pH 4.5) for 5 minutes, add the same buffer solu-tion to make exactly V mL so that each mL contains about 1.5 mg (potency) of Cefaclor according to the labeled po-tency of gastric-soluble granule, and centrifuge. Pipet 7 mL of the supernatant liquid, add exactly 10 mL of the internal test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.01 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 20 mg (potency) of Cefaclor accord-ing to the labeled total potency, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (potency), dissolve in the dissolution medium to make exactly 100 mL, and warm at 379C for 60 minutes. Pipet 2 mL of this solution, add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL, and use this solution as the standard solution. Deter- ‌644 mine the absorbances, AT and AS, at 265 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using 0.01 mol/L hydro-chloric acid TS as the blank. Dissolution rate (z) of cefaclor (C15H14ClN3O4S) with respect to the labeled potency ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefaclor RS taken C: Labeled total potency [mg (potency)] of Cefaclor in 1 package
(1) Total potency—Take out the total content of not less than 5 Cefaclor Combination Granules, add a small amount of 0.1 mol/L phosphate buffer solution (pH 4.5), grind well, add the same buffer solution so that each mL containing about 5 mg (potency) of Cefaclor according to the labeled total potency after shaking vigorously for 10 minutes, and centrifuge. Pipet 2 mL of the supernatant liq-uid, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 50 mg (po-tency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solu-tion, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solu-tion as the standard solution. Then, proceed as directed in the Assay under Cefaclor. Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) ＝ MS × QT/QS × 1/5 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700). (2) Potency of gastric-soluble granule—Stir gentry the total content of not less than 5 Cefaclor Combination Gran-ules with about 100 mL of 0.1 mol/L phosphate buffer solu-tion (pH 4.5) for 5 minutes, add the same buffer solution so that each mL containing about 2 mg (potency) of Cefaclor according to the labeled potency of gastric-soluble granule, and centrifuge. Pipet 5 mL of the supernatant liquid, add ex-actly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 50 mg (potency), and dis-solve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefaclor. Cefaclor Fine Granules
Cefaclor Fine Granules contain not less than 90.0z and not more than 110.0z of the labeled potency of cefaclor (C15H14ClN3O4S: 367.81). Method of preparation Prepare as directed under Gran-ules, with Cefaclor. Identification Shake vigorously a quantity of Cefaclor Fine Granules, equivalent to 20 mg (potency) of Cefaclor, with 10 mL of water, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 20 mg (potency) of Cefaclor RS in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of aceto-nitrile, water, ethyl acetate and formic acid (30:10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot from the sample solution and the spot from the standard solution show the same Rf value. Purity Related substances—Weigh accurately a quantity of Cefaclor Fine Granules after powdered if necessary, equiva-lent to about 0.1 g (potency) of Cefaclor, shake with 40 mL of 0.1 mol/L phosphate buffer solution (pH 4.5) for 10 minutes, add the same buffer solution to make exactly 50 mL, and filter through a 0.45-mm pore-size membrane filter. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (po-tency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Pipet 2 mL of this solution, add the same buffer solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas by the automatic integration method. Calculate the amount of each related substance by the following equa-tion: the amount of each related substance is not more than 0.5z, and the total amount of the related substances is not more than 3.0z. If necessary, correct the fluctuation of the base line by performing the test in the same manner with 50 mL of 0.1 mol/L phosphate buffer solution (pH 4.5). Amount (z) of each related substance ＝ MS/MT × AT/AS × 1/C × 5 Amount [mg (potency)] of cefaclor (C ＝ MS × QT/QS × 1/5 15H14ClN3O4S) Total amount (z) of the related substances ＝ MS/MT × SAT/AS × 1/C × 5 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700). Containers and storage Containers—Tight containers. Storage—Light-resistant. MS: Amount [mg (potency)] of Cefaclor RS taken MT: Amount (g) of Cefaclor Fine Granules taken AT: Area of the peak other than cefaclor and the solvent from the sample solution AS: Peak area of cefaclor from the standard solution C: Labeled potency [mg (potency)] of cefaclor (C15H14ClN3O4S) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefaclor. ‌JP XVIII Official Monographs / Cefadroxil 645 System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Confirm that the peak area of cefaclor obtained with 50 mL of this solution is equivalent to 3.5 to 6.5z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefaclor are not less than 40,000 and between 0.8 and 1.3, respectively. System repeatability: When the test is repeated 3 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefaclor is not more than 2.0z. Water <2.48> Not more than 1.5z (1 g, volumetric titra-tion, back titration). Uniformity of dosage units <6.02> The granules in single-dose packages meet the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefaclor Fine Granules is not less than 85z. Start the test with an accurately weighed amount of Cefaclor Fine Granules, equivalent to about 0.25 g (potency) of Cefaclor, withdraw not less than about 20 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make ex-actly V? mL so that each mL contains about 20 mg (potency) of Cefaclor, and use this solution as the sample solution.
Shake vigorously a quantity of Cefaclor Fine Granules, equivalent to 20 mg (potency) of Cefaclor, with 10 mL of water, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 20 mg (potency) of Cefaclor RS in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate with a mixture of aceto-nitrile, water, ethyl acetate and formic acid (30:10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot from the sample solution and the spot from the standard solution show the same Rf value.
Related substances—Weigh accurately a quantity of Cefaclor Fine Granules after powdered if necessary, equiva-lent to about 0.1 g (potency) of Cefaclor, shake with 40 mL of 0.1 mol/L phosphate buffer solution (pH 4.5) for 10 minutes, add the same buffer solution to make exactly 50 mL, and filter through a 0.45-mm pore-size membrane filter. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Cefaclor RS, equivalent to about 20 mg (po-tency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 20 mL. Pipet 2 mL of this solution, add the same buffer solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas by the automatic integration method. Calculate the amount of each related substance by the following equa-tion: the amount of each related substance is not more than 0.5z, and the total amount of the related substances is not more than 3.0z. If necessary, correct the fluctuation of the base line by performing the test in the same manner with 50 mL of 0.1 mol/L phosphate buffer solution (pH 4.5). Amount (z) of each related substance ＝ MS/MT × AT/AS × 1/C × 5 Amount [mg (potency)] of cefaclor (C ＝ MS × QT/QS × 1/5 15H14ClN3O4S) Total amount (z) of the related substances ＝ MS/MT × SAT/AS × 1/C × 5 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700).
Weigh accurately a quantity of Cefaclor Fine Gran-ules after powdered if necessary, equivalent to about 0.1 g (potency) of Cefaclor, shake vigorously with 60 mL of 0.1 mol/L phosphate buffer solution (pH 4.5) for 10 minutes, add the same buffer solution to make exactly 100 mL, and centrifuge. Pipet 10 mL of the supernatant liquid, add ex-actly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg (potency) of Cefaclor RS, and dissolve in 0.1 mol/L phos-phate buffer solution (pH 4.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and the same buffer solution to make 50 mL, and use this solution as standard solution. Proceed as directed in the Assay under Cefaclor. Amount [mg (potency)] of cefaclor (C15H14ClN3O4S) ＝ MS × QT/QS × 2 MS: Amount [mg (potency)] of Cefaclor RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 700).
Cefadroxil Capsules contain not less than 95.0z and not more than 105.0z of the labeled potency of cefadroxil (C16H17N3O5S: 363.39). Method of preparation Prepare as directed under Cap-sules, with Cefadroxil. Identification Dissolve the contents of Cefadroxil Cap-sules, equivalent to 10 mg (potency) of Cefadroxil, in 500 mL of water, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 228 nm and 232 nm, and between 261 nm and 265 nm. Water <2.48> Not more than 7.0z (0.15 g, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Place 1 capsule of Cefadroxil Capsules in 300 mL of water, disperse with the aid of ultrasonic waves, shake for 30 minutes, and add water to make exactly 500 mL. Pipet 5 mL of this solution, and add water to make exactly V mL so that each mL contains about 0.
Dissolve the contents of Cefadroxil Cap-sules, equivalent to 10 mg (potency) of Cefadroxil, in 500 mL of water, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 228 nm and 232 nm, and between 261 nm and 265 nm. Water <2.48> Not more than 7.0z (0.15 g, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Place 1 capsule of Cefadroxil Capsules in 300 mL of water, disperse with the aid of ultrasonic waves, shake for 30 minutes, and add water to make exactly 500 mL. Pipet 5 mL of this solution, and add water to make exactly V mL so that each mL contains about 0.1 mg (potency) of Cefadroxil. Filter the solution, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg (potency) of Cefadroxil RS, dissolve in water to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefadroxil. Amount [mg (potency)] of cefadroxil (C16H17N3O5S) ＝ MS × AT/AS × V/2 MS: Amount [mg (potency)] of Cefadroxil RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 90 minutes of Cefadroxil Capsules is not less than 80z. Start the test with 1 capsule of Cefadroxil Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefadroxil, and use this solution as the sample solution. Separately, weigh accurately about 22 mg (potency) of Cefadroxil RS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 263 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of cefadroxil (C16H17N3O5S) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefadroxil RS taken C: Labeled amount [mg (potency)] of cefadroxil in 1 cap-sule ‌JP XVIII Official Monographs / Cefalexin 647‌
Take out the contents of 20 Cefadroxil Capsules, and combine. Weigh accurately the mass of the combined contents, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg (potency) of Cefadroxil, add 300 mL of water, shake for 30 minutes, then add water to make exactly 500 mL, and filter. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Cefa-droxil RS, equivalent to about 20 mg (potency), dissolve in water to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefadroxil. Dissolution rate (z) with respect to the labeled amount of cefadroxil (C16H17N3O5S) ＝ MS/MT × AT/AS × 1/C × 450 MS: Amount [mg (potency)] of Cefadroxil RS taken MT: Amount (g) of Cefadroxil for Syrup taken C: Labeled amount [mg (potency)] of cefadroxil in 1 g Assay Weigh accurately an amount of powdered Cefa-droxil for Syrup, equivalent to about 50 mg (potency) of Cefadroxil, dissolve in water to make exactly 500 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefadroxil RS, equivalent to about Amount [mg (potency)] of cefadroxil (C ＝ MS × AT/AS × 5/2 16H17N3O5S) 20 mg (potency), dissolve in water to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefadroxil. MS: Amount [mg (potency)] of Cefadroxil RS taken
Cefadroxil for Syrup is a preparation for syrup, which is suspended before use. It contains not less than 95.
Dissolve an amount of Cefadroxil for Syrup, equivalent to 10 mg (potency) of Cefadroxil, in 500 mL of water, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 228 nm and 232 nm, and between 261 nm and 265 nm. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration). Uniformity of dosage units <6.02> The syrup in single-dose packages meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method (put the sample in the dissolution medium so that it disperses), using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefadroxil for Syrup is not less than 85z. Start the test with accurately weighed amount of Cefadroxil for Syrup, equivalent to about 0.1 g (potency) of Cefadroxil, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 4 mL of the subsequent filtrate, add water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg (potency) of Cefadroxil RS, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 263 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount [mg (potency)] of cefadroxil (C16H17N3O5S) ＝ MS × AT/AS × 5/2 MS: Amount [mg (potency)] of Cefadroxil RS taken
Cefalexin Capsules contain not less than 93.0z and not more than 107.0z of the labeled potency of cefalexin (C16H17N3O4S: 347.39). Method of preparation Prepare as directed under Cap-sules, with Cefalexin. Identification Take out the contents of Cefalexin Capsules, to a quantity of the contents, equivalent to 70 mg (potency) of Cefalexin, add 25 mL of water, shake vigorously for 5 minutes, and filter. To 1 mL of the filtrate add water to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 260 nm and 264 nm. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, back titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Open 1 capsule of Cefalexin Capsules, add 3V/5 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigor-ously for 10 minutes, add 0.1 mol/L phosphate buffer solu-tion (pH 4.5) to make exactly V mL so that each mL con-tains about 1.25 mg (potency) of Cefalexin. Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Cefalexin RS, equivalent to about 25 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solu-tion, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the standard solution. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefalexin to that of the internal standard. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 30 minutes of 125-mg (potency) capsule and in 60 minutes of 250-mg (potency) capsule are not less than 75z and 80z, respectively. Start the test with 1 capsule of Cefalexin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefalexin, and use this so-lution as the sample solution.
Take out the contents of Cefalexin Capsules, to a quantity of the contents, equivalent to 70 mg (potency) of Cefalexin, add 25 mL of water, shake vigorously for 5 minutes, and filter. To 1 mL of the filtrate add water to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 260 nm and 264 nm. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, back titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Open 1 capsule of Cefalexin Capsules, add 3V/5 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigor-ously for 10 minutes, add 0.1 mol/L phosphate buffer solu-tion (pH 4.5) to make exactly V mL so that each mL con-tains about 1.25 mg (potency) of Cefalexin. Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Cefalexin RS, equivalent to about 25 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solu-tion, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the standard solution. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefalexin to that of the internal standard. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 30 minutes of 125-mg (potency) capsule and in 60 minutes of 250-mg (potency) capsule are not less than 75z and 80z, respectively. Start the test with 1 capsule of Cefalexin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefalexin, and use this so-lution as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 22 mg (po-tency), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 262 nm. Dissolution rate (z) with respect to the labeled amount of cefalexin (C16H17N3O4S) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefalexin RS taken C: Labeled amount [mg (potency)] of cefalexin (C16H17N3O4S) in 1 capsule
Take out the contents of not less than 20 capsules of Cefalexin Capsules, weigh accurately the mass of the con-tents, and powder. Weigh accurately a portion of the pow-der, equivalent to about 0.1 g (potency) of Cefalexin, add 60 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigorously for 10 minutes, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL, and centrifuge. Pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 20 mg (po-tency), dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the standard solution. Perform the Amount [mg (potency)] of cefalexin (C H N O S) test with 10 mL each of the sample solution and standard ＝ MS × QT/QS × V/20 16 17 3 4 solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefalexin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cefalexin to that of the internal standard substance is not more than 1.0z. ratios, QT and QS, of the peak area of cefalexin to that of the internal standard. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × 5 MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. ‌650 Mobile phase: Dissolve 2.72 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 3.0 with diluted phosphoric acid (3 in 500). To 800 mL of this solution add 200 mL of methanol. Flow rate: Adjust so that the retention time of cefalexin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefalexin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). (2) Potency of gastric-soluble granules—To the total content of 1 package of Cefalexin Combination Granules, add 3 V/5 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake gently for 5 minutes, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly V mL so that each mL contains about 0.6 mg (potency) of Cefalexin according to the labeled potency of gastric-soluble granules, and centri-fuge. Pipet 7 mL of the supernatant liquid, add exactly 20 mL of the internal standard solution, add 0.1 mol/L phos-phate buffer solution (pH 4.5) to make 200 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay (1) Total potency. the peak area of cefalexin to that of the internal standard is not more than 1.0z. Amount [mg (potency)] of cefalexin (C ＝ MS × QT/QS × V/35 16H17N3O4S)
Cefalexin Combination Granules contain gastric-soluble granules and enteric-soluble granules in one package. It contains not less than 90.
Powder Cefalexin Combination Granules, weigh a portion of the powder, equivalent to 30 mg (po-tency) of Cefalexin according to the labeled total potency, shake vigorously for 5 minutes with 100 mL of water, and centrifuge. To 2 mL of the supernatant liquid add water to make 20 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 260 nm and 264 nm. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, direct titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Total potency—To the total content of 1 package of Cefalexin Combination Granules add a small amount of 0.1 mol/L phosphate buffer solution (pH 4.5), grind, add 3 V/5 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigorously for 10 minutes, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly V mL so that each mL contains about 2 mg (potency) of Cefalexin according to the labeled total potency, and centrifuge. Pipet 2 mL of the su-pernatant liquid, add exactly 20 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 200 mL, and use this solution as the sample solu-tion. Then, proceed as directed in the Assay (1) Total po-tency. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × V/10 MS: Amount [mg (potency)] of Cefalexin RS taken MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Cefalexin Combi-nation Granules is between 25z and 35z. Start the test with the total content of 1 package of Cefalexin Combination Granules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefalexin according to the labeled potency of gastric-soluble granules, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 22 mg (po-tency), and dissolve in the dissolution medium to make ex-actly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 262 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) of cefalexin (C16H17N3O4S) with respect to the labeled potency ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefalexin RS taken C: Labeled total potency [mg (potency)] of Cefalexin in 1 package When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolu-tion rate in 60 minutes of 200 mg (potency) preparation is not less than 80z, and the dissolution rate in 45 minutes of 500 mg (potency) preparation is not less than 75z. Start the test with the total content of 1 package of Cefalexin Combination Granules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefalexin according to the ‌JP XVIII Official Monographs / Cefalexin for Syrup 651 labeled total potency, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 22 mg (potency), dissolve in the dis-solution medium to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 262 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cefalexin to that of the internal standard is not more than 1.0z. Potency of gastric-soluble granules—Take out the content from not less than 20 packages of Cefalexin Combi-nation Granules, weigh accurately a quantity, equivalent to about 0.3 g (potency) of Cefalexin according to the labeled Dissolution rate (z) of cefalexin (C16H with respect to the labeled potency 17N3O4S) potency of gastric-soluble granules, shake gently for 5 minutes with 200 mL of 0.1 mol/L phosphate buffer solu-tion (pH 4.5), add 0.1 mol/L phosphate buffer solution (pH ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefalexin RS taken C: Labeled total potency [mg (potency)] of Cefalexin in 1 package
(1) Total potency—Powder the total content ob-tained from not less than 20 packages of Cefalexin Combi-nation Granules, weigh accurately a portion of the powder, equivalent to about 0.5 g (potency) of Cefalexin, shake vig-orously for 10 minutes with 150 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 250 mL, and centrifuge. Pipet 2 mL of this solution, add exactly 20 mL of the inter-nal standard solution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 200 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 20 mg (potency), and dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution, and add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefalexin to that of the inter-nal standard. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × 25 MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.72 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (3 in 500). To 800 mL of this solu-tion add 200 mL of methanol. Flow rate: Adjust so that the retention time of cefalexin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefalexin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. 4.5) to make exactly 300 mL, and centrifuge. Pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, and add 0.1 mol/L phosphate buffer so-lution (pH 4.5) to make 100 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay (1) Total potency. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × 15 MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000).
Cefalexin for Syrup is a preparation for syrup, which is dissolved or suspended before use. It contains not less than 90.
Dissolve a quantity of Cefalexin for Syrup, equivalent to 3 mg (potency) of Cefalexin, in water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Water <2.48> Not more than 5.0z (0.4 g, volumetric titra-tion, back titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Cefalexin for Syrup in single-dose packages meets the requirement of the Content unifor-mity test. Take out the total content of 1 package of Cefalexin for Syrup, add 3V/5 mL of 0.1 mol/L phosphate buffer solu-tion (pH 4.5), shake vigorously for 10 minutes, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly V mL so that each mL contains about 1 mg (potency) of Cefalexin, and centrifuge. Pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × V/20 ‌652 MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefalexin for Syrup is not less than 80z. Start the test with an accurately weighed amount of Cefalexin, equivalent to about 0.25 g (potency) of Cefalexin for Syrup, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 22 mg (potency), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 262 nm. Dissolution rate (z) with respect to the labeled amount of cefalexin (C16H17N3O4S) ＝ MS/MT × AT/AS × 1/C × 1125 MS: Amount [mg (potency)] of Cefalexin RS taken MT: Amount (g) of Cefalexin for Syrup taken C: Labeled amount [mg (potency)] of cefalexin (C16H17N3O4S) in 1 g
Powder Cefalexin for Syrup, if necessary, and weigh accurately a portion of the powder, equivalent to about 0.1 g (potency) of Cefalexin, add 60 mL of 0.1 mol/L phosphate buffer solution (pH 4.5), shake vigorously for 10 minutes, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL, and centrifuge. Pipet 2 mL of the superna-tant liquid, add exactly 10 mL of the internal standard solu-tion, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefalexin RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 4.5) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the inter-nal standard solution, add 0.1 mol/L phosphate buffer solu-tion (pH 4.5) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of cefalexin to that of the internal standard. Amount [mg (potency)] of cefalexin (C16H17N3O4S) ＝ MS × QT/QS × 5 MS: Amount [mg (potency)] of Cefalexin RS taken Internal standard solution—A solution of m-hydroxya-cetophenone in 0.1 mol/L phosphate buffer solution (pH 4.5) (1 in 15,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame- ter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.72 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 3.0 with diluted phosphoric acid (3 in 500). To 800 mL of this solution add 200 mL of methanol. Flow rate: Adjust so that the retention time of cefalexin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefalexin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cefalexin to that of the internal standard is not more than 1.0z.
Cefalotin Sodium for Injection is a preparation for injection, which is dissolved before use. It contains not less than 90.
Cefalotin Sodium for Injection occurs as white to light yellow-white, crystals or crystalline powder. Identiˆcation Determine the infrared absorption spectrum of Cefalotin Sodium for Injection as directed in the potas-sium bromide disk method under Infrared Spectrophotome- ‌654 try <2.25>, and compare the spectrum with the Reference Spectrum of Cefalotin Sodium or the spectrum of Cefalotin Sodium RS: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. pH <2.54> Dissolve an amount of Cefalotin Sodium for In-jection, equivalent to 0.5 g (potency) of Cefalotin Sodium, in 5 mL of water: the pH of the solution is between 4.5 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefalotin Sodium for Injection in 10 mL of water: the so-lution is clear. Perform the test with this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: the ab-sorbance at 450 nm is not more than 0.20. (2) Related substances—Dissolve an amount of Cefalo-tin Sodium for Injection, equivalent to 25 mg (potency), in the mobile phase to make 25 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than cefalotin obtained from the sample solution is not larger than the peak area of cefalotin from the standard so-lution, and the total area of the peaks other than cefalotin from the sample solution is not larger than 3 times the peak area of cefalotin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and ‰ow rate: Proceed as directed in the operating conditions in the Assay under Cefalotin Sodium. Time span of measurement: About 4 times as long as the retention time of cefalotin. System suitability— Proceed as directed in the system suitability in the Purity (4) under Cefalotin Sodium. Water <2.48> Not more than 1.0z (0.5 g, volumetric titra-tion, back titration). Bacterial endotoxins <4.01> Less than 0.2 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Cefalotin Sodium for Injection. Weigh accurately an amount of the contents, equivalent to about 25 mg (potency) of Cefalotin Sodium, dissolve in the mobile phase to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg (potency) of Cefalotin Sodium RS, and dissolve in the mo-bile phase to make exactly 25 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefalotin Sodium. Amount [mg(potency)] of cefalotin (C16H16N2O6S2) ＝ MS × AT/AS × 1000 MS: Amount [mg(potency)] of Cefalotin Sodium RS taken
Cefatrizine Propylene Glycolate for Syrup
シロップ用セファトリジンプロピレングリコール
Cefatrizine Propylene Glycolate for Syrup is a preparation for syrup, which is dissolved before use. It contains not less than 90.
Powder Cefatrizine Propylene Glycolate for Syrup, weigh a portion of the powder, equivalent to 10 mg (potency) of Cefatrizine Propylene Glycolate, and dissolve in 10 mL of water. To 2 mL of this solution add water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 225 nm and 229 nm, and between 266 nm and 271 nm. pH <2.54> Take an amount of Cefatrizine Propylene Glycolate for Syrup, equivalent to 0.4 g (potency) of Cefatri-zine Propylene Glycolate, and suspend in 10 mL of water: the pH of this suspension is between 4.0 and 6.0.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard Amount [ mg (potency)] of cefatrizine (C ＝ MS × AT/AS × 1000 18H18N6O5S2) solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area in each solution by the automatic integration method: MS: Amount [mg (potency)] of Cefatrizine Propylene Glycolate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of potassium dihy-drogenphosphate (17 in 12,500) and methanol (17:3). Flow rate: Adjust so that the retention time of cefatrizine is about 11 minutes. System suitability— System performance: Dissolve about 10 mg (potency) of Cefatrizine Propylene Glycolate and about 5 mg (potency) of Cefadroxil in 50 mL of water. When the procedure is run with 10 mL of this solution under the above operating condi-tions, cefadroxil and cefatrizine are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of peak areas of cefatrizine is not more than 1.0z.
Powder Cefatrizine Propylene Glycolate for Syrup, ‌656 weigh accurately a portion of the powder, equivalent to about 0.1 g (potency) of Cefatrizine Propylene Glycolate, dissolve in water to make exactly 500 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefatrizine Propylene Glycolate RS, equivalent to about 20 mg (potency), dissolve in water to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefatrizine Propylene Glycolate. Amount [mg (potency)] of cefatrizine (C18H18N6O5S2) ＝ MS × AT/AS × 5 MS: Amount [mg (potency)] of Cefatrizine Propylene Glycolate RS taken
Cefazolin Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Cefazolin Sodium for Injection occurs as white to light yellow-white, crystals or crystalline powder or masses.
(1) Determine the absorption spectrum of a solution of Cefazolin Sodium for Injection (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 270 nm and 274 nm. (2) Cefazolin Sodium for Injection responds to Qualita-tive Tests <1.09> (1) for chloride. Osmotic pressure ratio Being specified separately when the rected under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.35. (2) Related substances—Dissolve an amount of Cefazo-lin Sodium for Injection, equivalent to 0.10 g (potency) of Cefazolin Sodium, in 20 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and use this solution as the sample solu-tion. Prepare the sample solution before use. Perform the test with 5 mL of the sample solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method. Calculate the amount of each peak by the area percentage method: the amount of the peaks other than cefazolin is not more than 1.5z. Furthermore the total amount of the peaks other than cefazolin is not more than 2.5z. For the area of the peak, having the relative retention time of about 0.2 to cefazolin, multiply the correction fac-tor, 1.43. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Cefazolin Sodium. Time span of measurement: About 3 times as long as the retention time of cefazolin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Cefazolin Sodium. Test for required detectability: To 8 mL of the sample so-lution, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suita-bility test, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 20 mL. Confirm that the peak area of cefazolin obtained with 5 mL of this solution is equivalent to 3 to 7z of that with 5 mL of the solution for system suita-bility test. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of cefazolin is not more than 1.0z. Water <2.48> Not more than 3.0z (0.5 g, volumetric titra-tion, direct titration). Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination. Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem- ‌658 brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Cefazolin Sodium for Injection. Weigh accurately an amount of the contents, equivalent to about 50 mg (potency) of Cefazolin Sodium, dissolve in the internal standard solution to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Cefazolin RS, equivalent to about 50 mg (po- propionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits singlet signals, A and B, at around d 2.7 ppm and at around d 9.3 ppm. The ratio of integrated intensity of each signal, A:B, is about 3:1. (4) Cefazolin Sodium Hydrate responds to Qualitative Tests <1.09> (1) for sodium salt. tency), dissolve in the internal standard solution to make Absorbance <2.24> E 1z (272 nm): 272 – 292 (80 mg calcu- 1 cm exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefazolin Sodium. Amount [mg (potency)] of cefazolin (C14H14N8O4S3) ＝ MS × QT/QS MS: Amount [mg (potency)] of Cefazolin RS taken Internal standard solution—A solution of p-acetanisidide in 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000).
Cefcapene Pivoxil Hydrochloride Fine Granules
セフカペン ピボキシル塩酸塩細粒
Powder Cefcapene Pivoxil Hydrochloride Fine Granules. To a portion of the powder, equivalent to 10 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 40 mL of methanol, shake vigorously, and add metha-nol to make 50 mL. To 4 mL of this solution add methanol to make 50 mL, and filter through a membrane filter with a pore size of 0.45 mm. Determine the absorption spectrum of Amount [ mg (potency)] of cefcapene (C ＝ MS × QT/QS × 1000 17H19N5O6S2) the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 264 nm and 268 nm. MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-chloride RS taken Internal standard solution—A solution of p-benzylphenol in a mixture of water and methanol (1:1) (7 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.56 g of sodium dihydrogenphos-phate dihydrate and 1.22 g of sodium 1-decanesulfonate in water to make 1000 mL. To 700 mL of this solution add 300 mL of acetonitrile and 100 mL of methanol. Flow rate: Adjust so that the retention time of cefcapene pivoxil is about 5 minutes. System suitability— System performance: Dissolve 0.2 g of Cefcapene Pivoxil Hydrochloride Hydrate in 10 mL of methanol, and warm in a water bath at 609C for 20 minutes. After cooling, pipet 1 mL of this solution, and add exactly 10 mL of the internal standard solution and the mixture of water and methanol (1:1) to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, cefca-pene pivoxil, trans-cefcapene pivoxil and the internal stand-ard are eluted in this order, the relative retention time of trans-cefcapene pivoxil and the internal standard to that of cefcapene pivoxil are about 1.7 and about 2.0, respectively, and the resolution between the peaks of trans-cefcapene pivoxil and the internal standard is not less than 1.5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cefcapene pivoxil to that of the internal standard is not more than 1.0z.
(1) Related substances I—Powder Cefcapene Pivoxil Hydrochloride Fine Granules. To a portion of the powder, equivalent to 5 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 1 mL of methanol, and shake. Add 25 mL of a mixture of water and methanol (1:1), shake vigorously for 5 minutes, and filter through a membrane filter with a pore size of 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 30 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method. If necessary, compensate the base-line by performing in the same manner as the test with 30 mL of a mixture of water and methanol (1:1). Calculate the amount of the peaks other than the peak of cefcapene pivoxil by the area percentage method: the amount of the substance, having the relative retention time of about 1.3 to cefcapene pivoxil, is not more than 0.4z, the amount of the trans-isomer of cefcapene pivoxil, having the relative reten-tion time of about 1.5, is not more than 1.1z, the amount of the substance other than that mentioned above is not more than 0.3z, and the total amount of these substances is not more than 2.8z. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Cefcapene Pivoxil Hydrochloride Hydrate. System suitability— Proceed as directed in the system suitability in the Purity under Cefcapene Pivoxil Hydrochloride Hydrate. (2) Related substances II—Powder Cefcapene Pivoxil Hydrochloride Fine Granules. To a portion of the powder, equivalent to 2 mg (potency) of Cefcapene Pivoxil Hydro-chloride Hydrate, add 20 mL of N,N-dimethylformamide for liquid chromatography, shake vigorously for 10 minutes, and filter through a membrane filter with a pore size of 0.45 mm. Discard the first 3 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total area of the peaks eluted before that of cef-capene pivoxil is not more than 4.0z of the total area of all ‌JP XVIII Official Monographs / Cefcapene Pivoxil Hydrochloride Tablets 663 peaks other than the solvent peak. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefcapene Pivoxil Hydrochloride Hydrate. System suitability— Proceed as directed in the system suitability in the Purity under Cefcapene Pivoxil Hydrochloride Hydrate. Water <2.48> Not more than 1.4z (0.5 g, volumetric titra-tion, back titration). Perform the test without pulverizing the sample, and handling the sample under a relative hu-midity of less than 30z. Uniformity of dosage units <6.02> The granules in single-dose packages meet the requirement of the Mass variation test. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately an amount of Cefcapene Pivoxil Hydrochloride Fine Granules, equivalent to about 0.2 g (po-tency) of and Cefcapene Pivoxil Hydrochloride Hydrate, add 100 mL of the mixture of water and methanol (1:1), shake vigorously for 10 minutes, add the mixture of water and methanol (1:1) to make exactly 200 mL, and centrifuge at 3000 rpm for 5 minutes. Filter the supernatant liquid through a membrane filter with a pore size of 0.45 mm, dis-card the first 1 mL of the filtrate, pipet 2 mL of the subse-quent filtrate, add exactly 5 mL of the internal standard so-lution and the mixture of water and methanol (1:1) to make 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 20 mg (potency) of Cefcapene Pivoxil Hydrochloride RS, and dissolve in the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal stand-ard solution and the mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefcapene Pivoxil Hydrochloride Hydrate. Amount [mg (potency)] of cefcapene (C17H19N5O6S2) ＝ MS × QT/QS × 10 MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-chloride RS taken Internal standard solution—A solution of p-benzylphenol in the mixture of water and methanol (1:1) (7 in 4000).
To an amount of powdered Cefcapene Pivoxil Hydrochloride Tablets, equivalent to about 10 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 40 mL of methanol, shake vigorously, and add methanol to make 50 mL. To 4 mL of this solution add methanol to make 50 mL, filter through a membrane filter with pore size of 0.45 mm, and use the filtrate as the sample solution. De-termine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 263 nm and 267 nm.
(1) Related substances I—To an amount of pow-dered Cefcapene Pivoxil Hydrochloride Tables, equivalent to about 5 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 1 mL of methanol, and shake. Add 25 mL of a mixture of water and methanol (1:1), shake vigorously for 5 minutes, and filter through a membrane filter with pore size of 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 30 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method. If necessary, proceed with 30 mL of the mixture of water and methanol (1:1) in the same manner as the sample solution to compensate the base line. Calculate the amounts of the peaks other than cefcapene pivoxil by the area percentage method: the amount of the peak, having the relative retention time of about 1.3 to cefcapene pivoxil, is not more than 0.4z, the amount of the peak of cefcapene pivoxil trans-isomer, having the relative retention time of about 1.5, is not more than 0.5z, the amount of the peaks other than the peaks mentioned above are not more than 0.3z, respectively, and the total amount of these peaks is not more than 2.0z. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Cefcapene Pivoxil Hydrochloride Hydrate. System suitability— Proceed as directed in the system suitability in the Purity under Cefcapene Pivoxil Hydrochloride Hydrate. (2) Related substances II—To an amount of powdered Cefcapene Pivoxil Hydrochloride Tablets, equivalent to 2 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 20 mL of N,N-dimethylformamide for liquid chroma-tography, shake vigorously for 10 minutes, and filter through a membrane filter with pore size of 0.45 mm. Dis-card the first 3 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions, and deter-mine each peak area by the automatic integration method: the total area of the peaks which are eluted before cefcapene pivoxil is not more than 3.3z of the total area of the peaks other than the solvent peak. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefcapene Pivoxil Hydrochloride Hydrate. System suitability— Proceed as directed in the system suitability in the Purity under Cefcapene Pivoxil Hydrochloride Hydrate. Water <2.48> Not more than 3.9z (0.5 g, volumetric titra-tion, back titration). Powdering of the sample tablets and handling of the powder are performed under the relative humidity of not exceeding 30z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cefcapene Pivoxil Hydrochloride Tablets add 5 mL of water, and shake vigorously for 5 minutes to disintegrate. Add 20 mL of methanol, shake vigorously for 5 minutes, add a mixture of methanol and water (4:1) to make ‌664 exactly 50 mL, and centrifuge at 3000 rpm for 5 minutes. Filter the supernatant liquid through a membrane filter with pore size of 0.45 mm, and discard the first 1 mL of the fil-trate. Pipet V mL of the subsequent filtrate, equivalent to about 6 mg (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add exactly 15 mL of the internal standard solu-tion, add a mixture of water and methanol (1:1) to make 75 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of cefcapene (C17H19N5O6S2) ＝ MS × QT/QS × 15/V MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-chloride RS taken Internal standard solution—A solution of p-benzylphenol in a mixture of water and methanol (1:1) (7 in 4000). Dissolution Being specified separately when the drug is granted approval based on the Law.
To an amount of Cefcapene Pivoxil Hydrochloride Tablets, equivalent to about 0.6 g (potency) of Cefcapene Pivoxil Hydrochloride Hydrate, add 20 mL of water, and shake for 5 minutes to disintegrate. Add 80 mL of methanol, shake vigorously for 5 minutes, add a mixture of methanol and water (4:1) to make exactly 200 mL, and centrifuge at 3000 rpm for 5 minutes. Filter the supernatant liquid through a membrane filter with pore size of 0.45 mm, and discard the first 1 mL of the filtrate. Pipet 2 mL of the sub-sequent filtrate, add exactly 15 mL of the internal standard solution, add the mixture of water and methanol (1:1) to make 75 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefcapene Pivoxil Hydrochloride RS, equivalent to about 20 mg (po-tency), and dissolve in the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add the mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefcapene Pivoxil Hydrochloride Hydrate. Amount [mg (potency)] of cefcapene (C17H19N5O6S2) ＝ MS × QT/QS × 30 MS: Amount [mg (potency)] of Cefcapene Pivoxil Hydro-chloride RS taken Internal standard solution—A solution of p-benzylphenol in the mixture of water and methanol (1:1) (7 in 4000).
Cefdinir Capsules contain not less than 90.0z and not more than 110.0z of the labeled potency of cefdinir (C14H13N5O5S2: 395.41). Method of preparation Prepare as directed under Cap-sules, with Cefdinir. Identification To an amount of the contents of Cefdinir Capsules, equivalent to 10 mg (potency) of Cefdinir, add 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), ex-posure to ultrasonic waves for 1 minute, and filter. To 2 mL of the filtrate add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 221 nm and 225 nm and between 285 nm and 289 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate of a 50-mg capsule in 30 minutes is not less than 80z, and that of a 100-mg cap-sule in 45 minutes is not less than 75z. Start the test with 1 capsule of Cefdinir Capsules, with-draw not less than 20 mL of the medium at the specified ‌666 minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg (potency) of Cefdinir, and use this solution as the sample solution.
To an amount of the contents of Cefdinir Capsules, equivalent to 10 mg (potency) of Cefdinir, add 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), ex-posure to ultrasonic waves for 1 minute, and filter. To 2 mL of the filtrate add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 221 nm and 225 nm and between 285 nm and 289 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate of a 50-mg capsule in 30 minutes is not less than 80z, and that of a 100-mg cap-sule in 45 minutes is not less than 75z. Start the test with 1 capsule of Cefdinir Capsules, with-draw not less than 20 mL of the medium at the specified ‌666 minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg (potency) of Cefdinir, and use this solution as the sample solution. Separately, weigh accurately about 28 mg (potency) of Cefdinir RS, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01>, and determine the peak areas, AT and AS, of cefdinir in each solution. Dissolution rate (z) with respect to the labeled amount of cefdinir (C14H13N5O5S2) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount [mg (potency)] of Cefdinir RS taken C: Labeled amount [mg (potency)] of cefdinir (C14H13N5O5S2) in 1 capsule Operating conditions— Proceed as directed in the operating conditions in the Assay under Cefdinir. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefdinir are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefdinir is not more than 1.0z.
Weigh accurately not less than 5 Cefdinir Capsules, take out the contents, and powder. Wash the empty capsules with a little amount of diethyl ether, if necessary, allow to stand at a room temperature to vaporize the adhering diethyl ether, and weigh accurately the mass of the capsules to calcu-late the mass of the contents. Weigh accurately an amount of the contents, equivalent to about 0.1 g (potency) of Cefdinir, add 70 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30 minutes, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL. Centrifuge this solution at 3000 revolutions per minute for 10 minutes, pipet 4 mL of the supernatant liquid, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefdinir RS, equivalent to about 20 mg (po-tency), dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefdinir. Cefdinir Fine Granules
To an amount of Cefdinir Fine Granules, equivalent to 10 mg (potency) of Cefdinir, add 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), exposure to ultrasonic waves for 1 minute, and filter. To 2 mL of the fil-trate add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 221 nm and 225 nm and between 285 nm and 289 nm. Uniformity of dosage units <6.02> The granules in single-dose packages meet the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Cefdinir Fine Granules is not less than 75z. Start the test with an accurate amount of Cefdinir Fine Granules, equivalent to about 0.1 g (potency) of Cefdinir, withdraw not less than about 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, and use the sub-sequent filtrate as the sample solution. Separately, weigh ac-curately about 28 mg (potency) of Cefdinir RS, and dissolve in the dissolution medium to make exactly 50 mL. Pipet 4 mL of this solution, add the dissolution medium to make ex-actly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01>, and determine the peak areas, AT and AS, of cefdinir in each solution. Dissolution rate (z) with respect to the labeled amount of cefdinir (C14H13N5O5S2) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount [mg (potency)] of Cefdinir RS taken MT: Amount (g) of Cefdinir Fine Granules taken C: Labeled amount [mg (potency)] of cefdinir (C14H13N5O5S2) in 1 g Operating conditions— Amount [mg (potency)] of cefdinir (C ＝ MS × AT/AS × 5 14H13N5O5S2) Proceed as directed in the operating conditions in the Assay under Cefdinir. System suitability— MS: Amount [mg (potency)] of Cefdinir RS taken
Powder, if necessary, and weigh accurately an amount of Cefdinir Fine Granules, equivalent to about 0.1 g (potency) of Cefdinir, add 70 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30 minutes, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL. Centrifuge at 3000 revolutions per minute for 10 minutes, pipet 4 mL of the supernatant liquid, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefdinir RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefdinir. under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefditoren Pivoxil RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (4) Determine the 1H spectrum of a solution of Cefdito-ren Pivoxil in deuterated chloroform for nuclear magnetic resonance spectroscopy (1 in 50), using tetramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits singlet signals A, B and C, at around d 1.1 ppm, at around d 2.4 ppm and at around d 4.0 ppm, doublet signals D and E, at around d 6.4 ppm and at around d 6.7 ppm, and a singlet signal F at Amount [mg (potency)] of cefdinir (C ＝ MS × AT/AS × 5 14H13N5O5S2) around d 8.6 ppm. The ratio of integrated intensity of each signal A:B:C:D:E:F, is about 9:3:3:1:1:1. MS: Amount [mg (potency)] of Cefdinir RS taken Absorbance <2.24> E 1z 1 cm anol, 2500 mL). (231 nm): 340 – 360 (50 mg, meth-
To an amount of powdered Cefditoren Pivoxil Fine Granules, equivalent to 0.1 g (potency) of Cef-ditoren Pivoxil, add 10 mL of acetonitrile, shake vigorously, and filter. To 1 mL of the filtrate add acetonitrile to make 50 mL. To 1 mL of this solution add acetonitrile to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 230 nm and 234 nm.
Related substances—Being specified separately when the drug is granted approval based on the Law. Loss on drying <2.41> Not more than 4.5z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Uniformity of dosage units <6.02> The granules in single-dose packages meet the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 15 minutes of Cefditoren Pivoxil Fine Granules is not less than 80z. Start the test with an accurately weighed amount of Cef-ditoren Pivoxil Fine Granules, equivalent to about 0.1 g (po-tency) of Cefditoren Pivoxil, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 20 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Cefdito-ren Pivoxil RS, equivalent to about 22 mg (potency), dis-solve in 20 mL of diluted acetonitrile (3 in 4), and add the dissolution medium to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 272 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using water as the control. Dissolution rate (z) with respect to the labeled amount of cefditoren pivoxil (C25H28N6O7S3) ＝ MS/MT × AT/AS × 1/C × 450 MS: Amount [mg(potency)] of Cefditoren Pivoxil RS taken MT: Amount (g) of Cefditoren Pivoxil Fine Granules taken C: Labeled amount [mg(potency)] of cefditoren pivoxil (C25H28N6O7S3) in 1 g
Conduct this procedure using light-resistant vessels. Weigh accurately an amount of powdered Cefditoren Pivoxil Fine Granules, equivalent to about 40 mg (potency) of Cef-ditoren Pivoxil, add 70 mL of diluted acetonitrile (3 in 4), and shake vigorously. To this solution add exactly 10 mL of the internal standard solution, then add acetonitrile to make 100 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Cefditoren Pivoxil RS, equivalent to about 20 mg (potency), dissolve in 20 mL of acetonitrile, add exactly 5 mL of the internal standard solution, then add acetonitrile to make 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Cefditoren Pivoxil. Amount [mg (potency)] of cefditoren (C19H18N6O5S3) ＝ MS × QT/QS × 2 MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in acetonitrile (1 in 200).
Cefditoren Pivoxil Tablets contain not less than 90.0z and not more than 110.0z of the labeled potency of cefditoren (C19H18N6O5S3: 506.58). Method of preparation Prepare as directed under Tablets, with Cefditoren Pivoxil. Identification To an amount of powdered Cefditoren Pivoxil Tablets, equivalent to 35 mg (potency) of Cefditoren Pivoxil, add 100 mL of methanol, shake, and filter. To 5 mL of the filtrate add methanol to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 229 nm and 233 nm. Purity Related substances—Being specified separately when the drug is granted approval based on the Law. Loss on drying <2.41> Not more than 4.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Cefditoren Pivoxil Tablets add 12.5 mL of the 1st fluid for disintegration test, shake vigorously, add about 25 mL of acetonitrile, shake again, and add acetonitrile to make exactly 50 mL. Pipet V mL of this solution, equivalent to about 20 mg (potency) of Cefditoren Pivoxil, add exactly ‌JP XVIII Official Monographs / Cefepime Dihydrochloride Hydrate 669 5 mL of the internal standard solution, then add diluted acetonitrile (3 in 4) to make 50 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Cefditoren Pivoxil RS, equivalent to about 20 mg (potency), dissolve in 20 mL of acetonitrile, add exactly 5 mL of the internal standard solution, then add acetonitrile to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefditoren Pivoxil. Amount [mg (potency)] of cefditoren (C19H18N6O5S3) ＝ MS × QT/QS × 50/V MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken Internal standard solution—A solution of propyl para-hydroxybezoate in acetonitrile (1 in 200). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 20 minutes of Cefditoren Pivoxil Tablets is not less than 85z. Start the test with 1 tablet of Cefditoren Pivoxil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg (potency) of Cefditoren Pivoxil, and use this solution as the sample solution.
To an amount of powdered Cefditoren Pivoxil Tablets, equivalent to 35 mg (potency) of Cefditoren Pivoxil, add 100 mL of methanol, shake, and filter. To 5 mL of the filtrate add methanol to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 229 nm and 233 nm.
Related substances—Being specified separately when the drug is granted approval based on the Law. Loss on drying <2.41> Not more than 4.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Cefditoren Pivoxil Tablets add 12.5 mL of the 1st fluid for disintegration test, shake vigorously, add about 25 mL of acetonitrile, shake again, and add acetonitrile to make exactly 50 mL. Pipet V mL of this solution, equivalent to about 20 mg (potency) of Cefditoren Pivoxil, add exactly ‌JP XVIII Official Monographs / Cefepime Dihydrochloride Hydrate 669 5 mL of the internal standard solution, then add diluted acetonitrile (3 in 4) to make 50 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Cefditoren Pivoxil RS, equivalent to about 20 mg (potency), dissolve in 20 mL of acetonitrile, add exactly 5 mL of the internal standard solution, then add acetonitrile to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefditoren Pivoxil. Amount [mg (potency)] of cefditoren (C19H18N6O5S3) ＝ MS × QT/QS × 50/V MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken Internal standard solution—A solution of propyl para-hydroxybezoate in acetonitrile (1 in 200). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolution rate in 20 minutes of Cefditoren Pivoxil Tablets is not less than 85z. Start the test with 1 tablet of Cefditoren Pivoxil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg (potency) of Cefditoren Pivoxil, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefditoren Pivoxil RS, equivalent to about 22 mg (potency), dissolve in 20 mL of diluted aceto-nitrile (3 in 4), then add the dissolution medium to make ex-actly 200 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 272 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the control. Dissolution rate (z) with respect to the labeled amount of cefditoren pivoxil (C25H28N6O7S3) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount [mg (potency)] of Cefditoren Pivoxil RS taken C: Labeled amount [mg (potency)] of cefditoren pivoxil (C25H28N6O7S3) in 1 tablet
Conduct this procedure using light-resistant vessels. To an amount of Cefditoren Pivoxil Tablets, equivalent to 0.5 g (potency) of Cefditoren Pivoxil, add 63 mL of the 1st fluid for disintegration test, shake vigorously, add about 125 mL of acetonitrile, shake again, and add acetonitrile to make exactly 250 mL. Pipet 10 mL of this solution, add ex-actly 5 mL of the internal standard solution, then add diluted acetonitrile (3 in 4) to make 50 mL, filter, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Cefditoren Pivoxil RS, equivalent to about 20 mg (potency), dissolve in 20 mL of acetonitrile, add exactly 5 mL of the internal standard solution, then add acetonitrile to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefditoren Pivoxil. droxybenzoate in acetonitrile (1 in 200).
Cefepime Dihydrochloride for Injection is a prepa-ration for injection, which is dissolved before use. It contains not less than 95.
Cefepime Dihydrochloride for Injection occurs as a white to pale yellow powder.
(1) Dissolve 40 mg of Cefepime Dihydro-chloride in 2 mL of water, add 1 mL of a solution of hydrox-ylammonium chloride (1 in 10) and 2 mL of sodium hydrox-ide TS, allow to stand for 5 minutes, then add 3 mL of 1 mol/L hydrochloric acid TS and 3 drops of iron (III) chlo-ride TS: a red-brown color develops. (2) Determine the absorption spectrum of a solution of Cefepime Dihydrochloride for Injection (1 in 12,500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 233 nm and 237 nm and between 255 nm and 259 nm. pH <2.54> The pH of a solution obtained by dissolving an amount of Cefepime Dihydrochloride for Injection, equiva-lent to 0.5 g (potency) of Cefepime Dihydrochloride Hy-drate, in 5 mL of water is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve an amount of Cefepime Dihydrochloride for Injection, equiva-lent to 0.5 g (potency) of Cefepime Dihydrochloride Hy-drate, in 5 mL of water: the solution is clear and colorless or light yellow. The color is not darker than Matching Fluid I. N-Methylpyrrolidine—Weigh accurately an amount of Cefepime Dihydrochloride for Injection, equivalent to about 0.2 g (potency) of Cefepime Dihydrochloride Hydrate, dissolve in diluted nitric acid (2 in 625) to make exactly 20 mL, and use this solution as the sample solution. Separately, transfer 30 mL of water into a 100-mL volumetric flask, weigh accurately the mass of the flask, add about 0.125 g of N-methylpyrrolidine, then weigh accurately the mass, and add water to make exactly 100 mL. Pipet 4 mL of this solu-tion, add diluted nitric acid (2 in 3125) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas of N-methylpyrrolidine, AT and AS, by the automatic integration method within 20 minutes after the sample solution is prepared. Calculate the amount of N-methylpyrrolidine per mg (potency) of Cefepime Dihydro-chloride for Injection by the following formula: not more than 1.0z. Amount (z) of N-methylpyrrolidine ＝ (MS × f )/MT × AT/AS × 1/125 MS: Amount (mg) of N-methylpyrrolidine taken MT: Amount [mg (potency)] of Cefepime Dihydrochloride for Injection taken f: Purity (z) of N-methylpyrrolidine Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Cefepime Dihydrochloride Hydrate. System suitability— Proceed as directed in the system suitability in the Purity under Cefepime Dihydrochloride Hydrate. Water <2.48> Not more than 4.0z (Weigh accurately about 50 mg of Cefepime Dihydrochloride for Injection, dissolve in exactly 2 mL of methanol for water determination, and perform the test with exactly 0.5 mL of this solution: coulo-metric titration). Bacterial endotoxins <4.01> Less than 0.06 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Cefepime Dihydrochloride for Injection. Weigh ac-curately an amount of the content, equivalent to about 60 mg (potency) of Cefepime Dihydrochloride Hydrate, dis-solve in the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu- ‌672 rately an amount of Cefepime Dihydrochloride RS, equiva-lent to about 60 mg (potency), dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefepime Dihydrochloride Hydrate. Amount [ mg (potency)] of cefepime (C19H24N6O5S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefepime Dihydrochloride RS taken
Cefixime Capsules contain not less than 90.0z and not more than 105.0z of the labeled potency of cefixime (C16H15N5O7S2: 453.45). Method of preparation Prepare as directed under Cap-sules, with Cefixime Hydrate. Identification Take out the contents of Cefixime Capsules, solution, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the so-lution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Confirm that the peak area of cefixime obtained with 10 mL of this so-lution is equivalent to 7 to 13z of that with 10 mL of the so-lution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of cefixime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cefixime is not more than 2.0z. Water <2.48> Not more than 12.0z (0.1 g of the contents, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the contents of 1 capsule of Cefixime Capsules, and to the contents and the capsule shells add 7V/10 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30 minutes, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly V mL so that each mL contains about 1 mg (potency) of Cefixime Hydrate.
Take out the contents of Cefixime Capsules, solution, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the so-lution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Confirm that the peak area of cefixime obtained with 10 mL of this so-lution is equivalent to 7 to 13z of that with 10 mL of the so-lution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of cefixime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cefixime is not more than 2.0z. Water <2.48> Not more than 12.0z (0.1 g of the contents, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the contents of 1 capsule of Cefixime Capsules, and to the contents and the capsule shells add 7V/10 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30 minutes, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly V mL so that each mL contains about 1 mg (potency) of Cefixime Hydrate. Centrifuge this solu-tion, pipet 10 mL of the supernatant liquid, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefixime RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefixime Hydrate. to a quantity of the contents of Cefixime Capsules, equiva-lent to 70 mg (potency) of Cefixime Hydrate, add 100 mL of mol/L phosphate buffer solution (pH 7.0), shake for 30 Amount [mg (potency)] of cefixime (C ＝ MS × AT/AS × V/20 16H15N5O7S2) minutes, and filter. To 1 mL of the filtrate add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 50 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 286 nm and 290 nm.
Related substances—Take out the contents of Cefixime Capsules, to a quantity of the contents of Cefixime Capsules, equivalent to 0.1 g (potency) of Cefixime Hydrate, add 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0), shake for 30 minutes, filter, and use the filtrate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area from the sample solution by the automatic integration method, and calculate the amount of them by the area per-centage method: the amount of each peak other than cefixime is not more than 1.0z, and the total amount of the peaks other than cefixime is not more than 2.5z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Cefixime Hydrate. Time span for measurement: Proceed as directed in the operating conditions in the Purity under Cefixime Hydrate. System suitability— Test for required detectability: Pipet 1 mL of the sample MS: Amount [mg (potency)] of Cefixime RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of disodium hydrogen phosphate-citric acid buffer solution (pH 7.5) as the dissolution medium, the dissolution rates in 60 minutes of 50-mg (potency) capsule and in 90 minutes of 100-mg (potency) capsule are not less than 80z, respectively. Start the test with 1 capsule of Cefixime Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg (potency) of Cefixime Hydrate, and use this solution as the sample solution. Sepa-rately, weigh accurately an amount of Cefixime RS, equiva-lent to about 28 mg (potency), and dissolve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solu-tion, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefixime in each solution. Dissolution rate (z) with respect to the labeled amount of cefixime (C16H15N5O7S2) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount [mg (potency)] of Cefixime RS taken C: Labeled amount [mg (potency)] of Cefixime Hydrate in 1 capsule ‌Operating conditions— Proceed as directed in the operating conditions in the Assay under Cefixime Hydrate. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefixime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefixime is not more than 2.0z.
Take out the contents of not less than 20 Cefixime Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equiva-lent to about 0.1 g (potency) of Cefixime Hydrate, add 70 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and shake for 30 minutes, add 0.1 mol/L phosphate buffer solu-tion (pH 7.0) to make exactly 100 mL. Centrifuge this solu-tion, pipet 10 mL of the supernatant liquid, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefixime RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay under Cefixime Hydrate. Amount [mg (potency)] of cefixime (C16H15N5O7S2) ＝ MS × AT/AS × 5 MS: Amount [mg (potency)] of Cefixime RS taken
注射用セフメタゾールナトリウム
Cefmetazole Sodium for Injection is a preparation for injection which is dissolved before use. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Take 10 containers of Cefmetazole Sodium for In-jection, dissolve the contents of each in the mobile phase, rinse each of the containers with the mobile phase, combine the rinse with the respective previous solution, and add the mobile phase to make exactly 500 mL. Take exactly a volume of this solution equivalent to about 0.2 g (potency) of Cefmetazole Sodium, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefmetazole RS, equivalent to about 50 mg (potency), and dissolve in the mobile phase to make exactly 25 mL. Pipet 1 mL of this solution, add exactly 10 mL of the inter-nal standard solution, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefmetazole Sodium. It contains not less than 90.
Cefmetazole Sodium for Injection is a white to light yellow powder or masses. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Cefmetazole Sodium for Injection (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cef-metazole Sodium for Injection as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Take an amount of Cefmetazole Sodium for In-jection equivalent to 1.0 g (potency) of Cefmetazole Sodium, and dissolve in 10 mL of water: the pH of the solution is 4.2 to 6.2.
(1) Clarity and color of solution—Dissolve an amount of Cefmetazole Sodium for Injection, equivalent to 1.0 g (potency) of Cefmetazole Sodium, in 10 mL of water: the solution is clear and the color is not darker than the fol-lowing control solution. Control solution: Pipet 5 mL of Iron (III) Chloride CS and 0.5 mL of Cobalt (II) Chloride CS, and add water to make exactly 50 mL. Pipet 15 mL of this solution, and add water to make exactly 20 mL. (2) Related substances—Proceed as directed in the Purity (4) under Cefmetazole Sodium. Bacterial endotoxins <4.01> Less than 0.06 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign particulate matter <6.06> Perform the test accord-ing to Method 2: it meets the requirement. MS: Amount [mg (potency)] of Cefmetazole RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (1 in 10,000).
Take 10 containers of Cefmetazole Sodium for In-jection, dissolve the contents of each in the mobile phase, rinse each of the containers with the mobile phase, combine the rinse with the respective previous solution, and add the mobile phase to make exactly 500 mL. Take exactly a volume of this solution equivalent to about 0.2 g (potency) of Cefmetazole Sodium, and add the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefmetazole RS, equivalent to about 50 mg (potency), and dissolve in the mobile phase to make exactly 25 mL. Pipet 1 mL of this solution, add exactly 10 mL of the inter-nal standard solution, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefmetazole Sodium. It contains not less than 90.0z and not more than 110.0z of the labeled potency of cefmetazole Amount [mg (potency)] of cefmetazole (C 15H17N7O5S3) (C15H17N7O5S3: 471.53). ＝ MS × QT/QS × 4 Method of preparation Prepare as directed under Injec-tions, with Cefmetazole Sodium. Description Cefmetazole Sodium for Injection is a white to light yellow powder or masses. It is hygroscopic. Identification (1) Determine the absorption spectrum of a solution of Cefmetazole Sodium for Injection (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Cef-metazole Sodium for Injection as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. pH <2.54> Take an amount of Cefmetazole Sodium for In-jection equivalent to 1.0 g (potency) of Cefmetazole Sodium, and dissolve in 10 mL of water: the pH of the solution is 4.2 to 6.2. Purity (1) Clarity and color of solution—Dissolve an amount of Cefmetazole Sodium for Injection, equivalent to 1.0 g (potency) of Cefmetazole Sodium, in 10 mL of water: the solution is clear and the color is not darker than the fol-lowing control solution. Control solution: Pipet 5 mL of Iron (III) Chloride CS and 0.5 mL of Cobalt (II) Chloride CS, and add water to make exactly 50 mL. Pipet 15 mL of this solution, and add water to make exactly 20 mL. (2) Related substances—Proceed as directed in the Purity (4) under Cefmetazole Sodium. Bacterial endotoxins <4.01> Less than 0.06 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign particulate matter <6.06> Perform the test accord-ing to Method 2: it meets the requirement. MS: Amount [mg (potency)] of Cefmetazole RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (1 in 10,000).
Cefoperazone Sodium and Sulbactam Sodium for Injection
注射用セフォペラゾンナトリウム･スルバクタムナトリウム
Cefoperazone Sodium and Sulbactam Sodium for Injection is a preparation for injection which is dis-solved before use. It contains not less than 90.
Cefoperazone Sodium and Sulbactam Sodium for Injection occurs as white to pale yellowish white, masses or powder.
(1) The retention times of cefoperazone in the chromatogram obtained from the sample solution and the standard solution in the Assay are the same, and the peak area of cefoperazone obtained from the sample solu-tion in the Assay is 0.8 to 1.1 times the peak area of cefoper-azone obtained by the test performed with 10 mL of the sam-ple solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following condi-tions. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. (2) The retention times of sulbactam in the chromato-gram obtained from the sample solution and the standard solution in the Assay are the same, and the peak area of sul-bactam obtained from the sample solution in the Assay is 1.4 to 1.9 times the peak area of sulbactam obtained by the test performed with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. pH <2.54> The pH of a solution prepared by dissolving an amount of Cefoperazone Sodium and Sulbactam Sodium for Injection, equivalent to 1.0 g (potency) of Cefoperazone So-dium, in 20 mL of water is 4.5 to 6.5.
(1) Clarity and color of solution—A solution of an amount of Cefoperazone Sodium and Sulbactam Sodium for Injection, equivalent to 0.5 g (potency) of Cefoperazone So-dium, in 10 mL of water is clear. Perform the test with this solution as directed under Ultraviolet Spectrophotometry <2.24>: the absorbance at 425 nm is not more than 0.10. Related substances—Weigh accurately an amount of Cefoperazone Sodium and Sulbactam Sodium for Injection, equivalent to 0.1 g (potency) of Cefoperazone Sodium, dis-solve in the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sam-ple solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution (1). Weigh ac-curately about 40 mg of sulbactam sodium for sulbactam penicillamine, dissolve in 2 mL of water, add 0.5 mL of so-dium hydroxide TS, allow to stand at room temperature for 10 minutes, then add 0.5 mL of 1 mol/L hydrochloric acid TS, and add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with exactly 10 mL each of the sample solu-tion and the standard solutions (1) and (2) as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having a relative retention time of about 0.3 (related substance I) to cefopera-zone, obtained from the sample solution is not larger than 1.75 times the peak area of cefoperazone from the standard solution (1), the area of the peak, having a relative retention time of about 0.4 (related substance III) and about 1.3 (related substance II), obtained from the sample solution is not larger than 1/2 times the peak area of cefoperazone from the standard solution (1). When determine the peak areas, AT and AS, of sulbactam penicillamine with the sample solu-tion and the standard solution (2), and calculate the amount of sulbactam penicillamine by the following equation, it is not more than 1.0z. For the area of the peak of related sub-stance III, multiply the correction factor 0.4. Amount of sulbactam penicillamine (z) ＝ MS/MT × AT/AS × 5 MS: Amount (mg) of sulbactam sodium for sulbactam penicillamine taken MT: Amount (mg) of Cefoperazone Sodium and Sulbac-tam Sodium for Injection taken Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: To 1 mL of the standard solution (1) add 1 mL of the standard solution (2). When the procedure is run with 10 mL of this solution under the above operating conditions, sulbactam penicillamine, sulbactam and cefoper-azone are eluted in this order with the resolutions between the peaks, sulbactam penicillamine and sulbactam, and sul-bactam and cefoperazone, being not less than 4 and not less than 5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution (2) under the above operating conditions, the relative standard deviation of the peak area of sulbactam penicillamine is not more than 2.0z. Water <2.48> Not more than 1.0z (1 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.060 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement ‌684 of the Mass variation test (T: 105.0z). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the content of not less than 5 Cefoperazone Sodium and Sulbactam Sodium for In-jection. Weigh accurately a portion of the content, equiva-lent to about 50 mg (potency) of Cefoperazone Sodium, dis-solve in suitable amount of the mobile phase, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg (potency) each of Sulbactam RS and Cefoperazone RS, dissolve in suitable amount of the mobile phase, add exactly 5 mL of the inter-nal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QTa and QTb of the peak areas of sulbactam and cefopera-zone to that of the internal standard obtained from the sam-ple solution, and the ratios, QSa and QSb of the peak areas of sulbactam and cefoperazone to that of the internal standard obtained from the standard solution. Amount [mg (potency)] of sulbactam (C8H11NO5S) ＝ MS1 × QTa/QSa Cefotaxime Sodium
注射用セフォペラゾンナトリウム
Cefoperazone Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 93.
Cefoperazone Sodium for Injection occurs as a white to yellowish white, crystalline powder or masses.
Determine the absorption spectrum of a so-lution of Cefoperazone Sodium for Injection (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 226 nm and 230 nm, and between 263 nm and 267 nm. pH <2.54> The pH of a solution prepared by dissolving an amount of Cefoperazone Sodium for Injection, equivalent to 1.0 g (potency) of Cefoperazone Sodium, in 4 mL of water is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve an amount of Cefoperazone Sodium for Injection, equivalent to 1.0 g (potency) of Cefoperazone Sodium, in 10 mL of water: the solution is clear, and its absorbance at 400 nm, determined as directed under Ultraviolet-visible Spectropho-tometry <2.24>, is not more than 0.22. (2) Related substances—Dissolve an amount of Cefo-perazone Sodium for Injection, equivalent to 0.1 g (potency) of Cefoperazone Sodium, in 100 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add water to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of related substance I, having the relative retention time of about 0.8 to cefoperazone, obtained from the sample solution is not larger than 2.5 times the peak area of cefoperazone from the standard solution, the peak area of related substance II, hav-ing the relative retention time of about 1.7, from the sample solution is not larger than 3/4 times the peak area of cefoperazone from the standard solution. Furthermore, the total area of the peaks other than cefoperazone from the sample solution is not larger than 3.5 times the peak area of cefoperazone from the standard solution. For the peak areas of the related substances I and II, multiply their correction factors, 0.90 and 0.75, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Cefoperazone Sodium. Time span of measurement: About 3 times as long as the retention time of cefoperazone, beginning after the solvent peak. System suitability— Proceed as directed in the system suitability in the Purity (4) under Cefoperazone Sodium. Water <2.48> Not more than 1.0z (3 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Cefoperazone Sodium for Injection. Weigh accu-rately a portion of the content, equivalent to about 0.1 g (po-tency) of Cefoperazone Sodium, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add ex-actly 5 mL of the internal standard solution, and use this so-lution as the sample solution. Then, proceed as directed in the Assay under Cefoperazone Sodium. Amount [mg (potency)] of cefoperazone (C25H27N9O8S2) ＝ MS × QT/QS × 5 MS: Amount [mg (potency)] of Cefoperazone RS taken Internal standard solution—A solution of acetanilide in a mixture of water and acetonitrile (43:7) (3 in 8000).
Containers—Hermetic containers. Storage—In a cold place.
Cefotiam Hydrochloride for Injection is a prepara-tion for injection which is dissolved before use. It contains not less than 90.
Cefotiam Hydrochloride for Injection occurs as a white to light yellow powder.
(1) Determine the absorption spectrum of a solution of Cefotiam Hydrochloride for Injection (1 in 50,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 257 nm and 261 nm. (2) Determine the 1H spectrum of a solution of Cefotiam Hydrochloride for Injection in heavy water for nuclear mag-netic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using so-dium 3-trimethylsilylpropanesulfonate for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a singlet signal A between d 2.7 ppm and d 3.0 ppm, and a singlet signal B at around d 6.5 ppm. The ratio of the integrated intensity of each signal, A:B, is about 6:1. pH <2.54> The pH of a solution prepared by dissolving an amount of Cefotiam Hydrochloride for Injection, equivalent to 0.5 g (potency), in 5 mL of water is between 5.7 and 7.2.
Clarity and color of solution—Dissolve an amount of Cefotiam Hydrochloride for Injection, equivalent to 1.0 g (potency) of Cefotiam Hydrochloride, in 10 mL of water: the solution is clear, and the absorbance of this solution, determined at 450 nm 10 minutes after dissolving as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.20. Loss on drying <2.41> Not more than 6.0z (0.5 g, in vacu-um, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.125 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the contents of not less than 10 ‌JP XVIII Official Monographs / Cefozopran Hydrochloride 691 Cefotiam Hydrochloride for Injection. Weigh accurately an amount of the content, equivalent to about 50 mg (potency) of Cefotiam Hydrochloride, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg (potency) of Cefotiam Hydrochloride RS, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Cefotiam Hydrochloride. Amount [ mg (potency)] of cefotiam (C18H23N9O4S3) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefotiam Hydrochloride RS taken
Cefozopran Hydrochloride for Injection is a prepa-ration for injection which is dissolved before use. It contains not less than 90.
Cefozopran Hydrochloride for Injection oc-curs as a white to light yellow, powder or masses.
(1) Determine the absorption spectrum of a solution of Cefozopran Hydrochloride for Injection (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 236 nm and 241 nm. (2) To 50 mg of Cefozopran Hydrochloride for Injection add 0.8 mL of deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy, and filter after shaking, and determine the 1H spectrum of the filtrate as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a singlet signal A at around d 3.9 ppm, a doublet signal B at around d 5.0 ppm, and a quartet signal C at around d 8.0 ppm. The ratio of the integrated intensity of each signal, A:B:C, is about 3:1:1. pH <2.54> Dissolve an amount of Cefozopran Hydrochlo-ride for Injection, equivalent to 0.5 g (potency) of Cefozopran Hydrochloride, in 5 mL of water: the pH of this solution is between 7.5 and 9.0.
(1) Clarity and color of solution—Dissolve an amount of Cefozopran Hydrochloride for Injection, equiva-lent to 1 g (potency) of Cefozopran Hydrochloride, in 10 mL of water: the solution is clear and has no more color than Matching Fluid N. (2) Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po- MS: Amount [mg (potency)] of Cefozopran Hydrochlo-ride RS taken Internal standard solution—A solution 2,4-dihydroxyben-zoic acid in the mobile phase (1 in 1250).
Weigh accurately the mass of the contents of not less than 10 Cefozopran Hydrochloride for Injection. Weigh ac-curately an amount of the contents, equivalent to about 0.5 g (potency) of Cefozopran Hydrochloride, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add ex-actly 10 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Cefozopran Hydrochloride RS, equivalent to about 50 mg (potency), and dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the standard solution. Pro-ceed as directed in the Assay under Cefozopran Hydrochlo-ride. than 115.0z of the labeled potency of cefozopran Amount [mg (potency)] of cefozopran (C H N O S ) (C19H17N9O5S2: 515.53). ＝ MS × QT/QS × 10 19 17 9 5 2 Method of Preparation Prepare as directed under the In-jections, with Cefozopran Hydrochloride. Description Cefozopran Hydrochloride for Injection oc-curs as a white to light yellow, powder or masses. Identification (1) Determine the absorption spectrum of a solution of Cefozopran Hydrochloride for Injection (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 236 nm and 241 nm. (2) To 50 mg of Cefozopran Hydrochloride for Injection add 0.8 mL of deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy, and filter after shaking, and determine the 1H spectrum of the filtrate as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a singlet signal A at around d 3.9 ppm, a doublet signal B at around d 5.0 ppm, and a quartet signal C at around d 8.0 ppm. The ratio of the integrated intensity of each signal, A:B:C, is about 3:1:1. pH <2.54> Dissolve an amount of Cefozopran Hydrochlo-ride for Injection, equivalent to 0.5 g (potency) of Cefozopran Hydrochloride, in 5 mL of water: the pH of this solution is between 7.5 and 9.0. Purity (1) Clarity and color of solution—Dissolve an amount of Cefozopran Hydrochloride for Injection, equiva-lent to 1 g (potency) of Cefozopran Hydrochloride, in 10 mL of water: the solution is clear and has no more color than Matching Fluid N. (2) Related substances—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 2.5z (0.5 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Bacterial endotoxins <4.01> Less than 0.05 EU/mg (po- MS: Amount [mg (potency)] of Cefozopran Hydrochlo-ride RS taken Internal standard solution—A solution 2,4-dihydroxyben-zoic acid in the mobile phase (1 in 1250).
シロップ用セフポドキシム プロキセチル
Cefpodoxime Proxetil for Syrup is a preparation for syrups which is suspended before use. It contains not less than 93.
To an amount of Cefpodoxime Proxetil for Syrup, equivalent to 15 mg (potency) of Cefpodoxime Prox-etil, add 10 mL of 0.1 mol/L phosphate buffer solution (pH 8.0), treat with ultrasonic waves for 5 minutes while occa-sional shaking. Then, add 20 mL of ethyl acetate, shake for 5 minutes, and centrifuge. Take 3 mL of the supernatant liq-uid, evaporate the ethyl acetate by warming at 409C under reduced pressure. Dissolve the residue in acetonitrile to make 200 mL, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Cefpodoxime Proxetil for Syrup in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Cefpodoxime Proxetil for Syrup add exactly 30 mL of the internal standard solu-tion, treat with ultrasonic waves for 10 minutes while occa-sional shaking, and centrifuge. Take 3 mL of the superna-tant liquid, add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 50 mg (po-tency), dissolve in a suitable amount of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 15 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the standard solution. ‌JP XVIII Official Monographs / Cefpodoxime Proxetil Tablets 697 Then, proceed as directed in the Assay under Cefpodoxime Proxetil.
Weigh accurately an amount of Cefpodoxime Prox-etil and Cefpodoxime Proxetil RS, equivalent to about 60 mg (potency), dissolve in 80 mL of acetonitrile, add exactly 4 mL of the internal standard solution, add acetonitrile to make 100 mL, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT1, QS1, QT2 and QS2, of the areas of the two peaks of the isomers of cefpodoxime proxetil to the peak area of the internal standard. Amount [ mg (potency)] of cefpodoxime (C15H17N5O6S2) ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 1000 MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.3 g of ethyl parahy-droxybenzoate in a solution of citric acid in acetonitrile (1 in 2000) to make 100 mL. Method of preparation Prepare as directed under Syrups, with Cefpodoxime Proxetil. Identification To an amount of Cefpodoxime Proxetil for Syrup, equivalent to 15 mg (potency) of Cefpodoxime Prox-etil, add 10 mL of 0.1 mol/L phosphate buffer solution (pH 8.0), treat with ultrasonic waves for 5 minutes while occa-sional shaking. Then, add 20 mL of ethyl acetate, shake for 5 minutes, and centrifuge. Take 3 mL of the supernatant liq-uid, evaporate the ethyl acetate by warming at 409C under reduced pressure. Dissolve the residue in acetonitrile to make 200 mL, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Cefpodoxime Proxetil for Syrup in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Cefpodoxime Proxetil for Syrup add exactly 30 mL of the internal standard solu-tion, treat with ultrasonic waves for 10 minutes while occa-sional shaking, and centrifuge. Take 3 mL of the superna-tant liquid, add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 50 mg (po-tency), dissolve in a suitable amount of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 15 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the standard solution. ‌JP XVIII Official Monographs / Cefpodoxime Proxetil Tablets 697 Then, proceed as directed in the Assay under Cefpodoxime Proxetil. Assay Weigh accurately an amount of powdered Cef-podoxime Proxetil for Syrup, equivalent to about 0.1 g Amount [mg (potency)] of cefpodoxime (C ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 2 15H17N5O6S2) (potency) of Cefpodoxime Proxetil, add exactly 30 mL of the internal standard solution, treat with ultrasonic waves for 10 minutes while occasional shaking, and centrifuge. MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.2 g of ethyl parahy-droxybenzoate in a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 300 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of Cefpodoxime Proxetil for Syrup is not less than 85z. Take 3 mL of the supernatant liquid, add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 50 mg (potency), dissolve in a suitable amount of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 15 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefpodoxime Proxetil. Start the test with an accurately weighed amount of Cef-podoxime Proxetil for Syrup, equivalent to about 50 mg (po-tency) of Cefpodoxime Proxetil, withdraw not less than 20 Amount [mg (potency)] of cefpodoxime (C ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 2 15H17N5O6S2) mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add a so-lution of citric acid monohydrate in the mobile phase (1 in 2000) to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 22 mg (po-tency), dissolve in a solution of citric acid monohydrate in the mobile phase (1 in 2000) to make exactly 100 mL. Pipet 5 mL of this solution, add a solution of citric acid monohy-drate in the mobile phase (1 in 2000) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas, ATa and ASa, of the one peak which appears at the retention time of about 24 minutes among the two peaks ob-tainable from cefpodoxime proxetil, and the areas, ATb and ASb, of the peak which appears at the retention time of about 30 minutes, in each solution. Dissolution rate (z) with respect to the labeled amount of cefpodoxime proxetil (C21H27N5O9S2) ＝ MS/MT × (ATa ＋ ATb)/(ASa ＋ ASb) × 1/C × 225 MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken MT: Amount (g) of Cefpodoxime Proxetil for Syrup taken C: Labeled amount [mg (potency)] of cefpodoxime prox-etil (C21H27N5O9S2) in 1 g Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay under Cefpodoxime Proxetil. Flow rate: Adjust so that the retention time of the peak, which elutes faster among the two peaks obtained from cef-podoxime proxetil, is about 24 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the resolution between the two peaks obtained from cefpodoxime proxetil is not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the sum of the areas of the two peaks obtained from cefpodoxime prox-etil is not more than 2.0z. MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.2 g of ethyl parahy-droxybenzoate in a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 300 mL.
Cefpodoxime Proxetil Tablets contain not less than 93.0z and not more than 107.0z of the labeled po-tency of cefpodoxime (C15H17N5O6S2: 427.46). Method of preparation Prepare as directed under Tablets, with Cefpodoxime Proxetil. Identification Powder Cefpodoxime Proxetil Tablets. To a portion of the powder, equivalent to 65 mg (potency) of Cef-podoxime Proxetil, add 25 mL of acetonitrile, shake thor-oughly, and centrifuge. To 2 mL of the supernatant liquid add acetonitrile to make 50 mL. To 5 mL of this solution add acetonitrile to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cefpodoxime Proxetil Tablets, add exactly 20 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), agitate with the aid of ultrasonic waves for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet V mL of the subsequent filtrate, equivalent to 30 mg (potency) of Cefpodoxime Proxetil, add exactly 6 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 60 mg (potency), dissolve in 60 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 12 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the stand- ard solution. Then, proceed as directed in the Assay under Cefpodoxime Proxetil. ‌Amount [mg (potency)] of cefpodoxime (C15H17N5O6S2) ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 10/V MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.1 g of ethyl parahy-droxybenzoate in a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Cefpodoxime Proxetil Tablets is not less than 70z. Start the test with 1 tablet of Cefpodoxime Proxetil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a solution of citric acid monohy-drate in the mobile phase (1 in 2000) to make exactly V? mL so that each mL contains about 11 mg (potency) of Cef-podoxime Proxetil, and use this solution as the sample solu-tion.
Powder Cefpodoxime Proxetil Tablets. To a portion of the powder, equivalent to 65 mg (potency) of Cef-podoxime Proxetil, add 25 mL of acetonitrile, shake thor-oughly, and centrifuge. To 2 mL of the supernatant liquid add acetonitrile to make 50 mL. To 5 mL of this solution add acetonitrile to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cefpodoxime Proxetil Tablets, add exactly 20 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), agitate with the aid of ultrasonic waves for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet V mL of the subsequent filtrate, equivalent to 30 mg (potency) of Cefpodoxime Proxetil, add exactly 6 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 60 mg (potency), dissolve in 60 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 12 mL of the internal standard solution, then add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the stand- ard solution. Then, proceed as directed in the Assay under Cefpodoxime Proxetil. ‌Amount [mg (potency)] of cefpodoxime (C15H17N5O6S2) ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 10/V MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.1 g of ethyl parahy-droxybenzoate in a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Cefpodoxime Proxetil Tablets is not less than 70z. Start the test with 1 tablet of Cefpodoxime Proxetil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a solution of citric acid monohy-drate in the mobile phase (1 in 2000) to make exactly V? mL so that each mL contains about 11 mg (potency) of Cef-podoxime Proxetil, and use this solution as the sample solu-tion. Separately, weigh accurately an amount of Cef-podoxime Proxetil RS, equivalent to about 22 mg (potency), and dissolve in a solution of citric acid monohydrate in the mobile phase (1 in 2000) to make exactly 100 mL. Pipet 5 mL of this solution, add a solution of citric acid monohydrate in the mobile phase (1 in 2000) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas of separated two peaks, one has the retention time of about 24 minutes, ATa and ASa, and another one has the retention time of about 30 minutes, ATb and ASb, in each so-lution. Dissolution rate (z) with respect to the labeled amount of cefpodoxime proxetil (C21H27N5O9S2) ＝ MS × (ATa ＋ ATb)/(ASa ＋ ASb) × V?/V × 1/C × 45 MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken C: Labeled amount [mg (potency)] of cefpodoxime prox-etil (C21H27N5O9S2) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and methanol (11:9). Flow rate: Adjust so that the retention time of one of the two peaks that elutes firster is about 24 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, the resolution between the two peaks of cef-podoxime proxetil is not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the total area of the two peaks of cefpodoxime proxetil is not more than 2.0z.
Weigh accurately the mass of not less than 20 Cef-podoxime Proxetil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.3 g (potency) of Cefpodoxime Proxetil, add 80 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), agitate for 10 minutes with the aid of ultrasonic waves, and add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make exactly 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent fil-trate, add exactly 6 mL of the internal standard solution, then, add a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Cefpodoxime Proxetil RS, equivalent to about 60 mg (po-tency), dissolve in 60 mL of a mixture of water, acetonitrile and acetic acid (100) (99:99:2), add exactly 12 mL of the in-ternal standard solution, then add a mixture of water, aceto-nitrile and acetic acid (100) (99:99:2) to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefpodoxime Proxetil. Amount [mg (potency)] of cefpodoxime (C15H17N5O6S2) ＝ MS × (QT1 ＋ QT2)/(QS1 ＋ QS2) × 5 MS: Amount [mg (potency)] of Cefpodoxime Proxetil RS taken Internal standard solution—Dissolve 0.1 g of ethyl parahy-droxybenzoate in a mixture of water, acetonitrile and acetic acid (100) (99:99:2) to make 100 mL.
Cefroxadine for Syrup is a preparation for syrup, which is suspended before use. It contains not less than 93.
Powder Cefroxadine for Syrup, if necessary. To a portion of the powder, equivalent to 2 mg (potency) of Cefroxadine Hydrate, add 100 mL of 0.001 mol/L hydro-chloric acid TS, shake well, and filter. Determine the absorp-tion spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 267 nm and 271 nm. Water <2.48> Not more than 4.5z (0.1 g, volumetric titra-tion, direct titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Cefroxadine for Syrup in single-dose packages meet the requirement of the Content uniformity test. Take out the total contents of 1 package of Cefroxadine for Syrup, add 4V/5 mL of a mixture of dilute acetic acid and phosphoric acid (500:1), shake well for 15 minutes, add exactly 5 mL of the internal standard solution per 50 mg (po-tency) of Cefroxadine Hydrate, and add a mixture of dilute acetic acid and phosphoric acid (500:1) to make V mL so that each mL contains about 0.25 mg (potency) of Cefroxa-dine Hydrate. Filter this solution through a membrane filter with pore size of not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Cefroxadine RS, equivalent to about 50 mg (po-tency), dissolve in a mixture of dilute acetic acid and phos-phoric acid (500:1), add exactly 5 mL of the internal stand-ard solution, add a mixture of dilute acetic acid and phos-phoric acid (500:1) to make 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefroxadine Hydrate. Amount [mg (potency)] of cefroxadine (C16H19N3O5S) ＝ MS × QT/QS × V/200 MS: Amount [mg (potency)] of Cefroxadine RS taken Internal standard solution—Dissolve 1.6 g of vanillin in 5 mL of methanol, and add a mixture of dilute acetic acid and phosphoric acid (500:1) to make 100 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cefroxadine for Syrup is not less than 85z. Start the test with an accurately weighed amount of Cefroxadine for Syrup, equivalent to about 0.1 g (potency) of Cefroxadine Hydrate, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 5 mL of the first fil-trate, pipet 4 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 20 mL, and use this so-lution as the sample solution. Separately, weigh accurately an amount of Cefroxadine RS, equivalent to about 22 mg (potency), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add 10 mL of water, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 267 nm. Dissolution rate (z) with respect to the labeled amount of cefroxadine (C16H19N3O5S) ＝ MS/MT × AT/AS × 1/C × 450 MS: Amount [mg (potency)] of Cefroxadine RS taken MT: Amount (g) of Cefroxadine for Syrup taken C: Labeled amount [mg (potency)] of cefroxadine (C16H19N3O5S) in 1 g
Powder Cefroxadine for Syrup, if necessary, weigh accurately a portion of the powder, equivalent to about 50 mg (potency) of Cefroxadine Hydrate, add 160 mL of a mix-ture of dilute acetic acid and phosphoric acid (500:1), shake well for 15 minutes, add exactly 5 mL of the internal stand-ard solution, and add a mixture of dilute acetic acid and phosphoric acid (500:1) to make 200 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Sepa-rately, weigh accurately an amount of Cefroxadine RS, equivalent to about 50 mg (potency), dissolve in a mixture of dilute acetic acid and phosphoric acid (500:1), add exactly 5 mL of the internal standard solution, add a mixture of dilute acetic acid and phosphoric acid (500:1) to make 200 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Cefroxadine Hydrate. Amount [mg (potency)] of cefroxadine (C16H19N3O5S) ＝ MS × QT/QS MS: Amount [mg (potency)] of Cefroxadine RS taken Internal standard solution—Dissolve 1.6 g of vanillin in 5 mL of methanol, and add a mixture of dilute acetic acid and phosphoric acid (500:1) to make 100 mL.
Ceftazidime for Injection is a preparation for injec-tion which is dissolved before use. It contains not less than 93.
Ceftazidime for Injection is a white to pale yellow-white powder.
Determine the absorption spectrum of a so-lution of Ceftazidime for Injection (1 in 100,000) in phos-phate buffer solution (pH 6.0) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 255 nm and 259 nm. pH <2.54> Dissolve an amount of Ceftazidime for Injec-tion, equivalent to 1.0 g (potency) of Ceftazidime Hydrate, in 10 mL of water: the pH of this solution is 5.8 to 7.8.
Clarity and color of solution—Dissolve 5 g of diso-dium hydrogen phosphate and 1 g of potassium dihydrogen phosphate in water to make 100 mL. In 10 mL of this solu-tion dissolve an amount of Ceftazidime for Injection, equivalent to 1.0 g (potency) of Ceftazidime Hydrate: the so-lution is clear. Also, determine the absorption spectra of this solution as directed under Ultraviolet-visible Spectropho- tometry <2.24>: the absorbance at 420 nm is not more than 0.3. Loss on drying <2.41> Not more than 14.0z (0.1 g, in vacuum not exceeding 0.67 kPa, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.067 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filter method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Ceftazidime for Injection. Weigh accu-rately an amount of Ceftazidime Hydrate, equivalent to about 0.25 g (potency), and dissolve in 0.05 mol/L phos-phate buffer solution (pH 7.0) to make exactly 250 mL. Pipet 10 mL of this solution, add exactly 5 mL of the inter-nal standard solution, add more 0.05 mol/L phosphate buffer solution (pH 7.0) to make 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Ceftazidime RS, equivalent to about 25 mg (po-tency), and dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 25 mL. Pipet 10 mL of this solu-tion, add exactly 5 mL of the internal standard solution, then add 0.05 mol/L phosphate buffer solution (pH 7.0) to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Ceftazidime Hydrate. Amount [mg (potency)] of ceftazidime (C22H22N6O7S2) ＝ MS × QT/QS × 10 MS: Amount [mg(potency)] of Ceftazidime RS taken Internal standard solution—A solution of dimedon in 0.05 mol/L phosphate buffer solution (pH 7.0) (11 in 10,000).
Powder Cefteram Pivoxil Fine Granules. To a portion of the powder, equivalent to 0.1 g (potency) of Cefteram Pivoxil, add 20 mL of methanol, shake well, and filter. To 1 mL of the filtrate add 0.05 mol/L hydrochloric acid-methanol TS to make 500 mL, and determine the ab-sorption spectrum as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 262 nm and 266 nm.
Related substances—Powder Cefteram Pivoxil Fine Granules, if necessary. To a portion, equivalent to 0.1 g (po-tency) of Cefteram Pivoxil, add diluted acetonitrile (1 in 2) to make 100 mL, disperse the particle by sonicating, then filter, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.9 to cefteram pivoxil obtained from the sample solu-tion, is not larger than 1.75 times the peak area of cefteram pivoxil obtained from the standard solution, the area of the peak, having the relative retention time of about 0.1 from the sample solution, is not larger than 17/25 times the peak area of cefteram pivoxil from the standard solution, and the total area of the peaks other than cefteram pivoxil from the sample solution is not larger than 3.7 times the peak area of cefteram pivoxil from the standard solution. For the area of the peak, having the relative retention time of about 0.1 to cefteram pivoxil, multiply the correction factor, 0.74. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Cefteram Pivoxil. System suitability— Proceed as directed in the system suitability in the Purity (2) under Cefteram Pivoxil. Water <2.48> Not more than 0.3z (0.1 g (potency), coulo-metric titration). Uniformity of dosage units <6.02> The Granules in single-dose packages meet the requirement of the Mass variation test. Dissolution Being specified separately when the drug is granted approval based on the Law.
Powder Cefteram Pivoxil Fine Granules, if neces-sary. Weigh accurately an amount of the powder, equivalent to about 0.3 g (potency) of Cefteram Pivoxil, add exactly 30 mL of the internal standard solution and diluted acetonitrile (1 in 2) to make 300 mL. Disperse the particle by sonicating, then filter, and use the filtrate as the sample solution. Sepa-rately, weigh accurately an amount of Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to about 50 mg (poten-cy), dissolve in 20 mL of diluted acetonitrile (1 in 2), add ex- actly 5 mL of the internal standard solution and diluted acetonitrile (1 in 2) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefteram pivoxil to that of the internal standard. Amount [mg (potency)] of cefteram (C16H17N9O5S2) ＝ MS × QT/QS × 6 MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in diluted acetonitrile (1:2) (1 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Cefteram Pivoxil. System suitability— Proceed as directed in the system suitability in the Assay under Cefteram Pivoxil.
Cefteram Pivoxil Tablets contain not less than 90.0z and not more than 110.0z of the labeled potency of cefteram (C16H17N9O5S2: 479.49). Method of preparation Prepare as directed under Tablets, with Cefteram Pivoxil. Identification To a quantity of powdered Cefteram Pivoxil Tablets, equivalent to 0.1 g (potency) of Cefteram Pivoxil, add 20 mL of methanol, shake well, and filter. To 1 mL of the filtrate add 0.05 mol/L hydrochloric acid-methanol TS to make 500 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 262 nm and 266 nm. Purity Related substances—To a quantity of powdered Cefteram Pivoxil Tablets, equivalent to 0.1 g (potency) of Cefteram Pivoxil, add diluted acetonitrile (1 in 2) to make 100 mL. Disperse this solution by sonicating, filter, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.9 to cefteram pivoxil, obtained from the sample so-lution is not larger than 1.75 times the peak area of cefteram pivoxil from the standard solution, and the area of the peak, having the relative retention time of about 0.1 from the sam-ple solution is not larger than 17/25 times the peak area of cefteram pivoxil from the standard solution. Furthermore, the total area of the peaks other than cefteram pivoxil from the sample solution is not larger than 3.7 times the peak area of cefteram pivoxil from the standard solution. For the area of the peak, having the relative retention time of about 0.1 to cefteram pivoxil, multiply the correction factor, 0.74. ‌JP XVIII Official Monographs / Ceftibuten Hydrate 707 Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Cefteram Pivoxil. System suitability— Proceed as directed in the system suitability in the Purity (2) under Cefteram Pivoxil. Water <2.48> Not more than 4.0z (a quantity equivalent to 0.2 g (potency) of powdered Cefteram Pivoxil Tablets, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cefteram Pivoxil Tablets add exactly 5 mL of the internal standard solution per 50 mg (potency) of Cefteram Pivoxil, and add diluted acetonitrile (1 in 2) to make V mL so that each mL contains about 1 mg (potency) Assay To a number of tablet of Cefteram Pivoxil Tablets, equivalent to about 1.
To a quantity of powdered Cefteram Pivoxil Tablets, equivalent to 0.1 g (potency) of Cefteram Pivoxil, add 20 mL of methanol, shake well, and filter. To 1 mL of the filtrate add 0.05 mol/L hydrochloric acid-methanol TS to make 500 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 262 nm and 266 nm.
Related substances—To a quantity of powdered Cefteram Pivoxil Tablets, equivalent to 0.1 g (potency) of Cefteram Pivoxil, add diluted acetonitrile (1 in 2) to make 100 mL. Disperse this solution by sonicating, filter, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 50 mL and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.9 to cefteram pivoxil, obtained from the sample so-lution is not larger than 1.75 times the peak area of cefteram pivoxil from the standard solution, and the area of the peak, having the relative retention time of about 0.1 from the sam-ple solution is not larger than 17/25 times the peak area of cefteram pivoxil from the standard solution. Furthermore, the total area of the peaks other than cefteram pivoxil from the sample solution is not larger than 3.7 times the peak area of cefteram pivoxil from the standard solution. For the area of the peak, having the relative retention time of about 0.1 to cefteram pivoxil, multiply the correction factor, 0.74. ‌JP XVIII Official Monographs / Ceftibuten Hydrate 707 Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Cefteram Pivoxil. System suitability— Proceed as directed in the system suitability in the Purity (2) under Cefteram Pivoxil. Water <2.48> Not more than 4.0z (a quantity equivalent to 0.2 g (potency) of powdered Cefteram Pivoxil Tablets, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cefteram Pivoxil Tablets add exactly 5 mL of the internal standard solution per 50 mg (potency) of Cefteram Pivoxil, and add diluted acetonitrile (1 in 2) to make V mL so that each mL contains about 1 mg (potency)
To a number of tablet of Cefteram Pivoxil Tablets, equivalent to about 1.0 g (potency) of Cefteram Pivoxil, add 120 mL of diluted acetonitrile (1 in 2), disperse by sonicat-ing, and add diluted acetonitrile (1 in 2) to make exactly 200 mL. Centrifuge this solution, pipet 10 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, add diluted acetonitrile (1 in 2) to make 50 mL, filter through a membrane filter with pore size not exceeding 0.45 mm, discard the first 3 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately an amount of Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to about 50 mg (potency), dissolve in 20 mL of diluted acetonitrile (1 in 2), add exactly 5 mL of the inter-nal standard solution, add diluted acetonitrile (1 in 2) to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefteram Pivoxil. of Cefteram Pivoxil. Disperse this solution by sonicating, filter through a membrane filter with pore size of not exceeding 0.45 mm, discard the first 10 mL of the filtrate, Amount [mg (potency)] of cefteram (C ＝ MS × QT/QS × 20 16H17N9O5S2) and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately an amount of Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to about 50 mg (po-tency), dissolve in 20 mL of diluted acetonitrile (1 in 2), add exactly 5 mL of the internal standard solution, add diluted acetonitrile (1 in 2) to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Cefteram Pivoxil. Amount [mg (potency)] of cefteram (C16H17N9O5S2) ＝ MS × QT/QS × V/50 MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000). Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Cefteram Pivoxil Tablets is not less than 75z. Start the test with 1 tablet of Cefteram Pivoxil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg (potency) of Cefteram Pivoxil, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Cefteram Pivoxil Mesitylene Sulfonate RS, equivalent to about 22 mg (potency), and dissolve in 20 mL of methanol, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and determine the absorbances, AT and AS, at 300 nm. Dissolution rate (z) with respect to the labeled amount of cefteram (C16H17N9O5S2) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken C: Labeled amount [mg (potency)] of cefteram (C16H17N9O5S2) in 1 tablet MS: Amount [mg (potency)] of Cefteram Pivoxil Mesity-lene Sulfonate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 1000).
Cetirizine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of cetirizine hydrochloride (C21H25ClN2O3.2HCl: 461.81). Method of preparation Prepare as directed under Tablets, with Cetirizine Hydrochloride. Identification To a quantity of powdered Cetirizine Hydro-chloride Tablets, equivalent to 10 mg of Cetirizine Hydro-chloride, add about 70 mL of 0.1 mol/L hydrochloric acid TS, shake, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and filter. To 4 mL of the filtrate add 0.1 mol/L hydro-chloric acid TS to make 25 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 230 nm and 234 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Cetirizine Hydrochloride Tablets, add 4V/5 mL of sodium 1-heptanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, sonicate for 20 minutes, add sodium 1-heptanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS to exactly V mL so that each mL contains about 0.
To a quantity of powdered Cetirizine Hydro-chloride Tablets, equivalent to 10 mg of Cetirizine Hydro-chloride, add about 70 mL of 0.1 mol/L hydrochloric acid TS, shake, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and filter. To 4 mL of the filtrate add 0.1 mol/L hydro-chloric acid TS to make 25 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 230 nm and 234 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Cetirizine Hydrochloride Tablets, add 4V/5 mL of sodium 1-heptanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, sonicate for 20 minutes, add sodium 1-heptanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS to exactly V mL so that each mL contains about 0.2 mg of cetirizine hydrochloride (C21H25ClN2O3.2HCl), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 2 mL of the internal standard solution, add acetonitrile to make 10 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of cetirizine hydrochloride (C21H25ClN2O3.2HCl) of the sample solution and the standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of cetirizine hydrochloride (C21H25ClN2O3.2HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of cetirizine hydrochloride for assay taken C: Labeled amount (mg) of cetirizine hydrochloride (C21H25ClN2O3.2HCl) in 1 tablet
Weigh accurately not less than 20 Cetirizine Hydro-chloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of cetirizine hydro-chloride (C21H25ClN2O3.2HCl), add 40 mL of sodium 1-hep-tanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, treat with ultrasonic waves for 20 minutes, add sodium 1-heptanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, to make exactly 50 mL, and filter this solution through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 2 mL of the internal standard solution, add acetonitrile to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of cetirizine hydrochloride for assay, previously dried in vacuum at 609C for 3 hours, and add sodium 1-hep-tanesulfonate solution (1 in 5000) adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS, to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal stand-ard solution, add acetonitrile to make 10 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cetirizine to that of the internal standard. Amount (mg) of cetirizine hydrochloride (C21H25ClN2O3.2HCl) ＝ MS × QT/QS × 1/2 ＝ MS × QT/QS × V/100 MS: Amount (mg) of cetirizine hydrochloride for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 5-mg tablet and in 30 minutes of 10-mg tablet are not less than 85z and not less than 80z, respec-tively. Start the test with 1 tablet of Cetirizine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of cetirizine hydro-chloride (C21H25ClN2O3.2HCl), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of cetirizine hydrochloride for assay, previously dried in vacuum at 609C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 230 nm MS: Amount (mg) of cetirizine hydrochloride for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of sodium 1-hep-tansulfonate (1 in 2900) and acetonitrile (29:21), adjusted to pH 3.0 with 0.5 mol/L sulfuric acid TS. Flow rate: Adjust so that the retention time of cetirizine is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, cetirizine and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ‌JP XVIII Official Monographs / Cetotiamine Hydrochloride Hydrate 725 the peak area of cetirizine to that of the internal standard is not more than 1.0z.
Ceˆxime Fine Granules contain not less than 90.0z and not more than 105.0z of the labeled potency of ceˆxime (C16H15N5O7S2: 453.45). Method of preparation Prepare as directed under Gran-ules, with Ceˆxime Hydrate. Identiˆcation To a quantity of powdered Ceˆxime Fine Granules, equivalent to 2 mg (potency) of Ceˆxime Hydrate, add 150 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), and shake. If necessary, ˆlter or centrifuge. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maximum between 286 nm and 290 nm. Purity Related substances—To a quantity of powdered Ceˆxime Fine Granules, equivalent to 0.1 g (potency) of Ceˆxime Hydrate, add 100 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), shake, ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm, and use the ˆl-trate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than ceˆxime is not more than 1.0z, and the total amount of the peaks other than ceˆxime is not more than 2.5z. Operating conditions— Detector, column, column temperature, mobile phase and ‰ow rate: Proceed as directed in the operating conditions in the Assay under Ceˆxime Hydrate. Time span of measurement: Proceed as directed in the op-erating conditions in the Purity under Ceˆxime Hydrate. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 10 mL. Conˆrm that the peak area of ceˆxime obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of ceˆxime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ceˆxime is not more than 2.0z. Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ceˆxime Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Ceˆxime Fine Gran-ules add 7 V/10 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), shake, add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly V mL so that each mL contains about 1 mg (potency) of Ceˆxime Hydrate.
Related substances—To a quantity of powdered Ceˆxime Fine Granules, equivalent to 0.1 g (potency) of Ceˆxime Hydrate, add 100 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), shake, ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm, and use the ˆl-trate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than ceˆxime is not more than 1.0z, and the total amount of the peaks other than ceˆxime is not more than 2.5z. Operating conditions— Detector, column, column temperature, mobile phase and ‰ow rate: Proceed as directed in the operating conditions in the Assay under Ceˆxime Hydrate. Time span of measurement: Proceed as directed in the op-erating conditions in the Purity under Ceˆxime Hydrate. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 10 mL. Conˆrm that the peak area of ceˆxime obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of ceˆxime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ceˆxime is not more than 2.0z. Water <2.48> Not more than 3.0z (1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ceˆxime Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Ceˆxime Fine Gran-ules add 7 V/10 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), shake, add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly V mL so that each mL contains about 1 mg (potency) of Ceˆxime Hydrate. Centrifuge this solution, pipet 10 mL of the supernatant liquid, add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Ceˆxime RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay under Ceˆxime Hydrate. Amount [mg (potency)] of ceˆxime (C16H15N5O7S2) ＝ MS × AT/AS × V/20 MS: Amount [mg (potency)] of Ceˆxime RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd ‰uid for dissolution test as the dissolution me-dium, the dissolution rates in 30 minutes of Ceˆxime Fine Granules is not less than 75z. Start the test with an accurately weighed amount of Ceˆxime Fine Granules, equivalent to about 0.1 g (potency) of Ceˆxime Hydrate, withdraw not less than 20 mL of the ‌JP XVIII Official Monographs / Cefmenoxime Hydrochloride 675 medium at the speciˆed minute after starting the test, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the ˆrst ˆltrate, and use the subsequent ˆltrate as the sample solu-tion. Separately, weigh accurately an amount of Ceˆxime RS, equivalent to about 28 mg (potency), and dissolve in the dissolution medium to make exactly 50 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ceˆxime in each solution. Dissolution rate (z) with respect to the labeled amount of ceˆxime (C16H15N5O7S2) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount [mg (potency)] of Ceˆxime RS taken MT: Amount (g) of Ceˆxime Fine Granules taken C: Labeled amount [mg (potency)] of ceˆxime (C16H15N5O7S2) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Assay under Ceˆxime Hydrate. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ceˆxime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ceˆxime is not more than 2.0z.
Weigh accurately an amount of powdered Ceˆxime Fine Granules, equivalent to about 0.1 g (potency) of Ceˆxime Hydrate, add 70 mL of 0.1 mol/L phosphate buŠer solution (pH 7.0), shake, and add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 100 mL. Centrifuge this solution, pipet 10 mL of the supernatant liquid, add 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Ceˆxime RS, equivalent to about 20 mg (potency), dissolve in 0.1 mol/L phosphate buŠer solution (pH 7.0) to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay under Ceˆxime Hydrate. Cefmenoxime Hydrochloride
Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution
クロラムフェニコール･コリスチンメタンスルホン酸ナトリウム点眼液
Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution is a clear, colorless to pale yellow liquid.
(1) To a volume of Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution, equivalent to about 2.5 mg (potency) of Chloramphenicol, and add water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as a blank: it exhibits a maximum between 276 nm and 280 nm. (2) To a volume of Chloramphenicol and Colistin So-dium Methanesulfonate Ophthalmic Solution, equivalent to about 5 × 105 Units of Colistin Sodium Methanesulfonate add 0.5 mL of ninhydrin TS, boil for 1 minute, and cool: a blue color develops. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> 6.0 – 8.0 Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Chloramphenicol Test organism—Kocuria rhizophila ATCC 9341 Chloramphenicol RS, equivalent to about 20 mg (potency), dissolve in 2 mL of ethanol (95), add phosphate buffer solu-tion (pH 6.0) to make exactly 20 mL, and use this solution as the standard stock solution. Keep the standard stock solu-tion at 159C or below, and use within 30 days. Pipet a suita-ble amount of the standard stock solution before use, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 100 mg (potency) and 25 mg (potency), and use these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively. Sample solutions—Weigh accurately an amount of Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution, equivalent to about 10 mg (potency) of Chloramphenicol, add phosphate buffer solution (pH 6.0) to make exactly 100 mL, and filter, if necessary. Pipet a suitable amount of this solution, add phosphate buffer solu-tion (pH 6.0) to make solutions so that each mL contains 100 mg (potency) and 25 mg (potency), and use these solutions as the high concentration sample solution and the low concen-tration sample solution, respectively. Colistin Sodium Methanesulfonate Test organism—Bordetella bronchiseptica ATCC 4617 Agar medium for base layer—Casein peptone 17.0 g Sodium chloride 5.0 g Glucose 2.5 g Soybean peptone 3.0 g Dipotassium hydrogen phosphate 2.0 g Agar 20.0 g Water 1000 mL Mix all the ingredients, then add a suitable amount of sodium hydroxide TS so that the pH of the medium will be 7.2 to 7.3 after sterilization, and sterile. Agar medium for seed layer—Casein peptone 17.0 g Glucose 2.5 g Soybean peptone 3.0 g Sodium chloride 5.0 g Polysorbate 80 10.0 g Dipotassium hydrogen phosphate 2.5 g Agar 12.0 g Water 1000 mL Mix all the ingredients, then add a suitable amount of sodium hydroxide TS so that the pH of the medium will be 7.2 to 7.3 after sterilization, and sterile. Agar medium for transferring test organisms—Use the medium i in 2) under (2) Agar media for transferring test organisms. Preparation of test organism and seeded agar layer—Cultivate the test organism on the slant of the agar medium for transferring test organism at 32 to 379C for 16 to 24 hours. Subcultures at least three times. Cultivate the grown organism on the slant of the agar medium for trans-ferring test organism at 32 to 379C for 16 to 24 hours, add a suitable amount of water to the grown organism, and sus- ‌732 pend. Adjust the suspension so that the transmittance at 660 nm is 60z as directed under Ultraviolet-visible Spectropho-tometry <2.24> using a spectrophotometer or a photoelectric photometer, and use this suspension as the test organism sus-pension. Keep the test organism suspension at 159C or below, and use within 3 days. Before use, dissolve 0.13 mL of the test organism suspension, add it to 100 mL of agar medium for seed previously cooled at 489C, mix thoroughly, and use this as the seeded agar layer. Standard solutions—Weigh accurately an amount of Colistin Sodium Methanesulfonate RS, equivalent to about 1 × 106 Units, dissolve in phosphate buffer solution (pH 6.0) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 109C or below, and use within 7 days. Pipet a suitable amount of the standard stock solution before use, add phos-phate buffer solution (pH 6.0) to make solutions so that each mL contains 1000 Units and 250 Units, and use these solu-tions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution, equivalent to about 1 × 105 Units of Colistin Sodium Methanesulfonate, add phosphate buffer solution (pH 6.0) to make a solution so that each mL con-tains 1000 Units, and use this solution as the high concentra-tion sample solution. Pipet 5 mL of the high concentration sample solution, add phosphate buffer solution (pH 6.0) to make a solution so that each mL contains 250 Units, and use this solution as the low concentration sample solution.
Chlordiazepoxide Powder contains not less than 93.
(1) Weigh a portion of Chlordiazepoxide Powder, equivalent to 0.01 g of Chlordiazepoxide, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 5 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 244 nm and 248 nm and between 306 nm and 310 nm, and a minimum between 288 nm and 292 nm. (2) Weigh a portion of Chlordiazepoxide Powder, equivalent to 0.02 g of Chlordiazepoxide, add 10 mL of methanol, shake for 5 minutes, then filter by suction through a glass filter (G4), evaporate the filtrate with the aid of a current of air to dryness, and dry the residue in vacuum at 609C for 1 hour. Determine the infrared absorption spec-trum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 1625 cm－1, 1465 cm－1, 1265 cm－1, 850 cm－1 and 765 cm－1.
Conduct this procedure without exposure to light, using light-resistant vessels. To a portion of Chlordia-zepoxide Powder, equivalent to 50 mg of Chlordiazepoxide, add exactly 5 mL of a mixture of methanol and ammonia TS (97:3), shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 50 mg of Chlordia-zepoxide RS in a mixture of methanol and ammonia TS (97:3) to make exactly 50 mL, and use this solution as the standard solution (1). Dissolve 5.0 mg of 2-amino-5-chlorobenzophenone for thin-layer chromatography in methanol to make exactly 200 mL, and use this solution as the standard solution (2). Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL of the sample solution and 10 mL each of the standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Pro-ceed as directed in the Purity (2) under Chlordiazepoxide. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Chlordia-zepoxide Powder is not less than 70z. Start the test with an accurately weighed amount of Chlor-diazepoxide Powder, equivalent to about 3.3 mg of chlordia-zepoxide (C16H14ClN3O), withdraw not less than 15 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 12 mg of Chlordia-zepoxide RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 609C) for 4 hours, dissolve in 20 mL of 0.1 mol/L hydrochloric acid TS, and add the dissolution medium to make exactly 200 mL. Pipet 3 mL of this solu-tion, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 260 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of chlordiazepoxide (C16H14ClN3O) ＝ MS/MT × AT/AS × 1/C × 27 MS: Amount (mg) of Chlordiazepoxide RS taken MT: Amount (g) of Chlordiazepoxide Powder taken C: Labeled amount (mg) of chlordiazepoxide (C16H14ClN3O) in 1 g
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately a quantity of Chlordiazepoxide Powder, equivalent to about 0.1 g of chlordiazepoxide (C16H14ClN3O), transfer to a glass-stoppered flask, wet with exactly 10 mL of water, add ex-actly 90 mL of methanol, stopper, shake vigorously for 15 minutes, and centrifuge. Pipet 10 mL of the supernatant liq-uid, add exactly 5 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 0.1 g of Chlordiazepoxide RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 609C) for 4 hours, and dis-solve in exactly 10 mL of water and exactly 90 mL of metha-nol. Pipet 10 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of chlordiazepoxide to that of the inter-nal standard. Amount (mg) of chlordiazepoxide (C16H14ClN3O) ＝ MS × QT/QS MS: Amount (mg) of Chlordiazepoxide RS taken Internal standard solution—A solution of isobutyl salicylate in methanol (1 in 20). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and 0.02 mol/L am-monium dihydrogenphosphate TS (7:3). Flow rate: Adjust so that the retention time of chlordia-zepoxide is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating conditions, chlordiazepoxide and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of chlordiazepoxide to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage-Light-resistant.
Chlordiazepoxide Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of chlordiazepoxide (C16H14ClN3O: 299.75). Method of preparation Prepare as directed under Tablets, with Chlordiazepoxide. Identification (1) Weigh a portion of powdered Chlordia-zepoxide Tablets, equivalent to 0.01 g of Chlordiazepoxide, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 5 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 244 nm and 248 nm and between 306 nm and 310 nm, and a minimum between 288 nm and 292 nm. (2) Weigh a portion of powdered Chlordiazepoxide Tablets, equivalent to 0.01 g of Chlordiazepoxide, add 10 mL of diethyl ether, shake vigorously, and centrifuge. Evaporate 5 mL of the supernatant liquid by warming on a water bath to dryness. Determine the infrared absorption spectrum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 1625 cm－1, 1465 cm－1, 1265 cm－1, 850 cm－1 and 765 cm－1. Purity Related substances—Conduct this procedure with-out exposure to light, using light-resistant vessels. To a por-tion of powdered Chlordiazepoxide Tablets, equivalent to 50 mg of Chlordiazepoxide, add exactly 5 mL of a mixture of methanol and ammonia TS (97:3), shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 50 mg of Chlordiazepoxide RS in a mixture of meth-anol and ammonia TS (97:3) to make exactly 50 mL, and use this solution as the standard solution (1). Dissolve 5.0 mg of 2-amino-5-chlorobenzophenone for thin-layer chromatogra-phy in methanol to make exactly 200 mL, and use this solu-tion as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL of the sample solution and 10 mL each of the standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Pro-ceed as directed in the Purity (2) under Chlordiazepoxide. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To 1 tablet of Chlordiazepoxide Tablets add 1 mL of water, shake to disintegrate the tablet, then add 20 mL of methanol, shake, add methanol to make exactly 25 mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, take exactly V mL of the subsequent filtrate equiva-lent to about 2 mg of chlordiazepoxide (C16H14ClN3O), add exactly 1 mL of the internal standard solution, then add methanol to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. in methanol (1 in 20). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Chlordia-zepoxide Tablets is not less than 70z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 tablet of Chlordia-zepoxide Tablets, withdraw not less than 30 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 3.
(1) Weigh a portion of powdered Chlordia-zepoxide Tablets, equivalent to 0.01 g of Chlordiazepoxide, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 5 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 244 nm and 248 nm and between 306 nm and 310 nm, and a minimum between 288 nm and 292 nm. (2) Weigh a portion of powdered Chlordiazepoxide Tablets, equivalent to 0.01 g of Chlordiazepoxide, add 10 mL of diethyl ether, shake vigorously, and centrifuge. Evaporate 5 mL of the supernatant liquid by warming on a water bath to dryness. Determine the infrared absorption spectrum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 1625 cm－1, 1465 cm－1, 1265 cm－1, 850 cm－1 and 765 cm－1.
Related substances—Conduct this procedure with-out exposure to light, using light-resistant vessels. To a por-tion of powdered Chlordiazepoxide Tablets, equivalent to 50 mg of Chlordiazepoxide, add exactly 5 mL of a mixture of methanol and ammonia TS (97:3), shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 50 mg of Chlordiazepoxide RS in a mixture of meth-anol and ammonia TS (97:3) to make exactly 50 mL, and use this solution as the standard solution (1). Dissolve 5.0 mg of 2-amino-5-chlorobenzophenone for thin-layer chromatogra-phy in methanol to make exactly 200 mL, and use this solu-tion as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 25 mL of the sample solution and 10 mL each of the standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Pro-ceed as directed in the Purity (2) under Chlordiazepoxide. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To 1 tablet of Chlordiazepoxide Tablets add 1 mL of water, shake to disintegrate the tablet, then add 20 mL of methanol, shake, add methanol to make exactly 25 mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, take exactly V mL of the subsequent filtrate equiva-lent to about 2 mg of chlordiazepoxide (C16H14ClN3O), add exactly 1 mL of the internal standard solution, then add methanol to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. in methanol (1 in 20). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Chlordia-zepoxide Tablets is not less than 70z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 tablet of Chlordia-zepoxide Tablets, withdraw not less than 30 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 3.7 mg of chlordiazepoxide (C16H14ClN3O), and use this solution as the sample solution. Separately, weigh accu-rately about 12 mg of Chlordiazepoxide RS, previously dried under reduced pressure with phosphorus (V) oxide as a des-sicant at 609C for 4 hours, dissolve in 5 mL of 0.1 mol/L hy-drochloric acid TS, and add the dissolution medium to make exactly 200 mL. Pipet 3 mL of this solution, add the dissolu-tion medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 260 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of chlordiazepoxide (C16H14ClN3O) ＝ MS × AT/AS × V?/V × 1/C × 27 MS: Amount (mg) of Chlordiazepoxide RS taken C: Labeled amount (mg) of chlordiazepoxide (C16H14ClN3O) in 1 tablet
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately a quantity of Chlordiazepoxide Tablets, equivalent to about 0.1 g of chlordiazepoxide (C16H14ClN3O), add 10 mL of water, and shake well to disintegrate. Add 60 mL of methanol, shake well, add methanol to make exactly 100 mL, and centrifuge. Pipet 10 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, add methanol to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 10 mg of Chlordiazepoxide RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 609C) for 4 hours, dissolve in 1 mL of water and a suitable amount of methanol, add exactly 5 mL of the inter-nal standard solution, add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of chlordiazepoxide to that of the inter-nal standard. Amount (mg) of chlordiazepoxide (C16H14ClN3O) ＝ MS × QT/QS × 10 MS: Amount (mg) of Chlordiazepoxide RS taken Internal standard solution—A solution of isobutyl salicylate in methanol (1 in 20). Operating conditions— Amount (mg) of chlordiazepoxide (C ＝ MS × QT/QS × 5/V 16H 14ClN3O) Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter MS: Amount (mg) of Chlordiazepoxide RS taken Internal standard solution—A solution of isobutyl salicylate and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). ‌JP XVIII Official Monographs / Chlorhexidine Hydrochloride 735 ‌Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and 0.02 mol/L am-monium dihydrogenphosphate TS (7:3). Flow rate: Adjust so that the retention time of chlordia-zepoxide is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, chlordiazepoxide and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of chlordiazepoxide to that of the internal standard is not more than 1.0z.
クロルヘキシジングルコン酸塩液
Chlorhexidine Gluconate Solution is a solution of digluconate of chlorhexidine. It contains not less than 19.
Chlorhexidine Gluconate Solution is a clear, colorless or pale yellow liquid. It is odorless, and has a bitter Each mL of 0.1 mol/L perchloric acid VS ＝ 22.44 mg of C H Cl N .2C H O taste. 22 30 2 10 6 12 7 It is miscible with water and with acetic acid (100). 1 mL of Chlorhexidine Gluconate Solution is miscible with not more than 5 mL of ethanol (99.5) and with not more than 3 mL of acetone. By further addition of each of these solvents, a white turbidity is formed. It is gradually colored by light. Specific gravity d 20: 1.06 – 1.07
(1) To 0.05 mL of Chlorhexidine Gluconate Solution add 5 mL of methanol, 1 mL of bromine TS and 1 mL of 8 mol/L sodium hydroxide TS: a deep red color is produced. To 0.5 mL of Chlorhexidine Gluconate Solution add 10 mL of water and 0.5 mL of copper (II) sulfate TS: a white precipitate is formed. Heat to boiling: the precipitate changes to light purple. To 10 mL of Chlorhexidine Gluconate Solution add 5 mL of water, cool on ice, and add 5 mL of sodium hydrox-ide TS dropwise with stirring: a white precipitate is formed. Collect the precipitate by filtration, wash with water, recrystallize from diluted ethanol (95) (7 in 10), and dry at 1059C for 30 minutes: the crystals thus obtained melt <2.60> between 1309C and 1349C. Neutralize the filtrate obtained in (3) with 5 mol/L hydrochloric acid TS. To 5 mL of this solution add 0.65 mL of acetic acid (100) and 1 mL of freshly distilled phenyl-hydrazine, and heat on a water bath for 30 minutes. After cooling, scratch the inner wall of the vessel with a glass rod to induce crystallization. Collect the crystals, dissolve in 10 mL of hot water, add a small amount of activated charcoal, and filter. Cool the filtrate, scratch the inner side of the ves-sel, collect the formed crystals, and dry: the crystals thus ob-tained melt <2.60> at about 1959C (with decomposition).
4-Chloroaniline—To 2.0 mL of Chlorhexidine Gluconate Solution add water to make exactly 100 mL. Pipet 5 mL of the solution, and add 20 mL of water and 5 mL of 1 mol/L hydrochloric acid TS. Add 0.3 mL of sodium nitrite TS, shake, and allow to stand for 2 minutes. Add 4 mL of ammonium amidosulfate TS, and then allow to stand for 1 minute. Add 5 mL of N-(1-naphthyl)-N?-diethylethylenedia-mine oxalate-acetone TS, allow to stand for 10 minutes, add 1 mL of ethanol (95), and then add water to make 50 mL: the color of the solution is not more intense than the follow-ing control solution. Control solution: Dissolve 20 mg of 4-chloroaniline in 10 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of the solution, and add water to make exactly 100 mL. Pipet 5 mL of the solution, add 20 mL of water and 5 mL of 1 mol/L hydrochloric acid TS, and proceed as directed for the preparation of the sample solu-tion. Residue on ignition <2.44> Not more than 0.1z (2 g, after evaporation).
Pipet 2 mL of Chlorhexidine Gluconate Solution, evaporate to dryness on a water bath, dissolve the residue in 60 mL of acetic acid for nonaqueous titration, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man- 22 30 2 10 6 12 7 ner, and make any necessary correction. Description Chlorhexidine Gluconate Solution is a clear, colorless or pale yellow liquid. It is odorless, and has a bitter Each mL of 0.1 mol/L perchloric acid VS ＝ 22.44 mg of C H Cl N .2C H O taste. 22 30 2 10 6 12 7 It is miscible with water and with acetic acid (100). 1 mL of Chlorhexidine Gluconate Solution is miscible with not more than 5 mL of ethanol (99.5) and with not more than 3 mL of acetone. By further addition of each of these solvents, a white turbidity is formed. It is gradually colored by light. Specific gravity d 20: 1.06 – 1.07 Identification (1) To 0.05 mL of Chlorhexidine Gluconate Solution add 5 mL of methanol, 1 mL of bromine TS and 1 mL of 8 mol/L sodium hydroxide TS: a deep red color is produced. To 0.5 mL of Chlorhexidine Gluconate Solution add 10 mL of water and 0.5 mL of copper (II) sulfate TS: a white precipitate is formed. Heat to boiling: the precipitate changes to light purple. To 10 mL of Chlorhexidine Gluconate Solution add 5 mL of water, cool on ice, and add 5 mL of sodium hydrox-ide TS dropwise with stirring: a white precipitate is formed. Collect the precipitate by filtration, wash with water, recrystallize from diluted ethanol (95) (7 in 10), and dry at 1059C for 30 minutes: the crystals thus obtained melt <2.60> between 1309C and 1349C. Neutralize the filtrate obtained in (3) with 5 mol/L hydrochloric acid TS. To 5 mL of this solution add 0.65 mL of acetic acid (100) and 1 mL of freshly distilled phenyl-hydrazine, and heat on a water bath for 30 minutes. After cooling, scratch the inner wall of the vessel with a glass rod to induce crystallization. Collect the crystals, dissolve in 10 mL of hot water, add a small amount of activated charcoal, and filter. Cool the filtrate, scratch the inner side of the ves-sel, collect the formed crystals, and dry: the crystals thus ob-tained melt <2.60> at about 1959C (with decomposition).
クロルフェネシンカルバミン酸エステル錠
Chlorphenesin Carbamate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of chlorphenesin carbamate (C10H12ClNO4: 245.66). Method of preparation Prepare as directed under Tablets, with Chlorphenesin Carbamate. Identification To a quantity of powdered Chlorphenesin Carbamate Tablets, equivalent to 0.15 g of Chlorphenesin Carbamate, add 60 mL of ethanol (95), sonicate, and add ethanol (95) to make 100 mL. Centrifuge 20 mL of this solu-tion, add ethanol (95) to 1 mL of the supernatant liquid to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 226 nm and 230 nm, between 279 nm and 283 nm, and between 286 nm and 290 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Chlorphenesin Carbamate Tablets add 10 mL of water to disintegrate the tablet, add 70 mL of a mix-ture of water and methanol (1:1), sonicate for 15 minutes with occasional stirring, then add the mixture of water and methanol (1:1) to make exactly 100 mL. Centrifuge this solu-tion, pipet V mL of the supernatant liquid equivalent to about 2.5 mg of chlorphenesin carbamate (C10H12ClNO4), add the mixture of water and methanol (1:1) to make exactly 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of chlorphenesin carba-mate for assay, previously dried in a desiccator (in vacuum, silica gel) for 4 hours, and dissolve in the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 2 mL of this solution, add the mixture of water and methanol (1:1) to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances at 280 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of chlorphenesin carbamate (C10H12ClNO4) ＝ MS × AT/AS × 1/V × 5 MS: Amount (mg) of chlorphenesin carbamate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Chlorphenesin Carbamate Tablets is not less than 85z. Start the test with 1 tablet of Chlorphenesin Carbamate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.
To a quantity of powdered Chlorphenesin Carbamate Tablets, equivalent to 0.15 g of Chlorphenesin Carbamate, add 60 mL of ethanol (95), sonicate, and add ethanol (95) to make 100 mL. Centrifuge 20 mL of this solu-tion, add ethanol (95) to 1 mL of the supernatant liquid to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 226 nm and 230 nm, between 279 nm and 283 nm, and between 286 nm and 290 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Chlorphenesin Carbamate Tablets add 10 mL of water to disintegrate the tablet, add 70 mL of a mix-ture of water and methanol (1:1), sonicate for 15 minutes with occasional stirring, then add the mixture of water and methanol (1:1) to make exactly 100 mL. Centrifuge this solu-tion, pipet V mL of the supernatant liquid equivalent to about 2.5 mg of chlorphenesin carbamate (C10H12ClNO4), add the mixture of water and methanol (1:1) to make exactly 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of chlorphenesin carba-mate for assay, previously dried in a desiccator (in vacuum, silica gel) for 4 hours, and dissolve in the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 2 mL of this solution, add the mixture of water and methanol (1:1) to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances at 280 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of chlorphenesin carbamate (C10H12ClNO4) ＝ MS × AT/AS × 1/V × 5 MS: Amount (mg) of chlorphenesin carbamate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Chlorphenesin Carbamate Tablets is not less than 85z. Start the test with 1 tablet of Chlorphenesin Carbamate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.14 mg of chlorphenesin car-bamate (C10H12ClNO4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of chlor-phenesin carbamate for assay, previously dried in a desicca-tor (in vacuum, silica gel) for 4 hours, dissolve in 1 mL of methanol, and add water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 278 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of chlorphenesin carbamate (C10H12ClNO4) ＝ MS × AT/AS × V?/V × 1/C × 450 MS: Amount (mg) of chlorphenesin carbamate for assay taken C: Labeled amount (mg) of chlorphenesin carbamate (C10H12ClNO4) in 1 tablet
Weigh accurately the mass of not less than 20 Chlor-phenesin Carbamate Tablets, and powder them in an agate mortar. Weigh accurately a portion of the powder, equivalent to about 0.25 g of chlorphenesin carbamate (C10H12ClNO4), add 30 mL of ethyl acetate, disperse by sonicating, then add ethyl acetate to make exactly 50 mL. Centrifuge 20 mL of this solution, pipet 2 mL of the super-natant liquid, add exactly 2 mL of the internal standard solu-tion, add ethyl acetate to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of chlorphenesin carbamate for assay, previously dried in a desiccator (in vacuum, silica gel) for 4 hours, and dis-solve in ethyl acetate to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal standard solu-tion, then add ethyl acetate to make 20 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of chlorphenesin carbamate to that of the in-ternal standard. Amount (mg) of chlorphenesin carbamate (C10H12ClNO4) ＝ MS × QT/QS × 5/2 MS: Amount (mg) of chlorphenesin carbamate for assay taken Internal standard solution—A solution of ethenzamide in ethyl acetate (1 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with silica gel for liquid chro-matography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of hexane for liquid chromatog-raphy, 2-propanol and acetic acid (100) (700:300:1). Flow rate: Adjust so that the retention time of chlorphene-sin carbamate is about 9 minutes. System suitability— System performance: Proceed as directed in the system suitability in the Purity (3) under Chlorphenesin Carbamate. System repeatability: When the test is repeated 6 times ‌740 with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of chlorphenesin carbamate to that of the in-ternal standard is not more than 1.5z.
クロルフェニラミンマレイン酸塩注射液
Chlorpheniramine Maleate Injection is an aqueous injection. It contains not less than 95.
Chlorpheniramine Maleate Injection is a clear, colorless liquid. pH: 4.5 – 7.0
Take a volume of Chlorpheniramine Male-ate Injection, equivalent to 25 mg of Chlorpheniramine Maleate, add 5 mL of dilute sodium hydroxide TS, and ex-tract with 20 mL of hexane. Wash the hexane layer with 10 mL of water, shake with 0.5 g of anhydrous sodium sulfate for several minutes, and filter. Evaporate the filtrate in a water bath at 509C under a reduced pressure, and determine the infrared absorption spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, 1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. Bacterial endotoxins <4.01> Less than 8.8 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Transfer an exactly measured volume of Chlor-pheniramine Maleate Injection, equivalent to about 3 mg of Amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of Chlorpheniramine Maleate RS taken
クロルフェニラミンマレイン酸塩散
Chlorpheniramine Maleate Powder contains not less than 93.
Weigh a portion of Chlorpheniramine Maleate Powder, equivalent to 50 mg of Chlorpheniramine Maleate, shake with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. Transfer the filtrate to a separator, and wash with 40 mL of hexane. Add 10 mL of sodium hydroxide TS, and extract with 20 mL of hexane. Wash the hexane layer with 5 mL of water. Centrifuge, if necessary, shake the hexane extract with 0.5 g of anhydrous sodium sulfate for several minutes, and filter. Evaporate the filtrate in a water bath at about 509C under reduced pressure, and determine the infrared absorption spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave number of about 2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, 1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Chlorpheniramine Maleate Powder is not less than 85z. Start the test with an accurately weighed amount of Chlor-pheniramine Maleate Powder, equivalent to about 4 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4), withdraw not less than 20 mL of the medium at the specified minute chlorpheniramine maleate (C16H19 ClN2.C4H4O4), to a after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 100-mL separator, add 20 mL of water and 2 mL of sodium hydroxide TS, and extract with two 50-mL portions of diethyl ether. Combine the diethyl ether extracts, wash with 20 mL of water, and then extract with 20-mL, 20-mL and 5-mL portions of 0.25 mol/L sulfuric acid TS successively. Combine all acid extracts, and add 0.25 mol/L sulfuric acid TS to make exactly 50 mL. Pipet 10 mL of this solution, add 0.25 mol/L sulfuric acid TS to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 30 mg of Chlorpheniramine Maleate RS, previ-ously dried at 1059C for 3 hours, and dissolve in water to make exactly 200 mL. Pipet 20 mL of this solution, transfer to a 100-mL separator, add 2 mL of sodium hydroxide TS, and extract with two 50-mL portions of diethyl ether. Pro-ceed in the same manner as for the preparation of the sample solution, and use the solution so obtained as the standard so- 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 22 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of chlorpheniramine in each solution. Dissolution rate (z) with respect to the labeled amount of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS/MT × AT/AS × 1/C × 18 ‌742 MS: Amount (mg) of Chlorpheniramine Maleate RS taken MT: Amount (g) of Chlorpheniramine Maleate Powder taken C: Labeled amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry the peak area of chlorpheniramine to that of the internal standard is not more than 1.0z.
クロルフェニラミンマレイン酸塩錠
Chlorpheniramine Maleate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of dl-chlorpheniramine maleate factor of the peak of chlorpheniramine are not less than 2000 and not more than 2.5, respectively. (C16H19 ClN2.C4H4O4: 390.86). System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of chlorpheniramine is not more than 2.0z. Assay Weigh accurately an amount of Chlorpheniramine Maleate Powder, equivalent to about 4 mg of chlorphenira-mine maleate (C16H19ClN2.C4H4O4), add 70 mL of the inter-nal standard solution, shake for 15 minutes, then add the in-ternal standard solution to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 20 mg of Chlorpheniramine Maleate RS, previ-ously dried at 1059C for 3 hours, and add the internal stand-ard to make exactly 100 mL. Pipet 20 mL of this solution, add the internal standard solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solultion of methyl parahydroxybenzoate in methanol (1 in 1000) add water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100) and water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Method of preparation Prepare as directed under Tablets, with Chlorpheniramine Maleate. Identification Weigh a portion of powdered Chlorphenira-mine Maleate Tablets, equivalent to 50 mg of Chlorphenira-mine Maleate, shake with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. Transfer the filtrate to a separator, and wash with 40 mL of hexane. Add 10 mL of sodium hydrox-ide TS, and extract with 20 mL of hexane. Wash the hexane layer with 5 mL of water. Centrifuge, if necessary, shake the hexane extract with 0.5 g of anhydrous sodium sulfate for several minutes, and filter. Evaporate the filtrate in a water bath at about 509C under a reduced pressure, and determine the infrared absorption spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, 1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Chlorpheniramine Maleate Tablets add 10 mL of water, shake to disintegrate the tablet, then add water to make exactly V mL of a solution containing about 80 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4) per mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Pipet 5 mL of the filtrate, add exactly 2.5 mL of the internal standard solution, add water to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add ex-actly 25 mL of the internal standard solution, add water to make 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay, and calculate the ratios, QT and QS, of the peak area of chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) Flow rate: Adjust so that the retention time of chlor- 16 19 2 4 4 4 pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating conditions, the internal standard and chlorpheniramine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ＝ MS × QT/QS × V/250 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 250) add water to make 1000 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Chlorpheniramine Maleate Tablets is not JP XVIII Official Monographs / d-Chlorpheniramine Maleate 743 ‌less than 75z. Start the test with 1 tablet of Chlorpheniramine Maleate Tablets, withdraw not less than 20 mL of the medium at the chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) specified minute after starting the test, and filter through a 16 19 2 4 4 4 membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 4.
Weigh a portion of powdered Chlorphenira-mine Maleate Tablets, equivalent to 50 mg of Chlorphenira-mine Maleate, shake with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. Transfer the filtrate to a separator, and wash with 40 mL of hexane. Add 10 mL of sodium hydrox-ide TS, and extract with 20 mL of hexane. Wash the hexane layer with 5 mL of water. Centrifuge, if necessary, shake the hexane extract with 0.5 g of anhydrous sodium sulfate for several minutes, and filter. Evaporate the filtrate in a water bath at about 509C under a reduced pressure, and determine the infrared absorption spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, 1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Chlorpheniramine Maleate Tablets add 10 mL of water, shake to disintegrate the tablet, then add water to make exactly V mL of a solution containing about 80 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4) per mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Pipet 5 mL of the filtrate, add exactly 2.5 mL of the internal standard solution, add water to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add ex-actly 25 mL of the internal standard solution, add water to make 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay, and calculate the ratios, QT and QS, of the peak area of chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) Flow rate: Adjust so that the retention time of chlor- 16 19 2 4 4 4 pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating conditions, the internal standard and chlorpheniramine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ＝ MS × QT/QS × V/250 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 250) add water to make 1000 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Chlorpheniramine Maleate Tablets is not JP XVIII Official Monographs / d-Chlorpheniramine Maleate 743 ‌less than 75z. Start the test with 1 tablet of Chlorpheniramine Maleate Tablets, withdraw not less than 20 mL of the medium at the chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) specified minute after starting the test, and filter through a 16 19 2 4 4 4 membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 4.4 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of chlor- pheniramine in each solution. Dissolution rate (z) with respect to the labeled amount of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Chlorpheniramine Maleate RS taken C: Labeled amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100), and add water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of chlor-pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of chlorpheniramine are not less than 2000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 1000) add water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100) and water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of chlor-pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the internal standard and chlorpheniramine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of chlorpheniramine to that of the internal standard is not more than 1.0z.
Weigh accurately an amount of Chlorpheniramine Maleate Powder, equivalent to about 4 mg of chlorphenira-mine maleate (C16H19ClN2.C4H4O4), add 70 mL of the inter-nal standard solution, shake for 15 minutes, then add the in-ternal standard solution to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 20 mg of Chlorpheniramine Maleate RS, previ-ously dried at 1059C for 3 hours, and add the internal stand-ard to make exactly 100 mL. Pipet 20 mL of this solution, add the internal standard solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solultion of methyl parahydroxybenzoate in methanol (1 in 1000) add water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100) and water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Method of preparation Prepare as directed under Tablets, with Chlorpheniramine Maleate. Identification Weigh a portion of powdered Chlorphenira-mine Maleate Tablets, equivalent to 50 mg of Chlorphenira-mine Maleate, shake with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. Transfer the filtrate to a separator, and wash with 40 mL of hexane. Add 10 mL of sodium hydrox-ide TS, and extract with 20 mL of hexane. Wash the hexane layer with 5 mL of water. Centrifuge, if necessary, shake the hexane extract with 0.5 g of anhydrous sodium sulfate for several minutes, and filter. Evaporate the filtrate in a water bath at about 509C under a reduced pressure, and determine the infrared absorption spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 2940 cm－1, 2810 cm－1, 2770 cm－1, 1589 cm－1, 1491 cm－1, 1470 cm－1, 1434 cm－1, 1091 cm－1 and 1015 cm－1. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Chlorpheniramine Maleate Tablets add 10 mL of water, shake to disintegrate the tablet, then add water to make exactly V mL of a solution containing about 80 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4) per mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Pipet 5 mL of the filtrate, add exactly 2.5 mL of the internal standard solution, add water to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add ex-actly 25 mL of the internal standard solution, add water to make 100 mL, and use this solution as the standard solution. Perform the test with 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay, and calculate the ratios, QT and QS, of the peak area of chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) Flow rate: Adjust so that the retention time of chlor- 16 19 2 4 4 4 pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating conditions, the internal standard and chlorpheniramine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ＝ MS × QT/QS × V/250 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 250) add water to make 1000 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Chlorpheniramine Maleate Tablets is not JP XVIII Official Monographs / d-Chlorpheniramine Maleate 743 ‌less than 75z. Start the test with 1 tablet of Chlorpheniramine Maleate Tablets, withdraw not less than 20 mL of the medium at the chlorpheniramine to that of the internal standard. Amount (mg) of chlorpheniramine maleate (C H ClN .C H O ) specified minute after starting the test, and filter through a 16 19 2 4 4 4 membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 4.4 mg of chlorpheniramine maleate (C16H19ClN2.C4H4O4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Chlorpheniramine Maleate RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of chlor- pheniramine in each solution. Dissolution rate (z) with respect to the labeled amount of chlorpheniramine maleate (C16H19ClN2.C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Chlorpheniramine Maleate RS taken C: Labeled amount (mg) of chlorpheniramine maleate (C16H19ClN2.C4H4O4) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100), and add water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of chlor-pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of chlorpheniramine are not less than 2000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ＝ MS × QT/QS × 1/5 MS: Amount (mg) of Chlorpheniramine Maleate RS taken Internal standard solution—To 7 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 1000) add water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of sodium 1-heptane sulfonate in 900 mL of water, add 10 mL of acetic acid (100) and water to make 1000 mL. To 650 mL of this solution add 350 mL of acetonitrile. Flow rate: Adjust so that the retention time of chlor-pheniramine is about 8 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the internal standard and chlorpheniramine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of chlorpheniramine to that of the internal standard is not more than 1.0z.
Chlorpromazine Hydrochloride Injection is a clear, colorless or pale yellow liquid. pH: 4.0 – 6.5
(1) Proceed with a volume of Chlorproma-zine Hydrochloride Injection, equivalent to 5 mg of Chlor-promazine Hydrochloride, as directed in the Identification under Chlorpromazine Hydrochloride. Proceed with a volume of Chlorpromazine Hydro-chloride Injection, equivalent to 0.1 g of Chlorpromazine to a separator, add 30 mL of water and 10 mL of a solution of sodium hydroxide (1 in 5), and extract with two 30-mL portions and three 20-mL portions of diethyl ether. Wash the combined diethyl ether extracts with successive 10-mL portions of water until the last washing shows no red color upon the addition of phenolphthalein TS. Concentrate the diethyl ether extracts on a water bath to 20 mL, add 5 g of anhydrous sodium sulfate, allow to stand for 20 minutes, and filter through a pledget of absorbent cotton. Wash with diethyl ether, combine the washings with the filtrate, and evaporate the diethyl ether on a water bath. Dissolve the residue in 50 mL of acetone and 5 mL of acetic acid (100), and titrate <2.50> with 0.05 mol/L perchloric acid VS until the color of the solution changes from red-purple to blue-purple (indicator: 3 drops of bromocresol green-methyl-rosaniline chloride TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L perchloric acid VS ＝ 17.77 mg of C17H19ClN2S.HCl
Chlorpromazine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of chlorpromazine hydrochloride (C17H19ClN2S.HCl: 355.33). Method of preparation Prepare as directed under Tablets, with Chlorpromazine Hydrochloride. Identification (1) Shake a quantity of powdered Chlor-promazine Hydrochloride Tablets, equivalent to 0.2 g of Chlorpromazine Hydrochloride, with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. To 1 mL of the filtrate add 4 mL of water and 1 drop of iron (III) chloride TS: a red color develops. (2) To 20 mL of the filtrate obtained in (1) add 10 mL of 2,4,6-trinitrophenol TS dropwise, and proceed as directed in the Identification (2) under Chlorpromazine Hydrochloride. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedures using light-resistant vessels. To 1 tablet of Chlorpromazine Hydrochloride Tablets add an amount of a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) so that each mL contains about 0.
(1) Shake a quantity of powdered Chlor-promazine Hydrochloride Tablets, equivalent to 0.2 g of Chlorpromazine Hydrochloride, with 40 mL of 0.1 mol/L hydrochloric acid TS, and filter. To 1 mL of the filtrate add 4 mL of water and 1 drop of iron (III) chloride TS: a red color develops. (2) To 20 mL of the filtrate obtained in (1) add 10 mL of 2,4,6-trinitrophenol TS dropwise, and proceed as directed in the Identification (2) under Chlorpromazine Hydrochloride. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedures using light-resistant vessels. To 1 tablet of Chlorpromazine Hydrochloride Tablets add an amount of a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) so that each mL contains about 0.83 mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl), sonicate for 5 minutes, then shake vigorously for 20 minutes, and add the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make exactly V mL so that each mL contains about 0.5 mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl). Filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, pipet 2.5 mL of the subsequent filtrate, add ex-actly 5 mL of the internal standard solution, then add the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 25 mL, and use this solution as the sam-ple solution. Then, proceed as directed in the Assay. Amount (mg) of chlorpromazine hydrochloride (C17H19ClN2S.HCl) ＝ MS × QT/QS × V/50 MS: Amount (mg) of chlorpromazine hydrochloride for assay taken 0.45 mm, and discard the first 3 mL of the filtrate. To exactly 2.5 mL of the subsequent filtrate add exactly 5 mL of the in-ternal standard solution, then add the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 25 mg of chlorpromazine hy-drochloride for assay, previously dried at 1059C for 2 hours, and dissolve in the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution and the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 25 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of chlorpromazine to that of the internal standard. Amount (mg) of chlorpromazine hydrochloride (C17H19ClN2S.HCl) ＝ MS × QT/QS × 2 Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) (1 in 4500). Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 60 minutes of Chlorpromazine Hydrochloride Tablets is not less than 75z. Start the test with 1 tablet of Chlorpromazine Hydrochlo-ride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl), and use this solution as the sample solution. Separately, weigh accurately about 90 mg of chlorpromazine hydrochloride for assay, previously dried at 1059C for 2 hours, dissolve in the dissolution medium to make exactly 200 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, further pipet 5 mL of this solution, add the dissolu-tion medium to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and A S, of the sample solution and standard solution at 254 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to labeled amount of chlorpromazine hydrochloride (C17H19ClN2S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 45/8 MS: Amount (mg) of chlorpromazine hydrochloride for assay taken C: Labeled amount (mg) of chlorpromazine hydrochloride (C17H19ClN2S.HCl) in 1 tablet
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately, and powder not less than 20 Chlorpromazine Hydrochloride Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of chlorpromazine hydrochloride (C17H19ClN2S.HCl), add 60 mL of a mixture of diluted phos-phoric acid (1 in 500) and ethanol (99.5) (1:1), sonicate for 5 minutes, then shake vigorously for 20 minutes, and add the mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make exactly 100 mL. Filter the solution through a membrane filter with a pore size not exceeding MS: Amount (mg) of chlorpromazine hydrochloride for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) (1 in 4500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 256 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted 0.05 mol/L sodium di-hydrogen phosphate TS (1 in 2) and acetonitrile (27:13). Flow rate: Adjust so that the retention time of chlor-promazine is about 15 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and chlorpromazine are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of chlorpromazine to that of the internal standard is not more than 1.0z.
Chlorpropamide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of chlorpropamide (C10H13ClN2O3S: 276.74). Method of preparation Prepare as directed under Tablets, with Chlorpropamide. Identification Take a quantity of powdered Chlor-propamide Tablets, equivalent to 0.08 g of Chlorpropamide, add 50 mL of methanol, shake, and filter. To 1 mL of the filtrate add 0.01 mol/L hydrochloric acid TS to make 200 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 231 nm and 235 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Chlorpropamide Tablets add 75 mL of the mobile phase, sonicate for 20 minutes with occasional strong shaking, then add the mobile phase to make exactly V mL so that each mL contains about 2.
Take a quantity of powdered Chlor-propamide Tablets, equivalent to 0.08 g of Chlorpropamide, add 50 mL of methanol, shake, and filter. To 1 mL of the filtrate add 0.01 mol/L hydrochloric acid TS to make 200 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 231 nm and 235 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Chlorpropamide Tablets add 75 mL of the mobile phase, sonicate for 20 minutes with occasional strong shaking, then add the mobile phase to make exactly V mL so that each mL contains about 2.5 mg of Chlorpropamide. Centrifuge the solution, pipet 2 mL of the supernatant liq-uid, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. solution as the sample solution. Pipet 1 mL of the sample so-lution, add acetone to make exactly 300 mL, and use this so-lution as the standard solution (1). Separately, dissolve 60 Amount (mg) of chlorpropamide (C ＝ MS × AT/AS × V/20 10H13ClN2O3S) mg of 4-chlorobenzene sulfonamide in acetone to make ex-actly 300 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under MS: Amount (mg) of chlorpropamide for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 ‌748 mL of 2nd fluid for dissolution test as the dissolution medi-um, the dissolution rate in 45 minutes of Chlorpropamide Tablets is not less than 70z. Start the test with 1 tablet of Chlorpropamide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 10 mg of chlorpropamide (C10H13ClN2O3S), and use this solution as the sample solu-tion. Separately, weigh accurately about 50 mg of chlor-propamide for assay, previously dried at 1059C for 3 hours, dissolve in 10 mL of methanol, and add water to make ex-actly 50 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 232 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of chlorpropamide (C10H13ClN2O3S) ＝ MS × AT/AS × V?/V × 1/C × 18 with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of chlorpropamide is not more than 1.5z.
注射用ヒト絨毛性性腺刺激ホルモン
Human Chorionic Gonadotrophin for Injec-tion occurs as a white to light yellow-brown powder or masses.
Proceed as directed in the Identification under Human Chorionic Gonadotrophin. pH <2.54> Prepare a solution so that each mL of isotonic sodium chloride solution contains 2 mg of Human Chorionic Gonadotorophin for Injection: the pH of this solution is be-tween 5.0 and 7.0. Loss on drying <2.41> Not more than 5.0z (0.1 g, in vacu-um, phosphorus (V) oxide, 4 hours). Bacterial endotoxins <4.01> Less than 0.03 EU/unit. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test, when calculate the acceptance ‌JP XVIII Official Monographs / Human Menopausal Gonadotrophin 1087 value using the mean of estimated contents of the units tested as M. Foreign insoluble matter <6.06> Perform the test according to the Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Proceed as directed in the Assay under Human Chorionic Gonadotrophin. The ratio of the assayed Units to the labeled Units should be calculated by the following equa-tion. The ratio of the assayed Units to the labeled Units ＝ antilog M
Containers—Hermetic containers. Storage—Light-resistant, and in a cold place.
Cibenzoline Succinate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of cibenzoline succinate (C18H18N2.C4H6O4: 380.44). Method of preparation Prepare as directed under Tablets, with Cibenzoline Succinate. Identification To a quantity of powdered Cibenzoline Suc-cinate Tablets, equivalent to 50 mg of Cibenzoline Succinate, add 100 mL of water, shake for 10 minutes, and centrifuge. To 2 mL of the supernatant liquid add water to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cibenzoline Succinate Tablets add a suitable amount of water so that each mL contains about 10 mg of cibenzoline succinate (C18H18N2.
To a quantity of powdered Cibenzoline Suc-cinate Tablets, equivalent to 50 mg of Cibenzoline Succinate, add 100 mL of water, shake for 10 minutes, and centrifuge. To 2 mL of the supernatant liquid add water to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cibenzoline Succinate Tablets add a suitable amount of water so that each mL contains about 10 mg of cibenzoline succinate (C18H18N2.C4H6O4), and allow standing for 10 minutes while occasional shaking. To this solution add methanol so that each mL contains about 2 mg of ciben-zoline succinate (C18H18N2.C4H6O4), add exactly 1 mL of the internal standard solution per 10 mg of cibenzoline succinate (C18H18N2.C4H6O4), then add methanol so that each mL contains about 1 mg of cibenzoline succinate (C18H18N2.C4H6O4). Centrifuge the solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. as directed under Thin-layer Chromatography <2.03>. Spot Amount (mg) of cibenzoline succinate (C H N .C H O ) 10 mL each of the sample solution and standard solutions (1) ＝ MS × QT/QS × C/100 18 18 2 6 4 ‌JP XVIII Official Monographs / Ciclacillin 751 MS: Amount (mg) of cibenzoline succinate for assay taken C: Labeled amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4) in 1 tablet Internal standard solution—Dissolve 0.1 g of 2-ethylhexyl parahydroxybenzoate in methanol to make 100 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cibenzoline Succinate Tablets is not less than 80z. Start the test with 1 tablet of Cibenzoline Succinate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of cibenzoline suc-cinate (C18H18N2.C4H6O4), and use this solution as the sam-ple solution. Separately, weigh accurately about 28 mg of cibenzoline succinate for assay, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 222 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of cibenzoline succinate (C18H18N2.C4H6O4) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of cibenzoline succinate for assay taken C: Labeled amount (mg) of cibenzoline succinate (C18H18N2.C4H6O4) in 1 tablet
Weigh accurately not less than 20 Cibenzoline Suc-cinate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of cibenzoline suc-cinate (C18H18N2.C4H6O4), add 10 mL of water, shake, and add 40 mL of methanol and exactly 10 mL of the internal standard solution. Shake for 20 minutes, add methanol to make 100 mL, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 0.1 g of cibenzoline succinate for assay, previously dried at 1059C for 2 hours, add 10 mL of water and 40 mL of methanol to dissolve, then add exactly 10 mL of the internal standard so-lution and methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cibenzoline to that of the internal standard. sulfosuccinate in 2000 mL of a mixture of water, acetonitrile and diluted phosphoric acid (1 in 10) (1000:1000:1). Flow rate: Adjust so that the retention time of cibenzoline is about 3 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cibenzoline and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cibenzoline to that of the internal standard is not more than 1.0z.
Cilazapril Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of cilazapril (C H N O : 417.50). 22 31 3 5 Purity (1) Chloride <1.03>—Perform the test using 1.0 g of Cilazapril Hydrate. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.009z). Sulfate <1.14>—Dissolve 1.0 g of Cilazapril Hydrate in 40 mL of water and 1.5 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Heavy metals <1.07>—Proceed with 1.0 g of Cilazapril Hydrate according to Method 4, and perform the test. However, use 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 8). Prepare the control so-lution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Cilazapril Hydrate in 20 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 3 mL of the standard solu-tion (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (2). Separately, pipet 2 mL of the standard solution (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (3). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and three standard solutions on a plate of silica gel Method of preparation Prepare as directed under Tablets, with Cilazapril Hydrate. Identification To a quantity of powdered Cilazapril Tablets, equivalent to 2 mg of cilazapril (C22H31N3O5), add 2 mL of a mixture of acetonitrile and ethyl acetate (3:1), shake, sonicate for 30 seconds, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 5 mg of cilazapril in 5 mL of the mixture of acetonitrile and ethyl acetate (3:1), and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, acetic acid (100), hexane and water (62:15:10:10:3) to a dis-tance of about 15 cm, and air-dry the plate. Place the plate in iodine vapor for 2 hours, and immediately examine under ultraviolet light (main wavelength: 254 nm): the spots ob-tained from the sample and standard solutions are dark brown and they show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cilazapril Tablets add 5 mL of a mixture of water and acetonitrile (7:3), shake well until disintegration, add the mixture of water and acetonitrile (7:3) to make exactly V mL so that each mL contains about 25 mg of cilazapril (C22H31N3O5), and centrifuge.
To a quantity of powdered Cilazapril Tablets, equivalent to 2 mg of cilazapril (C22H31N3O5), add 2 mL of a mixture of acetonitrile and ethyl acetate (3:1), shake, sonicate for 30 seconds, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 5 mg of cilazapril in 5 mL of the mixture of acetonitrile and ethyl acetate (3:1), and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, acetic acid (100), hexane and water (62:15:10:10:3) to a dis-tance of about 15 cm, and air-dry the plate. Place the plate in iodine vapor for 2 hours, and immediately examine under ultraviolet light (main wavelength: 254 nm): the spots ob-tained from the sample and standard solutions are dark brown and they show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cilazapril Tablets add 5 mL of a mixture of water and acetonitrile (7:3), shake well until disintegration, add the mixture of water and acetonitrile (7:3) to make exactly V mL so that each mL contains about 25 mg of cilazapril (C22H31N3O5), and centrifuge. Pipet 4 mL of the supernatant liquid, add exactly 1 mL of the internal standard solution, add the mixture of water and acetonitrile (7:3) to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 26 mg of cilazapril for assay (separately determine the water <2.48> in the same manner as Cilazapril Hydrate), and dissolve in the mixture of water and acetonitrile (7:3) to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution, add the mixture of water and acetonitrile (7:3) to make 100 mL, and use this solution as the standard solution. Perform the test with 100 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilazapril to that of the internal standard. Amount (mg) of cilazapril (C22H31N3O5) ＝ MS × QT/QS × V/1000 MS: Amount (mg) of cilazapril for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of dimethyl phtha-late in a mixture of water and acetonitrile (7:3) (1 in 12,500). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above conditions, cilazapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of cilazapril to that of the internal standard is not more than 2.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cilazapril Tablets is not less than 85z. Start the test with 1 tablet of Cilazapril Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, and add water to make exactly V? mL so that each mL contains about 0.28 mg of cilazapril (C22H31N3O5). Pipet 10 mL of the solution, add exactly 5 mL of acetonitrile, and use this solution as the sample solution. Separately, weigh accu- MS: Amount (mg) of cilazapril for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of cilazapril (C22H31N3O5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To a solution consisting of 180 mL of tetra-hydrofuran for liquid chromatography, 120 mL of aceto-nitrile for liquid chromatography and 3 mL of triethylamine add water to make 1000 mL, and adjust the pH to 2.5 with phosphoric acid. Flow rate: Adjust so that the retention time of cilazapril is about 10 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cilazapril are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilazapril is not more than 2.0z.
(1) Chloride <1.03>—Perform the test using 1.0 g of Cilazapril Hydrate. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.009z). Sulfate <1.14>—Dissolve 1.0 g of Cilazapril Hydrate in 40 mL of water and 1.5 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.019z). Heavy metals <1.07>—Proceed with 1.0 g of Cilazapril Hydrate according to Method 4, and perform the test. However, use 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 8). Prepare the control so-lution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Cilazapril Hydrate in 20 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution (1). Pipet 3 mL of the standard solu-tion (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (2). Separately, pipet 2 mL of the standard solution (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (3). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and three standard solutions on a plate of silica gel Method of preparation Prepare as directed under Tablets, with Cilazapril Hydrate. Identification To a quantity of powdered Cilazapril Tablets, equivalent to 2 mg of cilazapril (C22H31N3O5), add 2 mL of a mixture of acetonitrile and ethyl acetate (3:1), shake, sonicate for 30 seconds, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 5 mg of cilazapril in 5 mL of the mixture of acetonitrile and ethyl acetate (3:1), and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol, acetic acid (100), hexane and water (62:15:10:10:3) to a dis-tance of about 15 cm, and air-dry the plate. Place the plate in iodine vapor for 2 hours, and immediately examine under ultraviolet light (main wavelength: 254 nm): the spots ob-tained from the sample and standard solutions are dark brown and they show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cilazapril Tablets add 5 mL of a mixture of water and acetonitrile (7:3), shake well until disintegration, add the mixture of water and acetonitrile (7:3) to make exactly V mL so that each mL contains about 25 mg of cilazapril (C22H31N3O5), and centrifuge. Pipet 4 mL of the supernatant liquid, add exactly 1 mL of the internal standard solution, add the mixture of water and acetonitrile (7:3) to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 26 mg of cilazapril for assay (separately determine the water <2.48> in the same manner as Cilazapril Hydrate), and dissolve in the mixture of water and acetonitrile (7:3) to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution, add the mixture of water and acetonitrile (7:3) to make 100 mL, and use this solution as the standard solution. Perform the test with 100 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilazapril to that of the internal standard. Amount (mg) of cilazapril (C22H31N3O5) ＝ MS × QT/QS × V/1000 MS: Amount (mg) of cilazapril for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of dimethyl phtha-late in a mixture of water and acetonitrile (7:3) (1 in 12,500). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above conditions, cilazapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of cilazapril to that of the internal standard is not more than 2.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Cilazapril Tablets is not less than 85z. Start the test with 1 tablet of Cilazapril Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, and add water to make exactly V? mL so that each mL contains about 0.28 mg of cilazapril (C22H31N3O5). Pipet 10 mL of the solution, add exactly 5 mL of acetonitrile, and use this solution as the sample solution. Separately, weigh accu- MS: Amount (mg) of cilazapril for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of cilazapril (C22H31N3O5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To a solution consisting of 180 mL of tetra-hydrofuran for liquid chromatography, 120 mL of aceto-nitrile for liquid chromatography and 3 mL of triethylamine add water to make 1000 mL, and adjust the pH to 2.5 with phosphoric acid. Flow rate: Adjust so that the retention time of cilazapril is about 10 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cilazapril are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilazapril is not more than 2.0z.
Weigh acurately the mass of not less than 20 Cilazapril Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 1 mg of cilazapril (C22H31N3O5), add 30 mL of a mixture of water and aceto-nitrile (7:3), and sonicate for 5 minutes. Next, add exactly 5 mL of the internal standard solution, add the mixture of water and acetonitrile (7:3) to make 50 mL, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 26 mg of cilazapril for assay (separately determine the water <2.48> in the same manner as Cilazapril Hydrate), and dissolve in the mixture of water and acetonitrile (7:3) to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add the mixture of water and aceto-nitrile (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of cilazapril to that of the internal standard. rately about 29 mg of cilazapril for assay (separately deter-mine the water <2.48> in the same manner as Cilazapril Hy-drate), and dissolve in water to make exactly 100 mL. Pipet 5 Amount (mg) of cilazapril (C ＝ MS × QT/QS × 1/25 22H31N3O5) mL of the solution, add water to make exactly 100 mL. Then, pipet 2 mL of this solution, add water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL of acetonitrile, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cilazapril in each so-lution. Dissolution rate (z) with respect to the labeled amount of cilazapril (C22H31N3O5) ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of cilazapril for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of dimethyl phtha-late in a mixture of water and acetonitrile (7:3) (1 in 12,500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 239C. ‌JP XVIII Official Monographs / Cilnidipine 757 Mobile phase: To a solution consisting of 180 mL of tetra-hydrofuran for liquid chromatography, 120 mL of aceto-nitrile for liquid chromatography and 3 mL of triethylamine add water to make 1000 mL, and adjust the pH to 2.5 with phosphoric acid. Flow rate: Adjust so that the retention time of cilazapril is about 10 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, cilazapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cilazapril to that of the internal standard is not more than 1.0z.
Cilnidipine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of cil-nidipine (C27H28N2O7: 492.52). Method of preparation Prepare as directed under Tablets, with Cilnidipine. Identification Powder Cilnidipine Tablets. To a portion of the powder, equivalent to 20 mg of Cilnidipine, add 20 mL of methanol, shake well, and centrifuge. To 1 mL of the su-pernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm and between 350 nm and 360 nm. Purity Related substances—Conduct this procedure using light-resistant vessels. Powder Cilnidipine Tablets. To a por-tion of the powder, equivalent to 25 mg of Cilnidipine, add 40 mL of the mobile phase, shake well, and add the mobile phase to make 50 mL. Centrifuge, and use the supernatant liquid as the sample solution. Pipet 3 mL of the sample solu-tion, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 1.09 to cilnidipine, obtained from the sample solution is not larger than 1/3 times the peak area of cilnidipine from the standard solution, the area of the peaks other than cilnidipine and the peak mentioned above from the sample solution is not larger than 2/15 times the peak area of cilnidipine from the stand-ard solution, and the total area of the peaks other than cil-nidipine from the sample solution is not larger than the peak area of cilnidipine from the standard solution. For the area of the peak, having the relative retention time of about 1.09 to cilnidipine, multiply the correction factor, 1.4. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of cilnidipine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 150 mL. Confirm that the peak area of cilnidipine obtained with 20 mL of this solution is equivalent to 2.4 to 4.3z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of cilnidipine are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilnidipine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Cilnidipine Tablets add V/10 mL of water, and shake to completely disintegrate the tablet. Add acetonitrile to make exactly V mL so that each mL contains about 0.
Powder Cilnidipine Tablets. To a portion of the powder, equivalent to 20 mg of Cilnidipine, add 20 mL of methanol, shake well, and centrifuge. To 1 mL of the su-pernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm and between 350 nm and 360 nm.
Related substances—Conduct this procedure using light-resistant vessels. Powder Cilnidipine Tablets. To a por-tion of the powder, equivalent to 25 mg of Cilnidipine, add 40 mL of the mobile phase, shake well, and add the mobile phase to make 50 mL. Centrifuge, and use the supernatant liquid as the sample solution. Pipet 3 mL of the sample solu-tion, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 1.09 to cilnidipine, obtained from the sample solution is not larger than 1/3 times the peak area of cilnidipine from the standard solution, the area of the peaks other than cilnidipine and the peak mentioned above from the sample solution is not larger than 2/15 times the peak area of cilnidipine from the stand-ard solution, and the total area of the peaks other than cil-nidipine from the sample solution is not larger than the peak area of cilnidipine from the standard solution. For the area of the peak, having the relative retention time of about 1.09 to cilnidipine, multiply the correction factor, 1.4. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of cilnidipine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 150 mL. Confirm that the peak area of cilnidipine obtained with 20 mL of this solution is equivalent to 2.4 to 4.3z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of cilnidipine are not less than 15,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilnidipine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Cilnidipine Tablets add V/10 mL of water, and shake to completely disintegrate the tablet. Add acetonitrile to make exactly V mL so that each mL contains about 0.2 mg of cilnidipine (C27H28N2O7), and centrifuge. Pipet 4 mL of the supernatant liquid, add a mixture of acetonitrile and water (9:1) to make exactly 20 mL, filter, if necessary, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Cilnidipine RS, previously dried in vacuum at 609C for 3 hours, dissolve in a mixture of acetonitrile and water (9:1) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of acetonitrile and water (9:1) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 355 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24> using a mixture of acetonitrile and water (9:1) as the control. Amount (mg) of cilnidipine (C27H28N2O7) ＝ MS × AT/AS × V/100 MS: Amount (mg) of Cilnidipine RS taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of polysorbate 80 (dissolving 1 g of poly-sorbate 80 in 1000 mL of 2nd fluid for dissolution test) as the dissolution medium, the dissolution rate in 90 minutes of Cilnidipine Tablets is not less than 70z. Start the test with 1 tablet of Cilnidipine Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent ‌JP XVIII Official Monographs / Cilostazol 759 filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of cilnidipine (C27H28N2O7), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Cilnidipine RS, previously dried in vacuum at 609C for 3 hours, dissolve in acetonitrile to make exactly 100 mL. Pipet 2 mL of this solu-tion, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cilnidipine in each solution. Dissolution rate (z) with respect to the labeled amount of cilnidipine (C27H28N2O7) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Cilnidipine RS taken C: Labeled amount (mg) of cilnidipine (C27H28N2O7) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.58 g of disodium hydrogen phos-phate dodecahydrate in 1000 mL of water, and adjust to pH 6.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of cilnidipine is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cilnidipine are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilnidipine is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 Cilnidipine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg of cilnidipine (C27H28N2O7), add 40 mL of the mobile phase, shake well, and add the mobile phase to make exactly 50 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 2.5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Cilnidipine RS, previously dried in vacuum at 609C for 3 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 2.5 mL of the inter-nal standard solution, add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilnidipine to that of the in-ternal standard. Internal standard solution—A solution of 4,4'-difluoroben-zophenone in the mobile phase (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 6 mm in inside diameter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.58 g of disodium hydrogen phos-phate dodecahydrate in 1000 mL of water, and adjust to pH 6.0 with phosphoric acid. To 400 mL of this solution add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of cilnidipine is about 23 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cilnidipine are eluted in this order with the resolution between these peaks being not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cilnidipine to that of the internal standard is not more than 1.0z.
Cilostazol Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of cilostazol (C20H27N5O2: 369.46). Method of preparation Prepare as directed under Tablets, with Cilostazol. Identification Mix well an amount of powdered Cilostazol Tablets, equivalent to 50 mg of Cilostazol, with 10 mL of acetone, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 25 mg of Cilostazol RS in 5 mL of acetone, and use this solution as the standard so-lution. Perform the test with these solutions as directed un-der Thin-layer Chromatography <2.03>. Spot 6 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, acetonitrile, methanol and formic acid (75:25:5:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the principal spot obtained from the sample solution and the spot from the standard solution are orange in color and have the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cilostazol Tablets add 2 mL of water to disintegrate the tablet, add the internal standard solution exactly 5 mL for a 50-mg tablet and exactly 10 mL for a 100-mg tablet, and add methanol to make 50 mL. Shake for ‌JP XVIII Official Monographs / Cimetidine 761 10 minutes for the 50-mg tablet and for 20 minutes for the 100-mg tablet. To 1 mL of the solution add methanol to make 10 mL for the 50-mg tablet and 20 mL for the 100-mg tablet, filter through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Proceed as directed in the Assay. Amount (mg) of cilostazol (C20H27N5O2) ＝ MS × QT/QS × C/50 MS: Amount (mg) of Cilostazol RS taken C: Labeled amount (mg) of cilostazol (C20H27N5O2) in 1 tablet Internal standard solution—A solution of benzophenone in methanol (1 in 250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of sodium lauryl sulfate (3 in 1000) as the dissolution medium, the dissolution rates of a 50-mg tablet in 45 minutes and a 100-mg tablet in 60 minutes are not less than 75z and not less than 70z, respectively. Start the test with 1 tablet of Cilostazol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.
Mix well an amount of powdered Cilostazol Tablets, equivalent to 50 mg of Cilostazol, with 10 mL of acetone, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 25 mg of Cilostazol RS in 5 mL of acetone, and use this solution as the standard so-lution. Perform the test with these solutions as directed un-der Thin-layer Chromatography <2.03>. Spot 6 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop the plate with a mixture of ethyl acetate, acetonitrile, methanol and formic acid (75:25:5:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: the principal spot obtained from the sample solution and the spot from the standard solution are orange in color and have the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Cilostazol Tablets add 2 mL of water to disintegrate the tablet, add the internal standard solution exactly 5 mL for a 50-mg tablet and exactly 10 mL for a 100-mg tablet, and add methanol to make 50 mL. Shake for ‌JP XVIII Official Monographs / Cimetidine 761 10 minutes for the 50-mg tablet and for 20 minutes for the 100-mg tablet. To 1 mL of the solution add methanol to make 10 mL for the 50-mg tablet and 20 mL for the 100-mg tablet, filter through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Proceed as directed in the Assay. Amount (mg) of cilostazol (C20H27N5O2) ＝ MS × QT/QS × C/50 MS: Amount (mg) of Cilostazol RS taken C: Labeled amount (mg) of cilostazol (C20H27N5O2) in 1 tablet Internal standard solution—A solution of benzophenone in methanol (1 in 250). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of sodium lauryl sulfate (3 in 1000) as the dissolution medium, the dissolution rates of a 50-mg tablet in 45 minutes and a 100-mg tablet in 60 minutes are not less than 75z and not less than 70z, respectively. Start the test with 1 tablet of Cilostazol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of cilostazol (C20H27N5O2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Cilostazol RS, previously dried at 1059C for 2 hours, and dissolve in metha-nol to make exactly 100 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 257 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24> using the dissolution medium as the control. Dissolution rate (z) with respect to the labeled amount of cilostazol (C20H27N5O2) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Cilostazol RS taken C: Labeled amount (mg) of cilostazol (C20H27N5O2) in 1 tablet
Weigh accurately the mass of not less than 20 Cilostazol Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of cilostazol (C20H27N5O2), add exactly 5 mL of the internal standard solution and methanol to make 50 mL, and shake well for 10 minutes. To 1 mL of this solution add methanol to make 10 mL, filter through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 50 mg of Cilostazol RS, previously dried at 1059C for 2 hours, dissolve in a suitable amount of methanol, and add exactly 5 mL of the internal standard solution, and add methanol to make 50 mL. To 1 mL of this solution add methanol to make 10 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cilostazol to that of the internal standard. MS: Amount (mg) of Cilostazol RS taken Internal standard solution—A solution of benzophenone in methanol (1 in 250). Operating conditions— Proceed as directed in the operating conditions in the Assay under Cilostazol. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Cilostazol. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cilostazol to that of the internal standard is not more than 1.5z.
Cinoxacin Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of cinox-acin (C12H10N2O5: 262.22). Method of preparation Prepare as directed under Cap-sules, with Cinoxacin. Identification To a quantity of the contents of Cinoxacin Capsules, equivalent to 10 mg of Cinoxacin, add 20 mL of acetone, shake well, and centrifuge. To 3 mL of the superna-tant liquid add acetone to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of cinoxa- JP XVIII Official Monographs / Ciprofloxacin 763 ‌cin for assay in 20 mL of acetone. To 3 mL of this solution add acetone to make 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of aceto-nitrile, water and ammonia solution (28) (14:4:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show a blue-purple color and the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Cinoxacin Capsules add 40 mL of dilute sodium hydroxide TS, and dissolve the capsule in lukewarm water with occasional shaking. After cooling, add water and shake well, add water to make exactly V mL so that each mL contains about 1 mg of cinoxacin (C H N O ), and filter.
To a quantity of the contents of Cinoxacin Capsules, equivalent to 10 mg of Cinoxacin, add 20 mL of acetone, shake well, and centrifuge. To 3 mL of the superna-tant liquid add acetone to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of cinoxa- JP XVIII Official Monographs / Ciprofloxacin 763 ‌cin for assay in 20 mL of acetone. To 3 mL of this solution add acetone to make 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of aceto-nitrile, water and ammonia solution (28) (14:4:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show a blue-purple color and the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Cinoxacin Capsules add 40 mL of dilute sodium hydroxide TS, and dissolve the capsule in lukewarm water with occasional shaking. After cooling, add water and shake well, add water to make exactly V mL so that each mL contains about 1 mg of cinoxacin (C H N O ), and filter.
Weigh accurately the mass of not less than 20 Cinoxacin Capsules, take out the contents, and powder. Wash the capsule shells with a small amount of diethyl ether, allow to stand at room temperature to vaporize the diethyl ether, weigh accurately the mass of the capsule shells, and calculate the mass of the contents. Weigh accurately a por-tion of the powder, equivalent to about 50 mg of cinoxacin (C12H10N2O5), add 10 mL of dilute sodium hydroxide TS, shake, add water to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet 1 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of cinoxacin for assay, previously dried at 1059C for 1 hour, dissolve in 10 mL of dilute sodium hydroxide TS, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add 0.1 mol/L hydro-chloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absor-bances, AT and AS, at 354 nm. 12 10 2 5 Discard the first 20 mL of the filtrate, pipet 1 mL of the sub-sequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 0.2 g of cinoxacin for assay, previously dried at 1059C for 1 hour, dissolve in 40 mL of dilute sodium hydroxide TS, and add water to make exactly 200 mL. Pipet 1 mL of this solution, add 0.1 mol/L of hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 354 nm. Amount (mg) of cinoxacin (C12H10N2O5) ＝ MS × AT/AS × V/200 MS: Amount (mg) of cinoxacin for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of 2nd solution for dissolution test as the dissolution medium, the dissolution rate in 90 minutes of Cinoxacin Capsules is not less than 70z. Start the test with 1 capsule of Cinoxacin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of cinoxacin (C12H10N2O5), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of cinoxacin for assay, previously dried at 1059C for 1 hour, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 351 nm. Dissolution rate (z) with respect to the labeled amount of cinoxacin (C12H10N2O5) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of cinoxacin for assay taken C: Labeled amount (mg) of cinoxacin (C12H10N2O5) in 1 capsule Amount (mg) of cinoxacin (C12H10N2O5) ＝ MS × AT/AS MS: Amount (mg) of cinoxacin for assay taken
Clarithromycin for Syrup is a preparation for syrup, which is suspended before use. It contains not less than 90.
Clarithromycin Tablets contain not less than 93.0z and not more than 107.0z of the labeled potency of with 100 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak clarithromycin (C 38H69 NO13 : 747.95). area of clarithromycin is not more than 2.0z. Assay Weigh accurately an amount of crushed Clarithro-mycin for Syrup, equivalent to about 50 mg (potency) of Clarithromycin, add 60 mL of ethanol (99.5), add exactly 10 mL of the internal standard solution, sonicate with occa-sional vigorous shaking, and add ethanol (99.5) to make 100 mL. Centrifuge this solution, and ˆlter the supernatant liq-uid through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard the ˆrst 3 mL of the ˆltrate, and use the subsequent ˆltrate as the sample solution. Separately, weigh accurately an amount of Clarithromycin RS, equivalent about 50 mg (potency), and dissolve in ethanol (99.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 2 mL of the internal standard solution, add ethanol (99.5) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clarithromycin to that of the internal standard. Amount [mg (potency)] of clarithromycin (C38H69NO13) ＝ MS × QT/QS MS: Amount [mg (potency)] of Clarithromycin RS taken Method of preparation Prepare as directed under Tablets, with Clarithromycin. Identification Shake a quantity of powdered Clarithromy-cin Tablets, equivalent to 60 mg (potency) of Clarithromy-cin, with 40 mL of acetone for 10 minutes, and centrifuge at 4000 rpm for 5 minutes. Evaporate 30 mL of the supernatant liquid, and determine the infrared absorption spectrum of the residue so obtained as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 2980 cm－1, 2940 cm－1, 1734 cm－1, 1693 cm－1, 1459 cm－1, 1379 cm－1 and 1171 cm－1. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Clarithromycin Tablets add exactly V/20 mL of the internal standard solution (1), then add the mobile phase so that each mL contains about 5 mg (potency) of clarithromycin (C38H69NO13) to make V mL, and disperse to fine particles by sonicating for 20 minutes while occasional vigorous shaking.
Shake a quantity of powdered Clarithromy-cin Tablets, equivalent to 60 mg (potency) of Clarithromy-cin, with 40 mL of acetone for 10 minutes, and centrifuge at 4000 rpm for 5 minutes. Evaporate 30 mL of the supernatant liquid, and determine the infrared absorption spectrum of the residue so obtained as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 2980 cm－1, 2940 cm－1, 1734 cm－1, 1693 cm－1, 1459 cm－1, 1379 cm－1 and 1171 cm－1. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Clarithromycin Tablets add exactly V/20 mL of the internal standard solution (1), then add the mobile phase so that each mL contains about 5 mg (potency) of clarithromycin (C38H69NO13) to make V mL, and disperse to fine particles by sonicating for 20 minutes while occasional vigorous shaking. Centrifuge this solution at 4000 rpm for 15 minutes, and filter the supernatant liquid through a mem-brane filter with a pore size not exceeding 0.45 mm. Then, proceed as directed in the Assay. Internal standard solution—A solution of butyl para- Amount [mg (potency)] of clarithromycin (C H NO ) hydroxybenzoate in ethanol (99.5) (1 in 12,500). Operating conditions— ＝ MS × QT/QS × V/10 38 69 13 Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in 3) and acetonitrile for liquid chromatography (13:7). Flow rate: Adjust so that the retention time of clarithromycin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, clarithromycin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of clarithromycin to that of the internal stand-ard is not more than 2.0z.
Weigh accurately an amount of crushed Clarithro-mycin for Syrup, equivalent to about 50 mg (potency) of Clarithromycin, add 60 mL of ethanol (99.5), add exactly 10 mL of the internal standard solution, sonicate with occa-sional vigorous shaking, and add ethanol (99.5) to make 100 mL. Centrifuge this solution, and ˆlter the supernatant liq-uid through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard the ˆrst 3 mL of the ˆltrate, and use the subsequent ˆltrate as the sample solution. Separately, weigh accurately an amount of Clarithromycin RS, equivalent about 50 mg (potency), and dissolve in ethanol (99.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 2 mL of the internal standard solution, add ethanol (99.5) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clarithromycin to that of the internal standard. Amount [mg (potency)] of clarithromycin (C38H69NO13) ＝ MS × QT/QS MS: Amount [mg (potency)] of Clarithromycin RS taken Method of preparation Prepare as directed under Tablets, with Clarithromycin. Identification Shake a quantity of powdered Clarithromy-cin Tablets, equivalent to 60 mg (potency) of Clarithromy-cin, with 40 mL of acetone for 10 minutes, and centrifuge at 4000 rpm for 5 minutes. Evaporate 30 mL of the supernatant liquid, and determine the infrared absorption spectrum of the residue so obtained as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 2980 cm－1, 2940 cm－1, 1734 cm－1, 1693 cm－1, 1459 cm－1, 1379 cm－1 and 1171 cm－1. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Clarithromycin Tablets add exactly V/20 mL of the internal standard solution (1), then add the mobile phase so that each mL contains about 5 mg (potency) of clarithromycin (C38H69NO13) to make V mL, and disperse to fine particles by sonicating for 20 minutes while occasional vigorous shaking. Centrifuge this solution at 4000 rpm for 15 minutes, and filter the supernatant liquid through a mem-brane filter with a pore size not exceeding 0.45 mm. Then, proceed as directed in the Assay. Internal standard solution—A solution of butyl para- Amount [mg (potency)] of clarithromycin (C H NO ) hydroxybenzoate in ethanol (99.5) (1 in 12,500). Operating conditions— ＝ MS × QT/QS × V/10 38 69 13 Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted 0.2 mol/L potassium dihydrogen phosphate TS (1 in 3) and acetonitrile for liquid chromatography (13:7). Flow rate: Adjust so that the retention time of clarithromycin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, clarithromycin and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of clarithromycin to that of the internal stand-ard is not more than 2.0z.
クリンダマイシン塩酸塩カプセル
Clindamycin Hydrochloride Capsules contain not less than 93.0z and not more than 107.0z of the labeled potency of clindamycin (C H ClN O S: ride. Centrifuge this solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of clindamycin (C18H33ClN2O5S) ＝ MS × AT/AS × V/100 MS: Amount [mg (potency)] of Clindamycin Hydrochlo-ride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate of a 75-mg capsule in 15 minutes and that of a 150-mg capsule in 30 minutes are not less than 80z, respectively. Start the test with 1 capsule of Clindamycin Hydrochlo-ride Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make ex-actly V? so that each mL contains about 83 mg (potency) of Clindamycin Hydrochloride, and use this solution as the sample solution.
To an amount of the contents of Clindamy-cin Hydrochloride Capsules, equivalent to 10 mg (potency) of Clindamycin Hydrochloride, add 2 mL of methanol, shake well, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of Clindamycin Hydrochloride RS in 2 mL of methanol, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol, toluene and ammonia solution (28) (140:60:3) to a distance of about 12 cm, and air-dry the plate. Spray evenly a mixture of 500 mL of a solution of L-tartaric acid (1 in 5) and 50 mL of bismuth subnitrate TS on the plate: the Rf values of the principal spot with the sample solution and the spot with the standard solution are not different each other. Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust the pH of 0.05 mol/L potassium di-hydrogen phosphate TS to 7.5 with 8 mol/L potassium hy-droxide TS. To 550 mL of this solution add 450 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of clindamy-cin is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clindamycin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times ‌JP XVIII Official Monographs / Clindamycin Phosphate 777 with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clindamycin is not more than 2.0z.
Take out the contents of not less than 20 Clindamy-cin Hydrochloride Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 75 mg (potency) of Clindamycin Hydrochloride, add the mobile phase, shake for 30 minutes, and add the mobile phase to make exactly 100 mL. Centri-fuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately an amount of Clindamycin Hydrochloride RS, equivalent to about 75 mg (potency), dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of clindamycin in each solution. Clindamycin Phosphate
クリンダマイシンリン酸エステル注射液
Clindamycin Phosphate Injection is an aqueous in-jection. It contains not less than 90.
Clindamycin Phosphate Injection is a clear, colorless or light yellow liquid.
To a volume of Clindamycin Phosphate Injection, equivalent to 0.15 g (potency) of Clindamycin Phosphate, add 4 mL of water, 2 mL of 8 mol/L sodium hy-droxide TS and 0.1 mL of sodium pentacyanonitrosylferrate (III) TS, mix, heat in a water bath for 10 minutes, and add 2 mL of hydrochloric acid: a blue-green color develops. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> 6.0 – 7.0 Bacterial endotoxins <4.01> Less than 0.1 EU/mg (po-tency). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Clindamycin Phos-phate Injection, equivalent to about 0.3 g (potency) of Clin-damycin Phosphate, and add the mobile phase to make ex-actly 100 mL. Pipet 7 mL of this solution, add exactly 25 mL of the internal standard solution and the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Clindamycin Phosphate RS, equivalent to about 20 mg (potency), dissolve in exactly 25 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Clindamycin Phosphate. Amount [mg (potency)] of clindamycin phosphate (C18H34ClN2O8PS) ＝ MS × QT/QS × 100/7 MS: Amount [mg (potency)] of Clindamycin Phosphate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (3 in 50,000).
Cut and open Clofibrate Capsules, and use the contents as the sample. Determine the absorption spectrum of a solution of the sample in ethanol (99.5) (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 278 nm and 282 nm, and it exhibits a maximum between 224 nm and 228 nm after diluting this solution 10 times with ethanol (99.5).
p-Chlorophenol—Cut and open not less than 20 Clofibrate Capsules, and proceed with 1.0 g of the well- mixed contents as directed in the Purity (4) under Clofibrate. Internal standard solution—A solution of 4-ethoxyphenol in the mobile phase (1 in 30,000).
Weigh accurately not less than 20 Clofibrate Cap-sules, cut and open the capsules, rinse the inside of the cap-sules with a small amount of diethyl ether after taking out the contents, evaporate the diethyl ether by allowing the cap-sules to stand at room temperature, and weigh the capsules accurately. Weigh accurately an amount of the contents, equivalent to about 0.1 g of clofibrate (C12H15ClO3), dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Clofibrate RS, proceed in the same manner as directed for the sample solution, and use the solution so obtained as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clofibrate to that of the internal standard. Amount (mg) of clofibrate (C12H15ClO3) ＝ MS × QT/QS MS: Amount (mg) of Clofibrate RS taken Internal standard solution—A solution of ibuprofen in the mobile phase (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column about 4 mm in inside diameter and about 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile and diluted phos-phoric acid (1 in 1000) (3:2). Flow rate: Adjust so that the retention time of clofibrate is about 10 minutes. Selection of column: Dissolve 0.05 g of clofibrate and 0.3 g of ibuprofen in 50 mL of acetonitrile. Proceed with 10 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of ibuprofen and clofibrate in this order with the resolution be-tween these peaks being not less than 6.
Clomifene Citrate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of the clomifene citrate (C26H28ClNO.C6H8O7: 598.08). Method of preparation Prepare as directed under Tablets, with Clomifene Citrate. Identification Weigh a portion of powdered Clomifene Citrate Tablets, equivalent to 50 mg of Clomifene Citrate, shake vigorously with 50 mL of methanol for 10 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of Clomifene Citrate RS in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 2-propanol, tolu-ene and diethylamine (10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clomifene Citrate Tablets add 10 mL of water, and shake until the tablets are disintegrated. To this ‌JP XVIII Official Monographs / Clomipramine Hydrochloride 785 solution add 50 mL of methanol, shake for 10 minutes, and add methanol to make exactly 100 mL. Centrifuge this solu-tion, pipet 4 mL of the supernatant liquid, add methanol to make exactly V mL so that each mL contains about 20 mg of clomifene citrate (C26H28ClNO.
Weigh a portion of powdered Clomifene Citrate Tablets, equivalent to 50 mg of Clomifene Citrate, shake vigorously with 50 mL of methanol for 10 minutes, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of Clomifene Citrate RS in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 2-propanol, tolu-ene and diethylamine (10:10:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots from the sample solution and standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clomifene Citrate Tablets add 10 mL of water, and shake until the tablets are disintegrated. To this ‌JP XVIII Official Monographs / Clomipramine Hydrochloride 785 solution add 50 mL of methanol, shake for 10 minutes, and add methanol to make exactly 100 mL. Centrifuge this solu-tion, pipet 4 mL of the supernatant liquid, add methanol to make exactly V mL so that each mL contains about 20 mg of clomifene citrate (C26H28ClNO.C6H8O7), and use this solu-tion as the sample solution. Proceed as directed in the Assay. Amount (mg) of clomifene citrate (C26H28ClNO.C6H8O7) ＝ MS × AT/AS × V/100 MS: Amount (mg) of Clomifene Citrate RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Clomifene Citrate Tablets is not less than 80z. Start the test with 1 tablet of Clomifene Citrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 28 mg of clomifene citrate (C26H28ClNO.C6H8O7), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Clomifene Citrate RS, previously dried in vacuum using phosphorus (V) oxide as a desiccant for 3 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 291 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of clomifene citrate (C26H28ClNO.C6H8O7) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Clomifene Citrate RS taken C: Labeled amount (mg) of clomifene citrate (C26H28ClNO.C6H8O7) in 1 tablet
Weigh accurately, and powder not less than 20 Clomifene Citrate Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of clomifene citrate (C26H28ClNO.C6H8O7), add 50 mL of methanol, shake for 10 minutes, and add methanol to make exactly 100 mL. Cen-trifuge a portion of this solution, pipet 4 mL of the superna-tant liquid, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of Clomifene Citrate RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 3 hours, and dissolve in methanol to make exactly 100 mL. Pipet 4 mL of this solution, and dilute with methanol to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and the standard solution, respectively, at 295 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of clomifene citrate (C26H28ClNO.C6H8O7) ＝ MS × AT/AS MS: Amount (mg) of Clomifene Citrate RS taken
To a portion of powdered Clomipramine Hydrochloride Tablets, equivalent to 50 mg of Clomipra-mine Hydrochloride, add a suitable amount of 0.1 mol/L hydrochloric acid TS, shake thoroughly, and add 0.1 mol/L hydrochloric acid TS to make 250 mL. Centrifuge this solu-tion, and to 10 mL of the supernatant liquid add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 250 nm and 254 nm. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clomipramine Hydrochloride Tablets add V/5 mL of a mixture of methanol and 0.1 mol/L hydrochlo- ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of clomipramine hydrochloride for assay taken C: Labeled amount (mg) of clomipramine hydrochloride (C19H23ClN2.HCl) in 1 tablet
Weigh accurately the mass of not less than 20 Clomipramine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg of clomipramine hydrochloride (C19H23ClN2.HCl), add 50 mL of a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1), sonicate, and shake thoroughly for 30 minutes. To this solution add 150 mL of methanol, shake for 15 minutes, and add methanol to make exactly 250 mL. Cen-trifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of clomipramine hydrochloride for assay, previously dried at 1059C for 3 hours, dissolve in 50 mL of a mixture of metha-nol and 0.1 mol/L hydrochloric acid TS (3:1), add methanol to make exactly 250 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of clomipramine in each solution. Amount (mg) of clomipramine hydrochloride (C19H23ClN2.HCl) ＝ MS × AT/AS MS: Amount (mg) of clomipramine hydrochloride for ‌JP XVIII Official Monographs / Clonazepam Fine Granules 787 assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2 g of sodium 1-octanesulfonate in 300 mL of water, and add 450 mL of methanol, 250 mL of acetonitrile and 1 mL of 0.5 mol/L sulfuric acid TS. Flow rate: Adjust so that the retention time of clomipra-mine is about 13 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clomipramine are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clomipramine is not more than 1.0z.
Powder Clonazepam Fine Granules. To a portion of the powder, equivalent to 1 mg of Clonazepam, add an appropriate volume of methanol and shake for 10 minutes, add methanol to make 100 mL, and filter. Deter-mine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 307 nm and 311 nm. Dissolution Being specified separately when the drug is granted approval based on the Law. ‌788
Powder Clonazepam Fine Granules. Weigh accu-rately a portion of the powder, equivalent to about 2.4 mg of clonazepam (C15H10ClN3O3), add exactly 30 mL of a mixture of methanol and water (7:3), and shake for 15 minutes. Cen-trifuge this solution, pipet 5 mL of the supernatant liquid, add a mixture of methanol and water (7:3) to make exactly 20 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 20 mg of clonazepam for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add a mixture of methanol and water (7:3) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of clonazepam in each solution. Amount (mg) of clonazepam (C15H10ClN3O3) ＝ MS × AT/AS × 3/25 MS: Amount (mg) of clonazepam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 310 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and metha-nol (4:3:3). Flow rate: Adjust so that the retention time of clonazepam is about 5 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clonazepam are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clonazepam is not more than 1.0z.
Clonazepam Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of clonazepam (C15H10ClN3O3: 315.71). Method of preparation Prepare as directed under Tablets, with Clonazepam. Identification Powder Clonazepam Tablets. To a portion of the powder, equivalent to 1 mg of Clonazepam, add an appropriate volume of methanol and shake for 10 minutes, then add methanol to make 100 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 307 nm and 311 nm. Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clonazepam Tablets, add V/10 mL of methanol, shake for 15 minutes, add 2-propanol to make exactly V mL so that each mL contains about 10 mg of clonazepam (C15H10ClN3O3).
Powder Clonazepam Tablets. To a portion of the powder, equivalent to 1 mg of Clonazepam, add an appropriate volume of methanol and shake for 10 minutes, then add methanol to make 100 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 307 nm and 311 nm. Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clonazepam Tablets, add V/10 mL of methanol, shake for 15 minutes, add 2-propanol to make exactly V mL so that each mL contains about 10 mg of clonazepam (C15H10ClN3O3). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of clonazepam for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 200 mL. Pipet 10 mL of this solution, add 2-propanol to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, at 312 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a mix-ture of 2-propanol and methanol (9:1) as the control. Amount (mg) of clonazepam (C15H10ClN3O3) ＝ MS × AT/AS × V/2000 MS: Amount (mg) of clonazepam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of 0.5-mg tablet and 1-mg tablet is not less than 80z, and that of 2-mg tablet is not less than 75z. Start the test with 1 tablet of Clonazepam Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less then 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.56 mg of clonazepam (C15H10ClN3O3), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of clonazepam for assay, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution and add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of clonazepam in each solution. Dissolution rate (z) with respect to the labeled amount of clonazepam (C15H10ClN3O3) ＝ MS × AT/AS × V?/V × 1/C × 9/4 MS: Amount (mg) of clonazepam for assay taken C: Labeled amount (mg) of clonazepam (C15H10ClN3O3) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clonazepam are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clonazepam is not more than 2.0z.
Weigh accurately the mass of not less than 20 ‌JP XVIII Official Monographs / Clonidine Hydrochloride 789 Clonazepam Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 2.5 mg of clonazep-am (C15H10ClN3O3), add exactly 50 mL of a mixture of meth-anol and water (7:3), and shake for 15 minutes. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of clonazepam for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add a mixture of methanol and water (7:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of clonazepam in each solution. Amount (mg) of clonazepam (C15H10ClN3O3) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of clonazepam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 310 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and metha-nol (4:3:3). Flow rate: Adjust so that the retention time of clonazepam is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clonazepam are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clonazepam is not more than 1.0z.
クロペラスチンフェンジゾ酸塩錠
To a quantity of powdered Cloperastine Fendizoate Tablets, equivalent to 1.5 mg of Cloperastine Fendizoate, add methanol, shake thoroughly, add methanol to make 100 mL, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 248 nm and 252 nm, and between 282 nm and 286 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Cloperastine Fendizoate Tablets add exactly Dissolution rate (z) with respect to the labeled amount of cloperastine fendizoate (C20H24ClNO.C20H14O4) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of cloperastine fendizoate for assay taken C: Labeled amount (mg) of cloperastine fendizoate (C20H24ClNO.C20H14O4) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, fendizoic acid and cloperastine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cloperastine is not more than 2.0z.
Weigh accurately the mass of not less than 20 tablets of Cloperastine Fendizoate Tablets, and powder. Weigh ac- curately a portion of the powder, equivalent to about 4.4 mg V/10 mL of the internal standard solution, add the mobile phase, shake vigorously until the tablet is disintegrated, add of cloperastine fendizoate (C 20H 24ClNO.C 20H 14O4), add ex- the mobile phase to make V mL so that each mL contains actly 5 mL of the internal standard solution, add 20 mL of about 88 mg of cloperastine fendizoate (C 20H 24ClNO. the mobile phase, shake vigorously for 10 minutes, then add the mobile phase to make 50 mL, and filter through a mem- C20H14O4), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of cloperastine fendizoate (C20H24ClNO.C20H14O4) ＝ MS × QT/QS × V/250 MS: Amount (mg) of cloperastine fendizoate for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mobile phase (3 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 90 minutes of Cloperastine Fen-dizoate Tablets is not less than 75z. Start the test with 1 tablet of Cloperastine Fendizoate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 4.9 mg of cloperastine fendizoate (C20H24ClNO.C20H14O4), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of cloperastine fendizoate for assay, previously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 200 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cloperastine in each solution. brane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 22 mg of cloperastine fendizoate for assay, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 50 mL. Pipet 10 mL of this solution, add ex-actly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01>, and calculate the ratios, QT and QS, of the peak area of cloperastine to that of the internal standard. Amount (mg) of cloperastine fendizoate (C20H24ClNO.C20H14O4) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of cloperastine fendizoate for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mobile phase (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 226 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.1 mol/L potassium dihydro-gen phosphate TS, acetonitrile for liquid chromatography and perchloric acid (400:320:1). Flow rate: Adjust so that the retention time of cloper-astine is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard, fendizoic acid and cloper- ‌792 astine are eluted in this order, and each resolution between these peaks is not less than 5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cloperastine to that of the internal standard is not more than 1.0z.
Clopidogrel Sulfate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of clopidogrel (C16H16ClNO2S: 321.82). Method of preparation Prepare as directed under Tablets, with Clopidogrel Sulfate. Identification To a quantity of powdered Clopidogrel Sulfate Tablets, equivalent to 75 mg of clopidogrel (C16H16ClNO2S), add 50 mL of methanol, and after sonicat-ing with ultrasonic waves with occasional shaking, add meth-anol to make 100 mL. To 10 mL of this solution add metha-nol to make 30 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 269 nm and 273 nm, and between 276 nm and 280 nm. Purity Related substances—Keep the sample solution and the standard solution at 59C or below and use within 24 hours. Take a quantity of Clopidogrel Sulfate Tablets equivalent to 0.15 g of clopidogrel (C16H16ClNO2S), add 120 mL of the mobile phase, sonicate with occasional shaking until the tablets are disintegrated, and add the mobile phase to make 200 mL. Centrifuge this solution, to 10 mL of the supernatant liquid add the mobile phase to make 30 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet Amount (mg) of clopidogrel sulfate (C H ClNO S.H SO ) 2 mL of the sample solution, add the mobile phase to make ＝ MS × AT/AS 16 16 2 2 4 exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 0.87 g of sodium 1-pen-tanesufonate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 950 mL of this solution add 50 mL of methanol. To 600 mL of this solution, add 400 mL of a mix-ture of acetonitrile for liquid chromatography and methanol (19:1). Flow rate: Adjust so that the retention time of clopidogrel solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the peaks, having the relative retention times of about 0.3, about 0.5 and about 0.9 to clopidogrel, obtained from the sample solution is not larger than 3/10 times the peak area of clopidogrel from the stand-ard solution. The area of the peak having the relative reten-tion time of about 2.0 from the sample solution is not larger than 1.2 times the peak area of clopidogrel from the stand-ard solution. The area of the peak other than clopidogrel and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of clopidogrel from the standard solution. The total area of the peaks other than clopidogrel from the sample solution is not larger than 1.7 times the peak area of clopidogrel from the standard solu-tion. JP XVIII Official Monographs / Clopidogrel Sulfate Tablets 795 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column of 4.6 mm in inside diameter and 15 cm in length, packed with ovomucoid-chemically bonded amino silica gel for liquid chromatogra-phy (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and to 750 mL of this solu-tion add 250 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of clopidogrel is about 6 minutes. Time span of measurement: About 2.5 times as long as the retention time of clopidogrel, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of clopidogrel obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- use this solution as the sample solution. Separately, weigh accurately about 30 mg of Clopidogrel Sulfate RS (separate-ly determine the water <2.48> in the same manner as Clopidogrel Sulfate), dissolve in 5 mL of methanol, and add water to make exactly 100 mL. Pipet 6 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 240 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as a blank. Dissolution rate (z) with respect to the labeled amount of clopidogrel (C16H16ClNO2S) ＝ MS × AT/AS × V?/V × 1/C × 108 × 0.766 MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of clopidogrel (C16H16ClNO2S) in 1 tablet Assay To 20 tablets of Clopidogrel Sulfate Tablets add 400 mL of the mobile phase, sonicate with occasional shaking until the tablets are disintegrated, add the mobile phase to make exactly 500 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add the mobile phase to make exactly V mL so that each mL con- ditions, the number of theoretical plates and the symmetry factor of the peak of clopidogrel are not less than 2500 and tains about 0.5 mg of clopidogrel (C 16H 16ClNO2S). Pipet 4 not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clopidogrel is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Clopidogrel Sulfate Tablets add a suitable amount of the mobile phase, sonicate with occasional shak-ing until the tablet is disintegrated, and add the mobile phase to make exactly 50 mL. Filter the solution through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add exactly V/5 mL of the internal standard solu-tion, and add the mobile phase to make V mL so that each mL contains about 0.
To a quantity of powdered Clopidogrel Sulfate Tablets, equivalent to 75 mg of clopidogrel (C16H16ClNO2S), add 50 mL of methanol, and after sonicat-ing with ultrasonic waves with occasional shaking, add meth-anol to make 100 mL. To 10 mL of this solution add metha-nol to make 30 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 269 nm and 273 nm, and between 276 nm and 280 nm.
Related substances—Keep the sample solution and the standard solution at 59C or below and use within 24 hours. Take a quantity of Clopidogrel Sulfate Tablets equivalent to 0.15 g of clopidogrel (C16H16ClNO2S), add 120 mL of the mobile phase, sonicate with occasional shaking until the tablets are disintegrated, and add the mobile phase to make 200 mL. Centrifuge this solution, to 10 mL of the supernatant liquid add the mobile phase to make 30 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet Amount (mg) of clopidogrel sulfate (C H ClNO S.H SO ) 2 mL of the sample solution, add the mobile phase to make ＝ MS × AT/AS 16 16 2 2 4 exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 0.87 g of sodium 1-pen-tanesufonate in 1000 mL of water, and adjust to pH 2.5 with phosphoric acid. To 950 mL of this solution add 50 mL of methanol. To 600 mL of this solution, add 400 mL of a mix-ture of acetonitrile for liquid chromatography and methanol (19:1). Flow rate: Adjust so that the retention time of clopidogrel solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the peaks, having the relative retention times of about 0.3, about 0.5 and about 0.9 to clopidogrel, obtained from the sample solution is not larger than 3/10 times the peak area of clopidogrel from the stand-ard solution. The area of the peak having the relative reten-tion time of about 2.0 from the sample solution is not larger than 1.2 times the peak area of clopidogrel from the stand-ard solution. The area of the peak other than clopidogrel and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of clopidogrel from the standard solution. The total area of the peaks other than clopidogrel from the sample solution is not larger than 1.7 times the peak area of clopidogrel from the standard solu-tion. JP XVIII Official Monographs / Clopidogrel Sulfate Tablets 795 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column of 4.6 mm in inside diameter and 15 cm in length, packed with ovomucoid-chemically bonded amino silica gel for liquid chromatogra-phy (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and to 750 mL of this solu-tion add 250 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of clopidogrel is about 6 minutes. Time span of measurement: About 2.5 times as long as the retention time of clopidogrel, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 100 mL. Confirm that the peak area of clopidogrel obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- use this solution as the sample solution. Separately, weigh accurately about 30 mg of Clopidogrel Sulfate RS (separate-ly determine the water <2.48> in the same manner as Clopidogrel Sulfate), dissolve in 5 mL of methanol, and add water to make exactly 100 mL. Pipet 6 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 240 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as a blank. Dissolution rate (z) with respect to the labeled amount of clopidogrel (C16H16ClNO2S) ＝ MS × AT/AS × V?/V × 1/C × 108 × 0.766 MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of clopidogrel (C16H16ClNO2S) in 1 tablet
To 20 tablets of Clopidogrel Sulfate Tablets add 400 mL of the mobile phase, sonicate with occasional shaking until the tablets are disintegrated, add the mobile phase to make exactly 500 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add the mobile phase to make exactly V mL so that each mL con- ditions, the number of theoretical plates and the symmetry factor of the peak of clopidogrel are not less than 2500 and tains about 0.5 mg of clopidogrel (C 16H 16ClNO2S). Pipet 4 not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clopidogrel is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Clopidogrel Sulfate Tablets add a suitable amount of the mobile phase, sonicate with occasional shak-ing until the tablet is disintegrated, and add the mobile phase to make exactly 50 mL. Filter the solution through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add exactly V/5 mL of the internal standard solu-tion, and add the mobile phase to make V mL so that each mL contains about 0.1 mg of clopidogrel (C H ClNO S). mL of this solution, add exactly 4 mL of the internal stand-ard solution and the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 33 mg of Clopidogrel Sulfate RS (separately de-termine the water <2.48> in the same manner as Clopidogrel Sulfate), and dissolve in the mobile phase to make exactly 50 mL. Pipet 4 mL of this solution, add exactly 4 mL of the in-ternal standard solution and the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of clopidogrel to that of the internal standard. Amount (mg) of clopidogrel (C16H16ClNO2S) in 1 tablet of Clopidogrel Sulfate Tablets ＝ MS × QT/QS × V/10 × 0.766 16 16 2 Use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of clopidogrel (C16H16ClNO2S) ＝ MS × QT/QS × V/10 × 0.766 MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of isopropyl para-hydroxybenzoate in the mobile phase (1 in 1500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate of 25-mg tablet in 30 minutes is not less than 70z, and that of 75-mg tablet in 45 minutes is not less than 80z. Start the test with 1 tablet of Clopidogrel Sulfate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of clopidogrel (C16H16ClNO2S), and MS: Amount (mg) of Clopidogrel Sulfate RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of isopropyl para-hydoxybenzoate in the mobile phase (1 in 1500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column of 3.9 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 0.87 g of sodium 1-pentanesul-fonate in 1000 mL of water, and adjust to pH 2.5 with phos-phoric acid. To 950 mL of this solution add 50 mL of metha-nol. To 600 mL of this solution add 400 mL of a mixture of acetonitrile for liquid chromatography and methanol (19:1). Flow rate: Adjust so that the retention time of clopidogrel is about 8 minutes. System suitability— System performance: When the procedure is run with 10 ‌796 mL of the standard solution under the above operating con-ditions, the internal standard and clopidogrel are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of clopidogrel to that of the internal standard is not more than 1.0z.
クロラゼプ酸二カリウムカプセル
Clorazepate Dipotassium Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of clorazepate dipotassium (C16H10ClKN2O3.KOH: 408.92). Method of preparation Prepare as directed under Cap-sules, with Clorazepate Dipotassium. Identification To 10 mL of the sample solution obtained in the Assay add water to make 20 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 228 nm and 232 nm. Purity Related substances—Take out the contents of Clorazepate Dipotassium Capsules, and powder. To a por-tion of the powder, equivalent to 15 mg of Clorazepate Dipotassium, add a mixture of water, potassium carbonate solution (97 in 1000) and acetonitrile (3:1:1) to make 25 mL, and shake for 10 minutes. Filter the solution through a mem-brane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water, potassium carbonate solution (97 in 1000) and acetonitrile (3:1:1) to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Purity (4) under Clorazepate Dipotassium: the peak area of nordiazepam, having the relative retention time of about 3.0 to clorazepic acid, obtained from the sam-ple solution is not larger than 3 times the peak area of clora-zepic acid from the standard solution, and the total area of the peaks other than clorazepic acid and nordiazepam is not larger than the peak area of clorazepic acid from the stand-ard solution. For the peak area of nordiazepam, multiply the correction factor, 0.64. MS: Amount (mg) of clorazepate dipotassium for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Clorazepate Dipotassium Capsules is not less than 80z. Start the test with 1 capsule of Clorazepate Dipotassium Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 8.
To 10 mL of the sample solution obtained in the Assay add water to make 20 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 228 nm and 232 nm.
Related substances—Take out the contents of Clorazepate Dipotassium Capsules, and powder. To a por-tion of the powder, equivalent to 15 mg of Clorazepate Dipotassium, add a mixture of water, potassium carbonate solution (97 in 1000) and acetonitrile (3:1:1) to make 25 mL, and shake for 10 minutes. Filter the solution through a mem-brane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water, potassium carbonate solution (97 in 1000) and acetonitrile (3:1:1) to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Purity (4) under Clorazepate Dipotassium: the peak area of nordiazepam, having the relative retention time of about 3.0 to clorazepic acid, obtained from the sam-ple solution is not larger than 3 times the peak area of clora-zepic acid from the standard solution, and the total area of the peaks other than clorazepic acid and nordiazepam is not larger than the peak area of clorazepic acid from the stand-ard solution. For the peak area of nordiazepam, multiply the correction factor, 0.64. MS: Amount (mg) of clorazepate dipotassium for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Clorazepate Dipotassium Capsules is not less than 80z. Start the test with 1 capsule of Clorazepate Dipotassium Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 8.3 mg of clorazepate dipotassium (C16H10ClKN2O3.KOH), and use this solution as the sample solution. Separately, weigh accurately about 21 mg of clorazepate dipotassium for assay, previously dried in vacuum over phosphorus (V) oxide at 609C for 5 hours, and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, at 252 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Dissolution rate (z) with respect to the labeled amount of clorazepate dipotassium (C16H10ClKN2O3.KOH) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of clorazepate dipotassium for assay taken C: Labeled amount (mg) of clorazepate dipotassium (C16H10ClKN2O3.KOH) in 1 capsule
Carefully take out the contents of not less than 20 Clorazepate Dipotassium Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a por-tion of the powder, equivalent to about 15 mg of clorazepate dipotassium (C16H10ClKN2O3.KOH), add 70 mL of water, shake for 15 minutes, and add water to make exactly 100 mL. Centrifuge the solution, pipet 4 mL of the supernatant liquid, add water to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 15 mg of clorazepate dipotassium for assay, previ-ously dried in vacuum over phosphorus (V) oxide at 609C for 5 hours, and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 252 nm. Amount (mg) of clorazepate dipotassium (C16H10ClKN2O3.KOH) Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Clorazepate Dipotassium Capsules add 70 mL of water, shake for 15 minutes, and add water to make exactly 100 mL. Centrifuge the solution, pipet V mL of the supernatant liquid, add water to make exactly V? mL so that each mL contains about 12 mg of clorazepate dipotassium (C16H10ClKN2O3.KOH), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of clorazepate dipotassium (C16H10ClKN2O3.KOH) ＝ MS × AT/AS × V?/V × 2/25 ＝ MS × AT/AS MS: Amount (mg) of clorazepate dipotassium for assay taken
Clotiazepam Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of clotiazepam (C16H15ClN2OS: 318.82). Method of preparation Prepare as directed under Tablets, with Clotiazepam. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clotiazepam Tablets add 35 mL of 0.1 mol/L hydrochloric acid TS, stir until the tablet is com-pletely disintegrated, stir for a further 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Centrifuge this solution, pipet V mL of the supernatant liq-uid, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 10 mg of clotiazepam (C16H15ClN2OS), and use this solution as the sample solu-tion.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Clotiazepam Tablets add 35 mL of 0.1 mol/L hydrochloric acid TS, stir until the tablet is com-pletely disintegrated, stir for a further 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Centrifuge this solution, pipet V mL of the supernatant liq-uid, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 10 mg of clotiazepam (C16H15ClN2OS), and use this solution as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of clotiazepam (C16H15ClN2OS) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of clotiazepam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 ‌JP XVIII Official Monographs / Clotrimazole 799 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of Clotiazepam Tablets is not less than 80z. Start the test with 1 tablet of Clotiazepam Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of clotiazepam (C16H 15ClN2OS), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of clotiazepam for assay, previously dried at 809C for 3 hours, and dissolve in ethanol (95) to make exactly 25 mL. Pipet 5 mL of this solu-tion, and add the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 262 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of clotiazepam (C16H15ClN2OS) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of clotiazepam for assay taken C: Labeled amount (mg) of clotiazepam (C16H15ClN2OS) in 1 tablet
To 20 Clotiazepam Tablets add 350 mL of 0.1 mol/L hydrochloric acid TS, stir until the tablets are com-pletely disintegrated, stir for a further 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 500 mL. Centrifuge this solution, pipet V mL of the supernatant liq-uid, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 10 mg of clotiazepam (C16H15ClN2OS), and use this solution as the sample solu-tion. Separately, weigh accurately about 25 mg of clotiazep-am for assay, previously dried at 809C for 3 hours, and dis-solve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 261 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of clotiazepam (C16H15ClN2OS) in 1 tablet ＝ MS × AT/AS × V?/V × 1/100 MS: Amount (mg) of clotiazepam for assay taken
To a quantity of powdered Codeine Phos-phate Tablets, equivalent to 0.1 g of Codeine Phosphate Hydrate, add 20 mL of water, shake, and filter. To 2 mL of the filtrate add water to make 100 mL, and determine the absorption spectrum as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Codeine Phosphate Tablets add 3V/25 mL of water to disintegrate, add 2V/25 mL of diluted dilute sul-furic acid (1 in 20), and sonicate for 10 minutes. To this solu-tion add exactly 2V/25 mL of the internal standard solution, add water to make V mL so that each mL contains about 0.2 mg of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2 H2O), filter, and use the filtrate as the sample solution. Separately, weigh accurately about 50 mg of codeine phosphate hydrate for assay (separately, deter-mine the water <2.48> in the same manner as Codeine Phos-phate Hydrate), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 2 mL of the internal standard solution, add water to make 25 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay. Amount (mg) of codeine phosphate hydrate factor of the peak of codeine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of codeine is not more than 2.0z.
Weigh accurately and powder not less than 20 Codeine Phosphate Tablets. Weigh accurately a portion of the powder, equivalent to about 0.1 g of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2 H2O), add 30 mL of water, shake, add 20 mL of diluted dilute sulfuric acid (1 in 20), sonicate the mixture for 10 minutes, and add water to make exactly 100 mL. Filter the solution, then pipet 5 mL of the filtrate, add exactly 10 mL of the internal standard solution and water to make 20 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 50 mg of codeine phosphate hydrate for assay (previously determine the water <2.48> in the same manner as Codeine Phosphate Hydrate), dissolve in water to make exactly 100 mL, then pipet 10 mL of this solution, add exactly 10 mL of the inter- (C18H21 NO3.H3PO4.1/2H2O) nal standard solution, and use this solution as the standard solution. Perform the test with 20 mL each of the sample so- ＝ MS × QT/QS × V/250 × 1.023 MS: Amount (mg) of codeine phosphate hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of ethylefurin hydro-chloride (3 in 2000). ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Codeine Phosphate Tablets is not less than 80z. Start the test with 1 tablet of Codeine Phosphate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2 H2O), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of codeine phosphate hydrate for assay (separately, deter-mine the water <2.48> in the same manner as Codeine Phos-phate Hydrate), dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of codeine in each solution. Dissolution rate (z) with respect to the labeled amount of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2H2O) ＝ MS × AT/AS × V?/V × 1/C × 18 × 1.023 MS: Amount (mg) of codeine phosphate hydrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of codeine to that of the internal standard. Amount (mg) of codeine phosphate hydrate (C18H21NO3.H3PO4.1/2H2O) ＝ MS × QT/QS × 2 × 1.023 MS: Amount (mg) of codeine phosphate hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of codeine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, codeine and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of codeine to that of the internal standard is not more than 1.0z.
Determine the infrared absorption spectrum of powdered Colestimide Granules as directed in the potas-sium chloride disk method under Infrared Spectrophotome-try <2.25>: it exhibits absorption at the wave numbers of about 1587 cm－1, 1528 cm－1 and 1262 cm－1. Uniformity of dosage units <6.02> Colestimide Granules in single-dose packages meet the requirement of the Mass varia-tion test. Disintegration <6.09> Carry out the test for 10 minutes with 0.09 – 0.11 g of Colestimide Granules in six glass tubes of the apparatus: it meets the requirement.
Weigh accurately about 4.5 g of sodium cholate hydrate (separately determine the water), dissolve in water to make exactly 1000 mL, and use this solution as the sodium cholate standard stock solution. Take out the contents of not less than 20 single-dose packages of Colestimide Granules, weigh accurately an amount of the contents, equivalent to about 0.2 g of colestimide, add exactly 200 mL of the sodium cholate standard stock solution, shake for 1 hour, and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Then, proceed as directed in the Assay (2) under Colestimide. Amount (mg) of colestimide ＝ MS × (QS － QT)/QS × 1/5 × 1/2.2 × 0.947 MS: Amount (mg) of sodium cholate hydrate taken, calcu-lated on the anhydrous basis 2.2: Quantity (g) of the cholic acid exchange per mg of colestimide Internal standard solution—A solution of butyl para-hydroxybenzoate in acetonitrile (1 in 80,000).
Powder Colestimide Tablets. Determine the infrared absorption spectrum of a portion of the powder as directed in the potassium chloride disk method under In-frared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 1587 cm－1, 1528 cm－1, 1262 cm－1, 1102 cm－1 and 1035 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Disintegration <6.09> When carry out the test for 10 minutes, it meets the requirement.
Weigh accurately about 0.45 g of sodium cholate hy-drate (separately determine the water), dissolve in water to make exactly 100 mL, and use this solution as the sodium cholate standard stock solution. Separately, weigh accurately the mass of not less than 20 Colestimide Tablets, and pow-der. Weigh accurately a portion of the powder, equivalent to about 30 mg of colestimide, add exactly 30 mL of the so-dium cholate standard stock solution, shake for 1 hour, and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, pipet 5 mL of the sodium cholate standard stock solution, add exactly 5 mL of the in-ternal standard solution, and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of cholic acid to that of the internal standard. Amount (mg) of colestimide ＝ MS × (QS － QT)/QS × 3/10 × 1/2.2 × 0.947 MS: Amount (mg) of sodium cholate hydrate taken, calcu-lated on the anhydrous basis 2.2: Exchanged amount (g) of cholic acid per g of coles-timide, calculated on the dried basis Internal standard solution—A solution of butyl parahy-droxybenzoate in acetonitrile (1 in 80,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica ‌810 gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of cholic acid is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cholic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cholic acid to that of the internal standard is not more than 1.0z.
Compound Diastase and Sodium Bicarbonate Powder
Compound Diastase and Sodium Bicarbonate Powder occurs as a slightly brownish, light yellow powder. It has a characteristic odor and a bitter taste.
Compound Oxycodone and Atropine Injection
複方オキシコドン・アトロピン注射液
Compound Oxycodone and Atropine Injection is an aqueous injection. It contains not less than 0.
Compound Oxycodone and Atropine Injection is a colorless or pale yellow, clear liquid. It is affected by light. pH: 2.5 – 4.0
(1) To 1 mL of Compound Oxycodone and Atropin Injection add 1 mL of 2,4-dinitrophenylhydrazine-ethanol TS: a yellow precipitate is formed (oxycodone). Evaporate 1 mL of Compound Oxycodone and Atropin Injection on a water bath, and dissolve the residue in 2 mL of sulfuric acid: a yellow color is produced. Heat the solution: it changes to red, and then to deep orange-red (hydrocotarnine). Evaporate 1 mL of Compound Oxycodone and Atropin Injection on a water bath. Dissolve the residue in 3 mL of sulfuric acid, add 2 drops of a solution of tannic acid in ethanol (95) (1 in 20), and allow to stand: a deep green color is produced (hydrocotarnine). To 1 mL of Compound Oxycodone and Atropine In-jection add 0.5 mL of 2,4-dinitrophenylhydrazine-ethanol TS, and allow to stand for 1 hour. Centrifuge, and add ace-tone to the supernatant liquid until no more precipitate is produced. Allow to stand for 20 minutes, and centrifuge. To the supernatant liquid add potassium hydroxide TS until the liquid is light purple. Shake the liquid with 5 mL of dichloro-methane, and separate the dichloromethane layer. Take 0.5 mL of the dichloromethane layer, and evaporate to dryness on a water bath. Add 5 drops of fuming nitric acid to the residue, and evaporate to dryness on a water bath. Cool, dis-solve the residue in 1 mL of N,N-dimethylformamide, and add 6 drops of tetraethylammonium hydroxide TS: a red-purple color is produced (atropine). Extractable volume <6.05> It meets the requirement.
(1) Oxycodone hydrochloride hydrate and hydrocotarnine hydrochloride hydrate—Pipet 2 mL of Com-pound Oxycodone and Atropine Injection, add exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 0.4 g of oxycodone hydrochloride hydrate for assay (separately determine the water <2.48> in the same manner as Oxyco-done Hydrochloride Hydrate) and about 0.1 g of hydrocotarnine hydrochloride hydrate for assay previously dried at 1059C for 3 hours, and dissolve in water to make ex-actly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq- Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized polyvinyl alcohol gel polymer for liquid chroma-tography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 500 mL of 0.05 mol/L disodium hydrogenphosphate TS add 0.05 mol/L sodium dihydrogen-phosphate TS, and adjust the pH to 8.0. To 300 mL of this solution add 200 mL of acetonitrile, and mix. Flow rate: Adjust so that the retention time of oxycodone hydrochloride is about 8 minutes. Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions, and use a column giving elution of the internal standard, oxycodone and hydrocotarine in this order with complete separation of these peaks. (2) Atropine sulfate hydrate—Pipet 2 mL of Compound Oxycodone and Atropine Injection, and add exactly 2 mL of the internal standard solution. To this solution add 10 mL of diluted dilute hydrochloric acid (1 in 10) and 2 mL of ammo-nia TS, immediately add 20 mL of dichloromethane, shake vigorously, filter the dichloromethane layer through filter paper on which 5 g of anhydrous sodium sulfate is placed, and evaporate the filtrate to dryness under reduced pressure. To the residue add 0.5 mL of 1,2-dichloroethane and 0.5 mL of bis-trimethyl silyl acetamide, stopper tightly, warm in a water bath at 609C for 15 minutes, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Atropine Sulfate RS (separately determine the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, and add exactly 2 mL of the in-ternal standard solution. Proceed with this solution in the same manner as directed for the sample solution, and use so obtained solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of atropine to that of the internal stand-ards. Amount (mg) of atropine sulfate hydrate [(C H NO ) .H SO .H O] uid Chromatography <2.01> according to the following con- 17 23 3 2 2 4 2 ditions. Calculate the ratios, QTa and Q Tb, of the peak area ＝ MS × QT/QS × 1/50 × 1.027 of oxycodone and hydrocotarnine to that of the internal standard obtained from the sample solution, and the ratios, QSa and QSb, of the peak area of oxycodone and hydrocotar-nine to that of the internal standard from the standard solu-tion. Amount (mg) of oxycodone hydrochloride hydrate (C18H21NO4.HCl.3H2O) ＝ MSa × QTa/QSa × 1/25 × 1.154 Amount (mg) of hydrocotarnine hydrochloride hydrate (C12H15NO3.HCl.H2O) ＝ MSb × QTb/QSb × 1/25 × 1.070 MSa: Amount (mg) of oxycodone hydrochloride hydrate for assay taken, calculated on the anhydrous basis MSb: Amount (mg) of hydrocotarnine hydrochloride hy-drate for assay taken Internal standard solution—Dissolve 0.02 g of phenacetin in 10 mL of ethanol (95), and add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of homatropine hydrobromide (1 in 4000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column about 3 mm in inside diameter and about 1.5 m in length, packed with 180- to 250-mm siliceous earth for gas chromatography coated with 1 to 3z of 50z phenyl-methylsilicone polymer. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen or helium. Flow rate: Adjust so that the retention time of atropine is about 5 minutes. Selection of column: Proceed with 2 mL of the standard solution under the above operating conditions, and calculate the resolution. Use a column giving elution of the internal standard and atropine in this order with the resolution be-tween these peaks being not less than 3.
Compound Oxycodone Injection is a clear, colorless to pale yellow liquid. It is affected by light. pH: 2.5 – 4.0
(1) To 1 mL of Compound Oxycodone In-jection add 1 mL of 2,4-dinitrophenylhydrazine-ethanol TS: a yellow precipitate is formed (oxycodone). Evaporate 1 mL of Compound Oxycodone Injection on a water bath. Dissolve the residue in 2 mL of sulfuric acid: a yellow color is produced. Heat the solution: it changes to red, and then to deep orange-red (hydrocotar-nine). Evaporate 1 mL of Compound Oxycodone Injection on a water bath. Dissolve the residue in 3 mL of sulfuric acid, add 2 drops of a solution of tannic acid in ethanol (95) (1 in 20), and allow to stand: a deep green color is produced (hydrocotarnine). Extractable volume <6.05> It meets the requirement.
Pipet 2 mL of Compound Oxycodone Injection, add exactly 10 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 0.4 g of oxycodone hydrochloride hydrate for assay (separately determine the water <2.48> in the same manner as Oxycodone Hydrochloride Hydrate) and about 0.1 g of hydrocotarnine hydrochloride hydrate for assay previously dried at 1059C for 3 hours, and dissolve in water to make ex-actly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Calculate the ratios, QTa and QTb, of the peak area of oxycodone and hydrocotarnine to that of the internal standard obtained from the sample solution, and the ratios, QSa and QSb, of the peak area of oxycodone and hydrocotar-nine to that of the internal standard from the standard solu-tion. Amount (mg) of oxycodone hydrochloride hydrate (C18H21NO4.HCl.3H2O) ＝ MSa × QTa/QSa × 1/25 × 1.154 Amount (mg) of hydrocotarnine hydrochloride hydrate (C12H15NO3.HCl.H2O) ＝ MSb × QTb/QSb × 1/25 × 1.070 MSa: Amount (mg) of oxycodone hydrochloride hydrate for assay taken, calculated on the anhydrous basis MSb: Amount (mg) of hydrocotarnine hydrochloride hy-drate for assay taken Internal standard solution—Dissolve 0.02 g of phenacetin in 10 mL of ethanol (95), and add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized polyvinyl alcohol gel polymer for liquid chroma-tography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 500 mL of 0.05 mol/L disodium hydro-gen phosphate TS add 0.05 mol/L sodium dihydrogen phos-phate TS, and adjust the pH to 8.0. To 300 mL of this solu-tion add 200 mL of acetonitrile, and mix. Flow rate: Adjust so that the retention time of oxycodone is about 8 minutes. Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions, and use a column giving elution of the internal standard, oxycodone and hydrocotarnine in this order, with complete separation of these peaks.
Compound Phellodendron Powder for Cataplasm occurs as a yellow-brown powder, having a char-acteristic odor.
Shake thoroughly 0.2 g of Compound Phel-lodendron Powder for Cataplasm with 5 mL of methanol, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of Berberine Chloride RS or berberine chloride hydrate for thin-layer chromatography in 1 mL of methanol, and use the solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 10 cm, air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): one of the several spots ob-tained from the sample solution has the same color tone and Rf value with the yellow to yellow-green fluorescent spot from the standard solution (phellodendron bark).
Compound Rhubarb and Senna Powder oc-curs as a yellow-brown powder, having a characteristic odor and a bitter taste.
To 2 g of Compound Rhubarb and Senna Powder add 50 mL of water, warm on a water bath for 30 minutes, and filter. Add 2 drops of dilute hydrochloric acid to the filtrate, shake with two 20-mL portions of diethyl ether, and remove the diethyl ether layer. Add 5 mL of hy-drochloric acid to the aqueous layer, and heat it on a water bath for 30 minutes. Cool, shake with 20 mL of diethyl ether, take the diethyl ether layer, add 10 mL of sodium hydrogen carbonate TS, and shake: the aqueous layer is red in color.
Compound Scopolia Extract and Diastase Powder is light yellow in color. It has a bitter taste.
Compound Thianthol and Salicylic Acid Solution
複方チアントール・サリチル酸液
Compound Thianthol and Salicylic Acid Solution contains not less than 1.
Compound Thianthol and Salicylic Acid Solu-tion is a light yellow liquid, having a characteristic odor.
(1) Place 1 mL of Compound Thianthol and Salicylic Acid Solution to a porcelain dish, and evapo-rate on a water bath to dryness. To the residue add cau-tiously 5 mL of sulfuric acid: a blue-purple color develops. Add 5 to 6 drops of nitric acid to the solution: the color of the solution changes to yellow-red with evolution of gas (thianthol). Shake 10 mL of Compound Thianthol and Salicylic Acid Solution with 10 mL of sodium hydrogen carbonate TS, and separate the water layer. To 0.5 mL of the water layer add hydrochloric acid-potassium chloride buffer solu-tion (pH 2.0) to make 50 mL, and to 5 mL of this solution add 5 mL of a solution of iron (III) nitrate enneahydrate (1 in 200): a red-purple color is produced (salicylic acid). Wash the upper phase obtained in (2) with 10 mL of sodium hydrogen carbonate TS, and extract with 10 mL of dilute sodium hydroxide TS. Shake 1 mL of the extract with 1 mL of sodium nitrate TS and 1 mL of dilute hydrochloric acid, and add 3 mL of sodium hydroxide TS: a yellow color is produced (phenol). To 1 mL of Compound Thianthol and Salicylic Acid Solution add 10 mL of ethanol (95), mix, and use this solu-tion as the sample solution. Dissolve 0.01 g each of salicylic acid, phenol and thianthol in 5 mL each of ethanol (95), and use each solution as standard solutions (1), (2) and (3). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): three spots obtained from the sample solution and the corresponding spots from the standard solu-tions (1), (2) and (3) show the same Rf value. Spray evenly iron (III) chloride TS on the plate: the spot from standard solution (1) and the corresponding spot from the sample so-lution reveal a purple color. ‌Assay Pipet 2 mL of Compound Thianthol and Salicylic Acid Solution, add exactly 10 mL of the internal standard solution, then add 70 mL of diluted methanol (1 in 2), mix well, and add diluted methanol (1 in 2) to make 100 mL. Filter, discard the first 10 mL of the filtrate, and use the sub-sequent filtrate as the sample solution. Weigh accurately about 0.2 g of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours, and about 0.2 g of phenol for assay, dissolve in diluted methanol (1 in 2) to make ex-actly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and diluted methanol (1 in 2) to make 100 mL, and use this solution as the standard solution. With 5 mL each of the sample solution and stand-ard solution, perform the test as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QTa and QTb, of the peak area of salicylic acid and phenol to that of the internal standard in the sample solution, and the ratios, QSa and QSb, of the peak area of salicylic acid and phenol to that of the internal stand-ard in the standard solution. Amount (mg) of salicylic acid (C H O ) Thiopental Sodium
Cresol Solution contains not less than 1.
Cresol Solution is a clear or slightly turbid, yellow solution. It has the odor of cresol.
Shake 0.5 mL of the oily layer obtained in the Assay with 30 mL of water, filter, and perform the fol-lowing tests using this filtrate as the sample solution: To 5 mL of the sample solution add 1 to 2 drops of iron (III) chloride TS: a blue-purple color develops. To 5 mL of the sample solution add 1 to 2 drops of bromine TS: a light yellow, flocculent precipitate is pro-duced.
Transfer 200 mL of Cresol Solution, exactly meas-ured, to a 500-mL distilling flask. Add 40 g of sodium chlo-ride and 3 mL of dilute sulfuric acid, and connect the distil-ling apparatus with the distilling flask, and distil into a cassia flask which contains 30 g of powdered sodium chloride and 3 mL of kerosene, exactly measured, until the distillate meas-ures 90 mL. Draw off the water from the condenser, and continue the distillation until water vapor begins to come out of the tip of the condenser. Shake often the cassia flask in warm water to dissolve the sodium chloride, and allow to stand for 15 minutes. After cooling to 159C, add a saturated solution of sodium chloride, and allow to stand for more than 3 hours with occasional shaking. Allow to stand for 1 to 2 minutes with gentle shaking to combine the separated oil drops with the oil layer. The difference between the number of mL of the oil layer measured and 3 mL represents the amount (mL) of cresol.
Cyclophosphamide Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O: 279.10). Method of preparation Prepare as directed under Tablets, with Cyclophosphamide Hydrate. Identification To Cyclophosphamide Tablets add 1 mL of water for every 53 mg of Cyclophosphamide Hydrate, shake vigorously for 5 minutes, add 6 mL of methanol for every 53 mg of Cyclophosphamide Hydrate, and shake vigorously for 10 minutes. To this solution add methanol so that each mL contains about 5.
To Cyclophosphamide Tablets add 1 mL of water for every 53 mg of Cyclophosphamide Hydrate, shake vigorously for 5 minutes, add 6 mL of methanol for every 53 mg of Cyclophosphamide Hydrate, and shake vigorously for 10 minutes. To this solution add methanol so that each mL contains about 5.3 mg of Cyclophosphamide Hydrate, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 3 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, dissolve 53 mg of cy-clophosphamide hydrate for assay in 10 mL of a mixture of methanol and water (9:1), and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-propanol and water (8:1) to a dis-tance of about 10 cm, and air-dry the plate. Heat the plate at 1309C for 15 minutes. After cooling, spray evenly ninhydrin-butanol TS on the plate, and after air-drying heat at 1309C for 10 minutes: the principal spot obtained from the sample solution and the spot from the standard solution show a red-purple color and the same Rf value. JP XVIII Official Monographs / Cycloserine 823 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. ‌To 1 tablet of Cyclophosphamide Tablets add 3V/5 mL of a mixture of water and methanol (3:2), and shake vigorously to homogenously disperse the tablet. To this solution add a mixture of water and methanol (3:2) to make exactly V mL so that each mL contains about 1.1 mg of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O), and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O) ＝ MS × AT/AS × V/50 MS: Amount (mg) of cyclophosphamide hydrate for assay taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Cyclophosphamide Tablets is not less than 80z. Start the test with 1 tablet of Cyclophosphamide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 59 mg of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O) and use this solution as the sample so-lution. Separately, weigh accurately about 30 mg of cy-clophosphamide hydrate for assay, and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions and determine the peak areas, AT and AS, of cyclophosphamide in each solution. Dissolution rate (z) with respect to the labeled amount of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O) this solution add a mixture of water and methanol (3:2) to make exactly V mL so that each mL contains about 2.7 mg of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O), and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, pipet 4 mL of the subsequent fil-trate, add a mixture of water and methanol (3:2) to make ex-actly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 53 mg of cyclophos-phamide hydrate for assay, dissolve in a mixture of water and methanol (3:2) to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak area, AT and AS, of cyclophosphamide in each solution. Amount (mg) of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O) ＝ MS × AT/AS × V/200 MS: Amount (mg) of cyclophosphamide hydrate for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (3:2). Flow rate: Adjust so that the retention time of cy- clophosphamide is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cyclophosphamide are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cyclophosphamide is not more than 1.0z. ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of cyclophosphamide hydrate for assay taken C: Labeled amount (mg) of cyclophosphamide hydrate (C7H15Cl2N2O2P.H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of clophosphamide are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clophosphamide is not more than 2.0z.
To 10 tablets of Cyclophosphamide Tablets add 13V/20 mL of a mixture of water and methanol (3:2), and shake vigorously to homogenously disperse the tablets. To
D-Mannitol Injection is an aqueous injection. It contains not less than 95.
D-Mannitol Injection is a clear, colorless liq-uid. It has a sweet taste. It may precipitate crystals.
Concentrate D-Mannitol Injection on a water bath to make a saturated solution. To 5 drops of this solution add 1 mL of iron (III) chloride TS and 5 drops of a solution of sodium hydroxide (1 in 5): a yellow precipitate is produced. Shake this solution vigorously: a clear solution is produced. On addition of a solution of sodium hydroxide (1
D-Sorbitol Solution contains not less than 97.
D-Sorbitol Solution is a clear, colorless liquid. It is odorless, and has a sweet taste. It is miscible with water, with ethanol (95), with glycerin and with propylene glycol. It sometimes separates crystalline masses.
(1) To a volume of D-Sorbitol Solution, equivalent to 0.7 g of D-Sorbitol, add 2 mL of iron (II) sul-fate TS and 1 mL of a solution of sodium hydroxide (1 in 5): a blue-green color develops, but no turbidity is produced. (2) To a volume of D-Sorbitol Solution, equivalent to 1 g of D-Sorbitol, add water to make 20 mL. To 1 mL of this so-lution add 1 mL of a freshly prepared solution of catechol (1 in 10), mix well, then add rapidly 2 mL of sulfuric acid, and mix: a reddish purple to red-purple color immediately devel- ‌ops.
(1) Acidity or alkalinity—D-Sorbitol Solution is neutral. Chloride <1.03>—Proceed with a volume of D- Sorbitol Solution, equivalent to 2.0 g of D-Sorbitol, and per-form the test. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.005z). Sulfate <1.14>—To a volume of D-Sorbitol Solution, equivalent to 4.0 g of D-Sorbitol, and perform the test. Pre-pare the control solution with 0.50 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.006z). Heavy metals <1.07>—Proceed with a volume of D- Sorbitol Solution, equivalent to 5.0 g of D-Sorbitol, and ac-cording to Method 1, perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 5 ppm). Nickel—Take a volume of D-Sorbitol Solution, equivalent to 0.5 g of D-Sorbitol, add 3 drops of dimethyl-glyoxime TS and 3 drops of ammonia TS, and allow to stand for 5 minutes: no red color develops. Arsenic <1.11>—Take a volume of D-Sorbitol Solu- tion, equivalent to 1.5 g of D-Sorbitol, dilute with water or concentrate to 5 mL on a water bath, if necessary, cool, and perform the test using this solution as the test solution (not more than 1.3 ppm). Glucose—Take a volume of D-Sorbitol Solution, equivalent to 20.0 g of D-Sorbitol, dilute with water or con-centrate to 40 mL on a water bath, if necessary, add 40 mL of Fehling's TS, and boil gently for 3 minutes. After cooling, filter the supernatant liquid cautiously through a glass filter (G4), leaving the precipitate in the flask as much as possible, wash the precipitate with hot water until the last washings no longer show alkalinity, and filter the washings through the glass filter. Dissolve the precipitate in the flask in 20 mL of iron (III) sulfate TS, filter through the glass filter, and wash the filter with water. Combine the filtrate and the washings, heat at 809C, and titrate <2.50> with 0.02 mol/L potassium permanganate VS: not more than 6.3 mL of 0.02 mol/L po-tassium permanganate VS is required. Sugars—Take a volume of D-Sorbitol Solution, equivalent to 20.0 g of D-Sorbitol, dilute with water or con-centrate to 40 mL of a water bath, if necessary, add 8 mL of dilute hydrochloric acid, and heat under a reflux condenser in a water bath for 3 hours. After cooling, add 2 drops of methyl orange TS, followed by sodium hydroxide TS until an orange color develops, and add water to make 100 mL. Boil gently 10 mL of this solution with 10 mL of water and 40 mL of Fehling's TS for 3 minutes and proceed as directed in (7). Residue on ignition <2.44> Measure exactly a volume of D-Sorbitol Solution, equivalent to 5 g of D-Sorbitol, add 3 to 4 drops of sulfuric acid, and heat gently to evaporate. Ignite to burn, cool, and perform the test with the residue: not more than 1 mg.
Measure exactly a volume of D-Sorbitol Solution, equivalent to about 5 g of D-sorbitol (C6H14O6), and add water to make exactly 250 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL. Pipet 10 mL of the solu-tion into an iodine flask, add exactly 50 mL of potassium periodate TS, and heat for 15 minutes in a water bath. Cool, add 2.5 g of potassium iodide, immediately stopper tightly, and shake well. Allow to stand for 5 minutes in a dark place, and titrate <2.50> with 0.1 mol/L sodium thiosulfate VS (in-dicator: 3 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 1.822 mg of C6H14O6
Dehydrocholic Acid Injection is an aqueous injec-tion. It contains not less than 95.
Dehydrocholic Acid Injection is a clear, color-less to light yellow liquid, and has a bitter taste. pH: 9 – 11
Transfer a volume of Dehydrocholic Acid Injection, equivalent to 0.1 g of Purified Dehydrocholic Acid, to a separator, and add 10 mL of water and 1 mL of dilute hydrochloric acid: a white precipitate is produced. Extract the mixture with three 15-mL portions of chlo-roform, combine all the chloroform extracts, evaporate the chloroform on a water bath, and dry the residue at 1059C for 1 hour: the residue so obtained melts <2.60> between 2359C and 2429C.
Heavy metals <1.07>—Evaporate a volume of Dehy-drocholic Acid Injection, equivalent to 1.0 g of Purified De-hydrocholic Acid, on a water bath to dryness. Proceed with the residue according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Transfer an exactly measured volume of Dehydro-cholic Acid Injection, equivalent to about 0.5 g of dehydro- wash the filter with water until 100 mL of the filtrate is ob- tained. To 10 mL of the filtrate add 1 mL of dilute sulfuric cholic acid (C 24H 34O5), to a 100-mL separator, and add, if acid: no turbidity is produced. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g). Assay Weigh accurately about 0.5 g of Purified Dehydro-cholic Acid , previously dried, add 40 mL of neutralized ethanol and 20 mL of water, and dissolve by warming. Add 2 drops of phenolphthalein TS, then titrate <2.50> with 0.1 mol/L sodium hydroxide VS, adding 100 mL of freshly necessary, water to make 25 mL. Add 2 mL of hydrochloric acid, and extract with 25-mL, 20-mL and 15-mL portions of chloroform successively. Combine the chloroform extracts, wash with cold water until the washings become negative to acid, and evaporate the chloroform on a water bath. Dis-solve the residue in 40 mL of neutralized ethanol and 20 mL of water by warming. Add 2 drops of phenolphthalein TS to this solution, titrate <2.50> with 0.1 mol/L sodium hydroxide VS, adding 100 mL of freshly boiled and cooled water as the end point is approached, and continue the titration. Each mL of 0.1 mol/L sodium hydroxide VS boiled and cooled water as the end point is approached, and continue the titration. ＝ 40.25 mg of C24 H34O5 Each mL of 0.1 mol/L sodium hydroxide VS
Dental Antiformin is a slightly light yellow-green, clear liquid. It has a slight odor of chlorine. It gradually changes by light.
(1) Dental Antiformin changes red litmus paper to blue, and then decolorizes it. To Dental Antiformin add dilute hydrochloric acid: it evolves the odor of chlorine, and the gas changes potas-sium iodide starch paper moistened with water to blue. Dental Antiformin responds to Qualitative Tests <1.09> (1) for sodium salt.
Measure exactly 3 mL of Dental Antiformin in a glass-stoppered flask, add 50 mL of water, 2 g of potassium iodide and 10 mL of acetic acid (31), and titrate <2.50> the liberated iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 3.722 mg of NaClO
Containers—Tight containers. Storage—Light-resistant, and not exceeding 109C.
Deslanoside Injection is an aqueous injection. It contains not less than 90.
Deslanoside Injection is a clear and colorless liquid. Dexamethasone
Dextran 40 Injection is an aqueous injection. It contains not less than 9.
Dextran 40 Injection is a clear and colorless liquid. It is slightly viscous.
(1) Dilute 1 mL of Dextran 40 Injection with water to 200 mL, and to 1 mL of the diluted solution add 2 mL of anthrone TS: a blue-green color develops and turns gradually dark blue-green. Add 1 mL of diluted sulfu-ric acid (1 in 2) or 1 mL of acetic acid (100) to this solution: the solution does not change in color. (2) Dextran 40 Injection responds to Qualitative Tests <1.09> for sodium salt and for chloride. pH <2.54> 4.5 – 7.0 Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Viscosity <2.53> To 2 to 5 mL of Dextran 40 Injection add isotonic sodiumchloride solution to make exactly 100 mL, and use this solution as the sample solution. Perform the test with the sample solution and with isotonic sodium chloride solution as directed in Method 1 at 259C: the intrinsic vis-cosity is between 0.16 and 0.19. Calculate the concentration of the sample solution (g/100 mL) as directed in the Assay.
To exactly 30 mL of Dextran 40 Injection add water to make exactly 50 mL, and use this solution as the sample solution. Determine the optical rotation aD with the sample ‌JP XVIII Official Monographs / Dextran 70 839 solution as directed under Optical Rotation Determination <2.49> in a 100-mm cell at 20 ± 19C. Amount (mg) of dextran 40 in 100 mL of Dextran 40 Injection ＝ aD × 846.0
Diagnostic Sodium Citrate Solution contains not less than 3.
Diagnostic Sodium Citrate Solution is a clear, colorless liquid.
Diagnostic Sodium Citrate Solution re-sponds to Qualitative Tests <1.09> for sodium salt and for citrate. pH <2.54> 7.0 – 8.5
Pipet 5 mL of Diagnostic Sodium Citrate Solution, evaporate on a water bath to dryness, dry the residue at 1809C for 2 hours, and dissolve in 30 mL of acetic acid (100) by warming. Cool, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (indicator: 3 drops of crystal violet TS). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 9.803 mg of C6H5Na3O7.2H2O
Diastase and Sodium Bicarbonate Powder oc-curs as a light yellow powder. It has a characteristic, salty taste.
To a portion of the powdered Diazepam Tablets, equivalent to 50 mg of Diazepam, add 50 mL of acetone, shake, and filter. Evaporate 1 mL of the filtrate on a water bath to dryness, and dissolve the residue with 100 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 240 nm and 244 nm, between 283 nm and 287 nm, and between 360 nm and 370 nm. ‌Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Diazepam Tablets add 5 mL of water, and disintegrate the tablet by shaking. Then add 30 mL of meth-anol, shake for 10 minutes, add methanol to make exactly 50 mL, and centrifuge. Pipet V mL of the supernatant liquid, equivalent to 0.4 mg of diazepam (C16H13ClN2O), add exactly 5 mL of the internal standard solution, then add methanol to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of dia-zepam for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 5 mL of the internal standard solu-tion and add methanol to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> under the following condi-tions, and calculate the ratios, QT and QS, of the peak area of diazepam to that of the internal standard. Amount (mg) of diazepam (C16H13ClN2O) ＝ MS × QT/QS × 1/V MS: Amount (mg) of diazepam for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in methanol (1 in 25,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and diazepam are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of diazepam to that of the internal standard is not more than 1.0z. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Diazepam Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of diazepam (C16H13ClN2O), add 10 mL of water, shake, then add 60 mL of methanol, shake for 10 minutes, add methanol to make exactly 100 mL, and centrifuge. Pipet 5 mL of the superna-tant liquid, add exactly 5 mL of the internal standard solu-tion, then add methanol to make 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg of diazepam for assay, previously dried at 1059C for 2 hours, and dissolve in 10 mL of water and add methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add exactly 5 mL of the internal standard solution and add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of diazepam to that of the internal standard. Amount (mg) of diazepam (C16H13ClN2O) ＝ MS × QT/QS MS: Amount (mg) of diazepam for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in methanol (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and water (13:7). Flow rate: Adjust so that the retention time of diazepam is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and diazepam are eluted in this order with the resolution between these peaks being not more than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of diazepam to that of the internal standard is not more than 1.0z.
Dibekacin Sulfate Ophthalmic Solution is an aque-ous ophthalmic preparation. It contains not less than 90.
Dibekacin Sulfate Ophthalmic Solution is a clear, colorless liquid.
To a volume of Dibekacin Sulfate Ophthal-mic Solution add water so that each mL contains about 2.5 mg (potency) of Dibekacin Sulfate, and use this solution as the sample solution. Separately, dissolve an amount of Dibekacin Sulfate RS, equivalent to 5 mg (potency), in 2 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Proceed as directed in the Iden-tification (1) under Dibekacin Sulfate. pH <2.54> 6.5 – 7.5 Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require- ‌846 ment. ‌Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium and standard solu-tions—Proceed as directed in the Assay under Dibekacin Sulfate. Sample solutions—Pipet a volume of Dibekacin Sul-fate Ophthalmic Solution, equivalent to about 12 mg (po-tency), and add water to make exactly 30 mL. Pipet a suita-ble volume of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 20 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low con-centration sample solution, respectively.
ジエチルカルバマジンクエン酸塩錠
Diethylcarbamazine Citrate occurs as a white, crystalline powder. It is odorless, and has an acid and bitter taste. It is very soluble in water, soluble in ethanol (95), and practically insoluble in acetone, in chloroform and in diethyl ether. A solution of Diethylcarbamazine Citrate (1 in 20) is acid. Diethylcarbamazine Citrate is hygroscopic.
To a quantity of the powdered Diethylcar-bamazine Citrate Tablets, equivalent to 0.1 g of Diethylcar-bamazine Citrate, add 10 mL of water, shake well, and filter. To the filtrate add 1 mL of Reinecke salt TS: a light red precipitate is formed. Description Diethylcarbamazine Citrate occurs as a white, crystalline powder. It is odorless, and has an acid and bitter taste. It is very soluble in water, soluble in ethanol (95), and practically insoluble in acetone, in chloroform and in diethyl ether. A solution of Diethylcarbamazine Citrate (1 in 20) is acid. Diethylcarbamazine Citrate is hygroscopic. Identification (1) Dissolve 0.5 g of Diethylcarbamazine Citrate in 2 mL of water, add 10 mL of sodium hydroxide TS, and extract with four 5-mL portions of chloroform. Wash the combined chloroform extracts with 10 mL of water, and evaporate the chloroform on a water bath. Add 1 mL of iodoethane to the residue, and boil gently under a Uniformity of dosage units <6.02> Perform the Mass varia- tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Diethylcarbamazine Citrate Tablets add 70 mL of the mobile phase, shake vigorously for 10 minutes, add the mobile phase to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, pipet V mL of the subsequent filtrate, equivalent to about 2.5 mg of diethylcarbamazine citrate (C10H21N3O.C6H8O7), add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of diethylcarbamazine citrate (C H N O.C H O ) reflux condenser for 5 minutes. Evaporate the excess iodo- 10 21 3 6 8 7 ethane with the aid of a current of air, and dissolve the residue in 4 mL of ethanol (95). Cool the ethanol solution in an ice bath, with continuous stirring, add diethyl ether until precipitates are formed, and stir until crystallization is evident. Allow to stand in the ice bath for 30 minutes, and collect the precipitate. Dissolve the precipitate in 4 mL of ethanol (95), repeat the recrystallization in the same manner, then dry at 1059C for 4 hours: the crystals so obtained melt <2.60> between 1519C and 1559C. (2) Neutralize the remaining aqueous layer obtained in (1) with dilute sulfuric acid: the solution responds to Qualitative Tests <1.09> (2) and (3) for citrate. Melting point <2.60> 135.5 – 138.59C
Heavy metals <1.07>—Proceed with 2.0 g of Diethylcarbamazine Citrate according to Method 4, and perform the test. Prepare the control solution with 4.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 1.0z (2 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). ＝ MS × QT/QS × 10/V MS: Amount (mg) of Diethylcarbamazine Citrate RS taken Internal standard solution—A solution of 2-aminoben-zimidazol in the mobile phase (1 in 12,500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Diethylcarbamazine Citrate Tablets is not less than 80z. Start the test with 1 tablet of Diethylcarbamazine Citrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of diethylcarbamazine ci-trate (C10H21N3O.C6H8O7), and use this solution as the sam-ple solution. Separately, weigh accurately about 22 mg of Diethylcarbamazine Citrate RS, previously dried at 1059C for 4 hours, and dissolve in water to make exactly 100 mL. ‌850 Pipet 25 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of diethylcarbamazine in each so-lution. Dissolution rate (z) with respect to the labeled amount of diethylcarbamazine citrate (C H N O.C H O ) 409C. Mobile phase: To 0.05 mol/L potassium dihydrogen phos-phate TS add phosphoric acid to adjust the pH to 2.5. To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of diethylcar-bamazine is about 14 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- 10 21 3 6 8 7 ditions, diethylcarbamazine and the internal standard are ＝ MS × AT/AS × V?/V × 1/C × 225 MS: Amount (mg) of Diethylcarbamazine Citrate RS taken C: Labeled amount (mg) of diethylcarbamazine citrate (C10H21N3O.C6H8O7) in 1 tablet Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of diethylcarbamazine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diethylcarbamazine is not more than 2.0z.
Weigh accurately the mass of not less than 20 eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of diethylcarbamazine to that of the internal standard is not more than 1.0z.
Digoxin Injection is an aqueous injection. It contains not less than 90.
Digoxin Injection is a clear, colorless liquid.
Dilute Digoxin Injection, if necessary, with methanol so that each mL contains about 0.25 mg of Digoxin, and use this solution as the sample solution. In case where ingredients are suspected to affect the test, remove them by means of a solid-phase extraction. Separately, dis-solve 0.5 mg of Digoxin RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of octadecylsilanized silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and water (7:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly a mixture of a solution of trichloroacetic acid in ethanol (99.5) (1 in 4) and a freshly prepared solution of sodium toluenesulfon-chloramide trihydrate (3 in 100) (4:1) on the plate, heat at 1109C for 10 minutes, and examine under ultraviolet light (main wavelength: 366 nm): the Rf values of the principal spots with the sample solution and the standard solution are not different each other. Alcohol number <1.01> 0.8 – 1.2 (Method 1). Amount (mg) of digoxin (C41 H64O 14) ＝ MS × QT/QS
Related substances—To a volume of Digoxin Injec-tion, equivalent to about 2.5 mg of Digoxin, add dilute MS: Amount (mg) of Digoxin RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in ethanol (95) (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ethanol to make 50 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of the sample solution by the automatic integration method and calculate the amounts of these peaks by the area percentage method: the total amount of the peaks other than digoxin is not more than 5z. Operating conditions— Detector, column, column temperature, mobile phase and ‌JP XVIII Official Monographs / Digoxin Tablets 855 flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of digoxin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 25 mg of digoxin in 50 mL of warm ethanol (95), cool, add ethanol (95) to make 100 mL. To 10 mL of this solution add 10 mL of water and dilute ethanol to make 50 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the so-lution for system suitability test, and add dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, and add dilute ethanol to make exactly 100 mL. Confirm that the peak area of digoxin obtained with 10 mL this solution is equivalent to 0.07 to 0.13z of that with 10 mL of the solu-tion for system suitability test. System performance: Dissolve 25 mg of digoxin in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make 100 mL. To 10 mL of this solution add 5 mL of a solution of propyl parahydroxybenzoate in ethanol (95) (1 in 4000), 10 mL of water and dilute ethanol to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, digoxin and propyl parahydroxyben-zoate are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of digoxin is not more than 2.5z. Bacterial endotoxins <4.01> Less than 200 EU/mg. Extractable volume <6.05> It meets the requirements. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> Perform the test ac-cording to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Digoxin Injection, equivalent to about 2.5 mg of digoxin (C41H64O14), add exactly 5 mL of the internal standard solution and dilute ethanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Digoxin RS, previously dried under reduced pressure at 1059C for 1 hour, dissolve in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, 10 mL of water and dilute ethanol to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of digoxin to that of the internal standard. Amount (mg) of digoxin (C41H64O14) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of Digoxin RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in ethanol (95) (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of digoxin is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, digoxin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of digoxin to that of the internal standard is not more than 1.0z.
Digoxin Tablets contain not less than 90.0z and not more than 105.0z of the labeled amount of digox-in (C41H64O14: 780.94). Method of preparation Prepare as directed under Tablets, with Digoxin. Identification To an amount of powdered Digoxin Tablets, equivalent to 0.5 mg of Digoxin, add 2 mL of methanol, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 0.5 mg of Digoxin RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of octadecyl-silanized silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and water (7:3) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly a mixture of a solution of trichloroacetic acid in ethanol (99.5) (1 in 4) and a freshly prepared solution of sodium tolu-enesulfonchloramide trihydrate (3 in 100) (4:1) on the plate, heat at 1109C for 10 minutes, and examine under ultraviolet light (main wavelength: 366 nm): the Rf values of the princi-pal spots with the sample solution and the standard solution are not different each other. Purity Related substances—Powder not less than 20 Digox-in Tablets. Weigh a portion of the powder equivalent to 2.5 mg of Digoxin, add 30 mL of dilute ethanol, sonicate for 20 minutes, and shake for 5 minutes. After cooling, add dilute ethanol to make 50 mL, filter, and use the filtrate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of the sample solution by the automatic integration method and calculate the amount of these peaks by the area percentage method: the total amount of the peaks other than digoxin is not more than 5z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of digoxin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 25 mg of digoxin in 50 mL of warm ethanol (95), cool, add ethanol (95) to make 100 mL. To 10 mL of this solution add 10 mL of water and dilute ethanol to make 50 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the so-lution for system suitability test, and add dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, and add dilute ethanol to make exactly 100 mL. Confirm that the peak area of digoxin obtained with 10 mL of this solution is equivalent to 0.07 to 0.13z of that with 10 mL of the solu-tion for system suitability test. System performance: Dissolve 25 mg of digoxin in 50 mL 1 hour, dissolve in a small portion of ethanol (95), and add a mixture of ethanol (95) and water (4:1) to make exactly 500 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 500 mL, and use this solution as the stand-ard solution. Pipet 2 mL each of the sample solution, the standard solution and the dissolution medium, and transfer to brown glass-stoppered test tubes. Add exactly 10 mL of 0.012 g/dL L-ascorbic acid-hydrochloric acid TS to these tubes, and shake. Immediately add exactly 1 mL of dilute hydrogen peroxide TS, shake well, and allow to stand at a constant temperature between 259C and 309C for 45 minutes. Determine the fluorescence intensities, FT, FS, and FB, of these solutions at 360 nm of the excitation wavelength and at 485 nm of the fluorescence wavelength as directed under Fluorometry <2.22>, respectively. Dissolution rate (z) with respect to the labeled amount of warm ethanol (95), cool, and add ethanol (95) to make 100 mL. To 10 mL of this solution add 5 mL of a solution of of digoxin (C 41H 64O14) propyl parahydroxybenzoate in ethanol (95) (1 in 4000), 10 mL of water and dilute ethanol to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, digoxin and propyl parahydroxyben-zoate are eluted in this order with the resolution between these peaks being not less than 5. ＝ MS × (FT － FB)/(FS － FB) × 1/C MS: Amount (mg) of Digoxin RS taken C: The labeled amount (mg) of digoxin (C41H64O14) in 1 tablet Assay Weigh accurately the mass of not less than 20 Digox-in Tablets, and powder. Weigh accurately a portion of the System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under powder, equivalent to about 2.5 mg of digoxin (C 41H 64O14), the above operating conditions, the relative standard devia-tion of the peak area of digoxin is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Digoxin Tablets add 0.5 mL of water to dis-integrate, then add exactly 0.5 mL of the internal standard solution, and add V mL of dilute ethanol so that each mL contains about 21 mg of digoxin (C41H64O14).
To an amount of powdered Digoxin Tablets, equivalent to 0.5 mg of Digoxin, add 2 mL of methanol, shake for 10 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 0.5 mg of Digoxin RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of octadecyl-silanized silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and water (7:3) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly a mixture of a solution of trichloroacetic acid in ethanol (99.5) (1 in 4) and a freshly prepared solution of sodium tolu-enesulfonchloramide trihydrate (3 in 100) (4:1) on the plate, heat at 1109C for 10 minutes, and examine under ultraviolet light (main wavelength: 366 nm): the Rf values of the princi-pal spots with the sample solution and the standard solution are not different each other.
Related substances—Powder not less than 20 Digox-in Tablets. Weigh a portion of the powder equivalent to 2.5 mg of Digoxin, add 30 mL of dilute ethanol, sonicate for 20 minutes, and shake for 5 minutes. After cooling, add dilute ethanol to make 50 mL, filter, and use the filtrate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of the sample solution by the automatic integration method and calculate the amount of these peaks by the area percentage method: the total amount of the peaks other than digoxin is not more than 5z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of digoxin, beginning after the solvent peak. System suitability— Test for required detectability: Dissolve 25 mg of digoxin in 50 mL of warm ethanol (95), cool, add ethanol (95) to make 100 mL. To 10 mL of this solution add 10 mL of water and dilute ethanol to make 50 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the so-lution for system suitability test, and add dilute ethanol to make exactly 100 mL. Pipet 5 mL of this solution, and add dilute ethanol to make exactly 100 mL. Confirm that the peak area of digoxin obtained with 10 mL of this solution is equivalent to 0.07 to 0.13z of that with 10 mL of the solu-tion for system suitability test. System performance: Dissolve 25 mg of digoxin in 50 mL 1 hour, dissolve in a small portion of ethanol (95), and add a mixture of ethanol (95) and water (4:1) to make exactly 500 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 500 mL, and use this solution as the stand-ard solution. Pipet 2 mL each of the sample solution, the standard solution and the dissolution medium, and transfer to brown glass-stoppered test tubes. Add exactly 10 mL of 0.012 g/dL L-ascorbic acid-hydrochloric acid TS to these tubes, and shake. Immediately add exactly 1 mL of dilute hydrogen peroxide TS, shake well, and allow to stand at a constant temperature between 259C and 309C for 45 minutes. Determine the fluorescence intensities, FT, FS, and FB, of these solutions at 360 nm of the excitation wavelength and at 485 nm of the fluorescence wavelength as directed under Fluorometry <2.22>, respectively. Dissolution rate (z) with respect to the labeled amount of warm ethanol (95), cool, and add ethanol (95) to make 100 mL. To 10 mL of this solution add 5 mL of a solution of of digoxin (C 41H 64O14) propyl parahydroxybenzoate in ethanol (95) (1 in 4000), 10 mL of water and dilute ethanol to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, digoxin and propyl parahydroxyben-zoate are eluted in this order with the resolution between these peaks being not less than 5. ＝ MS × (FT － FB)/(FS － FB) × 1/C MS: Amount (mg) of Digoxin RS taken C: The labeled amount (mg) of digoxin (C41H64O14) in 1 tablet
Weigh accurately the mass of not less than 20 Digox-in Tablets, and powder. Weigh accurately a portion of the System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under powder, equivalent to about 2.5 mg of digoxin (C 41H 64O14), the above operating conditions, the relative standard devia-tion of the peak area of digoxin is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Digoxin Tablets add 0.5 mL of water to dis-integrate, then add exactly 0.5 mL of the internal standard solution, and add V mL of dilute ethanol so that each mL contains about 21 mg of digoxin (C41H64O14). Sonicate for 20 minutes, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, weigh accurately about 25 mg of Digoxin RS, previously dried under reduced pressure at 1059C for 1 hour, dissolve in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make exactly 100 mL. Pipet add 30 mL of dilute ethanol, sonicate for 20 minutes, and shake for 5 minutes. Add exactly 5 mL of the internal stand-ard solution and dilute ethanol to make 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, weigh accurately about 25 mg of Digoxin RS, previ-ously dried under reduced pressure at 1059C for 1 hour , dis-solve in 50 mL of warm ethanol (95), cool, and add ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, 10 mL of water and dilute ethanol to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of digoxin to that of the internal standard. 10 mL of this solution, and add ethanol (95) to make exactly 20 mL. Pipet 1 mL of this solution, add exactly 0.5 mL of the internal standard solution, then add 1.5 mL of water and Amount (mg) of digoxin (C ＝ MS × QT/QS × 1/10 41H 64O 14) (V – 2) mL of dilute ethanol, and use this solution as the standard solution. Proceed with the sample solution and standard solution as directed in the Assay. Amount (mg) of digoxin (C41H64O14) ＝ MS × QT/QS × 1/200 MS: Amount (mg) of Digoxin RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in ethanol (95) (1 in 40,000/V ). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 500 mL of diluted hydrochloric acid (3 in 500) as the dissolu-tion medium, the dissolution rate in 60 minutes of Digoxin Tablets is not less than 65z. No retest requirement is ap-plied to Digoxin Tablets. Start the test with 1 tablet of Digoxin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of Digoxin RS, previously dried in vacuum at 1059C for MS: Amount (mg) of Digoxin RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in ethanol (95) (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (7:3). Flow rate: Adjust so that the retention time of digoxin is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, digoxin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of ‌JP XVIII Official Monographs / 1 Dihydrocodeine Phosphate Powder 857‌ the peak area of digoxin to that of the internal standard is not more than 1.0z.
Diltiazem Hydrochloride Extended-release Capsules
ジルチアゼム塩酸塩徐放カプセル
Diltiazem Hydrochloride Extended-release Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of diltiazem hydrochlo-ride (C22H26N2O4S.HCl: 450.98). Method of preparation Prepare as directed under Cap-sules, with Diltiazem Hydrochloride. Identification Take out the content of Diltiazem Hydro-chloride Extended-release Capsules, and powder. To a por-tion of the powder, equivalent to 0.1 g of Diltiazem Hydro-chloride, add 100 mL of 0.01 mol/L hydrochloric acid TS, shake thoroughly, and filter. To 1 mL of the filtrate add 0.01 mol/L hydrochloric acid TS to make 100 mL, and de-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 234 nm and 238 nm. Purity Related substances—Take out the content of Diltia-zem Hydrochloride Extended-release Capsules, and powder. To a portion of the powder, equivalent to 50 mg of Diltia-zem Hydrochloride, add 30 mL of methanol, shake vigor-ously for 20 minutes, then add methanol to make 50 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 3 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the total area of the peaks other than diltiazem obtained from the sample solution is not larger than the peak area of diltiazem from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of diltiazem, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, and add methanol to make exactly 30 mL. Confirm that the peak area of diltiazem obtained with 20 mL of this solution is equivalent to 4.7 to 8.6z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diltiazem is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the content of 1 capsule of Diltiazem Hydrochlo-ride Extended-release Capsules, add V/2 mL of methanol, then add exactly V/10 mL of the internal standard solution, and shake vigorously for 20 minutes. Add methanol to make V mL so that each mL contains about 1 mg of diltiazem hy-drochloride (C22H26N2O4S.
Take out the content of Diltiazem Hydro-chloride Extended-release Capsules, and powder. To a por-tion of the powder, equivalent to 0.1 g of Diltiazem Hydro-chloride, add 100 mL of 0.01 mol/L hydrochloric acid TS, shake thoroughly, and filter. To 1 mL of the filtrate add 0.01 mol/L hydrochloric acid TS to make 100 mL, and de-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 234 nm and 238 nm.
Related substances—Take out the content of Diltia-zem Hydrochloride Extended-release Capsules, and powder. To a portion of the powder, equivalent to 50 mg of Diltia-zem Hydrochloride, add 30 mL of methanol, shake vigor-ously for 20 minutes, then add methanol to make 50 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 3 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the total area of the peaks other than diltiazem obtained from the sample solution is not larger than the peak area of diltiazem from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of diltiazem, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, and add methanol to make exactly 30 mL. Confirm that the peak area of diltiazem obtained with 20 mL of this solution is equivalent to 4.7 to 8.6z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diltiazem is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the content of 1 capsule of Diltiazem Hydrochlo-ride Extended-release Capsules, add V/2 mL of methanol, then add exactly V/10 mL of the internal standard solution, and shake vigorously for 20 minutes. Add methanol to make V mL so that each mL contains about 1 mg of diltiazem hy-drochloride (C22H26N2O4S.HCl), and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet 3 mL of the subsequent filtrate, add methanol to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of diltiazem hydrochloride (C22H26N2O4S.HCl) ＝ MS × QT/QS × V/100 MS: Amount (mg) of diltiazem hydrochloride for assay taken Internal standard solution—A solution of phenyl benzoate in methanol (3 in 400). Dissolution Being specified separately when the drug is granted approval based on the Law.
Take out the content of not less than 20 Diltiazem Hydrochloride Extended-release Capsules, weigh the mass of the content accurately, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of diltiazem hydrochloride (C22H26N2O4S.HCl), add 50 mL of methanol, then add exactly 10 mL of the internal standard solution, shake vigorously for 20 minutes, and add methanol to make 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, to 3 mL of the subsequent filtrate add methanol to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of diltiazem hydro-chloride for assay, previously dried at 1059C for 2 hours, dissolve in 50 mL of methanol, add exactly 10 mL of the in-ternal standard solution, and add methanol to make 100 mL. To 3 mL of this solution add methanol to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of diltiazem to that of the internal standard. ‌JP XVIII Official Monographs / Dimemorfan Phosphate 865 Amount (mg) of diltiazem hydrochloride (C22H26N2O4S.HCl) ＝ MS × QT/QS MS: Amount (mg) of diltiazem hydrochloride for assay taken Internal standard solution—A solution of phenyl benzoate in methanol (3 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 8 g of sodium acetate trihydrate and 1.5 g of d-camphorsulfonic acid in 500 mL of water, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To the filtrate add 250 mL of acetonitrile and 250 mL of methanol. Flow rate: Adjust so that the retention time of diltiazem is about 9 minutes. System suitability— System performance: Dissolve 30 mg of diltiazem hydrochloride, 20 mg of d-3-hydroxy-cis-2,3-dihydro-5-[2-(dimethylamino)ethyl]-2-(4-methoxyphenyl)-1,5-benzothia-zepin-4(5H )-on hydrochloride (hereinafter referred to as de-acetyl substance) and 20 mg of phenyl benzoate in methanol to make 200 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, de-acetyl substance, diltiazem and phenyl benzoate are eluted in this order and the resolutions between the peaks of de-acetyl sub-stance and diltiazem and the peaks of diltiazem and phenyl benzoate are not less than 2.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of diltiazem to that of the internal standard is not more than 1.0z.
Diluted Opium Powder contains not less than 0.
Diluted Opium Powder occurs as a light brown powder.
(1) Proceed with 1 g of Diluted Opium Powder as directed in the Identification (1) under Powdered Opium. (2) Proceed with 1 g of Diluted Opium Powder as di-rected in the Identification (2) under Powdered Opium.
Place about 5 g of Powdered Opium, accurately weighed, in a mortar, and triturate it with exactly 10 mL of water. Add 2 g of calcium hydroxide and exactly 40 mL of water, and stir the mixture for 20 minutes. Filter, and shake 30 mL of the filtrate with 0.1 g of magnesium sulfate hepta-hydrate for 1 minute. To the mixture add 0.3 g of calcium hydroxide, shake for 1 minute, and allow to stand for 1 hour. Filter, place 20 mL of the filtrate, exactly measured, in a glass-stoppered flask, and add 10 mL of diethyl ether and 0.3 g of ammonium chloride. Shake vigorously with caution. When crystals begin to separate out, shake for 30 minutes with a mechanical shaker, and set aside overnight at a tem- Method of preparation Powdered Opium 100 g Starch or a suitable diluent a sufficient quantity To make 1000 g Prepare as directed under Powders, with the above ingre-dients. Lactose Hydrate should not be used. Description Diluted Opium Powder occurs as a light brown powder. Identification (1) Proceed with 1 g of Diluted Opium Powder as directed in the Identification (1) under Powdered Opium. (2) Proceed with 1 g of Diluted Opium Powder as di-rected in the Identification (2) under Powdered Opium. Assay Place about 50 g of Diluted Opium Powder, accu-rately weighed, in a glass-stoppered flask, and stir with 250 mL of dilute ethanol in a water bath at 409C for 1 hour. Filter the mixture through a glass filter (G3). Transfer the residue on the filter to the first glass-stoppered flask, and add 50 mL of dilute ethanol. Stir the mixture in a water bath at 409C for 10 minutes, and filter through the same glass filter. Repeat the extraction with three 50-mL portions of di-lute ethanol. Evaporate the combined filtrate in a mortar to dryness on a water bath. Add 10 mL of ethanol (99.5) to the residue, evaporate to dryness again, and, after cooling, triturate it with exactly 10 mL of water. Proceed with this so-lution as directed in Assay under Powdered Opium. Each mL of 0.05 mol/L sulfuric acid VS ＝ 28.53 mg of C17H19NO3
Dimenhydrinate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of dimenhydrinate (C17H21NO.C7H7ClN4O2: 469.96). Method of preparation Prepare as directed under Tablets, with Dimenhydrinate. Identification (1) Triturate a quantity of powdered Dimenhydrinate Tablets, equivalent to 0.5 g of Dimen-hydrinate, with 25 mL of warm ethanol (95), and filter. ‌JP XVIII Official Monographs / Dimercaprol 867 Dilute the filtrate with 40 mL of water, and filter again. Use the filtrate as the sample solution. Transfer 30 mL of the sample solution to a separator, and proceed as directed in the Identification (1) under Dimenhydrinate. (2) With 30 mL of the sample solution obtained in (1), proceed as directed in the Identification (2), (3) and (4) under Dimenhydrinate. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Dimenhydrinate Tablets is not less than 85z. Start the test with 1 tablet of Dimenhydrinate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of dimenhydrinate (C17H21NO.
(1) Triturate a quantity of powdered Dimenhydrinate Tablets, equivalent to 0.5 g of Dimen-hydrinate, with 25 mL of warm ethanol (95), and filter. ‌JP XVIII Official Monographs / Dimercaprol 867 Dilute the filtrate with 40 mL of water, and filter again. Use the filtrate as the sample solution. Transfer 30 mL of the sample solution to a separator, and proceed as directed in the Identification (1) under Dimenhydrinate. (2) With 30 mL of the sample solution obtained in (1), proceed as directed in the Identification (2), (3) and (4) under Dimenhydrinate. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Dimenhydrinate Tablets is not less than 85z. Start the test with 1 tablet of Dimenhydrinate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of dimenhydrinate (C17H21NO.C7H7ClN4O2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of dimenhydrinate for assay, previously dried in vacuum using phosphorous (V) oxide as the desiccant for 24 hours, and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 276 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Dissolution rate (z) with respect to the labeled amount of dimenhydrinate (C17H21NO.C7H7ClN4O2) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of dimenhydrinate for assay taken C: Labeled amount (mg) of dimenhydrinate (C17H21NO.C7H7ClN4O2) in 1 tablet
Weigh accurately, and powder not less than 20 Dimenhydrinate Tablets. Weigh accurately a portion of the powder, equivalent to about 0.5 g of dimenhydrinate (C H NO.C H ClN O ), transfer to a flask, add 40 mL of Dimercaprol
Dimercaprol Injection is an oily solution for injec-tion. It contains not less than 95.
Dimercaprol Injection is a clear, colorless or light yellow liquid. It has an unpleasant odor.
Measure a volume of Dimercaprol Injection, equivalent to 30 mg of Dimercaprol, and proceed as directed in the Identification (1) under Dimercaprol. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> Perform the test ac-cording to Method 2: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Dimercaprol Injection, equivalent to about 0.1 g of dimercaprol (C3H8OS2), into a flask, and rinse the pipet several times with a mixture of methanol and diethyl ether (3:1), adding the rinsings to the flask. Add the mixture of methanol and diethyl ether (3:1) to make 50 mL, and titrate <2.50> with 0.05 mol/L iodine VS until a yellow color persists. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L iodine VS ＝ 6.212 mg of C3H8OS2
Dimorpholamine Injection is an aqueous injection. It contains not less than 95.
Dimorpholamine Injection is a clear, colorless liquid. pH: 3.0 – 5.5
(1) To a volume of Dimorpholamine Injec-tion, equivalent to 0.1 g of Dimorpholamine, add 3 drops of Dragendorff's TS: an orange color develops. To a volume of Dimorpholamine Injection, equiva-lent to 50 mg of Dimorpholamine, add 1 mL of dilute hydro-chloric acid, and evaporate on a water bath to dryness. Dis-solve the residue in 2 mL of hydrochloric acid, boil for 10 minutes under a reflux condenser, and evaporate to dryness on a water bath. Dissolve the residue with 1 mL of water, neurtralize with sodium hydroxide TS, and add 0.2 mL of a solution of acetaldehyde (1 in 20), 0.1 mL of sodium pentac-yanonitrosyl ferrate (III) TS and 0.5 mL of sodium carbon-ate TS: a blue color develops. Bacterial endotoxins <4.01> Less than 5.0 EU/mg. Perform the test with the sample diluted to 0.15 w/vz with water for bacterial endotoxins test. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Dimorpholamine Injection, equivalent to about 30 mg of dimorpholamine (C20H38N4O4), and add water to make exactly 200 mL. Pipet 1 mL of this solution, shake with exactly 4 mL of the internal standard solution for 5 minutes, and use this solution as the sample solution. Separately, weigh accurately about 0.15 g of dimorpholamine for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 8 hours, and dissolve in water to make exactly 1000 mL. Pipet 1 mL of this solu-tion, shake with exactly 4 mL of the internal standard solu-tion for 5 minutes, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of dimorphola-mine to that of the internal standard. length: 216 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of dimor- pholamine is about 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, dimorpholamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of dimorpholamine to that of the internal standard is not more than 1.0z.
Diphenhydramine and Bromovalerylurea Powder
ジフェンヒドラミン・バレリル尿素散
Diphenhydramine and Bromovalerylurea Pow-der occurs as a slightly grayish white powder.
(1) To 0.1 g of Diphenhydramine and Bromovalerylurea Powder add 5 mL of dilute hydrochloric acid, 1 mL of ethanol (95) and 10 mL of water, shake, and filter. To the filtrate add 10 mL of sodium hydroxide TS, and extract with 10 mL of chloroform. Separate the chlo-roform layer, add 1 mL of bromophenol blue TS, and shake: a yellow color develops in the chloroform layer (diphen-hydramine tannate). Shake 0.02 g of Diphenhydramine and Bromo-valerylurea Powder with 10 mL of diethyl ether, filter, and evaporate the filtrate on a water bath. Dissolve the residue in 2 mL of sodium hydroxide TS, and add 5 mL of dimethyl-glyoxime-thiosemicarbazide TS, and heat on a water bath for 30 minutes: a red color develops (bromovalerylurea). Shake 0.3 g of Diphenhydramine and Bromo-valerylurea Powder with 5 mL of methanol, filter, and use the filtrate as the sample solution. Dissolve 0.15 of bromo-valerylurea and 0.03 g of diphenhydramine tannate in 5 mL each of methanol, and use the solutions as the standard solu-tion (1) and standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solutions (1) and (2) on a plate of silica gel with fluores- ‌872‌ cent indicator for thin-layer chromatography. Develop the plate in a mixture of ethyl acetate, ethanol (99.5) and ammo-nia solution (28) (50:5:1) to a distance of about 10 cm. Air-dry the plate, and examine under ultraviolet light (main wavelength: 254 nm): 3 spots obtained from the sample solu-tion and the corresponding spot from standard solutions (1) and (2) show the same Rf value. Spray Dragendorff's TS for spraying evenly on the plate: the spot from the standard so-lution (2) and the corresponding spot from the sample solu-tion reveal an orange color.
Determine the absorption spectrum of the sample solution obtained in the Assay, as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 268 nm and 272 nm, and a minimum be-tween 239 nm and 243 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Distigmine Bromide Tablets add 30 mL of 0.1 mol/L hydrochloric acid TS, shake for 1 hour, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate, and add 0.1 mol/L hydrochloric acid (C22H32Br2N4O4) in 1 tablet
Weigh accurately and powder not less than 20 tablets of Distigmine Bromide Tablets. Weigh accurately a portion of the powder, equivalent to about 15 mg of Distig-mine Bromide (C22H32Br2N4O4), add 30 mL of 0.1 mol/L hy-drochloric acid TS, shake for 1 hour, add 0.1 mol/L hydro-chloric acid TS to make exactly 50 mL, and filter. Discard the first 20 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 30 mg of distigmine bromide for assay (previously determine the water <2.48> in the same manner as Distigmine Bromide), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 10 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances of the sample solution and standard solution, AT2 and AS2, at 270 nm and, AT1 and AS1, at 241 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>, respectively. Amount (mg) of distigmine bromide (C22H32Br2N4O4) ＝ MS × (AT2 － AT1)/(AS2 － AS1) × 1/2 MS: Amount (mg) of distigmine bromide for assay taken, calculated on the anhydrous basis
Docetaxel for Injection is a preparation for injection which is dissolved before use. It contains not less than 93.
Docetaxel for Injection occurs as a clear and yellow to orange-yellow, viscous liquid.
To an amount of Docetaxel for Injection, tion of the peak area of docetaxel is not more than 2.0z. equivalent to 20 mg of docetaxel (C 43H 53NO14 ), add 50 mL Bacterial endotoxins <4.01> Less than 2.5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Related substances—Perform the test with 20 mL of the sample solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of them by the ‌JP XVIII Official Monographs / Domperidone 881 area percentage method: the amount of each peak, having the relative retention time of about 0.27, about 1.05, about 1.08, about 1.13, and about 1.18 to docetaxel, is not more than 0.30z, not more than 1.3z, not more than 1.5z, not more than 0.50z, and not more than 0.50z, respectively, the amount of each peak other than docetaxel, the peak hav-ing the relative retention time of about 0.97 and the peaks mentioned above is not more than 0.20z, and the total amount of the peaks other than docetaxel and the peak hav-ing the relative retention time of about 0.97 is not more than 3.5z. For the area of the peak, having the relative retention time of about 0.27 to docetaxel, multiply the correction factor 0.67. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Docetaxel Hydrate. Time span of measurement: For 39 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water, acetonitrile for liquid chroma-tography and acetic acid (100) (1000:1000:1) to make 100 mL, and use this solution as the solution for system suitabil-ity test. Pipet 5 mL of the solution for system suitability test, add a mixture of water, acetonitrile for liquid chromatogra-phy and acetic acid (100) (1000:1000:1) to make exactly 100 mL. Confirm that the peak area of docetaxel obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of docetaxel is not more than 2.0z. Bacterial endotoxins <4.01> Less than 2.5 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. (T: 120.0z). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To exactly a volume of Docetaxel Injection, equiva-lent to about 20 mg of docetaxel (C H NO ), add 5 mL of of methanol, and use this solution as the sample solution. Separately, dissolve 4 mg of docetaxel hydrate in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Then develop the plate with a mixture of ethyl acetate, heptane and ethanol (99.5) (12:3:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the spot obtained from the sample solution and the standard solution is the same. 43 53 14 ethanol (99.5), further add a mixture of water, acetonitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Docetaxel RS (separately determine the water <2.48> and the residual solvent in the same manner as Docetaxel Hydrate), dissolve in 20 mL of ethanol (99.5), add pH Being specified separately when the drug is granted ap-proval based on the Law. Purity Related substances—Perform the test with 20 mL of the sample solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of them by the ‌JP XVIII Official Monographs / Domperidone 881 area percentage method: the amount of each peak, having the relative retention time of about 0.27, about 1.05, about 1.08, about 1.13, and about 1.18 to docetaxel, is not more than 0.30z, not more than 1.3z, not more than 1.5z, not more than 0.50z, and not more than 0.50z, respectively, the amount of each peak other than docetaxel, the peak hav-ing the relative retention time of about 0.97 and the peaks mentioned above is not more than 0.20z, and the total amount of the peaks other than docetaxel and the peak hav-ing the relative retention time of about 0.97 is not more than 3.5z. For the area of the peak, having the relative retention time of about 0.27 to docetaxel, multiply the correction factor 0.67. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Docetaxel Hydrate. Time span of measurement: For 39 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water, acetonitrile for liquid chroma-tography and acetic acid (100) (1000:1000:1) to make 100 mL, and use this solution as the solution for system suitabil-ity test. Pipet 5 mL of the solution for system suitability test, add a mixture of water, acetonitrile for liquid chromatogra-phy and acetic acid (100) (1000:1000:1) to make exactly 100 mL. Confirm that the peak area of docetaxel obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of docetaxel is not more than 2.0z. Bacterial endotoxins <4.01> Less than 2.5 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. (T: 120.0z). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Weigh accurately an amount of Docetaxel for Injec-tion, equivalent to about 20 mg of docetaxel (C43H53NO14), add 5 mL of ethanol (99.5), further add a mixture of water, acetonitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Docetaxel RS (separately determine the water <2.48> and the residual solvent in the same manner as Docetaxel Hydrate), dissolve in 20 mL of ethanol (99.5), add a mixture of water, acetonitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of docetaxel in each solution. Amount (mg) of docetaxel (C43H53NO14) in 1 mL of Docetaxel for Injection ＝ MS/MT × AT/AS × d × 1/2 MS: Amount (mg) of Docetaxel RS taken, calculated on the anhydrous and residual solvent-free basis MT: Amount (mg) of Docetaxel for Injection taken d: Density (g/mL) of Docetaxel for Injection Operating conditions— Proceed as directed in the operating conditions in the Assay under Docetaxel Hydrate. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of docetaxel is not more than 1.0z.
Docetaxel Injection is a hydrophilic injection. It contains not less than 93.
Docetaxel Injection occurs as a clear and pale yellow to orange-yellow, liquid.
To a volume of Docetaxel Injection, equiva-lent to 20 mg of docetaxel (C43H53NO14), add 50 mL of methanol, and use this solution as the sample solution. Separately, dissolve 4 mg of docetaxel hydrate in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Then develop the plate with a mixture of ethyl acetate, heptane and ethanol (99.5) (12:3:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf value of the spot from the sample solution and the standard solution is the same. ‌880 pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—Perform the test with 20 mL of the sample solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of them by the area percentage method: the amount of each peak, having the relative retention time of about 0.27, about 1.05, about 1.08, about 1.13, and about 1.18 to docetaxel, is not more than 0.30z, not more than 1.3z, not more than 1.5z, not more than 0.50z, and not more than 0.50z, respectively, the amount of each peak other than docetaxel, the peak hav-ing the relative retention time of about 0.97 and the peaks mentioned above is not more than 0.20z, and the total amount of the peaks other than docetaxel and the peak hav-ing the relative retention time of about 0.97 is not more than 3.5z. For the area of the peak, having the relative retention time of about 0.27 to docetaxel, multiply the correction factor 0.67. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Docetaxel Hydrate. Time span of measurement: For 39 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of water, acetonitrile for liquid chroma-tography and acetic acid (100) (1000:1000:1) to make 100 mL, and use this solution as the solution for system suitabil-ity test. Pipet 5 mL of the solution for system suitability test, add a mixture of water, acetonitrile for liquid chroma-tography and acetic acid (100) (1000:1000:1) to make exactly 100 mL. Confirm that the peak area of docetaxel obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia- a mixture of water, acetonitrile for liquid chromatography and acetic acid (100) (1000:1000:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of docetaxel in each solution. Amount (mg) of docetaxel (C43H53NO14) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Docetaxel RS taken, calculated on the anhydrous and residual solvent-free basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Docetaxel Hydrate. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of docetaxel are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of docetaxel is not more than 1.0z.
Donepezil Hydrochloride Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of donepezil hydrochloride (C24H29NO3.HCl: 415.95). Method of preparation Prepare as directed under Gran-ules, with Donepezil Hydrochloride. Identification To 2.5 mL of the sample solution obtained in the Assay add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 228 nm and 232 nm, between 269 nm and 273 nm, and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Donepezil Hydrochloride Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Donepezil Hydrochloride Fine Granule add exactly V mL of 0.1 mol/L hydrochloric acid TS so that each mL contains about 0.
To 2.5 mL of the sample solution obtained in the Assay add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 228 nm and 232 nm, between 269 nm and 273 nm, and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Donepezil Hydrochloride Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Donepezil Hydrochloride Fine Granule add exactly V mL of 0.1 mol/L hydrochloric acid TS so that each mL contains about 0.2 mg of donepezil hydrochloride (C24H29NO3.HCl), disperse the particles by sonicating with occasional shaking, and sonicate for a further 10 minutes. Centrifuge this solu-tion, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of Donepezil Hy-drochloride RS, (separately determine the water <2.48> in the same manner as Donepezil Hydrochloride), dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 25 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of donepezil in each solution. length: 271 nm). Amount (mg) of donepezil hydrochloride (C H NO .HCl) Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ＝ MS × AT/AS × V/250 24 29 3 gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 2.5 g of sodium 1-decansulfonate in 650 mL of water, and add 350 mL of acetonitrile and 1 mL of perchloric acid. Flow rate: Adjust so that the retention time of donepezil is about 11 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌area of donepezil is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Donepezil Hydro-chloride Fine Granules is not less than 80z. Start the test with an accurately weighed amount of Donepezil Hydrochloride Fine Granules, equivalent to about 3 mg of donepezil hydrochloride (C H NO .HCl), with- chloride), and dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 25 mL. Pipet 10 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of donepezil in each solution. 24 29 3 draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 55 mg of Donepezil Hydrochloride RS (separately de-termine the water <2.48> in the same manner as Donepezil Hydrochloride), and dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid (3:1) to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL. Pipet 3 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of donepezil in each solution. Dissolution rate (z) with respect to the labeled amount of donepezil hydrochloride (C24H29NO3.HCl) ＝ MS/MT × AT/AS × 1/C × 27/5 MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (mg) of Donepezil Hydrochloride Fine Granules taken C: Labeled amount (mg) of donepezil hydrochloride (C24H29NO3.HCl) in 1 g Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. Mobile phase: A mixture of water, acetonitrile and per-chloric acid (650:350:1). Flow rate: Adjust so that the retention time of donepezil is about 4 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of throretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of donepezil is not more than 1.0z.
Powder Donepezil Hydrochloride Fine Granules, if necessary. Weigh accurately a portion of the powder, equivalent to about 20 mg of donepezil hydrochloride (C24H29NO3.HCl), add 30 mL of 0.1 mol/L hydrochloric acid TS, disperse into the fine particles by sonicating with occasional shaking, and sonicate for a further 15 minutes. Add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of Donepezil Hydrochloride RS (separately determine the water <2.48> in the same manner as Donepezil Hydro- Amount (mg) of donepezil hydrochloride (C24H29NO3.HCl) ＝ MS × AT/AS × 2/5 MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of donepezil is not more than 1.0z.
Donepezil Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of donepezil hydrochloride (C24H29NO3.HCl: 415.95). Method of preparation Prepare as directed under Tablets, with Donepezil Hydrochloride. Identification To 2.5 mL of the sample solution obtained in the Assay add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 228 nm and 232 nm, between 269 nm and 273 nm, and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Donepezil Hydrochloride Tablets add ex-actly V mL of a mixture of methanol and 0.1 mol/L hydro-chloric acid TS (3:1) so that each mL contains about 0.
To 2.5 mL of the sample solution obtained in the Assay add water to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 228 nm and 232 nm, between 269 nm and 273 nm, and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Donepezil Hydrochloride Tablets add ex-actly V mL of a mixture of methanol and 0.1 mol/L hydro-chloric acid TS (3:1) so that each mL contains about 0.2 mg of donepezil hydrochloride (C24H29NO3.HCl), disperse by sonicating. Shake until the tablet is disintegrated, and soni-cate for a further 10 minutes. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of Donepezil Hydrochloride RS (separately, determine the water <2.48> in the same man-ner as Donepezil Hydrochloride), dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 25 mL. Pipet 5 mL of this solution, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make JP XVIII Official Monographs / Donepezil Hydrochloride Tablets 885 exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of donepezil in each solution. about 4 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and Amount (mg) of donepezil hydrochloride (C ＝ MS × AT/AS × V/250 24H 29NO3.HCl) not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat- MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of donepezil is not more than 1.0z. ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Donepezil Hydro-chloride Tablets is not less than 80z. Start the test with 1 tablet of Donepezil Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 3.3 mg of donepezil hydrochloride (C24H29NO3.HCl), and use this so-lution as the sample solution. Separately, weigh accurately about 55 mg of Donepezil Hydrochloride RS (separately de-termine the water <2.48> in the same manner as Donepezil Hydrochloride), and dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add the dissolution medium to make exactly 50 mL. Further, pipet 3 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of donepezil in each solution. Dissolution rate (z) with respect to the labeled amount of donepezil hydrochloride (C24H29NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 27/5 MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of donepezil hydrochloride (C24H29NO3.HCl) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. Mobil phase: A mixture of water, acetonitrile and perchlo-ric acid (650:350:1). Flow rate: Adjust so that the retention time of donepezil is ing conditions, the relative standard deviation of the peak area of donepezil is not more than 1.0z.
Accurately weigh the mass of not less than 20 Donepezil Hydrochloride Tablets and powder. Weigh accu-rately a portion of the powder, equivalent to about 20 mg of donepezil hydrochloride (C24H29NO3.HCl), add 30 mL of a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1), disperse by sonicating, and add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of Donepezil Hydrochloride RS (separately, deter-mine the water <2.48> in the same manner as Donepezil Hy-drochloride), dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 25 mL. Pipet 10 mL of this solution, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of donepezil in each solution. Amount (mg) of donepezil hydrochloride (C24H29NO3.HCl) ＝ MS × AT/AS × 2/5 MS: Amount (mg) of Donepezil Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Donepezil Hydrochloride. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of donepezil are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of donepezil is not more than 1.0z.
Dopamine Hydrochloride Injection is an aqueous injection. It contains not less than 97.
Dopamine Hydrochloride Injection occurs as a clear, colorless liquid.
To a volume of Dopamine Hydrochloride Injection, equivalent to 0.04 g of Dopamine Hydrochloride, add 0.1 mol/L hydrochloric acid TS to make 100 mL. To 5 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 278 nm and 282 nm. pH <2.54> 3.0 – 5.0 Bacterial endotoxins <4.01> Less than 4.2 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Dopamine Hydrochloride In-jection, equivalent to about 30 mg of dopamine hydrochlo-ride (C8H11NO2.HCl), add the mobile phase to make exactly 50 mL. Pipet 2.5 mL of this solution, add exactly 2.5 mL of the internal standard solution and the mobile phase to make 20 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 30 mg of dopamine hydro-chloride for assay, previously dried at 1059C for 3 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 2.5 mL of this solution, add exactly 2.5 mL of the internal standard solution and the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution ‌JP XVIII Official Monographs / Doripenem Hydrate 887 as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of dopamine to that of the internal standard. solution of Doripenem Hydrate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Doripenem RS prepared in the Amount (mg) of dopamine hydrochloride (C H NO .HCl) same manner as the sample solution: both spectra exhibit ＝ MS × QT/QS 8 11 2 similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of MS: Amount (mg) of dopamine hydrochloride for assay taken Internal standard solution—A solution of uracil in the mo-bile phase (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Disodium hydrogen phosphate-citric acid buffer solution (pH 3.0). Flow rate: Adjust so that the retention time of dopamine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and dopamine are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of peak area of dopamine to that of the internal standard is not more than 1.0z.
Doripenem for Injection is a preparation for injec-tion, which is dissolved before use. It contains not less than 95.
Doripenem for Injection occurs as a white to pale yellow-brown-white crystalline powder. Identiˆcation Proceed as directed in the Identiˆcation (2) under Doripenem Hydrate. pH <2.54> Dissolve an amount of Doripenem for Injection, equivalent to 0.3 g (potency) of Doripenem Hydrate, in 30 mL of water: the pH of the solution is between 4.5 and 6.0.
(1) Clarity and color of solution—Dissolve an amount of Doripenem for Injection, equivalent to 0.2 g (potency) of Doripenem Hydrate in 20 mL of water, and proceed as directed in the Purity (1) under Doripenem Hy-drate. (2) Related substances—(i) Dissolve an amount of Doripenem for Injection, equivalent to 20 mg (potency) of Doripenem Hydrate, in 10 mL of water, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than doripenem and the peak having the relative retention time of about 2.1 to doripenem, related substance A having the relative retention time of about 2.2, related substance B having the relative retention time of about 2.5 and related substance C having the relative retention time of about 3.2, obtained from the sample solution, is not larger than 1/10 times the peak area of doripenem from the standard solu-tion, and the total area of the peaks other than doripenem and the peak mentioned above from the sample solution is not larger than 1/2 times the peak area of doripenem from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) (i) under Doripenem Hydrate. System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, and add water to make exactly 50 mL. Conˆrm that the peak area of doripenem obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- Operating conditions— Proceed as directed in the operating conditions in the Purity (3) (ii) under Doripenem Hydrate. System suitability— System performance: To 1 mL of the sample solution add 1 mL of 0.1 mol/L hydrochloric acid TS, allow to stand at 25 ± 59C for 15 minutes, and add water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, related substance D and doripenem are eluted in this order with the resolution be-tween these peaks being not less than 5. The number of theo-retical plates and the symmetry factor of the peak of related substance D are not less than 300 and 0.7 to 1.3, respec-tively, and those of the peak of doripenem are not less than 5000 and 0.7 to 1.3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 2.0z. Water <2.48> 4.0 – 5.0z (0.3 g, volumetric titration, back titration). Bacterial endotoxins <4.01> Less than 0.25 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Doripenem for Injection. Weigh accu-rately an amount of the contents, equivalent to about 25 mg (potency) of Doripenem Hydrate, dissolve in water to make exactly 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg (potency) of Doripenem RS (separately determine the water <2.48> in the same manner as Doripenem Hydrate), dissolve in water to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Doripenem Hydrate. ditions, the number of theoretical plates and the symmetry Amount [mg (potency)] of doripenem (C H N O S ) factor of the peak of doripenem are not less than 5000 and not more than 1.3, respectively. ＝ MS × AT/AS × 1000 15 24 4 6 2 System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doripenem is not more than 0.95z. (ii) Dissolve an amount of Doripenem for Injection, equivalent to 20 mg (potency) of Doripenem Hydrate, in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the peak area of related substance D, having the relative retention time of about 0.5 to doripenem, obtained from the sample solution is not larger than the peak area of doripen-em from the standard solution. MS: Amount [mg (potency)] of Doripenem RS taken, cal-culated on the anhydrous basis
Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution contains not less than 93.
Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution is a clear, colorless, and slightly viscous liquid.
(1) Perform the test with 20 mL each of the sample solution and the standard solution obtained in the Assay (1) as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay (1): the retention times of the peak of dorzolamide in the chromato-grams obtained from the sample solution and the standard solution are the same. (2) Perform the test with 20 mL each of the sample solu-tion and the standard solution obtained in the Assay (2) as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay (2): the retention times of the peak of timolol in the chromatograms obtained from the sample solution and the standard solution are the same. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. Viscosity Being specified separately when the drug is grant-ed approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Related substance 1—Use the sample solution obtained in the Assay (1) as the sample solution. Pipet 1 mL of the sample solution, add a mixture of diluted phosphoric acid (1 in 500) and acetonitrile (19:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of related substance OA having the relative reten-tion time of about 0.8 to dorzolamide obtained from the sample solution is not larger than 1/5 times the peak area of dorzolamide from the standard solution, and the peak area of related substance OB having the relative retention time of about 1.2 to dorzolamide from the sample solution is not larger than 2.4 times the peak area of dorzolamide from the standard solution. The area of the peak other than dor-zolamide and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of dor-zolamide from the standard solution. Furthermore, the total area of the peaks other than dorzolamide from the sample solution is not larger than 2.5 times the peak area of dor-zolamide from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (1). Time span of measurement: For 18 minutes after injection of the sample solution. System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). Test for required detectability: Pipet 2 mL of the standard solution, add a mixture of diluted phosphoric acid (1 in 500 mL) and acetonitrile (19:1) to make exactly 20 mL. Confirm that the peak area of dorzolamide obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of dorzolamide is not more than 5.0z. (2) Related substance 2—Use the sample solution ob-tained in the Assay (2) as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the area of the peak other than timolol and the peak having the relative retention time of about 0.49 to timolol obtained from the sample solution is not larger than 2/5 times the peak area of timolol from the standard solution. Further- length: 253 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile (19:1). Mobile phase B: A mixture of acetonitrile and diluted phosphoric acid (1 in 500) (19:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. more, the total area of the peaks other than timolol and the peak having the relative retention time of about 0.49 to timolol, from the sample solution is not larger than 1/2 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) times the peak area of timolol from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (2). Time span of measurement: For 10 minutes after injection of the sample solution. System suitability— ‌System performance and system repeatability: Proceed as directed in the system suitability in the Assay (2). Test for required detectability: Pipet 10 mL of the stand-ard solution, add the mobile phase to make exactly 100 mL. Confirm that the peak area of timolol obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> It meets the requirement.
(1) Dorzolamide hydrochloride—Pipet a volume of Dorzolamide Hydrochloride and Timolol Maleate Oph-thalmic Solution, equivalent to about 2.5 mg of dorzolamide (C10H16N2O4S3), add a mixture of diluted phosphoric acid (1 in 500) and acetonitrile (19:1) to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Dorzolamide Hydrochloride RS (separately determine the water <2.48> in the same manner as Dorzolamide Hydrochloride), dissolve in a mixture of diluted phosphoric acid (1 in 500) and acetonitrile (19:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of dor-zolamide in each solution. Amount (mg) of dorzolamide (C10H16N2O4S3) in 1 mL of Dorzolamide Hydrochloride and Timolol Maleate Ophthal-mic Solution ＝ MS/MT × AT/AS × 1/8 × d × 0.899 MS: Amount (mg) of Dorzolamide Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (g) of Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution taken d: Density (g/mL) of Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution Operating conditions— Detector: An ultraviolet absorption photometer (wave- 0 – 15.0 100 0 15.0 – 15.1 100 → 0 0 → 100 15.1 – 20.0 0 100 Flow rate: 1.2 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution, the number of theoretical plates and the symmetry factor of the peak of dorzolamide are not less than 5000 and not more than 3.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution, the relative standard de-viation of the peak area of dorzolamide is not more than 2.0z. (2) Timolol maleate—Pipet a volume of Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution, equivalent to about 6.5 mg of timolol (C13H24N4O3S), add the mobile phase to make exactly 25 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 34 mg of Timolol Maleate RS, previously dried at 1009C under reduced pressure for 3 hours, dissolve in the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of timolol in each solution. Amount (mg) of timolol (C13H24N4O3S) in 1 mL of Dor-zolamide Hydrochloride and Timolol Maleate Ophthalmic Solution ＝ MS/MT × AT/AS × 1/4 × d × 0.732 MS: Amount (mg) of Timolol Maleate RS taken MT: Amount (g) of Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution taken d: Density (g/mL) of Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 25 g of sodium dihydrogen phos-phate dihydrate in water to make 2000 mL, and adjust to pH 2.8 with phosphoric acid. To 600 mL of this solution add 400 mL of methanol. Flow rate: 1.0 mL per minute. JP XVIII Official Monographs / Doxapram Hydrochloride Hydrate 895 System suitability— System performance: Dissolve 44 mg of Timolol Maleate RS in 4 mL of sodium hydroxide solution (1 in 250), warm at 709C for 15 hours, and add the mobile phase to make 25 mL. To 5 mL of this solution, add 28 mg of Dorzolamide Hydrochloride RS to dissolve, add the mobile phase to make 25 mL, and use this solution as the solution for system suitability test. When the procedure is run with 20 mL of the solution for system suitability test according to the above op-erating conditions, the number of theoretical plates and the symmetry factor of the peak of timolol are not less than 3000 and not more than 2.0, respectively. The resolution between the co-eluting peak of dorzolamide and maleate, having the relative retention time of about 0.49 to timolol, and the peak, having the relative retention time of about 0.58 to timolol, is not less than 1.5, and the resolution between the peaks having the relative retention times of about 0.58 and about 0.70 to timolol is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test, the rel-ative standard deviation of the peak area of timolol is not more than 2.0z.
Dorzolamide Hydrochloride Ophthalmic Solution
Dorzolamide Hydrochloride Ophthalmic Solution is an aqueous ophthalmic preparation. It contains not less than 95.
Dorzolamide Hydrochloride Ophthalmic Solu-tion occurs as a clear and colorless liquid.
To a volume of Dorzolamide Hydrochloride Ophthalmic Solution, equivalent to about 1.2 mg of dor-zolamide (C10H16N2O4S3), add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 252 nm and 256 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
cis-Isomer—Use the sample solution obtained in the Assay as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions, and deter-mine the peak area of dorzolamide, A2, and that of cis-isomer, having the relative retention time of about 1.1 to dorzolamide, A1, by the automatic integration method: A1/ (A1 ＋ A2) is not larger than 0.020. Diluting solution: To 2 mL of phosphoric acid add 900 mL of water, adjust to pH 3.0 with triethylamine, then add water to make 1000 mL. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the sample solution add the diluting solution to make exactly 100 mL. Pipet 1 mL of this solution, add the diluting solu-tion to make exactly 20 mL, and use this solution as the solu-tion for system suitability test. Confirm that the peak area of dorzolamide obtained with 20 mL of the solution for system suitability test is equivalent to 0.07 to 0.13z of that with 20 mL of the sample solution. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of dorzolamide is not more than 7z. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Direct in-oculation method, using the culture medium containing 0.7z polysorbate 80 and 0.1z of lecithin: it meets the re-quirement. JP XVIII Official Monographs / Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution 893
Weigh accurately a portion of Dorzolamide Hydro-chloride Ophthalmic Solution, equivalent to about 5 mg of dorzolamide (C10H16N2O4S3), add the diluting solution to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 20 mg of Dor-zolamide Hydrochloride RS (separately determine the water <2.48> in the same manner as Dorzolamide Hydrochloride), dissolve in the diluting solution to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of dorzolamide in each solution. ‌Diluting solution: To 2 mL of phosphoric acid add 900 mL of water, adjust to pH 3.0 with triethylamine, then add water to make 1000 mL. Amount (mg/mL) of dorzolamide (C10H16N2O4S3) ＝ MS/MT × AT/AS × 1/4 × d × 0.899 MS: Amount (mg) of Dorzolamide Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (g) of Dorzolamide Hydrochloride Ophthal-mic Solution taken d: Density (g/mL) of Dorzolamide Hydrochloride Oph-thalmic Solution Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 253 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of the diluting solution and acetonitrile (19:1). Flow rate: Adjust so that the retention time of dorzola-mide is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of dorzolamide are not less than 6000 and not more than 1.8, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of dorzolamide is not more than 1.0z.
Doxazosin Mesilate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of doxazosin (C23H25N5O5: 451.48). Method of preparation Prepare as directed under Tablets, with Doxazosin Mesilate. Identification To a quantity of powdered Doxazosin Mesi-late Tablets, equivalent to 5 mg of doxazosin (C23H25N5O5), add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake vigorously, and centrifuge. To 4 mL of the superna-tant liquid add 0.01 mol/L hydrochloric acid-methanol TS to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 244 nm and 248 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Doxazosin Mesilate Tablets add 1 mL of water, disintegrate the tablet by shaking, add 0.01 mol/L hy-drochloric acid-methanol TS to make exactly 100 mL, and shake for 30 minutes. Centrifuge, pipet V mL of the super-natant liquid, add 0.01 mol/L hydrochloric acid-methanol TS to make exactly V? mL so that each mL contains about 5 mg of doxazosin (C23H25N5O5), and use this solution as the sample solution.
To a quantity of powdered Doxazosin Mesi-late Tablets, equivalent to 5 mg of doxazosin (C23H25N5O5), add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake vigorously, and centrifuge. To 4 mL of the superna-tant liquid add 0.01 mol/L hydrochloric acid-methanol TS to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 244 nm and 248 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Doxazosin Mesilate Tablets add 1 mL of water, disintegrate the tablet by shaking, add 0.01 mol/L hy-drochloric acid-methanol TS to make exactly 100 mL, and shake for 30 minutes. Centrifuge, pipet V mL of the super-natant liquid, add 0.01 mol/L hydrochloric acid-methanol TS to make exactly V? mL so that each mL contains about 5 mg of doxazosin (C23H25N5O5), and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Doxazosin Mesilate RS, previously dried at 1059C for 4 hours, and dissolve in 0.01 mol/L hydrochloric acid-methanol TS to make exactly 100 mL. Pipet 2 mL of this so-lution, add 0.01 mol/L hydrochloric acid-methanol TS to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of doxazosin (C23H25N5O5) ＝ MS × AT/AS × V?/V × 1/50 × 0.825 MS: Amount (mg) of Doxazosin Mesilate RS taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 15 minutes of Doxazosin Mesilate Tablets is not less than 75z. Start the test with 1 tablet of Doxazosin Mesilate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add the dissolution medium to make exactly V? mL so that each mL contains about 0.56 mg of doxazosin (C23H25N5O5). Pipet 5 mL of this solution, add exactly 5 mL of methanol, and use this solution as the sample solution. Separately, weigh accurately about 21 mg of Doxazosin Mesilate RS, previously dried at 1059C for 4 hours, and dis-solve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL. Then, pipet 2 mL of this solution, add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the dissolu-tion medium, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of doxazosin in each solution. Dissolution rate (z) with respect to the labeled amount of doxazosin (C23H25N5O5) ＝ MS × AT/AS × V?/V × 1/C × 72/25 × 0.825 MS: Amount (mg) of Doxazosin Mesilate RS taken C: Labeled amount (mg) of doxazosin (C23H25N5O5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 246 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 500 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (1 in 10). To 450 mL of this solution add 550 mL of methanol. Flow rate: Adjust so that the retention time of doxazosin is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of doxazosin are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of doxazosin is not more than 2.0z.
Weigh accurately the mass of not less than 20 Dox-azosin Mesilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 5 mg of doxazo-sin (C23H25N5O5), add 0.01 mol/L hydrochloric acid-methanol TS to make exactly 100 mL, and stir for 30 minutes. Centrifuge, pipet 4 mL of the supernatant liquid, add 0.01 mol/L hydrochloric acid-methanol TS to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 24 mg of Doxazosin Mesilate RS, previously dried at 1059C for 4 hours, dissolve in 0.01 mol/L hydrochloric acid-methanol TS to make ex-actly 100 mL. Pipet 2 mL of this solution, add 0.01 mol/L hydrochloric acid-methanol TS to make exactly 100 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, of the sample solution and standard solution at 246 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of doxazosin (C23H25N5O5) ＝ MS × AT/AS × 1/4 × 0.825 MS: Amount (mg) of Doxazosin Mesilate RS taken
(1) Dissolve an amount of the contents of Doxifluridine Capsules, equivalent to 20 mg of Doxifluri-dine, in 0.1 mol/L hydrochloric acid TS to make 100 mL, and filter. To 1 mL of the filtrate add 0.1 mol/L hydrochlo-ric acid TS to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using 0.1 mol/L hydro-chloric acid TS as the blank: it exhibits a maximum between 267 nm and 271 nm. (2) To an amount of powdered contents of Doxifluridine Capsules, equivalent to 20 mg of Doxifluridine, add 2 mL of methanol, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 20 mg of doxi-fluridine in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer ‌JP XVIII Official Monographs / Doxorubicin Hydrochloride 899 chromatography. Develop the plate with a mixture of ethyl acetate, acetic acid (100) and water (17:2:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show a dark purple color and these Rf values are the same. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Doxifluridine Capsules is not less than 85z. Start the test with 1 capsule of Doxifluridine Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 13 mg of doxifluridine (C9H11FN2O5), and use this solution as the sample solution. Separately, weigh accu-rately about 26 mg of doxifluridine for assay, previously dried at 1059C for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 269 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of doxifluridine (C9H11FN2O5) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of doxifluridine for assay taken C: Labeled amount (mg) of doxifluridine (C9H11FN2O5) in 1 capsule
Weigh accurately the mass and powder the contents of not less than 20 Doxifluridine Capsules. Weigh accurately a portion of the powder, equivalent to about 50 mg of doxi-fluridine (C9H11FN2O5), add 40 mL of water, shake for 10 minutes, add water to make exactly 50 mL, and filter. Dis-card the first 10 mL of the filtrate, pipet 5 mL of the subse-quent filtrate, add exactly 10 mL of the internal standard so-lution, then add a mixture of water and methanol (5:3) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of doxifluridine for assay, previously dried at 1059C for 4 hours, and dis- Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (13:7). Flow rate: Adjust so that the retention time of doxiflu- ridine is about 2.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, doxifluridine and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak height of doxifluridine to that of the internal stand-ard is not more than 1.0z.
Doxorubicin Hydrochloride for Injection is a prepa-ration for injection, which is dissolved before use. It contains not less than 90.
Doxorubicin Hydrochloride for Injection oc-curs as a red-orange, powder or masses.
Dissolve an amount of Doxorubicin Hydro-chloride for Injection, equivalent to 10 mg (potency) of Doxorubicin Hydrochloride, in methanol to make 100 mL. To 5 mL of this solution add methanol to make 50 mL, and determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 231 nm and 235 nm, between 250 nm and 254 nm, between 477 nm and 481 nm, and between 493 nm and 497 nm, and exhibits a shoulder between 528 nm and 538 nm. ‌JP XVIII Official Monographs / Doxycycline Hydrochloride Hydrate 901 pH <2.54> The pH of a solution, prepared by dissolving an amount of Doxorubicin Hydrochloride for Injection equiva-lent to 10 mg (potency) of Doxorubicin Hydrochloride, in 2 mL of water, is 5.0 to 6.0.
Clarity and color of solution—Dissolve an amount of Doxorubicin Hydrochloride for Injection, equivalent to 50 mg (potency) of Doxorubicin Hydrochloride, in 10 mL of water: the solution is clear and red. Water <2.48> Not more than 4.0z (0.25 g, volumetric titration, direct titration). Bacterial endotoxins <4.01> Less than 2.50 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately amounts of Doxorubicin Hydro-chloride and Doxorubicin Hydrochloride RS, equivalent to about 10 mg (potency), add exactly 5 mL of the internal standard solution to each, dissolve each in the mobile phase to make 100 mL, and use these solutions as the sample solu-tion and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according Method of preparation Prepare as directed under Injec-tions, with Doxorubicin Hydrochloride. Description Doxorubicin Hydrochloride for Injection oc-curs as a red-orange, powder or masses. Identification Dissolve an amount of Doxorubicin Hydro-chloride for Injection, equivalent to 10 mg (potency) of Doxorubicin Hydrochloride, in methanol to make 100 mL. To 5 mL of this solution add methanol to make 50 mL, and determine the absorption spectrum of the solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 231 nm and 235 nm, between 250 nm and 254 nm, between 477 nm and 481 nm, and between 493 nm and 497 nm, and exhibits a shoulder between 528 nm and 538 nm. ‌JP XVIII Official Monographs / Doxycycline Hydrochloride Hydrate 901 pH <2.54> The pH of a solution, prepared by dissolving an amount of Doxorubicin Hydrochloride for Injection equiva-lent to 10 mg (potency) of Doxorubicin Hydrochloride, in 2 mL of water, is 5.0 to 6.0. Purity Clarity and color of solution—Dissolve an amount of Doxorubicin Hydrochloride for Injection, equivalent to 50 mg (potency) of Doxorubicin Hydrochloride, in 10 mL of water: the solution is clear and red. Water <2.48> Not more than 4.0z (0.25 g, volumetric titration, direct titration). Bacterial endotoxins <4.01> Less than 2.50 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Weigh accurately the mass of the contents of not less than 10 containers of Doxorubicin Hydrochloride for Injec-tion. Weigh accurately an amount of the contents, equiva-lent to about 10 mg (potency) of Doxorubicin Hydrochlo-ride, add exactly 5 mL of the internal standard solution and the mobile phase to make 100 mL, and use the solution as the sample solution. Separately, weigh accurately an amount of Doxorubicin Hydrochloride RS, equivalent to 10 mg (po-tency), add exactly 5 mL of the internal standard solution and the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and calculate the ratios, QT and QS, of the peak area of doxorubicin to that of the inter-nal standard. Amount [mg (potency)] of doxorubicin hydrochloride (C27H29NO11.HCl) ＝ MS × QT/QS MS: Amount [mg (potency)] of Doxorubicin Hydrochlo-ride RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 3 g of sodium lauryl sulfate in 1000 mL of diluted phosphoric acid (7 in 5000). To this solu-tion add 1000 mL of acetonitrile. Flow rate: Adjust so that the retention time of doxoru-bicin is about 8 minutes. System suitability— bicin is between 0.8 and 1.2. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of doxorubicin to that of the internal standard is not more than 1.0z.
Doxycycline Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled potency of doxycycline (C22H24N2O8: 444.43). Method of preparation Prepare as directed under Tablets, with Doxycycline Hydrochloride Hydrate. Identification Weigh a portion of powdered Doxycycline Hydrochloride Tablets, equivalent to 1 mg (potency) of Doxycycline Hydrochloride Hydrate, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake thoroughly, and filter. Determine the absorption spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 266 nm and 271 nm and between 347 nm and 353 nm. Purity 4-Epidoxycycline—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 2 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con- Test for required detectability: To exactly 2 mL of the standard solution add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Confirm that the peak area of doxycy-cline obtained with 10 mL of this solution is equivalent to 7 to 13z of the peak area of doxycycline with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of doxycycline are not less than 2200 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of doxycycline is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Doxycycline Hydrochloride Tablets add 0.01 mol/L hydrochloric acid TS, disperse the tablet by sonicating, shake for 15 minutes, then add 0.01 mol/L hy-drochloric acid TS to make exactly V mL so that each mL contains about 1 mg (potency) of Doxycycline Hydrochlo-ride Hydrate.
Weigh a portion of powdered Doxycycline Hydrochloride Tablets, equivalent to 1 mg (potency) of Doxycycline Hydrochloride Hydrate, add 100 mL of 0.01 mol/L hydrochloric acid-methanol TS, shake thoroughly, and filter. Determine the absorption spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 266 nm and 271 nm and between 347 nm and 353 nm.
4-Epidoxycycline—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 2 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con- Test for required detectability: To exactly 2 mL of the standard solution add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Confirm that the peak area of doxycy-cline obtained with 10 mL of this solution is equivalent to 7 to 13z of the peak area of doxycycline with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of doxycycline are not less than 2200 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of doxycycline is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Doxycycline Hydrochloride Tablets add 0.01 mol/L hydrochloric acid TS, disperse the tablet by sonicating, shake for 15 minutes, then add 0.01 mol/L hy-drochloric acid TS to make exactly V mL so that each mL contains about 1 mg (potency) of Doxycycline Hydrochlo-ride Hydrate. Centrifuge this solution, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of doxycycline (C22H24N2O8) ＝ MS × AT/AS × V/20 MS: Amount [mg (potency)] of Doxycycline Hydrochlo-ride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Doxycycline Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Doxycycline Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg (potency) of Doxycycline Hydrochloride Hydrate, and use this solution as the sample solution. Separately, weigh accurately about 22 mg (potency) of Doxycycline Hydrochloride RS, dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 274 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of doxycycline (C22H24N2O8) ＝ MS × AT/AS × V?/V × 1/C × 45 ‌904 MS: Amount [mg (potency)] of Doxycycline Hydrochlo-ride RS taken C: Labeled amount [mg(potency)] of doxycycline (C22H24N2O8) in 1 tablet
To 10 Doxycycline Hydrochloride Tablets add 0.01 mol/L hydrochloric acid TS, disperse them by sonicating, shake for 15 minutes, and add 0.01 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 2 mg (potency) of Doxycycline Hydrochloride Hydrate. Cen-trifuge, if necessary, pipet 10 mL of the supernatant liquid, add 0.01 mol/L hydrochloric acid TS to make exactly 20 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 20 mg (potency) of Doxycycline Hydrochloride RS, dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT Droperidol
Dried Aluminum Hydroxide Gel Fine Granules
乾燥水酸化アルミニウムゲル細粒
To 0.2 g of Dried Aluminum Hydroxide Gel Fine Granules add 20 mL of dilute hydrochloric acid, warm and centrifuge: the supernatant liquid responds to Qualita-tive Tests <1.09> for aluminum salt. Acid-consuming capacity Proceed as directed for Acid-consuming capacity under Dried Aluminum Hydroxide Gel: the volume of 0.1 mol/L hydrochloric acid VS consumed is not less than 235 mL per g of Dried Aluminum Hydroxide Gel Fine Granules.
Proceed as directed in the Assay under Dried Alumi-num Hydroxide Gel. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.549 mg of Al O Perform the test with this solution as the test solution. 2 3 Prepare the control solution as follows: evaporate 10 mL of dilute hydrochloric acid to dryness, and add 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—To 0.8 g of Dried Aluminum Hy- droxide Gel add 10 mL of dilute sulfuric acid, heat gently to boil while shaking, cool, and filter. Take 5 mL of the fil-trate, use this solution as the test solution, and perform the
(1) To an amount of the contents of Drox-idopa Capsules, equivalent to 50 mg of Droxidopa, add 50 mL of water, shake for 10 minutes, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and heat in a water bath for 3 minutes: a blue-purple color develops. To an amount of the contents of Droxidopa Cap-sules, equivalent to 20 mg of Droxidopa, add 20 mL of diluted acetic acid (100) (1 in 500), shake for 10 minutes, and filter. To 1 mL of the filtrate add 4 mL of water and 1 drop of iron (III) chloride TS: a deep green color is produced, and it gradually changes to light brown. To an amount of the contents of Droxidopa Cap-sules, equivalent to 50 mg of Droxidopa, add 50 mL of 0.1 mol/L hydrochloric acid TS, shake well, add 0.1 mol/L hy-drochloric acid TS to make 100 mL, and filter. Discard the first 10 mL of the filtrate, and to 2 mL of the subsequent fil-trate add 0.1 mol/L hydrochloric acid TS to make 25 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 278 nm and 282 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To the contents of 1 capsule of Droxidopa Capsules, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake well, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 0.5 mg of droxidopa (C9H11NO5). Filter this solution, discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of droxidopa for assay, previously dried in vacuum at 609C for 3 hours, dissolve in 0.1 mol/L hydro-chloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make ex-actly 25 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 280 nm. Amount (mg) of droxidopa (C9H11NO5) ＝ MS × AT/AS × V/100 MS: Amount (mg) of droxidopa for assay taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Droxidopa Capsules is not less than 70z. Start the test with 1 capsule of Droxidopa Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of droxidopa (C9H11NO5), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of droxidopa for assay, previously dried in vacuum at 609C for 3 hours, dissolve in water to make ex-actly 100 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT1 and AS1, at 280 nm, and AT2 and AS2, at 350 nm. Dissolution rate (z) with respect to the labeled amount of droxidopa (C9H11NO5) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 180 MS: Amount (mg) of droxidopa for assay taken C: Labeled amount (mg) of droxidopa (C9H11NO5) in 1 capsule
Take out the contents of not less than 20 Droxidopa Capsules, weigh accurately the mass of the contents, and mix uniformly. Weigh accurately an amount equivalent to about 50 mg of droxidopa (C9H11NO5), add 50 mL of 0.1 mol/L hydrochloric acid TS, shake well, add 0.1 mol/L hydrochlo-ric acid TS to make exactly 100 mL, and filter. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent fil-trate, add 0.1 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of droxidopa for assay, previously dried in vacuum at 609C for 3 hours, dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the stand-ard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and determine the absorbances, AT and AS, at 280 nm. Amount (mg) of droxidopa (C9H11NO5) ＝ MS × AT/AS MS: Amount (mg) of droxidopa for assay taken
(1) To a quantity of powdered Droxidopa Fine Granules, equivalent to 50 mg of Droxidopa, add 50 mL of water, shake for 10 minutes, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, heat in a water bath for 3 minutes: a blue-purple color develops. To a quantity of powdered Droxidopa Fine Granules, equivalent to 20 mg of Droxidopa, add 20 mL of diluted ace-tic acid (100) (1 in 500), shake for 10 minutes, and filter. To 1 mL of the filtrate add 4 mL of water and 1 drop of iron (III) chloride TS: a deep green color is produced, and it gradually changes to light brown. To a quantity of powdered Droxidopa Fine Granules, ‌JP XVIII Official Monographs / Dydrogesterone 907 equivalent to 50 mg of Droxidopa, add 50 mL of 0.1 mol/L hydrochloric acid TS, shake well, add 0.1 mol/L hydrochlo-ric acid TS to make 100 mL, and filter. Discard the first 10 mL of the filtrate, to 2 mL of the subsequent filtrate add 0.1 mol/L hydrochloric acid TS to make 25 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 278 nm and 282 nm. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Droxidopa Fine Granules is not less than 70z. Start the test with an accurately weighed amount of Drox-idopa Fine Granules, equivalent to about 0.1 g of droxidopa (C9H11NO5), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of water, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of droxidopa for assay, previously dried in vacuum at 609C for 3 hours, dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and determine the absorbances, AT1 and AS1, at 280 nm, and AT2 and AS2, at 350 nm. Dissolution rate (z) with respect to the labeled amount of droxidopa (C9H11NO5) ＝ MS/MT × (AT1 － AT2)/(AS1 － AS2) × 1/C × 360 MS: Amount (mg) of droxidopa for assay taken MT: Amount (g) of Droxidopa Fine Granules taken C: Labeled amount (mg) of droxidopa (C9H11NO5) in 1 g
Powder not less than 20 g of Droxidopa Fine Gran-ules. Weigh accurately a portion of the powder, equivalent to about 50 mg of droxidopa (C9H11NO5), add 50 mL of 0.1 mol/L hydrochloric acid TS, shake well, add 0.1 mol/L hy-drochloric acid TS to make exactly 100 mL, and filter. Dis-card the first 10 mL of the filtrate, pipet 2 mL of the subse-quent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of droxidopa for assay, previously dried in vacuum at 609C for 3 hours, dis-solve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 25 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 280 nm. Amount (mg) of droxidopa (C9H11NO5) ＝ MS × AT/AS MS: Amount (mg) of droxidopa for assay taken
Dydrogesterone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of dydrogesterone (C21H28O2: 312.45). Method of preparation Prepare as directed under Tablets, with Dydrogesterone. Identification (1) To a quantity of powdered Dydroges-terone Tablets, equivalent to 0.05 g of Dydrogesterone, add 50 mL of methanol, shake well, and filter. Evaporate 5 mL of the filtrate on a water bath to dryness. Proceed with the residue as directed in the Identification (1) under Dydroges-terone. (2) To 1 mL of the filtrate obtained in (1) add methanol MS: Amount (mg) of dydrogesterone for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Dydrogesterone Tablets is not less than 80z. Start the test with 1 tablet of Dydrogesterone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of dydrogesterone (C21H28O2), and use this solution as the sample solution.
(1) To a quantity of powdered Dydroges-terone Tablets, equivalent to 0.05 g of Dydrogesterone, add 50 mL of methanol, shake well, and filter. Evaporate 5 mL of the filtrate on a water bath to dryness. Proceed with the residue as directed in the Identification (1) under Dydroges-terone. (2) To 1 mL of the filtrate obtained in (1) add methanol MS: Amount (mg) of dydrogesterone for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Dydrogesterone Tablets is not less than 80z. Start the test with 1 tablet of Dydrogesterone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of dydrogesterone (C21H28O2), and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of dydrogesterone for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 24 hours, and dissolve in methanol to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 296 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>, using water as the con-trol. Dissolution rate (z) with respect to the labeled amount of dydrogesterone (C21H28O2) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of dydrogesterone for assay taken C: Labeled amount (mg) of dydrogesterone (C21H28O2) in 1 tablet.
Weigh accurately and powder not less than 20 Dydrogesterone Tablets. Weigh accurately a portion of the powder, equivalent to about 10 mg of dydrogesterone (C21H28O2), shake with 50 mL of methanol, and add metha-nol to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of dydrogesterone for assay, previously dried in vacuum for 24 hours using phosphorus (V) oxide as a des-sicant, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 100 mL, and use the solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 286 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. to make 200 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 284 nm and Amount (mg) of dydrogesterone (C ＝ MS × AT/AS 21H28O2) 288 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Crush 1 tablet of Dydrogesterone Tablets, and add metha-nol to make exactly 100 mL. Shake until the tablet is com-pletely disintegrated, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 20 MS: Amount (mg) of dydrogesterone for assay taken
Ebastine Orally Disintegrating Tablets contain not less than 95.0z and not more than 105.0z of the with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> labeled amount of ebastine (C32 H39 NO2: 469.66). according to the following conditions, and determine each peak area by the automatic integration method: each peak area other than ebastine obtained from the sample solution is not larger than the peak area of ebastine from the stand-ard solution, and the total area of the peaks other than ebastine from the sample solution is not larger than 4 times the peak area of ebastine from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame- Method of preparation Prepare as directed under Tablets, with Ebastine. Identification Powder Ebastine Orally Disintegrating Tablets. To a portion of the powder, equivalent to 30 mg of Ebastine, add 70 mL of methanol, shake for 10 minutes, then add methanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 251 nm and 255 nm. Purity Related substances—Powder Ebastine Orally Disin-tegrating Tablets. To a portion of the powder, equivalent to 50 mg of Ebastine, add 30 mL of methanol for liquid chro-matography, shake for 10 minutes, and add the mobile phase to make 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ebastine obtained from the sam-ple solution is not larger than the peak area of ebastine from the standard solution, and the total area of the peaks other than ebastine from the sample solution is not larger than 2 times the peak area of ebastine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of ebastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of ebastine obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ebastine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ebastine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ebastine Orally Disintegrating Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disperse the particles by sonicating with occasional shaking. Add Start the test with 1 tablet of Ebastine Orally Disintegrat-ing Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.
Powder Ebastine Orally Disintegrating Tablets. To a portion of the powder, equivalent to 30 mg of Ebastine, add 70 mL of methanol, shake for 10 minutes, then add methanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 251 nm and 255 nm.
Related substances—Powder Ebastine Orally Disin-tegrating Tablets. To a portion of the powder, equivalent to 50 mg of Ebastine, add 30 mL of methanol for liquid chro-matography, shake for 10 minutes, and add the mobile phase to make 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than ebastine obtained from the sam-ple solution is not larger than the peak area of ebastine from the standard solution, and the total area of the peaks other than ebastine from the sample solution is not larger than 2 times the peak area of ebastine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of ebastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of ebastine obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ebastine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ebastine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ebastine Orally Disintegrating Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disperse the particles by sonicating with occasional shaking. Add Start the test with 1 tablet of Ebastine Orally Disintegrat-ing Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of ebastine (C32H39NO2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of ebastine for assay, previously dried at 609C under reduced pressure with phosphorous (V) oxide for 2 hours, and dis-solve in methanol to make exactly 50 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 258 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of ebastine (C32H39NO2) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of ebastine for assay taken C: Labeled amount (mg) of ebastine (C32H39NO2) in 1 tablet
Weigh accurately the mass of not less than 20 Ebastine Orally Disintegrating Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of ebastine (C32H39NO2), add 20 mL of 0.1 mol/L hydro-chloric acid TS, and disperse the particles by sonicating. Add 120 mL of methanol, shake for 10 minutes, add methanol to make exactly 200 mL, and centrifuge. Pipet 5 mL of the su-pernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of ebastine for assay, previously dried at 609C under reduced pressure with phos-phorous (V) oxide for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add 5 mL of 0.1 mol/L hydrochloric acid TS, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ebastine to that of the internal standard. 3V/5 mL of methanol, shake for 10 minutes, then add meth- anol to make exactly V mL so that each mL contains about Amount (mg) of ebastine (C 32H39NO2) 0.1 mg of ebastine (C 32H 39NO2), and centrifuge. Pipet 5 mL ＝ MS × QT/QS × 2/5 of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of ebastine (C32H39NO2) ＝ MS × QT/QS × V/500 MS: Amount (mg) of ebastine for assay taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 40,000). Disintegration Being specified separately when the drug is granted approval based on the Law. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Ebastine Orally Disintegrating Tablets is not less than 80z. MS: Amount (mg) of ebastine for assay taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust to pH 3.0 with diluted phosphoric acid (1 in 5), and add water to make 1000 mL. To 375 mL of this solution add 625 mL of acetonitrile for liquid chromatography, and dissolve 0.72 g of sodium lauryl sulfate in this solution. ‌JP XVIII Official Monographs / Ebastine Tablets 911 Flow rate: Adjust so that the retention time of ebastine is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ebastine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ebastine to that of the internal standard is not more than 1.0z.
Ebastine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of ebastine (C32H39NO2: 469.66). Method of preparation Prepare as directed under Tablets, with Ebastine. Identification Powder Ebastine Tablets. To a portion of the powder, equivalent to 30 mg of Ebastine, add 70 mL of methanol, shake for 10 minutes, then add methanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 251 nm and 255 nm. Purity Related substances—Powder Ebastine Tablets. To a portion of the powder, equivalent to 50 mg of Ebastine, add 30 mL of methanol for liquid chromatography, shake for 10 minutes, and add the mobile phase to make 50 mL. Centri-fuge this solution, and use the supernatant liquid as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than ebastine obtained from the sample solution is not larger than the peak area of ebastine from the standard solution, and the total area of the peaks other than ebastine from the sample solution is not larger than 2 times the peak area of ebastine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of ebastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of ebastine obtained with 10 mL of this solution is equivalent to 15 to 25z of that ob- tained with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ebastine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ebastine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ebastine Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disperse the particles by sonicat-ing with occasional shaking. Add 3V/5 mL of methanol, shake for 10 minutes, then add methanol to make exactly V mL so that each mL contains about 0.
Powder Ebastine Tablets. To a portion of the powder, equivalent to 30 mg of Ebastine, add 70 mL of methanol, shake for 10 minutes, then add methanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 251 nm and 255 nm.
Related substances—Powder Ebastine Tablets. To a portion of the powder, equivalent to 50 mg of Ebastine, add 30 mL of methanol for liquid chromatography, shake for 10 minutes, and add the mobile phase to make 50 mL. Centri-fuge this solution, and use the supernatant liquid as the sam-ple solution. Pipet 1 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than ebastine obtained from the sample solution is not larger than the peak area of ebastine from the standard solution, and the total area of the peaks other than ebastine from the sample solution is not larger than 2 times the peak area of ebastine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of ebastine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of ebastine obtained with 10 mL of this solution is equivalent to 15 to 25z of that ob- tained with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ebastine are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ebastine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ebastine Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disperse the particles by sonicat-ing with occasional shaking. Add 3V/5 mL of methanol, shake for 10 minutes, then add methanol to make exactly V mL so that each mL contains about 0.1 mg of ebastine (C32H39NO2), and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of ebastine (C32H39NO2) ＝ MS × QT/QS × V/500 MS: Amount (mg) of ebastine for assay taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 40,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Ebastine Tablets is not less than 75z. Start the test with 1 tablet of Ebastine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of ebastine (C32H39NO2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of ebastine for assay, previ-ously dried at 609C under reduced pressure with phos-phorous (V) oxide for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 1 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 258 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of ebastine (C32H39NO2) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of ebastine for assay taken C: Labeled amount (mg) of ebastine (C32H39NO2) in 1 tablet
Weigh accurately the mass of not less than 20 Ebastine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of ebastine (C32H39NO2), add 20 mL of 0.1 mol/L hydrochloric acid TS, and disperse the particles by sonicating. Add 120 mL of methanol, shake for 10 minutes, add methanol to make ex-actly 200 mL, and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, ‌912 and use this solution as the sample solution. Separately, weigh accurately about 50 mg of ebastine for assay, previ-ously dried at 609C under reduced pressure with phos-phorous (V) oxide for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add 5 mL of 0.1 mol/L hydrochloric acid TS, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ebastine to that of the internal standard. Amount (mg) of ebastine (C32H39NO2) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of ebastine for assay taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.8 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust to pH 3.0 with diluted phosphoric acid (1 in 5), and add water to make 1000 mL. To 375 mL of this solution add 625 mL of acetonitrile for liquid chromatography, and dissolve 0.72 g of sodium lauryl sulfate in this solution. Flow rate: Adjust so that the retention time of ebastine is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ebastine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ebastine to that of the internal standard is not more than 1.0z.
Ecabet Sodium Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of ecabet sodium hydrate (C20H27NaO5S.5H2O: 492.56). Method of preparation Prepare as directed under Gran-ules, with Ecabet Sodium Hydrate. Identification To a quantity of Ecabet Sodium Granules, equivalent to 50 mg of Ecabet Sodium Hydrate, add 25 mL of dilute sodium hydroxide TS, shake, and filter. Discard the first 10 mL of the filtrate, and to 3 mL of the subsequent fil-trate add dilute sodium hydroxide TS to make 20 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 269 nm and 273 nm, and between 278 nm and 282 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ecabet Sodium Granules in single-dose packages meet the requirement of the Content uniformity test. Take out the total amount of the content of 1 package of Ecabet Sodium Granules, add 70 mL of dilute sodium hy-droxide TS, sonicate for 5 minutes with occasional shaking, add dilute sodium hydroxide TS to make exactly V mL so that each mL contains about 10 mg of ecabet sodium hydrate (C20H27NaO5S.
To a quantity of Ecabet Sodium Granules, equivalent to 50 mg of Ecabet Sodium Hydrate, add 25 mL of dilute sodium hydroxide TS, shake, and filter. Discard the first 10 mL of the filtrate, and to 3 mL of the subsequent fil-trate add dilute sodium hydroxide TS to make 20 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 269 nm and 273 nm, and between 278 nm and 282 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ecabet Sodium Granules in single-dose packages meet the requirement of the Content uniformity test. Take out the total amount of the content of 1 package of Ecabet Sodium Granules, add 70 mL of dilute sodium hy-droxide TS, sonicate for 5 minutes with occasional shaking, add dilute sodium hydroxide TS to make exactly V mL so that each mL contains about 10 mg of ecabet sodium hydrate (C20H27NaO5S.5H2O), and filter. Discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of ecabet sodium hydrate for assay (separately, determine the water <2.48> in the same manner as Ecabet Sodium Hydrate), dis- solve in 2 mL of dilute sodium hydroxide TS, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 271 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Amount (mg) of ecabet sodium hydrate (C20H27NaO5S.5H2O) ＝ MS × AT/AS × V/2 × 1.224 MS: Amount (mg) of ecabet sodium hydrate for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Ecabet Sodium Granules is not less than 80z. Start the test with an accurately weighed amount of Ecabet Sodium Granules, equivalent to about 1 g of Ecabet Sodium Hydrate, withdraw not less than 20 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of ecabet sodium hydrate for assay (separately, determine the water <2.48> in the same manner as Ecabet Sodium Hydrate), dissolve in 1 mL of methanol, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 271 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of ecabet sodium hydrate (C20H27NaO5S.5H2O) ＝ MS/MT × AT/AS × 1/C × 4500 × 1.224 MS: Amount (mg) of ecabet sodium hydrate for assay taken, calculated on the anhydrous basis MT: Amount (g) of Ecabet Sodium Granules taken C: Labeled amount (mg) of ecabet sodium hydrate (C20H27NaO5S.5H2O) in 1 g
Weigh accurately an amount of Ecabet Sodium Granules, equivalent to about 30 mg of ecabet sodium hy-drate (C20H27NaO5S.5H2O), add exactly 5 mL of the internal standard solution, add 25 mL of diluted methanol (1 in 2), shake vigorously for 20 minutes, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, to 3 mL of the subsequent fil-trate add the mobile phase to make 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 30 mg of ecabet sodium hydrate for assay (separately, determine the water <2.48> in the same manner as Ecabet So-dium Hydrate), add exactly 5 mL of the internal standard so-lution, and dissolve in dilute methanol (1 in 2) to make 30 mL. To 3 mL of this solution add the mobile phase to make ‌914 50 mL, and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ecabet to that of the internal standard. Amount (mg) of ecabet sodium hydrate (C20H27NaO5S.5H2O) ＝ MS × QT/QS × 1.224 MS: Amount (mg) of ecabet sodium hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in diluted methanol (1 in 2) (3 in 400). ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 0.1 mol/L potassium dihydrogen phos-phate TS add phosphoric acid to adjust the pH to 3.0. To 730 mL of this solution add 270 mL of acetonitrile. Flow rate: Adjust so that the retention time of ecabet is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, ecabet and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ecabet to that of the internal standard is not more than 1.0z.
Edaravone Injection is an aqueous injection. It contains not less than 95.
Edaravone Injection occurs as a clear and col-orless liquid.
To a volume of Edaravone Injection, equivalent to 1.5 mg of Edaravone, add water to make 50 mL. To 5 mL of this solution add water to make 25 mL. De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 238 nm and 242 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substance—(i) Perform the following test 1). For preparations to which the test 2) can be applied, the test 2) may be performed instead of 1). To a suitable amount of Edaravone Injection add the mobile phase so that each mL contains 0.3 mg of edaravone (C10H10N2O), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak other than edaravone obtained from the sample solution is not larger than 2 times the peak area of edaravone from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Edaravone. Time span of measurement: About 7 times as long as the retention time of edaravone, beginning after the peak of edaravone. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of edaravone are not less than 1500 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of edaravone is not more than 2.0z. Use Edaravone Injection as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak other than edaravone obtained from the sample solution is not larger than 2 times the peak area of edaravone from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Edaravone. Time span of measurement: About 7 times as long as the retention time of edaravone, beginning after the peak of edaravone. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of edaravone are not less than 1500 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of edaravone is not more than 2.0z. Perform the following test 1). For preparations to which the test 2) can be applied, the test 2) may be per-formed instead of 1). To a suitable amount of Edaravone Injection add the mobile phase so that each mL contains 0.3 mg of edaravone (C10H10N2O), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak, having the relative retention time of about 0.3 to edaravone, obtained from the sample solution is not larger than 4 times the peak area of edaravone from the standard solution, the area of the peak, having the relative retention time of about 0.4, ob- tained from the sample solution is not larger than the peak area of edaravone from the standard solution, and the area of the peak other than edaravone and the peaks mentioned above from the sample solution is not larger than 2 times the peak area of edaravone from the standard solution. Operating conditions— Detector, column, and mobile phase: Proceed as directed in the operating conditions in the Assay 1). Column temperature: A constant temperature of about 409C. Flow rate: Adjust so that the retention time of edaravone is about 11 minutes. Time span of measurement: About 2.5 times as long as the retention time of edaravone, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of edaravone are not less than 2000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of edaravone is not more than 2.0z. Use Edaravone Injection as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak, having the relative retention time of about 0.3 to edaravone, obtained from the sample solution is not larger than 4 times the peak area of edaravone from the standard solution, the area of the peak, having the relative retention time of about 0.4, from the sample solution is not larger than the peak area of edaravone from the standard solution, and the area of the peak other than edaravone and the peaks mentioned above from the sample solution is not larger than 2 times the peak area of edaravone from the standard solution. Operating conditions— Detector, column, and mobile phase: Proceed as directed in the operating conditions in the Assay 1). Column temperature: A constant temperature of about 409C. Flow rate: Adjust so that the retention time of edaravone is about 11 minutes. Time span of measurement: About 2.5 times as long as the retention time of edaravone, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of edaravone are not less than 2000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of edaravone is not more than 2.0z. Bacterial endotoxins <4.01> Less than 5.0 EU/mg. Extractable volume <6.05> It meets the requirement. JP XVIII Official Monographs / Edrophonium Chloride 917 ‌Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. the test with 2 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of edaravone to that of Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. the internal standard. Amount (mg) of edaravone (C ＝ MS × QT/QS × 1/25 10H 10N2O)
Perform the following test 1). For preparations to which the test 2) can be applied, the test 2) may be per-formed instead of 1). To exactly V mL of Edaravone Injection add metha-nol to make exactly V? mL so that each mL contains about 0.3 mg of edaravone (C10H10N2O). Pipet 2 mL of this solu-tion, add exactly 10 mL of the internal standard solution, then add methanol to make 20 ml, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of edaravone for assay, previously dried in vacuum using phosphorus (V) oxide as a desiccant for 3 hours, and dissolve in methanol to make exactly 100 mL. Pipet 2 mL of this so-lution, add exactly 10 mL of the internal standard solution, then add methanol to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of edaravone to that of the internal standard. Amount (mg) of edaravone (C10H10N2O) ＝ MS × QT/QS × V?/V × 1/100 MS: Amount (mg) of edaravone for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of diluted dilute acetic acid (1 in 100) and methanol (3:1), adjusted to pH 5.5 with diluted am-monia solution (28) (1 in 20). Flow rate: Adjust so that the retention time of edaravone is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, edaravone and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of edaravone to that of the internal standard is not more than 1.0z. To an exact volume of Edaravone Injection, equiva-lent to about 3 mg of edaravone (C10H10N2O) add exactly 10 mL of the internal standard solution, add methanol to make 20 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 75 mg of edaravone for assay, previously dried in vacuum using phosphorus (V) oxide as a desiccant for 3 hours, and dissolve in methanol to make ex-actly 50 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution, add methanol to make 20 mL, and use this solution as the standard solution. Perform MS: Amount (mg) of edaravone for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 500). Operating conditions— Proceed as directed in the operating conditions in the Assay 1). System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, edaravone and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of edaravone to that of the internal standard is not more than 1.0z.
Edrophonium Chloride Injection is an aqueous in-jection. It contains not less than 95.
Edrophonium Chloride Injection is a clear and colorless liquid.
(1) To a volume of Edrophonium Chloride Injection, equivalent to 0.04 g of Edrophonium Chloride, add 4 mL of barium nitrate TS, shake, and filter. Proceed with the filtrate as directed in the Identification (1) under <2.24>. Amount (mg) of edrophonium chloride (C10H16ClNO) ＝ MS × AT/AS MS: Amount (mg) of Edrophonium Chloride RS taken
Emedastine Fumarate Extended-release Capsules
エメダスチンフマル酸塩徐放カプセル
Emedastine Fumarate Extended-release Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of emedastine fumarate (C17H26N4O.2C4H4O4: 534.56). Method of preparation Prepare as directed under Cap-sules, with Emedastine Fumarate. Identification (1) Powder the content of Emedastine Fumarate Extended-release Capsules. To a portion of the powder, equivalent to 10 mg of Emedastine Fumarate, add 10 mL of water, shake thoroughly, and filter. Spot 1 drop of the filtrate on a filter paper, and spray Dragendorff's TS for spraying on the filter: the spot shows an orange color. (2) To 2 mL of the filtrate obtained in (1) add 1 mol/L hydrochloric acid TS to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 278 nm and 282 nm, and between 284 nm and 288 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Emedastine Fumarate Extended-release Capsules add 40 mL of the mobile phase, sonicate for 30 minutes while occasional vigorous shaking, and add the mo-bile phase to make exactly V mL so that each mL contains about 20 mg of emedastine fumarate (C17H26N4O.
(1) Powder the content of Emedastine Fumarate Extended-release Capsules. To a portion of the powder, equivalent to 10 mg of Emedastine Fumarate, add 10 mL of water, shake thoroughly, and filter. Spot 1 drop of the filtrate on a filter paper, and spray Dragendorff's TS for spraying on the filter: the spot shows an orange color. (2) To 2 mL of the filtrate obtained in (1) add 1 mol/L hydrochloric acid TS to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 278 nm and 282 nm, and between 284 nm and 288 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Emedastine Fumarate Extended-release Capsules add 40 mL of the mobile phase, sonicate for 30 minutes while occasional vigorous shaking, and add the mo-bile phase to make exactly V mL so that each mL contains about 20 mg of emedastine fumarate (C17H26N4O.2C4H4O4). Centrifuge this solution, pipet 10 mL of the supernatant liq-uid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of emedastine fumarate (C17H26N4O.2C4H4O4) ＝ MS × QT/QS × V/1000 MS: Amount (mg) of emedastine fumarate for assay taken Internal standard solution—A solution of 4-methyl-ben-zophenone in the mobile phase (1 in 40,000). Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the content of not less than 20 Emedastine Fumarate Extended-release Capsules, and pow-der. Weigh accurately a portion of the powder, equivalent to about 2 mg of emedastine fumarate (C17H26N4O.2C4H4O4), add 10 mL of the mobile phase, sonicate for 30 minutes while occasional vigorous shaking, and add the mobile phase to make exactly 100 mL. Centrifuge this solution, pipet 10 mL of the supernatant liquid, add exactly 5 mL of the inter-nal standard solution, and use this solution as the sample so-lution. Separately, weigh accurately about 20 mg of emedastine fumarate for assay, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make 100 mL. Pipet 5 mL of this solution, and add the mobile phase to make exactly 50 mL. Then, pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01>, according to the following conditions, and calculate the ratios, QT and QS, of the peak area of emedastine to that of the internal standard. Amount (mg) of emedastine fumarate (C17H26N4O.2C4H4O4) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of emedastine fumarate for assay taken Internal standard solution—A solution of 4-methylben-zophenone in the mobile phase (1 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.9 g of sodium dihydrogen phos- ‌JP XVIII Official Monographs / Emorfazone 923 phate dihydrate and 2.5 g of sodium lauryl sulfate in 1000 mL of water, and adjust to pH 2.4 with phosphoric acid. To 500 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of emedastine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, emedastine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of emedastine to that of the internal standard is not more than 1.0z.
Emorfazone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of emorfazone (C11H17N3O3: 239.27). Method of preparation Prepare as directed under Tablets, with Emorfazone. Identification To a quantity of powdered Emorfazone Tablets, equivalent to 0.1 g of Emorfazone, add 100 mL of water, shake well, and centrifuge. Filter the supernatant liq-uid, and to 1 mL of the filtrate add water to make 100 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 237 nm and 241 nm, and between 310 nm and 314 nm, and a shoulder between 288 nm and 298 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Emorfazone Tablets add methanol to make exactly V mL so that each mL contains about 4 mg of emor-fazone (C11H17N3O3), and shake well to disintegrate.
To a quantity of powdered Emorfazone Tablets, equivalent to 0.1 g of Emorfazone, add 100 mL of water, shake well, and centrifuge. Filter the supernatant liq-uid, and to 1 mL of the filtrate add water to make 100 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 237 nm and 241 nm, and between 310 nm and 314 nm, and a shoulder between 288 nm and 298 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Emorfazone Tablets add methanol to make exactly V mL so that each mL contains about 4 mg of emor-fazone (C11H17N3O3), and shake well to disintegrate. Centri-fuge this solution, pipet 2 mL of the supernatant liquid, add exactly 10 mL of the internal standard solution, add metha-nol to make 50 mL, and use this solution as the sample solu-tion. Proceed as directed in the Assay. Amount (mg) of emorfazone (C11H17N3O3) ＝ MS × QT/QS × V/5 MS: Amount (mg) of emorfazone for assay taken Internal standard solution—A solution of 2,4-dinitrophenyl-hidrazine in methanol (3 in 2000). Prepare before use. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Emorfazone Tablets is not less than 80z. Start the test with 1 tablet of Emorfazone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of emorfazone (C11H17N3O3), and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of emorfazone for assay, previously dried in vacuum at 609C for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 239 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of emorfazone (C11H17N3O3) exactly 250 mL, and centrifuge. Pipet a volume of the supernatant liquid, equivalent to about 8 mg of emorfazone (C11H17N3O3), add exactly 10 mL of the internal standard so-lution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of emorfazone for assay, previously dried in vacuum at 609C for 4 hours, and dissolve in methanol to make ex-actly 25 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of emorfazone to that of the internal standard. Amount (mg) of emorfazone (C11H17N3O3) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of emorfazone for assay taken Internal standard solution—A solution of 2,4-dinitrophenyl-hidrazine in methanol (3 in 2000). Prepare before use. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 313 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (11:10). Flow rate: Adjust so that the retention time of emorfazone is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, emorfazone and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of emorfazone to that of the internal standard is not more than 1.0z.
Enalapril Maleate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of enalapril maleate (C20H28N2O5.C4H4O4: 492.52). Method of preparation Prepare as directed under Tablets, with Enalapril Maleate. Identification To a quantity of powdered Enalapril Male-ate Tablets, equivalent to 50 mg of Enalapril Maleate, add 20 mL of methanol, shake, centrifuge, and then use the su-pernatant liquid as the sample solution. Separately, dissolve 25 mg of enalapril maleate in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-Layer Chromatogra-phy <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of water, acetone, 1-butanol, acetic acid (100) and toluene (1:1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the Rf values of the 2 spots obtained from the sample solution and the 2 spots from the standard solu-tion are equivalent. Purity Enalaprilat and enalapril diketopiperazine—Use the sample solution obtained in the Assay as the sample solu-tion. Pipet 1 mL of the sample solution, add sodium dihy-drogen phosphate TS (pH 2.2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the peak area of enalaprilat, having the relative retention time of about 0.5 to enalapril obtained from the sample solution, is not larger than 2 times the peak area of enalapril from the standard solution. Also, the peak area of enalapril diketopiperazine, having the relative retention time of about 1.5 to enalapril, from the sample solution is not larger than the peak area of enalapril from the standard so-lution. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 1 mL of the standard solution, and add sodium dihydrogen phosphate TS (pH 2.2) to make exactly 10 mL. Confirm that the peak area of enalapril obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Enalapril Maleate Tablets, add V/2 mL of sodium dihydrogen phosphate TS (pH 2.2), sonicate for 15 minutes, shake for 30 minutes, and add sodium dihydro-gen phosphate TS (pH 2.2) to make exactly V mL so that each mL contains about 0.
To a quantity of powdered Enalapril Male-ate Tablets, equivalent to 50 mg of Enalapril Maleate, add 20 mL of methanol, shake, centrifuge, and then use the su-pernatant liquid as the sample solution. Separately, dissolve 25 mg of enalapril maleate in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-Layer Chromatogra-phy <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of water, acetone, 1-butanol, acetic acid (100) and toluene (1:1:1:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the Rf values of the 2 spots obtained from the sample solution and the 2 spots from the standard solu-tion are equivalent.
Enalaprilat and enalapril diketopiperazine—Use the sample solution obtained in the Assay as the sample solu-tion. Pipet 1 mL of the sample solution, add sodium dihy-drogen phosphate TS (pH 2.2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of both solutions by the automatic integration method: the peak area of enalaprilat, having the relative retention time of about 0.5 to enalapril obtained from the sample solution, is not larger than 2 times the peak area of enalapril from the standard solution. Also, the peak area of enalapril diketopiperazine, having the relative retention time of about 1.5 to enalapril, from the sample solution is not larger than the peak area of enalapril from the standard so-lution. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 1 mL of the standard solution, and add sodium dihydrogen phosphate TS (pH 2.2) to make exactly 10 mL. Confirm that the peak area of enalapril obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Enalapril Maleate Tablets, add V/2 mL of sodium dihydrogen phosphate TS (pH 2.2), sonicate for 15 minutes, shake for 30 minutes, and add sodium dihydro-gen phosphate TS (pH 2.2) to make exactly V mL so that each mL contains about 0.1 mg of enalapril maleate (C20H28 N2O5.C4H4O4). Sonicate this solution for 15 minutes, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4) ＝ MS × AT/AS × V/200 MS: Amount (mg) of Enalapril Maleate RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of a 2.5- and 5-mg tablet and in 30 minutes of a 10-mg tablet are not less than 85z, respectively. Start the test with 1 tablet of Enalapril Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.8 mg of enalapril maleate (C20H28N2O5. C4H4O4), and use this solution as the sample solution. Sepa-rately, weigh accurately about 14 mg of Enalapril Maleate RS, previously dried in vacuum at 609C for 2 hours, and dis-solve in water to make exactly 500 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of enalapril in each solution. Dissolution rate (z) with respect to the labeled amount of enalapril maleate (C20H28N2O5.C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Enalapril Maleate RS taken C: Labeled amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4) in 1 tablet Operating conditions— Detector, column, column temperature, and flow rate: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 1.88 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust the pH to 2.2 with phosphoric acid, and add water to make 1000 mL. To 750 mL of this solution add 250 mL of acetonitrile. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of enalapril are not less than 300 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of enalapril is not more than 2.0z.
Weigh accurately not less than 20 Enalapril Maleate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of enalapril maleate (C20H28N2O5.C4H4O4), add 50 mL of sodium dihydrogen phosphate TS (pH 2.2), sonicate for 15 minutes, shake for 30 minutes, and then add sodium dihydrogen phosphate TS (pH 2.2) to make exactly 100 mL. Sonicate this solution for 15 minutes, filter through a membrane filter with a pore size ‌JP XVIII Official Monographs / Enflurane 927 not exceeding 0.45 mm, and use the filtrate as the sample so-lution. Separately, weigh accurately about 20 mg of Enalapril Maleate RS, previously dried in vacuum at 609C for 2 hours, dissolve in sodium dihydrogen phosphate TS (pH 2.2) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of enalapril in each solution. Amount (mg) of enalapril maleate (C20H28N2O5.C4H4O4) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Enalapril Maleate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of sodium dihydrogen phosphate TS (pH 2.2) and acetonitrile (3:1). Flow rate: Adjust so that the retention time of enalapril is about 5 minutes. System suitability— System performance: Heat to fusion about 20 mg of enalapril maleate. After cooling, add 50 mL of acetonitrile, and sonicate to dissolve. To 1 mL of this solution, add the standard solution to make 50 mL, and use this solution as the solution for system suitability test. When the procedure is run with 50 mL of the solution for system suitability test under the above operaring conditions, enalapril and enalapril diketopiperazine, which has a relative retention time of about 1.5 to enalapril, are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of enalapril is not more than 1.0z.
Entacapone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of en-tacapone (C14H15N3O5: 305.29). Method of preparation Prepare as directed under Tablets, with Entacapone. Identification To 1 mL of the sample solution obtained in the Assay add methanol to make 50 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 301 nm and 305 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Entacapone Tablets add 70 mL of methanol, shake for 5 minutes, and add 60 mL of tetrahydrofuran. Sonicate for 3 minutes, shake for 5 minutes, and add methanol to make exactly 200 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL so that each mL contains about 0.
To 1 mL of the sample solution obtained in the Assay add methanol to make 50 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 301 nm and 305 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Entacapone Tablets add 70 mL of methanol, shake for 5 minutes, and add 60 mL of tetrahydrofuran. Sonicate for 3 minutes, shake for 5 minutes, and add methanol to make exactly 200 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL so that each mL contains about 0.5 mg of entacapone (C14H15N3O5), and use this solution as the sample solution. Then, proceed as directed in the Assay. ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Entacapone RS taken, calculated on the dried basis C: Labeled amount (mg) of entacapone (C14H15N3O5) in 1 tablet
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 Entacapone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of entacapone (C14H15N3O5), add 60 mL of tetrahydrofuran, and sonicate for 3 minutes. Add 60 mL of methanol, shake for 5 minutes, and add methanol to make exactly 200 mL. Centrifuge this solution, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 50 mg of Entaca-pone RS (separately determine the loss on drying <2.41> under the same conditions as Entacapone), dissolve in 30 mL of tetrahydrofuran, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of entacapone in each solution. Amount (mg) of entacapone (C14H15N3O5) ＝ MS × AT/AS × 2 MS: Amount (mg) of Entacapone RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 300 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Amount (mg) of entacapone (C14H ＝ MS × AT/AS × V?/V × 2 15N3O5) 259C. Mobile phase: Dissolve 2.34 g of sodium dihydrogen phos-phate dihydrate in water, add 2 mL of phosphoric acid, and MS: Amount (mg) of Entacapone RS taken, calculated on the dried basis add water to make 1000 mL. To 540 mL of this solution add 440 mL of methanol and 20 mL of tetrahydrofuran. ‌JP XVIII Official Monographs / Enviomycin Sulfate 931 Flow rate: Adjust so that the retention time of entacapone is about 12 minutes. System suitability— System performance: To 20 mL of the standard solution add a mixture of methanol and tetrahydrofuran (7:3) to make 50 mL, and use this solution as the solution for system suitability test. Separately, dissolve 5 mg of Entacapone Related Substance A for System Suitability RS in a mixture of methanol and tetrahydrofuran (7:3) to make 25 mL. To 15 mL of this solution and 15 mL of the solution for system suitability test add a mixture of methanol and tetrahydrofu-ran (7:3) to make 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the related substance A, having the relative retention time of about 0.8 to entacapone, and entacapone are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of entacapone is not more than 1.0z.
Epalrestat Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of epal-restat (C15H13NO3S2: 319.40). Method of preparation Prepare as directed under Tablets, with Epalrestat. Identification (1) Powder Epalrestat Tablets. To a por-tion of the powder, equivalent to 50 mg of Epalrestat, add 100 mL of methanol, shake thoroughly, and filter. To 1 mL of the filtrate add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, between 290 nm and 294 nm, and between 387 nm and 391 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. this solution, add N,N-dimethylformamide to make exactly 100 mL. Pipet 5 mL of this solution, add N,N-dimethylfor-mamide to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 392 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of epalrestat (C15H13NO3S2) ＝ MS × AT/AS × V?/V × 1/4 MS: Amount (mg) of Epalrestat RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of Epalrestat Tablets is not less than 70z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Epalrestat Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.
(1) Powder Epalrestat Tablets. To a por-tion of the powder, equivalent to 50 mg of Epalrestat, add 100 mL of methanol, shake thoroughly, and filter. To 1 mL of the filtrate add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, between 290 nm and 294 nm, and between 387 nm and 391 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. this solution, add N,N-dimethylformamide to make exactly 100 mL. Pipet 5 mL of this solution, add N,N-dimethylfor-mamide to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 392 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of epalrestat (C15H13NO3S2) ＝ MS × AT/AS × V?/V × 1/4 MS: Amount (mg) of Epalrestat RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of Epalrestat Tablets is not less than 70z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Epalrestat Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of epalrestat (C15H13NO3S2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Epalrestat RS, previously dried in vacuum at 609C for 3 hours with silica gel as a desiccant, dissolve in 10 mL of N,N-dimethylformamide, and add the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 398 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the control. Dissolution rate (z) with respect to the labeled amount of epalrestat (C15H13NO3S2) ＝ MS × AT/AS × V?/V × 1/C ×45/2 MS: Amount (mg) of Epalrestat RS taken C: Labeled amount (mg) of epalrestat (C15H13NO3S2) in 1 tablet
Conduct this procedure using light-resistant ves-sels. Weigh accurately the mass of not less than 20 Epalrestat Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of epalrestat (C15H13NO3S2), add 20 mL of N,N-dimethylformamide, add exactly 5 mL of the internal standard solution, shake, and centrifuge. To 2 mL of the supernatant liquid add N,N-dimethylformamide to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Epalrestat RS, previously dried in vacuum at 609C for 3 hours with silica gel as a desiccant, dissolve in 8 mL of N,N-dimethylformamide, add exactly 2 mL of the internal standard solution, and shake. To 2 mL of this solution add ‌934 N,N-dimethylformamide to make 20 mL, and use this solu-tion as the standard solution. Then, proceed as directed in the Assay under Epalrestat. each of the sample solution and standard solution add 10 mL of a mixture of isopropylether and carbon disulfide (3:1), shake for 30 seconds, allow them to stand for 2 minutes, and Amount (mg) of epalrestat (C ＝ MS ×QT/QS × 5/2 15H13NO3S2) compare the color of the upper layer: the color obtained from the sample solution is not more darker than that ob-tained from the standard solution. MS: Amount (mg) of Epalrestat RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in N,N-dimethylformamide (1 in 100).
Ephedrine Hydrochloride Injection is an aqueous injection. It contains not less than 95.
Ephedrine Hydrochloride Injection is a clear, colorless liquid. pH: 4.5 – 6.5 ‌936
To a volume of Ephedrine Hydrochloride Injection, equivalent to 0.05 g of Ephedrine Hydrochloride, add water to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 249 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 265 nm. Bacterial endotoxins <4.01> Less than 7.5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Ephedrine Hydrochloride In-jection, equivalent to about 40 mg of ephedrine hydrochlo-ride (C10H15NO.HCl), add exactly 10 mL of the internal standard solution and water to make 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of ephedrine hydrochloride for assay, previ-ously dried at 1059C for 3 hours, add exactly 10 mL of the internal standard solution to dissolve, add water to make 200 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ephedrine to that of the internal standard. Amount (mg) of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS × QT/QS MS: Amount (mg) of ephedrine hydrochloride for assay taken Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (4) under Ephedrine Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ephedrine are eluted in this order with the resolution between these peaks being not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ephedrine to that of the internal standard is not more than 1.0z.
Ephedrine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of ephedrine hydrochloride (C10H15NO.HCl: 201.69). Method of preparation Prepare as directed under Tablets, with Ephedrine Hydrochloride. Identification To an amount of powdered Ephedrine Hy-drochloride Tablets, equivalent to 0.05 g of Ephedrine Hy-drochloride, add 100 mL of water, shake for 20 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 249 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 265 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ephedrine Hydrochloride Tablets add V mL of water so that each mL contains about 0.
To an amount of powdered Ephedrine Hy-drochloride Tablets, equivalent to 0.05 g of Ephedrine Hy-drochloride, add 100 mL of water, shake for 20 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 249 nm and 253 nm, between 255 nm and 259 nm, and between 261 nm and 265 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ephedrine Hydrochloride Tablets add V mL of water so that each mL contains about 0.25 mg of ephe-drine hydrochloride (C10H15NO.HCl), then add exactly V/4 mL of the internal standard solution, disperse the tablet into small particles by sonicating, then sonicate for a further 10 minutes. Shake this solution for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of ephedrine hydrochloride for assay, previously dried at 1059C for 3 hours, dissolve in water to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Then, proceed as di-rected in the Assay. Amount (mg) of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS × QT/QS × V/100 MS: Amount (mg) of ephedrine hydrochloride for assay taken Internal standard solution—A solution of etilefrine hydro-chloride (1 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Ephedrine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Ephedrine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, and use the sub-sequent filtrate as the sample solution. Separately, weigh ac-curately about 28 mg of ephedrine hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed un-der Liquid Chromatography <2.01> according to the follow-ing conditions, and determine the peak areas, AT and AS, of ephedrine in each solution. Dissolution rate (z) with respect to the labeled amount of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS × AT/AS × 1/C × 90 MS: Amount (mg) of ephedrine hydrochloride for assay taken C: Labeled amount (mg) of ephedrine hydrochloride (C10H15NO.HCl) in 1 tablet Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Purity (4) under Ephedrine Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ephedrine are not less than 10,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ephedrine is not more than 2.0z.
Weigh accurately not less than 20 tablets of Ephe-drine Hydrochloride Tablets, and powder. Weigh accurately an amount of the powder, equivalent to about 40 mg of ‌ephedrine hydrochloride (C10H15NO.HCl), add 150 mL of water, and extract by sonicating for 10 minutes with occa-sional shaking. Shake more for 10 minutes, then add exactly 10 mL of the internal standard solution and water to make 200 mL, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 40 mg of ephedrine hydrochloride for assay, previously dried at 105 9C for 3 hours, add exactly 10 mL of the internal standard solution to dissolve, add water to make 200 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and Q S, of the peak area of ephedrine to that of the internal stand-ard. Amount (mg) of ephedrine hydrochloride (C10H15NO.HCl) ＝ MS × QT/QS MS: Amount (mg) of ephedrine hydrochloride for assay taken Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (4) under Ephedrine Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ephedrine are eluted in this order with the resolution between these peaks being not less than 15. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ephedrine to that of the internal standard is not more than 1.0z.
Adrenaline Injection is a colorless, clear liq-uid. It changes gradually to pale red and then to brown on exposure to air and light. pH: 2.3 – 5.0
(1) To 1 mL of Adrenaline Injection add 4 mL of water and 1 drop of iron (III) chloride TS: a deep green color is produced, and it gradually changes to red. (2) Place 1 mL each of Adrenaline Injection in test tubes A and B, and proceed as directed in the Identification (2) under Adrenaline. Extractable volume <6.05> It meets the requirement. D
Pipet 30 mL of Adrenaline Injection into a separa-tor, add 25 mL of carbon tetrachloride, shake vigorously for 1 minute, allow the liquids to separate, and discard the car-bon tetrachloride. Repeat this procedure three times. Rinse the stopper and mouth of the separator with a small amount of water. Add 0.2 mL of starch TS, then while swirling the separator add iodine TS dropwise until a persistent blue color develops, and immediately add sodium thiosulfate TS to discharge the blue color. Add 2.1 g of sodium hydrogen carbonate to the liquid in the separator, preventing it from coming in contact with the mouth of the separator, and shake until most of the sodium hydrogen carbonate dis-solves. Rapidly inject 1.0 mL of acetic anhydride into the contents of the separator. Immediately stopper the separator loosely, and allow to stand until the evolution of gas ceases. Shake vigorously, allow to stand for 5 minutes, extract with six 25-mL portions of chloroform, and filter each chlo-roform extract through a pledget of absorbent cotton. Evaporate the combined chloroform extracts on a water bath in a current of air to 3 mL, completely transfer this residue by means of small portions of chloroform to a tared beaker, and heat again to evaporate to dryness. Dry the residue at 1059C for 30 minutes, cool in a desiccator (silica gel), and accurately measure the mass M (mg) of the dried residue. Dissolve in chloroform to make exactly 5 mL, and determine the optical rotation <2.49> [a]20 using a 100-mm cell. Amount (mg) of adrenaline (C9H13NO3) D ＝ M ×{0.5 ＋ (0.5 ×|[a]20|)/93}× 0.592
Adrenaline Solution is clear, colorless or ‌JP XVIII Official Monographs / Afloqualone 419 slightly reddish liquid. It changes gradually to pale red and then to brown on exposure to air and light. pH: 2.3 – 5.0
Proceed as directed in the Identification under Adrenaline Injection.
Proceed as directed in the Assay under Adrenaline Injection. Amount (mg) of adrenaline (C9H13NO3) D ＝ M ×{0.5 ＋ (0.5 ×|[a]20|)/93}× 0.592
Eplerenone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of eplerenone (C24H30O6: 414.49). Method of preparation Prepare as directed under Tablets, with Eplerenone. Identification Determine the absorption spectrum of the sample solution obtained in the Uniformity of dosage units as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol- lowing method: it meets the requirement. To 1 tablet of Eplerenone Tablets add a suitable amount of a mixture of acetonitrile and water (3:2), shake, disinte-grate the tablet by sonicating, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and centrifuge. Take exactly V mL of the supernatant liquid, add a mixture of acetonitrile and water (3:2) to make exactly V? mL so that each mL contains about 25 mg of eplerenone (C24H30O6), and use this solution as the sample solution.
Determine the absorption spectrum of the sample solution obtained in the Uniformity of dosage units as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol- lowing method: it meets the requirement. To 1 tablet of Eplerenone Tablets add a suitable amount of a mixture of acetonitrile and water (3:2), shake, disinte-grate the tablet by sonicating, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and centrifuge. Take exactly V mL of the supernatant liquid, add a mixture of acetonitrile and water (3:2) to make exactly V? mL so that each mL contains about 25 mg of eplerenone (C24H30O6), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Eplerenone RS (separately deter-mine the loss on drying <2.41> under the same conditions as Eplerenone), and dissolve in a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Pipet 5 mL of this solu-tion, add a mixture of acetonitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, at 243 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of eplerenone (C24H30O6) ＝ MS × AT/AS × V?/V × 1/10 MS: Amount (mg) of Eplerenone RS taken, calculated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Eplerenone Tablets is not less than 75z. Start the test with 1 tablet of Eplerenone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of eplerenone (C24H30 O6), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Eplerenone RS (separately determine the loss on drying <2.41> under the same condi-tions as Eplerenone), dissolve in 5 mL of acetonitrile, and add the dissolution medium to make exactly 500 mL. Pipet 10 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 243 nm of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24> using the dissolu-tion medium as the blank. Dissolution rate (z) with respect to the labeled amount of eplerenone (C24H30O6) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of Eplerenone RS taken, calculated on the dried basis C: Labeled amount (mg) of eplerenone (C24H30O6) in 1 tablet
Weigh accurately the mass of not less than 20 Eplerenone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of eplerenone (C24H30O6), add a suitable amount of a mixture of aceto-nitrile and water (3:2), sonicate to disperse the particles, and add a mixture of acetonitrile and water (3:2) to make exactly 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of Eplerenone RS (separately determine the loss on drying <2.41> under the same conditions as Eplerenone), dissolve in a mixture of acetonitrile and water (3:2) to make exactly 50 mL, and use this solution as the standard solution. ‌942 Perform the test with exactly 15 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of eplerenone in each solution. Amount (mg) of eplerenone (C24H30O6) ＝ MS × AT/AS × 2 MS: Amount (mg) of Eplerenone RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 243 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.4 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.0 with phosphoric acid. To 550 mL of this solution add 360 mL of methanol and 90 mL of acetonitrile. Flow rate: Adjust so that the retention time of eplerenone is about 12 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of eplerenone are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of eplerenone is not more than 1.0z.
エルゴメトリンマレイン酸塩注射液
Ergometrine Maleate Injection is an aqueous injec-tion. It contains not less than 90.
Ergometrine Maleate Injection is a clear, col-orless to pale yellow liquid. pH <2.54> Dissolve 0.10 g of Ergometrine Maleate in 10 mL of water. The pH of the solution is between 3.0 and 5.0.
(1) Measure a volume of Ergometrine Maleate Injection, equivalent to 3 mg of Ergometrine Male-ate, if necessary, dilute with water or evaporate on a water bath to make 15 mL, and use this solution as the sample so-lution. The sample solution shows a blue fluorescence. To 1 mL of the sample solution obtained in (1) add 1 mL of ammonia TS, and extract with 20 mL of diethyl ether. To the diethyl ether extract add 1 mL of dilute sulfuric acid, shake, and warm to remove diethyl ether in a water bath. Cool, to the residue obtained add 2 mL of 4-dimethylamino-benzaldehyde-iron (III) chloride TS, and allow to stand for 5 to 10 minutes: a deep blue color develops. To 5 mL of the sample solution obtained in (1) add 1 drop of potassium permanganate TS: a red color disappears immediately. Bacterial endotoxins <4.01> Less than 1500 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. ‌950 Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay Transfer an exactly measured volume of Ergome-trine Maleate Injection, equivalent to about 2 mg of ergome-trine maleate (C19H23N3O2.C4H4O4), and add sodium chlo-ride in a ratio of 0.3 g to 1 mL of the solution. To this mixture add 20 mL of diethyl ether and 2 mL of ammonia TS, shake, and extract. Further, extract with three 15-mL portions of diethyl ether, combine all the extracts, add 5 g of anhydrous sodium sulfate, filter through a pledget of absor-bent cotton, and wash with three 5-mL portions of diethyl ether. Add the washings to the filtrate, shake with 5 mL of dilute sulfuric acid, evaporate the diethyl ether by warming in a current of nitrogen, to the remaining solution add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 2 mg of Ergo-metrine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, add water to make exactly 50 mL, and use this solution as the standard solution. Transfer 2 mL each of the sample solution and standard solution, accurately meas-ured, to separate glass-stoppered test tubes, and proceed as directed in the Assay under Ergometrine Maleate. and allow to stand for 1 hour at ordinary temperature. Per-form the test with these solutions as directed under Ultravio-let-visible Spectrophotometry <2.24>, using a solution, pre-pared with 4 mL of water in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 550 nm, respectively. Amount (mg) of ergometrine maleate (C19H23N3O2.C4H4O4) ＝ MS × AT/AS × V/100 MS: Amount (mg) of Ergometrine Maleate RS taken
(1) Clarity and color of solution—Dissolve 0.10 g of Ergometrine Maleate in 10 mL of water: the solution is clear and colorless to light yellow. Ergotamine and ergotoxine—To 0.02 g of Ergome-trine Maleate add 2 mL of a solution of sodium hydroxide (1 in 10), and heat to boiling: the gas evolved does not change moistened red litmus paper to blue. Related substances—Dissolve 5.0 mg each of Ergo-metrine Maleate and Ergometrine Maleate RS in 1.0 mL of methanol, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate, prepared with silica gel for thin-layer chromatography and dilute sodium hydroxide TS. Develop the plate with a mixture of chloroform and metha-nol (4:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly 4-dimethylaminobenzaldehyde TS on the plate: the spots obtained from the sample solution and the stand-ard solution show a red-purple color and the same Rf value, and any spot from the sample solution other than that cor-responding to the spot from the standard solution does not appear. pH: 2.7 – 3.5 Identification (1) Measure a volume of Ergometrine Maleate Injection, equivalent to 3 mg of Ergometrine Male-ate, if necessary, dilute with water or evaporate on a water bath to make 15 mL, and use this solution as the sample so-lution. The sample solution shows a blue fluorescence. To 1 mL of the sample solution obtained in (1) add 1 mL of ammonia TS, and extract with 20 mL of diethyl ether. To the diethyl ether extract add 1 mL of dilute sulfuric acid, shake, and warm to remove diethyl ether in a water bath. Cool, to the residue obtained add 2 mL of 4-dimethylamino-benzaldehyde-iron (III) chloride TS, and allow to stand for 5 to 10 minutes: a deep blue color develops. To 5 mL of the sample solution obtained in (1) add 1 drop of potassium permanganate TS: a red color disappears immediately. Bacterial endotoxins <4.01> Less than 1500 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. ‌950 Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay Transfer an exactly measured volume of Ergome-trine Maleate Injection, equivalent to about 2 mg of ergome-trine maleate (C19H23N3O2.C4H4O4), and add sodium chlo-ride in a ratio of 0.3 g to 1 mL of the solution. To this mixture add 20 mL of diethyl ether and 2 mL of ammonia TS, shake, and extract. Further, extract with three 15-mL portions of diethyl ether, combine all the extracts, add 5 g of anhydrous sodium sulfate, filter through a pledget of absor-bent cotton, and wash with three 5-mL portions of diethyl ether. Add the washings to the filtrate, shake with 5 mL of dilute sulfuric acid, evaporate the diethyl ether by warming in a current of nitrogen, to the remaining solution add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 2 mg of Ergo-metrine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, add water to make exactly 50 mL, and use this solution as the standard solution. Transfer 2 mL each of the sample solution and standard solution, accurately meas-ured, to separate glass-stoppered test tubes, and proceed as directed in the Assay under Ergometrine Maleate. and allow to stand for 1 hour at ordinary temperature. Per-form the test with these solutions as directed under Ultravio-let-visible Spectrophotometry <2.24>, using a solution, pre-pared with 4 mL of water in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 550 nm, respectively. Amount (mg) of ergometrine maleate (C19H23N3O2.C4H4O4) ＝ MS × AT/AS × V/100 MS: Amount (mg) of Ergometrine Maleate RS taken
Weigh accurately, and powder not less the 20 Ergo-metrine Maleate Tablets. Weigh accurately a portion of the powder, equivalent to about 2 mg of ergometrine maleate (C19H23N3O2.C4H4O4), transfer to a glass filter (G4), add 10 mL of a solution of L-tartaric acid (1 in 100), and filter with thorough shaking. Repeat the procedures 3 times, combine the filtrates, add a solution of L-tartaric acid (1 in 100) to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 2 mg of Ergome-trine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, dissolve in a solution of L-tartaric acid (1 in 100) Amount (mg) of ergometrine maleate (C ＝ MS × AT/AS 19H23N3O2.C4H4O4) to make exactly 50 mL, and use this solution as the standard solution. Pipet 2 mL each of the sample solution and stand-ard solution, and proceed as directed in the Assay under MS: Amount (mg) of Ergometrine Maleate RS taken
Ergometrine Maleate Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of ergometrine maleate (C19H23N3O2.C4H4O4: 441.48). Method of preparation Prepare as directed under Tablets, with Ergometrine Maleate. Identification To a quantity of powdered Ergometrine Maleate Tablets, equivalent to 3 mg of Ergometrine Maleate, add 15 mL of warm water, shake, and filter: the filtrate shows a blue fluorescence. Proceed with this solution as di-rected in the Identification (2) and (3) under Ergometrine Maleate. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Ergometrine Maleate Tablets to a brown glass-stoppered centrifuge tube, and add exactly V mL of a solution of L-tartaric acid (1 in 100) so that each mL contains about 40 mg of ergometrine maleate (C19H23N3O2.
To a quantity of powdered Ergometrine Maleate Tablets, equivalent to 3 mg of Ergometrine Maleate, add 15 mL of warm water, shake, and filter: the filtrate shows a blue fluorescence. Proceed with this solution as di-rected in the Identification (2) and (3) under Ergometrine Maleate. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Ergometrine Maleate Tablets to a brown glass-stoppered centrifuge tube, and add exactly V mL of a solution of L-tartaric acid (1 in 100) so that each mL contains about 40 mg of ergometrine maleate (C19H23N3O2.C4H4O4). Stopper the tube, shake for 30 minutes vigorously, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 4 mg of Ergometrine Maleate RS, previously dried in a desiccator (silica gel) for 4 hours, dissolve in water to make exactly 100 mL, and use this solution as the standard solu-tion. Pipet 4 mL each of the sample solution and standard solution into separate brown glass-stoppered test tubes, add exactly 8 mL each of 4-dimethylaminobenzaldehyde-iron (III) chloride TS while cooling in an ice bath, after shaking, Ergometrine Maleate. Amount (mg) of ergometrine maleate (C19H23N3O2.C4H4O4) ＝ MS × AT/AS MS: Amount (mg) of Ergometrine Maleate RS taken
To a quantity of powdered Erythromycin Delayed-release Tablets, equivalent to 10 mg (potency) of Erythromycin, add 1 mL of methanol, shake well, filter, and use the filtrate as the sample solution. Separately, dissolve 10 mg of Erythromycin RS in 1 mL of methanol, and use this solution as the standard solution. Then, proceed as directed in the Identification (2) under Erythromycin. Loss on drying <2.41> Not more than 10.0z (0.2 g, in vacuum not exceeding 0.67 kPa, 609C, 3 hours). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Disintegration <6.09> It meets the requirement. For the test with 2nd fluid for disintegration test, use the disk.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, and standard solu-tions—Proceed as directed in the Assay under Erythromy-cin. Sample solutions—Weigh accurately the mass of not less than 20 Erythromycin Delayed-release Tablets, and powder. Weigh accurately a portion of the powder, equiva-lent to about 25 mg (potency) of Erythromycin, add 25 mL of methanol, shake vigorously, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make exactly 100 mL, and filter. Take exactly an appropriate volume of the filtrate, add 0.1 mol/L phosphate buffer solution (pH 8.0) to prepare solu-tions containing 20 mg (potency) and 5 mg (potency) per mL, and use these solutions as the high and the low concentration sample solutions, respectively.
Estradiol Benzoate Injection (Aqueous Suspension)
エストラジオール安息香酸エステル水性懸濁注射液
Estradiol Benzoate Injection (Aqueous Suspension) is an aqueous suspension for injection. It contains not less than 90.
Estradiol Benzoate Injection (Aqueous Sus-pension) produces a white turbidity on shaking.
Extract a volume of Estradiol Benzoate In-jection (Aqueous Suspension), equivalent to 1 mg of Estra-diol Benzoate, with 5 mL of chloroform, and use this extract as the sample solution. Separately, dissolve 1 mg of Es-tradiol Benzoate RS in 5 mL of chloroform, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 50 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and methanol (99:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Direct in-oculation method: it meets the requirement. MS: Amount (mg) of Estradiol Benzoate RS taken Internal standard solution—A solution of progesterone in methanol (13 in 100,000).
Estriol Injection (Aqueous Suspension) is an aque-ous suspension for injection. It contains not less than 90.
Shake Estriol Injection (Aqueous Suspension): a white turbidity is produced.
(1) Shake well, take a volume of Estriol In-jection (Aqueous Suspension), equivalent to 2 mg of Estriol, add ethanol (95) to make 20 mL, and use this solution as the sample solution. Proceed with the sample solution as di-rected in the Identification (1) under Estriol. (2) Determine the absorption spectrum of the sample so-lution obrtained in (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 279 nm and 283 nm. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Direct in-oculation method: it meets the requirement.
Shake well, pipet a volume of Estriol Injection (Aqueous Suspension), equivalent to about 5 mg of estriol (C18H24O3), and dissolve in methanol to make exactly 20 mL. Pipet 4 mL of this solution, add exactly 5 mL of the internal Amount (mg) of estriol (C 18H 24O3) ＝ MS × QT/QS standard solution, then add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh MS: Amount (mg) of Estriol RS taken Internal standard solution—A solution of methyl benzoate for estriol test in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (51:49). Flow rate: Adjust so that the retention time of estriol is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, estriol and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of estriol to that of the internal standard is not more than 1.0z.
Estriol Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of estriol (C18H24O3: 288.38). Method of preparation Prepare as directed under Tablets, with Estriol. Identification (1) Weigh a portion of powdered Estriol ‌962 Tablets, equivalent to 2 mg of Estriol, add 20 mL of ethanol (95), shake for 10 minutes, centrifuge, and use the superna-tant liquid as the sample solution. Proceed with the sample solution as directed in the Identification (1) under Estriol. (2) Determine the absorption spectrum of the sample so-lution obtained in (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 279 nm and 283 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Estriol Tablets add exactly 5 mL of water, disperse the fine particles by sonicating, add exactly 15 mL of methanol, and shake for 15 minutes. Centrifuge this solu-tion for 10 minutes, pipet a definite amount of the superna-tant liquid, and add methanol to make exactly a definite amount of solution so that each ml of the solution contains about 5 mg of estriol (C18H24O3).
(1) Weigh a portion of powdered Estriol ‌962 Tablets, equivalent to 2 mg of Estriol, add 20 mL of ethanol (95), shake for 10 minutes, centrifuge, and use the superna-tant liquid as the sample solution. Proceed with the sample solution as directed in the Identification (1) under Estriol. (2) Determine the absorption spectrum of the sample so-lution obtained in (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 279 nm and 283 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Estriol Tablets add exactly 5 mL of water, disperse the fine particles by sonicating, add exactly 15 mL of methanol, and shake for 15 minutes. Centrifuge this solu-tion for 10 minutes, pipet a definite amount of the superna-tant liquid, and add methanol to make exactly a definite amount of solution so that each ml of the solution contains about 5 mg of estriol (C18H24O3). Pipet 5 mL of this solution, add exactly 1 mL of the internal standard solution, and use this solution as the sample solution. Proceed with 20 mL of System suitability— Proceed as directed in the system suitability in the Assay under Estriol.
Weigh accurately and powder not less than 20 Estriol Tablets. Weigh accurately a portion of the powder, equivalent to about 1 mg of estriol (C18H24O3), add exactly 5 mL of water, disperse the fine particles by sonicating, shake with 25 mL of methanol for 10 minutes, centrifuge, and take the supernatant liquid. Add 25 mL of methanol, repeat the above procedure twice, combine the supernatant liquid, add exactly 5 mL of the internal standard solution, then add methanol to make 100 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of Estriol RS, previously dried at 1059C for 3 hours, and dis-solve in methanol to make exactly 100 mL. Pipet 4 mL of this solution, add exactly 5 mL of the internal standard solu-tion, then add methanol to make 100 mL, and use this solu-tion as the standard solution. Proceed with 20 mL each of the sample solution and standard solution as directed in the Assay under Estriol. the sample solution as directed in the Assay under Estriol. Use a solution of methyl benzoate for estriol test in methanol (1 in 40,000) as the internal standard solution. Calculate the Amount (mg) of estriol (C ＝ MS × QT/QS × 1/25 18H 24O3) mean value from each ratio of peak areas of 10 samples: the samples conform to the requirements if the deviation (z) of the mean value and each ratio of peak areas is within 15z. If the deviation (z) exceeds 15z, and 1 sample shows devia-tion within 25z, repeat the test with 20 samples. Calculate the deviation (z) of the mean value from each ratio of peak areas of the 30 samples used in the 2 tests and each ratio of peak areas: the samples conform to the requirements if the deviation exceeds 15z, not more than 1 sample shows devia-tion within 25z, and no sample shows deviation exceeding 25z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Estriol Tablets is not less than 80z. Start the test with 1 tablet of Estriol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.1 mg of estriol (C18H24O3), and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Estriol RS, previously dried at 1059C for 3 hours, dis-solve in methanol to make exactly 100 mL, then pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of estriol. Dissolution rate (z) with respect to the labeled amount of estriol (C18H24O3) ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of Estriol RS taken C: Labeled amount (mg) of estriol (C18H24O3) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay under Estriol. MS: Amount (mg) of Estriol RS taken Internal standard solution—A solution of methyl benzoate for estriol test in methanol (1 in 5000).
Etacrynic Acid Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of etacrynic acid (C13H12Cl2O4: 303.14). Method of preparation Prepare as directed under Tablets, with Etacrynic Acid. Identification (1) Weigh a quantity of powdered Eta-crynic Acid Tablets, equivalent to 0.3 g of Etacrynic Acid, add 25 mL of 0.1 mol/L hydrochloric acid TS, and extract with 50 mL of dichloromethane. Filter the dichloromethane extract, and evaporate the filtrate on a water bath to dry-ness. Proceed with the residue as directed in the Identifica-tion (1), (2) and (4) under Etacrynic Acid. (2) Prepare a solution of the residue obtained in (1), equivalent to a solution of Etacrynic Acid in methanol (1 in 20,000), and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 268 nm and 272 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Etacrynic Acid Tablets is not less than 70z. Start the test with 1 tablet of Etacrynic Acid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of etacrynic acid (C13H12Cl2O4), and use this solution as the sample solution.
(1) Weigh a quantity of powdered Eta-crynic Acid Tablets, equivalent to 0.3 g of Etacrynic Acid, add 25 mL of 0.1 mol/L hydrochloric acid TS, and extract with 50 mL of dichloromethane. Filter the dichloromethane extract, and evaporate the filtrate on a water bath to dry-ness. Proceed with the residue as directed in the Identifica-tion (1), (2) and (4) under Etacrynic Acid. (2) Prepare a solution of the residue obtained in (1), equivalent to a solution of Etacrynic Acid in methanol (1 in 20,000), and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 268 nm and 272 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Etacrynic Acid Tablets is not less than 70z. Start the test with 1 tablet of Etacrynic Acid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of etacrynic acid (C13H12Cl2O4), and use this solution as the sample solution. Separately, weigh accurately about 55 mg of etacrynic acid for assay, previous-ly dried in vacuum at 609C for 2 hours, dissolve in 10 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, at 277 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec- trophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of etacrynic acid (C13H12Cl2O4) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of etacrynic acid for assay taken C: Labeled amount (mg) of etacrynic acid (C13H12Cl2O4) in 1 tablet
Weigh accurately and powder not less than 20 Etacrynic Acid Tablets. Weigh accurately a portion of the powder, equivalent to about 0.1 g of etacrynic acid (C13H12Cl2O4), add 25 mL of 0.1 mol/L hydrochloric acid TS, and extract with three 30-mL portions of dichloro-methane. Filter the dichloromethane extracts through a pledget of absorbent cotton into an iodine bottle. Wash the pledget of absorbent cotton with a small amount of dichloromethane, and combine the washing with the extracts. Evaporate this solution on a water bath to dryness in a current of air, to the residue add 20 mL of acetic acid (100), and proceed as directed in the Assay under Etacrynic Acid. Each mL of 0.05 mol/L bromine VS ＝ 15.16 mg of C13H12Cl2O4
(1) Evaporate to dryness 5 mL of the sam-ple solution obtained in Assay, and add 2 mL of a mixture of sulfuric acid and ethanol (95) (2:1) to the residue: a light red color with a yellow fluorescence develops. To the solution add carefully 4 mL of water: the color of the solution changes to red-purple. (2) Evaporate to dryness 10 mL of the sample solution obtained in Assay, add 0.2 mL of acetic acid (31) and 2 mL of phosphoric acid to the residue, and heat on a water bath for 5 minutes: a red color with a yellow-green fluorescence develops. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Place 1 tablet of Ethinylestradiol Tablets in a separator, add 10 mL of 2nd fluid for disintegration test, and shake until the tablet is disintegrated. Add 10 mL of dilute sulfuric acid and 20 mL of chloroform, shake vigorously for 5 minutes, and filter the chloroform layer into a conical flask through filter paper on which 5 g of anhydrous sodium sul-fate is placed. Extract the aqueous layer with two 20-mL portions of chloroform, proceed with the extracts in the same manner as before, and combine the filtrates with the previous one. Evaporate gently the combined filtrate on a water bath with the aid of a current of nitrogen, dissolve the residue in exactly 100 mL of methanol, and centrifuge, if necessary. Pipet x mL of the supernatant liquid, add metha-nol to make exactly V mL of a solution containing about 40 ng of ethinylestradiol (C20H24O2) per mL, and use this so-lution as the sample solution. Separately, weigh accurately about 10 mg of Ethinylestradiol RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, dissolve in methanol, dilute to a volume containing about 40 ng of ethinylestradiol (C20H24O2) per mL, and use this solution as the standard solution. Pipet 4 mL each of sulfu-ric acid-methanol TS into three glass-stoppered test tubes, T, S and B, cool in ice, to each tube add exactly 1 mL each of the sample solution, the standard solution and methanol, shake immediately, and allow to stand in a water bath at 309C for 40 minutes, then allow to stand in a water bath at 209C for 5 minutes. Perform the test with these solutions as directed under Fluorometry <2.22>. Determine the fluores- cence intensities, FT, FS and FB, of these solutions using the fluorophotometer, at about 460 nm of the excitation and at about 493 nm of the fluorescence. Amount (mg) of ethinylestradiol (C20H24O2) ＝ MS × (FT － FB)/(FS － FB) × V/2500 × 1/x MS: Amount (mg) of Ethinylestradiol RS taken Dissolution Being specified separately when the drug is granted approval based on the Law.
(i) Chromatographic tube: Pack a pledget of glass wool in the bottom of a tube 25 mm in inside diameter and 300 mm in length, and place 5 g of anhydrous sodium sulfate on the glass wool. Chromatographic column: Place 5 g of siliceous earth for chromatography in a 200-mL beaker, soak well in 4 mL of 1 mol/L hydrochloric acid TS, and mix uniformly. Put the siliceous earth into the chromatographic tube in small portions to make 60 to 80 mm in height in proper hard-ness with a tamping rod. Standard solution: Weigh accurately about 10 mg of Ethinylestradiol RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in chloroform to make exactly 100 mL. Pipet 5 mL of this solu-tion, and add chloroform to make exactly 100 mL. Sample: Weigh accurately not less than 20 Ethinyles-tradiol Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.5 mg of ethinylestradiol (C20H24O2), place in a 50-mL beaker, add 2 mL of water, shake well, add 3 mL of chloroform, and shake well again. Add 4 g of siliceous earth for chromatography, mix well until the contents do not stick to the inner wall of the beaker, and use the substance as the sample. Procedure: To the chromatographic column add the sample with a funnel, and pack in proper hardness. Mix well the sample sticking to the beaker with 0.5 g of siliceous earth for chromatography, and place in the chromatographic tube. Wipe off the sample solution sticking to the beaker and the tamping rod with glass wool, and place it in the chromato-graphic tube. Push down the sample, and press lightly on the chromatographic column to make the height of the column 110 mm to 130 mm. Take 70 mL of chloroform, rinse the in-ner wall of the chromatographic tube with a portion of the chloroform, and transfer the remaining portion to the chro-matographic tube. Collect the effluent solution at a flow rate not more than 0.8 mL per minute. After completing the elu-tion, rinse the lower end of the chromatographic tube with a small quantity of chloroform, add chloroform to make exactly 100 mL, and use this solution as the sample solution. Transfer 6 mL each of the sample solution and standard so-lution to each separators, and add 20 mL each of isooctane. Add exactly 10 mL of a mixture of sulfuric acid and metha-nol (7:3), shake vigorously for 5 minutes, allow to stand in a dark place for 15 minutes, and centrifuge. Perform the test with the resulting color solutions as directed under Ultravio-let-visible Spectrophotometry <2.24>, using a solution, pre-pared with 6 mL of chloroform in the same manner, as the blank. Determine the absorbances, AT and AS, of the subse-quent solutions obtained from the sample solution and standard solution at 540 nm, respectively. Amount (mg) of ethinylestradiol (C20H24O2) ＝ MS × AT/AS × 1/20 MS: Amount (mg) of Ethinylestradiol RS taken
Take out the content of Ethyl Icosapentate Capsules, to a quantity of the contents, equivalent to 20 mg of Ethyl Icosapentate, add 3 mL of a solution of potassium hydroxide in ethylene glycol (21 in 100), stopper the vessel tightly while passing a current of nitrogen, and heat at 1809C for 15 minutes. After cooling, add methanol to make 100 mL. To 1 mL of this solution add methanol to make 25 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution, obtained by proceeding as above with 3 mL of a solution of potassium hydroxide in ethylene glycol (21 in 100), as a blank: it exhibits maxima between 298 nm and 302 nm, between 311 nm and 315 nm, between 325 nm and 329 nm, and between 343 nm and 347 nm.
Peroxide—Take out the content of Ethyl Icosapen-tate Capsules. Weigh accurately about 1 g of the content, dissolve in 25 mL of a mixture of acetic acid (100) and isooc-tane (3:2), replace the air of the inside gently with Nitrogen, then add 1 mL of saturated potassium iodide TS under a current of Nitrogen, stopper immediately and shake gently, and allow to stand in a dark place for 10 minutes. Then, add ‌976 30 mL of water, shake vigorously, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. The amount of peroxide calculated by the following formula is not more than 20 mEq/kg. Amount (mEq/kg) of peroxide ＝ V/M × 10 V: Amount (mL) of 0.01 mol/L sodium thiosulfate VS consumed M: Amount (g) of Ethyl Icosapentate Capsules taken Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Disintegration <6.09> Perform the test using the disk: it meets the requirement. However, for the preparations in single-dose packages, carry out the test for 10 minutes.
Weigh accurately the mass of an amount of not less than 20 Ethyl Icosapentate Capsules, then open the capsules and take out the contents. Wash the empty capsules with a little amount of hexane, volatilize the hexane by allowing them to stand at the room temperature, and weigh the mass of the total empty capsules accurately. Weigh accurately a portion of the content, equivalent to about 0.4 g of ethyl icosapentate (C22H34O2), add exactly 40 mL of the internal standard solution, then add hexane to make 200 mL, and use this solution as the sample solution. For the preparations in single-dose packages, weigh accurately the mass of the total capsules of not less than 20 packages, and mix them well. Weigh accurately a portion of the capsules, equivalent to about 0.4 g of ethyl icosapentate (C22H34O2), add 15 mL of hexane, then extract the content by opening the capsules. Separate the hexane extract from the residual solids, wash the residues with three 10-mL portions of hexane, combine the washings and the hexane extract, add exactly 40 mL of the internal standard solution, then add hexane to make 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Ethyl Icosapentate RS, add exactly 5 mL of the internal standard solution, then add hexane to make 25 mL, and use this solution as the standard solution. Perform the test with 4 mL each of the sample solu-tion and standard solution as directed under Gas Chroma-tography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethyl icosapentate to that of the internal standard. Amount (mg) of ethyl icosapentate (C22H34O2) ＝ MS × QT/QS × 8 MS: Amount (mg) of Ethyl Icosapentate RS taken Internal standard solution—A solution of methyl docosanate in hexane (1 in 200). Operating conditions— Proceed as directed in the operating conditions in the Assay under Ethyl Icosapentate. System suitability— System performance: When the procedure is run with 4 mL of the standard solution under the above operating condi-tions, the internal standard and ethyl icosapentate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 4 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ethyl icosapentate to that of the internal stand-ard is not more than 1.0z.
Ethyl Loflazepate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of ethyl loflazepate (C18H14ClFN2O3: 360.77). Method of preparation Prepare as directed under Tablets, with Ethyl Loflazepate. Identification To a quantity of powdered Ethyl Loflaze-pate Tablets, equivalent to 1 mg of Ethyl Loflazepate, add 10 mL of acetonitrile, shake for 15 minutes, and centrifuge. To 1 mL of the supernatant liquid add acetonitrile to make 10 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 227 nm and 231 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ethyl Loflazepate Tablets add exactly 0.5 mL of water, sonicate to disintegrate the tablet, add exactly 10 mL of the internal standard solution, shake for 20 minutes, and centrifuge. Pipet V mL of the supernatant liquid, add water so that each mL contains 48 mL of water, add the internal standard solution to make exactly V? mL so that each mL contains about 95 mg of ethyl loflazepate (C18H14ClFN2O3), and use this solution as the sample solu-tion.
To a quantity of powdered Ethyl Loflaze-pate Tablets, equivalent to 1 mg of Ethyl Loflazepate, add 10 mL of acetonitrile, shake for 15 minutes, and centrifuge. To 1 mL of the supernatant liquid add acetonitrile to make 10 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 227 nm and 231 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ethyl Loflazepate Tablets add exactly 0.5 mL of water, sonicate to disintegrate the tablet, add exactly 10 mL of the internal standard solution, shake for 20 minutes, and centrifuge. Pipet V mL of the supernatant liquid, add water so that each mL contains 48 mL of water, add the internal standard solution to make exactly V? mL so that each mL contains about 95 mg of ethyl loflazepate (C18H14ClFN2O3), and use this solution as the sample solu-tion. Then, proceed as directed in the Assay. (C18H14ClFN2O3) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and ethanol (99.5) (2:1:1). Flow rate: Adjust so that the retention time of ethyl loflazepate is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ethyl loflazepate are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ethyl loflazepate is not more than 3.0z.
Weigh accurately the mass of not less than 20 tablets of Ethyl Loflazepate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 1 mg of ethyl loflazepate (C18H14ClFN2O3), add 0.5 mL of water, and sonicate. Add exactly 10 mL of the internal standard, shake, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 10 mg of Ethyl Amount (mg) of ethyl loflazepate (C ＝ MS × QT/QS × V?/V × 1/10 18H14ClFN2O3) Loflazepate RS, previously dried at 1059C for 3 hours, and add the internal standard solution to make exactly 100 mL. To 10 mL of this solution add 0.5 mL of water, and use this MS: Amount (mg) of Ethyl Loflazepate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile for liquid chromatography (1 in 3000). Dissolution <6.10> When the test is performed at 50 revolu- solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of ethyl loflazepate to that of the internal standard. tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Ethyl Loflazepate Tablets is not less than Amount (mg) of ethyl loflazepate (C ＝ MS × QT/QS × 1/10 18H14ClFN2O3) 80z. Start the test with 1 tablet of Ethyl Loflazepate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.1 mg of ethyl loflazepate (C18H14ClFN2O3), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Ethyl Loflazepate RS, pre-viously dried at 1059C for 3 hours, and dissolve in ethanol (95) to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of ethyl loflazepate in each solution. Dissolution rate (z) with respect to the labeled amount of ethyl loflazepate (C18H14ClFN2O3) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of Ethyl Loflazepate RS taken C: Labeled amount (mg) of ethyl loflazepate MS: Amount (mg) of Ethyl Loflazepate RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile for liquid chromatography (1 in 3000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 229 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile for liquid chromatography and ethanol (95) (2:1:1). Flow rate: Adjust so that the retention time of ethyl loflazepate is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ethyl loflazepate are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times ‌JP XVIII Official Monographs / Ethyl Parahydroxybenzoate 979‌ with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ethyl loflazepate to that of the internal standard is not more than 1.0z.
Etidronate Disodium Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of etidronate disodium (C2H6Na2O7P2: 249.99). Method of preparation Prepare as directed under Tablets, with Etidronate Disodium. Identification (1) Shake an amount of powdered Etidronate Disodium Tablets, equivalent to 0.2 g of Etidronate Disodium, with 20 mL of water, and filter. Pro-ceed with the filtrate as directed in the Identification (1) under Etidronate Disodium. (2) Shake an amount of powdered Etidronate Disodium Tablets, equivalent to 0.4 g of Etidronate Disodium, with 10 mL of water, and filter. Evaporate total amount of the fil-trate to dryness under reduced pressure, shake the residue with 15 mL of ethanol (99.5), centrifuge, and dry the precipi-tate at 1509C for 4 hours. Determine the infrared absorption spectrum of the precipitate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 1170 cm－1, 1056 cm－1, 916 cm－1 and 811 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Etidronate Disodium Tablets is not less than 85z. ‌Start the test with 1 tablet of Etidronate Disodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, take exactly V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.
(1) Shake an amount of powdered Etidronate Disodium Tablets, equivalent to 0.2 g of Etidronate Disodium, with 20 mL of water, and filter. Pro-ceed with the filtrate as directed in the Identification (1) under Etidronate Disodium. (2) Shake an amount of powdered Etidronate Disodium Tablets, equivalent to 0.4 g of Etidronate Disodium, with 10 mL of water, and filter. Evaporate total amount of the fil-trate to dryness under reduced pressure, shake the residue with 15 mL of ethanol (99.5), centrifuge, and dry the precipi-tate at 1509C for 4 hours. Determine the infrared absorption spectrum of the precipitate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 1170 cm－1, 1056 cm－1, 916 cm－1 and 811 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Etidronate Disodium Tablets is not less than 85z. ‌Start the test with 1 tablet of Etidronate Disodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, take exactly V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.22 mg of etidronate disodium (C2H6Na2O7P2), and use this solution as the sample solution. Separately, weigh accurately about 30 mg of etidronate disodium for assay, previously dried at 2109C for 2 hours, and dissolve in water to make exactly 100 mL. Di-lute exactly a suitable amount of this solution with water to make solutions so that each mL contains about 0.12 mg, about 0.21 mg and about 0.24 mg of etidronate disodium (C2H6Na2O7P2), and use these solutions as the standard solu-tions. Pipet 2 mL each of the sample solution and standard solutions, add exactly 2 mL of a solution of copper (II) sul-fate (7 in 10,000) and water to make exactly 10 mL. Deter-mine the absorbances of these solutions at 233 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared by diluting exactly 2 mL of the so-lution of copper (II) sulfate (7 in 10,000) with water to make exactly 10 mL as the control. From the calibration curve ob-tained with the standard solutions calculate the concentra-tion of etidronate disodium, CT, in the sample solution. Dissolution rate (z) with respect to the labeled amount of etidronate disodium (C2H6Na2O7P2) ＝ CT × V?/V × 1/C × 90 CT: Concentration (mg/mL) of etidronate disodium (C2H6Na2O7P2) in the sample solution C: Labeled amount (mg) of etidronate disodium (C2H6Na2O7P2) in 1 tablet
Weigh accurately and powder not less than 20 Etidronate Disodium Tablets. Weigh accurately a portion of the powder, equivalent to about 0.5 g of etidronate disodium (C2H6Na2O7P2), add 30 mL of water, shake vigorously for 10 minutes, add water to make exactly 50 mL, and filter. Proceed with exactly 15 mL of the filtrate as directed in the Assay under Etidronate Disodium.
Etilefrine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of etilefrine hydrochloride (C10H15NO2.HCl: 217.69). Method of preparation Prepare as directed under Tablets, with Etilefrine Hydrochloride. Identification To a quantity of powdered Etilefrine Hydro-chloride Tablets, equivalent to 5 mg of Etilefrine Hydrochlo-ride, add 60 mL of diluted hydrochloric acid (1 in 1000), shake well, add 40 mL of diluted hydrochloric acid (1 in 1000), and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotome-try <2.24>, using diluted hydrochloric acid (1 in 1000) as the blank: it exhibits a maximum between 271 nm and 275 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Etilefrine Hydrochloride Tablets add 60 mL of diluted hydrochloric acid (1 in 1000), and proceed as di-rected in the Assay. Amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl) ＝ MS × AT/AS × 1/10 ‌MS: Amount (mg) of etilefrine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Etilefrine Hydrochloride Tablets is not less than 70z. Start the test with 1 tablet of Etilefrine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5 mg of etilefrine hydrochlo-ride (C10H15NO2.
To a quantity of powdered Etilefrine Hydro-chloride Tablets, equivalent to 5 mg of Etilefrine Hydrochlo-ride, add 60 mL of diluted hydrochloric acid (1 in 1000), shake well, add 40 mL of diluted hydrochloric acid (1 in 1000), and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotome-try <2.24>, using diluted hydrochloric acid (1 in 1000) as the blank: it exhibits a maximum between 271 nm and 275 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Etilefrine Hydrochloride Tablets add 60 mL of diluted hydrochloric acid (1 in 1000), and proceed as di-rected in the Assay. Amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl) ＝ MS × AT/AS × 1/10 ‌MS: Amount (mg) of etilefrine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Etilefrine Hydrochloride Tablets is not less than 70z. Start the test with 1 tablet of Etilefrine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5 mg of etilefrine hydrochlo-ride (C10H15NO2.HCl), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of etilefrine hydrochloride for assay, previously dried at 1059C for 4 hours, and dissolve in water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of etilefrine in each solution. Dissolution rate (z) with respect to the labeled amount of etilefrine hydrochloride (C10H15NO2.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of etilefrine hydrochloride for assay taken C: Labeled amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl) in 1 tablet. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of etilefrine are not less than 8000 and 0.9 – 1.2, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of etilefrine is not more than 2.0z.
Weigh accurately the mass of not less than 20 Etilefrine Hydrochloride Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 5 mg of etilefrine hydrochloride (C10H15NO2.HCl), add 60 mL of diluted hydrochloric acid (1 in 1000), shake for 10 minutes, add diluted hydrochloric acid (1 in 1000) to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 50 mg of etilefrine hydrochlo-ride for assay, previously dried at 1059C for 4 hours, and dissolve in diluted hydrochloric acid (1 in 1000) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted hy-drochloric acid (1 in 1000) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as direct under Liquid Chromatography <2.01> accord-ing to the following conditions, and determine the peak areas, AT and AS, of etilefrine in each solution. Amount (mg) of etilefrine hydrochloride (C10H15NO2.HCl) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of etilefrine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 5 g of sodium lauryl sulfate in 940 mL of water and 500 mL of acetonitrile, and adjust the pH to 2.3 with phosphoric acid. Flow rate: Adjust so that the retention time of etilefrine is about 6 minutes. System suitability— System performance: Dissolve 4 mg of bamethan sulfate and 4 mg of etilefrine hydrochloride in the mobile phase to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, etilefrine and bamethan are eluted in this order with the resolution be-tween these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of etilefrine is not more than 1.0z.
(1) To a quantity of powdered Etizolam Fine Granules, equivalent to 5 mg of Etizolam, add 10 mL of methanol, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the fil-trate to dryness on a water bath, cool, and then dissolve the residue in 2 mL of sulfuric acid. The solution gives off a light yellow-green fluorescent when exposed to ultraviolet light (main wavelength: 365 nm). (2) To a quantity of powdered Etizolam Fine Granules, equivalent to 1 mg of Etizolam, add 80 mL of 0.1 mol/L hy-drochloric acid TS, shake, and then filter. Determine the absorption spectrum of the filtrate as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits absorption maxima between 249 nm and 253 nm, and between 292 nm and 296 nm, when perform the measurement within 10 minutes. ‌JP XVIII Official Monographs / Etizolam Tablets 985 Uniformity of dosage units <6.02> The Granules in single-dose packages meet the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Etizolam Fine Granules is not less than 75z. Start the test with an accurately weighed amount of Etizol-am Fine Granules, equivalent to about 1 mg of etizolam (C17H15ClN4S), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of first filtrate, pipet 2 mL of the subsequent filtrate, add exactly 2 mL of aceto-nitrile, and use this solution as the sample solution. Sepa-rately, weigh accurately about 28 mg of etizolam for assay, previously dried at 1059C for 3 hours, and dissolve in metha-nol to make exactly 50 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 4 mL of this solu-tion, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of etizolam in each solution. Dissolution rate (z) with respect to the labeled amount make exactly 100 mL. Pipet 2 mL of this solution, and add diluted methanol (7 in 10) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal stand-ard solution, add diluted methanol (7 in 10) to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of etizolam to that of the inter-nal standard. Amount (mg) of etizolam (C17H15ClN4S) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of etizolam for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in diluted methanol (7 in 10) (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust the pH to 3.5 with diluted phosphoric acid (1 in 10). To 550 mL of this solution add 450 mL of acetonitrile. of etizolam (C 17H 15ClN4S) Flow rate: Adjust so that the retention time of etizolam is about 6 minutes. ＝ MS/MT × AT/AS × 1/C × 18/5 MS: Amount (mg) of etizolam for assay taken MT: Amount (g) of Etizolam Fine Granules taken C: Labeled amount (mg) of etizolam (C17H15ClN4S) in 1 g Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 243 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of etizolam is about 7 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of etizolam are not less than 3000 and not more than 2.0, respectively. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and etizolam are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of etizolam to that of the internal standard is not more than 1.0z.
Etizolam Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat- etizolam (C 17H15 ClN4S: 342.85). ing conditions, the relative standard deviation of the peak area of etizolam is not more than 2.0z. Assay Weigh accurately an amount of powdered Etizolam Fine Granules, equivalent to about 4 mg of etizolam (C17H15ClN4S), add 30 mL of water, and stir. Add 60 mL of methanol, stir for 20 minutes, add methanol to make exactly 100 mL, and centrifuge. Pipet 5 mL of the supernatant liq-uid, add exactly 10 mL of the internal standard solution, add diluted methanol (7 in 10) to make 25 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 0.1 g of etizolam for assay, previously dried at 1059C for 3 hours, and dissolve in diluted methanol (7 in 10) to Method of preparation Prepare as directed under Tablets, with Etizolam. Identification (1) To a quantity of powdered Etizolam Tablets, equivalent to 5 mg of Etizolam, add 10 mL of meth-anol, shake, and filter. Evaporate the filtrate to dryness on a water bath, and dissolve the residue in 2 mL of sulfuric acid. The solution gives off a light yellow-green fluorescence when exposed to ultraviolet light (main wavelength: 365 nm). (2) To a quantity of powdered Etizolam Tablets, equivalent to 1 mg of Etizolam, add 80 mL of 0.1 mol/L hy-drochloric acid TS, shake, and then filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Deter- mine the absorption spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its absorption maxima between 249 nm and 253 nm, and be-tween 292 nm and 296 nm when perform the measurement within 10 minutes. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Etizolam Tablets add 2.5 mL of water, and stir until the tablet is disintegrated. Add 20 mL of methanol, stir for 20 minutes, add methanol to make exactly 25 mL, and centrifuge. Pipet V mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add diluted methanol (9 in 10) to make 25 mL so that each mL contains about 8 mg of etizolam (C17H15ClN4S), and use this solution as the sample solution.
(1) To a quantity of powdered Etizolam Tablets, equivalent to 5 mg of Etizolam, add 10 mL of meth-anol, shake, and filter. Evaporate the filtrate to dryness on a water bath, and dissolve the residue in 2 mL of sulfuric acid. The solution gives off a light yellow-green fluorescence when exposed to ultraviolet light (main wavelength: 365 nm). (2) To a quantity of powdered Etizolam Tablets, equivalent to 1 mg of Etizolam, add 80 mL of 0.1 mol/L hy-drochloric acid TS, shake, and then filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Deter- mine the absorption spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its absorption maxima between 249 nm and 253 nm, and be-tween 292 nm and 296 nm when perform the measurement within 10 minutes. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Etizolam Tablets add 2.5 mL of water, and stir until the tablet is disintegrated. Add 20 mL of methanol, stir for 20 minutes, add methanol to make exactly 25 mL, and centrifuge. Pipet V mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add diluted methanol (9 in 10) to make 25 mL so that each mL contains about 8 mg of etizolam (C17H15ClN4S), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of etizolam (C17H15ClN4S) ＝ MS × QT/QS × 1/V × 1/20 MS: Amount (mg) of etizolam for assay taken Internal standard solution—A solution of ethyl para-hydroxybenzoate in diluted methanol (9 in 10) (1 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Etizolam Tablets is not less than 70z. Start the test with 1 tablet of Etizolam Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 0.28 mg of etizolam (C17H15ClN4S). Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile, and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of etizolam for assay, previously dried at 1059C for 3 hours, dissolve in 50 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL. Pipet 2 mL of this solu-tion, add water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile, and use this solu-tion as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of etizolam in each solution. Dissolution rate (z) with respect to the labeled amount of etizolam (C17H15ClN4S) ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of etizolam for assay taken C: Labeled amount (mg) of etizolam (C17H15ClN4S) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 243 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of etizolam is about 7 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of etizolam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of etizolam is not more than 2.0z.
Weigh accurately an amount of powdered Etizolam Fine Granules, equivalent to about 4 mg of etizolam (C17H15ClN4S), add 30 mL of water, and stir. Add 60 mL of methanol, stir for 20 minutes, add methanol to make exactly 100 mL, and centrifuge. Pipet 5 mL of the supernatant liq-uid, add exactly 10 mL of the internal standard solution, add diluted methanol (7 in 10) to make 25 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 0.1 g of etizolam for assay, previously dried at 1059C for 3 hours, and dissolve in diluted methanol (7 in 10) to Method of preparation Prepare as directed under Tablets, with Etizolam. Identification (1) To a quantity of powdered Etizolam Tablets, equivalent to 5 mg of Etizolam, add 10 mL of meth-anol, shake, and filter. Evaporate the filtrate to dryness on a water bath, and dissolve the residue in 2 mL of sulfuric acid. The solution gives off a light yellow-green fluorescence when exposed to ultraviolet light (main wavelength: 365 nm). (2) To a quantity of powdered Etizolam Tablets, equivalent to 1 mg of Etizolam, add 80 mL of 0.1 mol/L hy-drochloric acid TS, shake, and then filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Deter- mine the absorption spectrum of this filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its absorption maxima between 249 nm and 253 nm, and be-tween 292 nm and 296 nm when perform the measurement within 10 minutes. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Etizolam Tablets add 2.5 mL of water, and stir until the tablet is disintegrated. Add 20 mL of methanol, stir for 20 minutes, add methanol to make exactly 25 mL, and centrifuge. Pipet V mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add diluted methanol (9 in 10) to make 25 mL so that each mL contains about 8 mg of etizolam (C17H15ClN4S), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of etizolam (C17H15ClN4S) ＝ MS × QT/QS × 1/V × 1/20 MS: Amount (mg) of etizolam for assay taken Internal standard solution—A solution of ethyl para-hydroxybenzoate in diluted methanol (9 in 10) (1 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Etizolam Tablets is not less than 70z. Start the test with 1 tablet of Etizolam Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 0.28 mg of etizolam (C17H15ClN4S). Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile, and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of etizolam for assay, previously dried at 1059C for 3 hours, dissolve in 50 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL. Pipet 2 mL of this solu-tion, add water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of acetonitrile, and use this solu-tion as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of etizolam in each solution. Dissolution rate (z) with respect to the labeled amount of etizolam (C17H15ClN4S) ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of etizolam for assay taken C: Labeled amount (mg) of etizolam (C17H15ClN4S) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 243 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of etizolam is about 7 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of etizolam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of etizolam is not more than 2.0z. Assay To 20 Etizolam Tablets add 50 mL of water, and stir until they disintegrate. Add 400 mL of methanol, stir for 20 minutes, add methanol to make exactly 500 mL, and centri-fuge. Pipet an amount of the supernatant liquid, equivalent to about 0.2 mg of etizolam (C17H15ClN4S), add exactly 2 mL of the internal standard solution, add diluted methanol (9 in 10) to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 100 mg of etizolam for assay, previously dried at 1059C for 3 hours, and dissolve in diluted methanol (9 in 10) to make exactly 100 mL. Pipet 2 mL of this solution, and add diluted metha-nol (9 in 10) to make exactly 100 mL. Pipet 10 mL of this so-lution, add exactly 2 mL of the internal standard solution, add diluted methanol (9 in 10) to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of etizolam to that of the internal standard. Amount (mg) of etizolam (C17H15ClN4S) ＝ MS × QT/QS × 1/500 MS: Amount (mg) of etizolam for assay taken Internal standard solution—A solution of ethyl para-hydroxybenzoate in diluted methanol (9 in 10) (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust the pH to 3.5 with diluted phosphoric acid (1 in 10). To 550 mL of this solution add 450 mL of acetonitrile. Flow rate: Adjust so that the retention time of etizolam is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and etizolam are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of etizolam to that of the internal standard is not more than 1.0z.
Famotidine for Injection is a preparation for injec-tion which is dissolved before use. It contains not less than 94.
Famotidine for Injection occurs as white, porous masses or powder.
Dissolve an amount of Famotidine for Injec-tion, equivalent to 0.01 g of Famotidine, in 50 mL of 0.05 mol/L potassium dihydrogen phosphate TS. To 5 mL of this solution add 0.05 mol/L potassium dihydrogen phosphate TS to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 263 nm and 267 nm. pH <2.54> Dissolve an amount of Famotidine for Injection, equivalent to 0.02 g of Famotidine, in 1 mL of water: the pH of this solution is between 4.9 and 5.5.
(1) Clarity and color of solution—Dissolve an amount of Famotidine for Injection, equivalent to 0.02 g of Famotidine, in 1 mL of water: the solution is clear and col-orless. (2) Related substances—Take a number of Famotidine for Injection, equivalent to about 0.1 g of famotidine (C8H15N7O2S3), dissolve each content in water, wash the in-side of the container with water, combine the solutions of the contents with the washings, add water to the combined solution to make exactly 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area of these solutions by the automatic integration method: the total area of the peaks other than famotidine obtained from the sample solu-tion is not larger than peak area of famotidine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of famotidine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the water to make exactly 20 mL. Confirm that the peak area of famotidine obtained with 5 mL of this solution is equivalent to 8 to 12z of that with 5 mL of the standard solution. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of famotidine is not more than 2.0z. ‌JP XVIII Official Monographs / Famotidine Powder 991 Water <2.48> Not more than 1.5z (0.1 g, coulometric titration). Bacterial endotoxins <4.01> Not more than 15 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take a number of Famotidine for Injection, equiva-lent to about 0.1 g of famotidine (C8H15N7O2S3), dissolve each content in water, wash the inside of each container with water, combine the solutions of the contents with the wash-ings, add water to the combined solution to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of famotidine for assay, previ-ously dried in vacuum with phosphorus (V) oxide at 809C for 4 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of famotidine to that of the internal standard. Amount (mg) of famotidine (C8H15N7O2S3) ＝ MS × QT/QS × 2 MS: Amount (mg) of famotidine for assay taken Internal standard solution—To 5 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 500) add water to make 50 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2 g of sodium 1-heptane sulfonate in 900 mL of water, adjust to pH 3.0 with acetic acid (100), and add water to make 1000 mL. To this solution add 240 mL of acetonitrile and 40 mL of methanol. Flow rate: Adjust so that the retention time of famotidine is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, famotidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 11. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of famotidine to that of the internal standard is not more than 1.0z.
Famotidine Injection is an aqueous injection. It contains not less than 92.
Famotidine Injection is a colorless or light yel-low, clear liquid.
To an amount of Famotidine Injection, equivalent to 10 mg of Famotidine, add water to make 100 mL. Run 1 mL of this solution on a column prepared by fill-ing about 1 cm inside diameter chromatography tube with about 0.4 g of 55 – 105 mm octadecylsilanized silica gel for pretreatment. Wash the column with 15 mL of water, fol-lowed by elution with 5 mL of methanol. To the eluate add methanol to make 10 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 285 nm and 289 nm. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—To an exact amount of Famotidine Injection, equivalent to 25 mg of Famotidine, add the mobile phase to make exactly 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 10 mg of famotidine for assay, dissolve in methanol, and add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak area of each solution by the automatic integration method, and calculate the amounts of the related substances by the following equation: the amounts of related sub-stances, having the relative retention time about 1.3 and about 1.5 to famotidine are not more than 3.0z respec-tively, and the amount of other related substances except the above substances is not more than 0.5z, and the total amount of the related substances is not more than 5.0z. Amount (z) of related substance ＝ MS × AT/AS × 1/10 Total amount (z) of related substances ＝ MS × SAT/AS × 1/10 MS: Amount (mg) of famotidine for assay taken AS: Peak area of famotidine in the standard solution AT: Peak area of related substances in the sample solution SAT: Total peak area of the related substances in the sam-ple solution Operating conditions— Detector, column and column temperature: Proceed as di-rected in the operating conditions in the Assay. Mobile phase: Dissolve 1.74 g of sodium 1-pentane sul-fonate in 900 mL of water, adjust to pH 4.0 with diluted acetic acid (100) (1 in 10), and add water to make 1000 mL. To 840 mL of this solution add 80 mL of methanol and 40 mL of acetonitrile. Flow rate: Adjust so that the retention time of famotidine is about 17 minutes. Time span of measurement: About 4 times as long as the retention time of famotidine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add the mobile phase to make exactly 50 mL. Con-firm that the peak area of famotidine obtained with 20 mL of this solution is equivalent to 8 to 12z of that with 20 mL of the standard solution. System performance: To 20 mg of famotidine for assay add 2 mL of a solution of methyl parahydroxybenzoate in acetonitrile (1 in 500), and add methanol to make 20 mL. To 5 mL of this solution add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, famotidine and methyl par-ahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 19. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above opera-tions conditions, the relative standard deviation of the peak area of famotidine is not more than 2.0z. Bacterial endotoxins <4.01> Less than 15 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according ‌990 to Method I: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to Membrane filtration method: it meets the requirement.
To an exactly measured volume of Famotidine Injection, equivalent to about 25 mg of famotidine (C8H15N7O2S3), add exactly 2.5 mL of the internal standard solution, and add the mobile phase to make 50 mL. Pipet 10 mL of this solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of famotidine for assay, previ-ously dried in vacuum with phosphorus (V) oxide at 809C for 4 hours, dissolve in methanol, add exactly 5 mL of the internal standard solution, and add methanol to make 50 mL. Pipet 5 mL of this solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QT and QS of the peak area of famotidine to that of the internal standard. Amount (mg) of famotidine (C8H15N7O2S3) ＝ MS × QT/QS × 1/2 MS: Amount (mg) of famotidine for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in acetonitrile (1 in 500). Operating conditions— Ditector: An ultraviolet absorption photometer (Wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.74 g of sodium 1-pentane sul-fonate in 900 mL of water, adjust to pH 4.0 with diluted acetic acid (100) (1 in 10), and add water to make 1000 mL. To 750 mL of this solution add 200 mL of methanol and 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of famotidine is about 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, famotidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 26. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of famotidine to that of the internal standard is not more than 1.0z.
Famotidine Powder contains not less than 94.
Weigh a portion of Famotidine Powder, equivalent to 0.01 g of Famotidine, add 50 mL of 0.05 mol/L potassium dihydrogen phosphate TS, shake well, and centrifuge. To 5 mL of the supernatant liquid add 0.05 mol/L potassium dihydrogen phosphate TS to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 263 nm and 267 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Famotidine Powder in single-dose packages meets the requirement of the Content unifor-mity test. Take out the total content of 1 package of Famotidine Powder, add 10 mL of water per 10 mg of famotidine (C8H15N7O2S3), shake well, add 10 mL of methanol, shake well, add methanol to make exactly V mL so that each mL contains about 0.4 mg of famotidine (C8H15N7O2S3), and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of famotidine (C8H15N7O2S3) ＝ MS × QT/QS × V/250 MS: Amount (mg) of famotidine for assay taken Internal standard solution—To 5 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 500) add water to make 50 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rates in 15 minutes of a 20-mg/g powder and a 100-mg/g powder are not less than 80z and not less than 85z, respectively. Start the test with an accurately weighed amount of Famo-tidine Powder, equivalent to about 20 mg of famotidine (C8H15N7O2S3), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of famotidine for assay, previ-ously dried in vacuum on phosphorus (V) oxide at 809C for 4 hours, dissolve in the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 266 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of famotidine (C8H15N7O2S3) ＝ MS/MT × AT/AS × 1/C × 45 MS: Amount (mg) of famotidine for assay taken ‌992 MT: Amount (g) of Famotidine Powder taken C: Labeled amount (mg) of famotidine (C8H15N7O2S3) in 1 g
Weigh accurately a portion of Famotidine Powder, equivalent to about 20 mg of famotidine (C8H15N7O2S3), add 20 mL of water, and shake well. Add 20 mL of methanol, then shake well, add methanol to make exactly 50 mL, and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of famotidine for assay, previously dried in vacuum with phosphorus (V) oxide at 809C for 4 hours, dissolve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of famotidine to that of the internal standard. Amount (mg) of famotidine (C8H15N7O2S3) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of famotidine for assay taken Internal standard solution—To 5 mL of a solution of methyl parahydroxybenzoate in methanol (1 in 500) add water to make 50 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2 g of sodium 1-heptane sulfonate in 900 mL of water, adjust to pH 3.0 with acetic acid (100), and add water to make 1000 mL. To this solution add 240 mL of acetonitrile and 40 mL of methanol. Flow rate: Adjust so that the retention time of famotidine is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, famotidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 11. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of famotidine to that of the internal standard is not more than 1.0z. Famotidine Tablets
Weigh a portion of powdered Famotidine Tablets, equivalent to 0.01 g of Famotidine, add 50 mL of 0.05 mol/L potassium dihydrogen phosphate TS, shake well, and centrifuge. To 5 mL of the supernatant liquid add 0.05 mol/L potassium dihydrogen phosphate TS to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 263 nm and 267 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Famotidine Tablets add 2 mL of water, shake to disintegrate, then add a suitable amount of methanol, and shake well. Add methanol to make exactly V mL of a solution containing about 0.2 mg of famotidine (C8H15N7O2S3) per mL, and centrifuge. Pipet 10 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of famotidine for assay, previously dried in vacuum with phosphorus (V) oxide at 809C for 4 hours, dis-solve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, and add methanol to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 2 mL of the inter-nal standard solution, add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the operating conditions described in the Assay, and calculate the ratios, QT and QS, of the peak area of famotidine to that of the internal standard. Amount (mg) of famotidine (C8H15N7O2S3) ＝ MS × QT/QS × V/500 MS: Amount (mg) of famotidine for assay taken Internal standard solution—To 5 mL of a solution of methyl paraphydroxybenzoate in methanol (1 in 500) add water to make 50 mL. Dissolution Being specified separately when the drug is granted approval based on the Law.
Take a number of Famotidine Tablets, equivalent to 0.2 g of famotidine (C8H15N7O2S3), add 50 mL of water, and
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Faropenem Sodium for Syrup is a preparation for syrup, which is dissolved before use. It contains not less than 93.
Dissolve an amount of powdered Faropenem Sodium for Syrup, equivalent to 25 mg (potency) of Faro-penem Sodium Hydrate, in water to make 50 mL. To 5 mL of this solution add water to make 50 mL, filter, if neces-sary, and determine the absorption spectrum of the solution so obtained as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 254 nm and 258 0 – 54 84 → 30 16 → 70 Flow rate: 1.5 mL per minute. Time span of measurement: 2.5 times as long as the reten-tion time of faropenem, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add water to make exactly 20 mL. Con-firm that the peak area of faropenem obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution obtained in the Assay under the above operating conditions, the internal standard and faro-penem are eluted in this order with the resolution between these peaks being not less than 11. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of faropenem is not more than 3.0z. Water <2.48> Not less than 1.5z and not more than 2.1z (80 mg, coulometric titration). ‌JP XVIII Official Monographs / Faropenem Sodium Tablets 995 Uniformity of dosage units <6.02> Faropenem Sodium for Syrup in single-dose packages meet the requirement of the Mass variation test.
Powder, if necessary, and weigh accurately an amount of Faropenem Sodium for Syrup, equivalent to lution. For the area of the peak, having the relative retention time of about 0.71 to faropenem, multiply its correction fac-tor 0.37. Operating conditions— Detector: An ultraviolet absorption photometer (wave- about 25 mg (potency) of faropenem (C 12H 15NO5S), add length: 240 nm). Column: A stainless steel column 4 mm in inside diameter exactly 10 mL of the internal standard solution and a suita-ble amount of water, shake well, and add water to make 50 mL. Filter, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg (potency) of Faropenem Sodium RS, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Preceed as directed in the Assay under Faropenem Sodium Hydrate. Amount [mg (potency)] of faropenem (C12H15NO5S) ＝ MS × QT/QS MS: Amount [mg (potency)] of Faropenem Sodium RS taken and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 6.12 g of potassium dihydrogen phosphate, 1.79 g of disodium hydrogen phosphate dodeca-hydrate and 1.61 g of tetra n-butylammonium bromide in water to make 1000 mL. Mobile phase B: A mixture of the mobile phase A and acetonitrile (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Internal standard solution—Dissolve 0.5 g of m-hydroxya-cetophenone in 20 mL of acetonitrile, and add water to make Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 200 mL.
To powdered Faropenem Sodium Tablets, equivalent to 70 mg (potency) of Faropenem Sodium Hy-drate, add water to make 100 mL. To 5 mL of this solution add water to make 100 mL, filter, if necessary, and deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 254 nm and 258 nm and between 304 nm and 308 nm.
Related substances—Powder not less than 5 Faro-penem Sodium Tablets. To a part of the powder, equivalent to about 25 mg (potency) of Faropenem Sodium Hydrate, add about 10 mL of water, shake well, then add water to make exactly 50 mL, and filter. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Pipet 2 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area of both solutions by the automatic integration method: the area of the peak of cleaved derivative, having the relative retention time of about 0.71 to faropenem, obtained from the sample solution is not larger than 1.5 times the peak area of faropenem from the standard solution, and the total area of the peaks other than faropenem from the sample solution is not larger than 2.5 times the peak area of faropenem from the standard so- 0 – 54 84 → 30 16 → 70 Flow rate: 1.5 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of faropenem, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add water to make exactly 20 mL. Con-firm that the peak area of faropenem obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution obtained in the Assay under the above operating conditions, the internal standard and faro-penem are eluted in this order with the resolution between these peaks being not less than 11. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of faropenem is not more than 3.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Faropenem Sodium Tablets add 130 mL of water, shake vigorously until the tablets are disintegrated, and add water to make exactly V mL so that each mL con-tains about 1 mg (potency) of Faropenem Sodium Hydrate. Pipet 5 mL of this solution, add water to make exactly 100 mL, and filter. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 25 mg (potency) of Faro-penem Sodium RS, and dissolve in water to make exactly 50 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT275, AT305, AT354, AS275, AS305 and AS354, of the sample solution and standard solution at 275 nm, 305 nm and 354 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, and calculate AT and AS, using the following equations. AT ＝ AT305 － (49 × AT275 ＋ 30 × AT354)/79 AS ＝ AS305 － (49 × AS275 ＋ 30 × AS354)/79 ‌996 Amount [mg (potency)] of faropenem (C12H15NO5S) ＝ MS × AT/AS × V/25 MS: Amount [mg (potency)] of Faropenem Sodium RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Faropenem Sodium Tablets is not less than 85z. Start the test with 1 tablet of Faropenem Sodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less Felbinac
Felodipine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of felodipine (C18H19Cl2NO4: 384.25). Method of preparation Prepare as directed under Tablets, with Felodipine. Identiˆcation To a quantity of powdered Felodipine Tablets, equivalent to 4 mg of Felodipine, add 200 mL of methanol, shake thoroughly, add methanol to make 250 mL, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 235 nm and 239 nm, and between 357 nm and 363 nm. Uniformity of dosage units <6.02> Perform the Mass varia- tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Felodipine Tablets add 1 mL of water per 2.5 mg of felodipine (C18H19Cl2NO4), and shake thoroughly until the tablet is completely disintegrated. Add exactly 1 mL of the internal standard solution per 2.5 mg of felodipine (C 18H19Cl2NO4), and add methanol to make V mL so that each mL contains about 0.
Weigh accurately the mass of not less than 20 Felodipine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of felodipine (C18H19Cl2NO4), add 20 mL of water, add exactly 4 mL of the internal standard solution, add methanol to make 100 mL and shake. Centrifuge this solution, ˆlter the superna-tant liquid through a membrane ˆlter with a pore size not exceeding 0.45 mm, and use the ˆltrate as the sample solu-tion. Separately, weigh accurately about 10 mg of felodipine for assay (separately determine the loss on drying <2.41> in the same conditions as Felodipine), add 20 mL of water, add exactly 4 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of felodipine to that of the internal standard. Fenbufen
Fenofibrate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of fenofibrate (C20H21ClO4: 360.83). Method of preparation Prepare as directed under Tablets, with Fenofibrate. Identification To a quantity of powdered Fenofibrate Tablets, equivalent to 10 mg of Fenofibrate, add 10 mL of a mixture of acetonitrile and water (7:3), shake, and centri-fuge. To 1 mL of the supernatant liquid add a mixture of Operating conditions— Detector, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay under Fenofibrate. Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust so that the retention time of fenofibrate is about 21 minutes. Time span of measurement: About 2 times as long as the retention time of fenofibrate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of acetonitrile and water (7:3) to make exactly 10 mL. Confirm that the peak area of fenofibrate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fenofibrate are not less than 10,000 and 0.8 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fenofibrate is not more than 3.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Fenofibrate Tablets add exactly 20 mL of a mixture of acetonitrile and water (7:3), and shake until the tablet is disintegrated. Centrifuge this solution, pipet V mL of the su-pernatant liquid, equivalent to about 20 mg of fenofibrate (C20H21ClO4), and add a mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, add a mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. acetonitrile and water (7:3) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi- Amount (mg) of fenofibrate (C ＝ MS × QT/QS × 8/V 20H21ClO4) mum between 285 nm and 289 nm. Purity Related substances—Conduct this procedure using light-resistant vessels. To 4 mL of the supernatant liquid ob-tained in the Assay add a mixture of acetonitrile and water (7:3) to make 10 mL, and use this solution as the sample so-lution. Pipet 1 mL of this solution, and add a mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 5 mL of this solution, add a mixture of acetonitrile and water (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than fenofibrate and related substance A having the relative reten-tion time of about 1.4 to fenofibrate obtained from the sam-ple solution, is not larger than 2/5 times the peak area of fenofibrate from the standard solution, and the total area of the peaks other than fenofibrate from the sample solution is not larger than the peak area of fenofibrate from the stand-ard solution. MS: Amount (mg) of Fenofibrate RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 1 g of polysorbate 80 in water to make 100 mL as the dissolution medium, the dissolution rate in 30 minutes of Fenofibrate Tablets is not less than 75z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Fenofibrate Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 59 mg of fenofibrate (C20H21ClO4), and use this solution as the sample solution.
To a quantity of powdered Fenofibrate Tablets, equivalent to 10 mg of Fenofibrate, add 10 mL of a mixture of acetonitrile and water (7:3), shake, and centri-fuge. To 1 mL of the supernatant liquid add a mixture of Operating conditions— Detector, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay under Fenofibrate. Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust so that the retention time of fenofibrate is about 21 minutes. Time span of measurement: About 2 times as long as the retention time of fenofibrate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of acetonitrile and water (7:3) to make exactly 10 mL. Confirm that the peak area of fenofibrate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fenofibrate are not less than 10,000 and 0.8 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fenofibrate is not more than 3.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. To 1 tablet of Fenofibrate Tablets add exactly 20 mL of a mixture of acetonitrile and water (7:3), and shake until the tablet is disintegrated. Centrifuge this solution, pipet V mL of the su-pernatant liquid, equivalent to about 20 mg of fenofibrate (C20H21ClO4), and add a mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, add a mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. acetonitrile and water (7:3) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi- Amount (mg) of fenofibrate (C ＝ MS × QT/QS × 8/V 20H21ClO4) mum between 285 nm and 289 nm.
Related substances—Conduct this procedure using light-resistant vessels. To 4 mL of the supernatant liquid ob-tained in the Assay add a mixture of acetonitrile and water (7:3) to make 10 mL, and use this solution as the sample so-lution. Pipet 1 mL of this solution, and add a mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 5 mL of this solution, add a mixture of acetonitrile and water (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than fenofibrate and related substance A having the relative reten-tion time of about 1.4 to fenofibrate obtained from the sam-ple solution, is not larger than 2/5 times the peak area of fenofibrate from the standard solution, and the total area of the peaks other than fenofibrate from the sample solution is not larger than the peak area of fenofibrate from the stand-ard solution. MS: Amount (mg) of Fenofibrate RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 1 g of polysorbate 80 in water to make 100 mL as the dissolution medium, the dissolution rate in 30 minutes of Fenofibrate Tablets is not less than 75z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Fenofibrate Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 59 mg of fenofibrate (C20H21ClO4), and use this solution as the sample solution. Separately, weigh accurately about 12 mg of Fenofibrate RS, previously dried in vacuum over phosphorus (V) oxide at 609C for 4 hours, and dissolve in a mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 2 mL of this solution add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with ex- ‌JP XVIII Official Monographs / Fentanyl Citrate 1003 actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of fenofibrate in each solution. Dissolution rate (z) with respect to the labeled amount of fenofibrate (C20H21ClO4) ＝ MS × AT/AS × V?/V × 1/C × 450 MS: Amount (mg) of Fenofibrate RS taken C: Labeled amount (mg) of fenofibrate (C20H21ClO4) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay under Fenofibrate. Mobile phase: A mixture of acetonitrile and 0.02 mol/L phosphate buffer solution (pH 3.0) (4:1). Storage—Light-resistant. Others Related substance A: Refer to it described in Fenofibrate. Fentanyl Citrate
Fexofenadine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of fexofenadine hydrochloride (C32H39NO4.HCl: 538.12). Method of preparation Prepare as directed under Tablets, with Fexofenadine Hydrochloride. Identification To an amount of powdered Fexofenadine Hydrochloride Tablets, equivalent to 40 mg of Fexofenadine Hydrochloride, add 100 mL of methanol, and shake well. Filter, discard the first 10 mL of the filtrate, and determine the absorption spectrum of the subsequent filtrate as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 257 nm and 261 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Fexofenadine Hydrochloride Tablets add V/5 mL of diluted acetic acid (100) (17 in 10,000), shake until the tablet is disintegrated. Add 3V/5 mL of acetonitrile for liquid chromatography, shake well, add a mixture of acetonitrile for liquid chromatography and diluted acetic acid (100) (17 in 10,000) (3:1) to make exactly V mL so that each mL contains about 0.
To an amount of powdered Fexofenadine Hydrochloride Tablets, equivalent to 40 mg of Fexofenadine Hydrochloride, add 100 mL of methanol, and shake well. Filter, discard the first 10 mL of the filtrate, and determine the absorption spectrum of the subsequent filtrate as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 257 nm and 261 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Fexofenadine Hydrochloride Tablets add V/5 mL of diluted acetic acid (100) (17 in 10,000), shake until the tablet is disintegrated. Add 3V/5 mL of acetonitrile for liquid chromatography, shake well, add a mixture of acetonitrile for liquid chromatography and diluted acetic acid (100) (17 in 10,000) (3:1) to make exactly V mL so that each mL contains about 0.3 mg of fexofenadine hydrochlo-ride (C32H39NO4.HCl). Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of Fexofenadine Hydrochloride RS (separately determine the water <2.48> in the same manner as Fexofenadine Hydrochloride), and dissolve in a mixture of acetonitrile for liquid chromatography and diluted acetic acid (100) (17 in 10,000) (3:1) to make exactly 200 mL. Pipet 6 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of fexofenadine hydrochloride (C32H39NO4.HCl) ＝ MS × AT/AS × 3V/500 MS: Amount (mg) of Fexofenadine Hydrochloride RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Fexofenadine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Fexofenadine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 30 mg of fexofenadine hydro-chloride (C32H39NO4.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 30 mg of Fexofenadine Hydrate RS (separately determine the water <2.48> in the same manner as Fexofenadine Hydrochloride), dissolve in 5 mL of methanol, add water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of fexofenadine in each solution. Dissolution rate (z) with respect to the labeled amount of fexofenadine hydrochloride (C32H39NO4.HCl) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Fexofenadine Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of fexofenadine hydrochloride (C32H39NO4.HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.1 g of sodium dihydrogen phos-phate dihydrate, 0.3 mL of phosphoric acid and 0.5 g of so-dium perchlorate in 300 mL of water, add 700 mL of aceto-nitrile for liquid chromatography. Flow rate: Adjust so that the retention time of fexofena-dine is about 3.5 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fexofenadine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fexofenadine is not more than 2.0z.
To 20 Fexofenadine Hydrochloride Tablets add V/5 mL of diluted acetic acid (100) (17 in 10,000), and shake until the tablets are disintegrated. Then, add 3V/5 mL of acetonitrile for liquid chromatography, shake well, and add a mixture of acetonitrile for liquid chromatography and diluted acetic acid (100) (17 in 10,000) (3:1) to make exactly V mL so that each mL contains about 1.2 mg of fexofena-dine hydrochloride (C32H39NO4.HCl). Pipet 15 mL of this solution, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 45 mg of Fex-ofenadine Hydrochloride RS (separately determine the water <2.48> in the same manner as Fexofenadine Hydrochloride), and dissolve in a mixture of acetonitrile for liquid chroma-tography and diluted acetic acid (100) (17 in 10,000) (3:1) to make exactly 200 mL. Pipet 20 mL of this solution, add the mobile phase to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of fexofenadine in each solution. Amount (mg) of fexofenadine hydrochloride (C32H39NO4.HCl) in 1 tablet ＝ MS × AT/AS × V/750 MS: Amount (mg) of Fexofenadine Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 1000 mL of diluted acetic acid (100) (17 in 10,000) add 15 mL of a mixture of triethylamine and acetonitrile for liquid chromatography (1:1), and adjust to pH 5.25 with phosphoric acid. To 16 volumes of this solu-tion add 9 volumes of acetonitrile for liquid chromato-graphy. Flow rate: Adjust so that the retention time of fexofena-dine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fexofenadine are not less than 7000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fexofenadine is not more than 1.0z.
Filgrastim (Genetical Recombination) Injection
フィルグラスチム（遺伝子組換え）注射液
Filgrastim (Genetical Recombination) Injection is an aqueous injection. It contains not less than 90.
Filgrastim (Genetical Recombination) Injec-tion is a clear and colorless liquid.
Take a volume of Filgrastim (Genetical Recombination) Injection, equivalent to 5 to 10 mg of Fil-grastim (Genetical Recombination) depending on the size of polyacrylamide gel for filgrastim, and add 0 to 16 mL of water. To 3 volumes of this solution add 1 volume of buffer solution for filgrastim sample so that each mL contains about 0.19 mg of protein, and use this solution as the sample solution. Then, proceed as directed in the Identification (1) under Filgrastim (Genetical Recombination). Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law.
Multimers—Proceed as directed in the Purity (1) under Filgrastim (Genetical Recombination) using a volume of Filgrastim (Genetical Recombination) Injection, equiva-lent to about 125 mg of Filgrastim (Genetical Recombina-tion). Where, the test for required detectability and the sys-tem repeatability under the system suitability are tested using Filgrastim RS. Bacterial endotoxins <4.01> Less than 0.25 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according ‌1010 to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Biological activity Calculate the biological activity in 1 am-poule or syringe of Filgrastim (Genetical Recombination) Injection by the following equation, using the biological ac-tively in 1 mL of Filgrastim (Genetical Recombination) In-jection determined as directed in the Assay (2) under Fil-grastim (Genetical Recombination) and the labeled volume of Filgrastim (Genetical Recombination) Injection: it is not less than 70z and not more than 140z of the target biologi-cal activity (unit). Biological activity (unit) in 1 ampoule or syringe of Fil-grastim (Genetical Recombination) Injection ＝ antilog M × biological activity (unit/mL) of Filgrastim RS × dilution factor for UH/dilution factor for SH × UH/SH × labeled volume (mL) of Filgrastim (Genetical Recombination) Injection where, the target biological activity (unit) is calculated by the following formula. Target biological activity (unit) ＝ 1.5 × 108 (unit/mg) × labeled amount (mg) of Filgrastim (Genetical Recombination) in labeled volume (mL)
Perform the test with an exact volume each of Fil-grastim (Genetical Recombination) Injection and Filgrastim RS, equivalent to about 100 mg of Filgrastim (Genetical Recombination), as directed in the Assay (1) under Fil-grastim (Genetical Recombination). Calculate the amount of filgrastim in 1 mL of Filgrastim (Genetical Recombination) Injection by following formula. Amount (mg) of filgrastim in 1 mL ＝ C × AT/AS × VS/VT C: Protein concentration (mg/mL) of Filgrastim RS VS: Amount (mL) of Filgrastim RS taken VT: Amount (mL) of Filgrastim (Genetical Recombina-tion) Injection taken
Flecainide Acetate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of flecainide acetate (C17H20F6N2O3.C2H4O2: 474.39). Method of preparation Prepare as directed under Tablets, with Flecainide Acetate. Identification To an amount of powdered Flecainide Ace-tate Tablets, equivalent to 0.2 g of Flecainide Acetate, add 4 mL of methanol, shake for 20 minutes, then centrifuge and use the supernatant liquid as the sample solution. Separately, dissolve 0.1 g of flecainide acetate in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and ‌1014 standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of acetone and ammonia solution (28) (20:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Completely disintegrate 1 tablet of Flecainide Acetate Tablets in 4V/5 mL of a solution of lactic acid (1 in 500) by sonicating. After allowing to stand for 30 minutes while swirling occasionally, add a solution of lactic acid (1 in 500) to make exactly V mL so that each mL contains about 1 mg of flecainide acetate (C17H20F6N2O3.
To an amount of powdered Flecainide Ace-tate Tablets, equivalent to 0.2 g of Flecainide Acetate, add 4 mL of methanol, shake for 20 minutes, then centrifuge and use the supernatant liquid as the sample solution. Separately, dissolve 0.1 g of flecainide acetate in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and ‌1014 standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of acetone and ammonia solution (28) (20:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Completely disintegrate 1 tablet of Flecainide Acetate Tablets in 4V/5 mL of a solution of lactic acid (1 in 500) by sonicating. After allowing to stand for 30 minutes while swirling occasionally, add a solution of lactic acid (1 in 500) to make exactly V mL so that each mL contains about 1 mg of flecainide acetate (C17H20F6N2O3.C2H4O2), and filter. Dis-card the first 10 mL of the filtrate, pipet 5 mL of the subse-quent filtrate, add a solution of lactic acid (1 in 500) to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of flecainide acetate (C17H20F6N2O3.C2H4O2) ＝ MS × AT/AS × V/25 MS: Amount (mg) of flecainide acetate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Flecainide Acetate Tablets is not less than 70z. Start the test with 1 tablet of Flecainide Acetate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of flecainide acetate (C17H20F6N2O3.C2 H4O2), and use this solution as the sample solution. Sepa-rately, weigh accurately about 28 mg of flecainide acetate for assay, previously dried under reduced pressure not exceeding 0.67 kPa at 609C for 2 hours, and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 296 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of flecainide acetate (C17H20F6N2O3.C2H4O2) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of flecainide acetate for assay taken C: Labeled amount (mg) of flecainide acetate (C17H20F6N2O3.C2H4O2) in 1 tablet
Accurately weigh the mass of not less than 20 Flecainide Acetate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of flecainide acetate (C17H20F6N2O3.C2H4O2), add 80 mL of a solution of lactic acid (1 in 500), sonicate for 5 minutes, then add a solution of lactic acid (1 in 500) to make exactly 100 mL, and filter. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add a solution of lactic acid (1 in 500) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of flecainide acetate for assay, previously dried under reduced pressure not exceeding 0.67 kPa at 609C for 2 hours, dissolve in a solution of lactic acid (1 in 500) to make exactly 50 mL. Pipet 10 mL of this solution, add a solution of lactic acid (1 in 500) to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 296 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of flecainide acetate (C17H20F6N2O3.C2H4O2) ＝ MS × AT/AS × 4 MS: Amount (mg) of flecainide acetate for assay taken
Flomoxef Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Flomoxef Sodium for Injection occurs as white to light yellow-white, friable masses or powder.
Proceed as directed in the Identification (3) under Flomoxef Sodium. pH <2.54> The pH of a solution obtained by dissolving an amount of Flomoxef Sodium for Injection, equivalent to 0.5 g (potency) of Flomoxef Sodium, in 5 mL of water is be-tween 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve an amount of Flomoxef Sodium for Injection, equivalent to 1.0 g (potency) of Flomoxef Sodium, in 10 mL of water: the solution is clear and colorless or pale yellow. (2) 1-(2-Hydroxyethyl)-1H-tetrazol-5-thiol—Use the sample solution obtained in the Assay as the sample solu-tion. Weigh accurately about 20 mg of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 25 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internal standard. Calculate the amount of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol per 1 g (potency) of Flomoxef Sodium for Injection by the following formula: not more than 10 mg. Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol (C H N OS) System repeatability: When the test is repeated 3 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internal standard is not more than 1.0z. Water <2.48> Not more than 1.5z (0.5 g, volumetric titra-tion, back titration). Bacterial endotoxins <4.01> Less than 0.025 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Flomoxef Sodium for Injection, and calculate the average mass of the content. Spread out thinly about 1 g of the content in a petri dish, allow the dish to stand in a desic-cator containing a saturated solution of magnesium bromide without light exposure to equilibrate the sample to constant water content. Determine the water content, separately, with about 0.1 g of the sample according to the method described in Water. Weigh accurately an amount of the sample, equivalent to about 50 mg (potency) of Flomoxef Sodium, add exactly 50 mL of the internal standard solution to dis-solve, add water to make 100 mL, and use this solution as the sample solution. Separately weigh accurately about 50 mg (potency) of Flomoxef Triethylammonium RS, add exactly 50 mL of the internal standard solution to dissolve, add water to make 100 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Flomoxef Sodium. Amount [ mg (potency)] of flomoxef (C15H18F2N6O7S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Flomoxef Triethylammo-nium RS taken Internal standard solution—A solution of m-cresol (3 in 1000). 3 6 4 ＝ MS × QT/QS × 1/10 MS: Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol taken Internal standard solution—A solution of m-cresol (3 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Flomoxef Sodium. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add water to make exactly 20 mL. Confirm that the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol obtained with 5 mL of this solution is equivalent to 3.5 – 6.5z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol and the internal standard are eluted in this order with the resolution between these peaks being not less than 20.
Containers—Hermetic containers. Plastic containers for aqueous injection may be used.
Flopropione Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of flopropione (C9H10O4: 182.17). Method of preparation Prepare as directed under the Cap-sules, with Flopropione. Identification (1) Powder the contents of Flopropione Capsules. To a portion of the powder, equivalent to 60 mg of Flopropione, add 40 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of iron (III) nitrate TS: a red-purple color appears. (2) Powder the contents of Flopropione Capsules. To a portion of the powder, equivalent to 90 mg of Flopropione, add 100 mL of ethanol (99.5), shake well, and filter. To 5 mL of the filtrate add ethanol (99.5) to make 50 mL. To 5 mL of this solution add ethanol (99.5) to make 100 mL, and use this solution as the sample solution. Determine the ab-sorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Flopropione Capsules add 43 mL of a mixture of water and phosphoric acid (86:1), and disinte-grate the capsule in a water bath at 509C. After cooling, add ‌1018 a suitable amount of acetonitrile to make exactly V mL of a solution containing about 0.4 mg of flopropione (C9H10O4) per mL. Stir the solution for 10 minutes, centrifuge a part of the solution at 3000 rpm for 5 minutes, and use the superna-tant liquid as the sample solution. Proceed as directed in the Assay. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 267 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Amount (mg) of flopropione (C9H ＝ MS × AT/AS × V/100 10O4) Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, water and phos- MS: Amount (mg) of flopropione for assay taken, calcu-lated on the anhydrous basis Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Flopropione Capsules is not less than 80z. Start the test with 1 capsule of Flopropione Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 8.
(1) Powder the contents of Flopropione Capsules. To a portion of the powder, equivalent to 60 mg of Flopropione, add 40 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of iron (III) nitrate TS: a red-purple color appears. (2) Powder the contents of Flopropione Capsules. To a portion of the powder, equivalent to 90 mg of Flopropione, add 100 mL of ethanol (99.5), shake well, and filter. To 5 mL of the filtrate add ethanol (99.5) to make 50 mL. To 5 mL of this solution add ethanol (99.5) to make 100 mL, and use this solution as the sample solution. Determine the ab-sorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Flopropione Capsules add 43 mL of a mixture of water and phosphoric acid (86:1), and disinte-grate the capsule in a water bath at 509C. After cooling, add ‌1018 a suitable amount of acetonitrile to make exactly V mL of a solution containing about 0.4 mg of flopropione (C9H10O4) per mL. Stir the solution for 10 minutes, centrifuge a part of the solution at 3000 rpm for 5 minutes, and use the superna-tant liquid as the sample solution. Proceed as directed in the Assay. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 267 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Amount (mg) of flopropione (C9H ＝ MS × AT/AS × V/100 10O4) Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile, water and phos- MS: Amount (mg) of flopropione for assay taken, calcu-lated on the anhydrous basis Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Flopropione Capsules is not less than 80z. Start the test with 1 capsule of Flopropione Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 8.8 mg of flopropione (C9H10O4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of flopropione for assay (separately determine the water <2.48> in the same manner as Flopropione), and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 284 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using 0.1 mol/L hydrochloric acid TS as the blank. Dissolution rate (z) with respect to the labeled amount of flopropione (C9H10O4) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of flopropione for assay taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of flopropione (C9H10O4) in 1 capsule
Take out the contents of not less than 20 Flopropione Capsules, weigh accurately the mass of the con-tents, and power. Weigh accurately a part of the powder, equivalent to about 40 mg of flopropione (C9H10O4), and add the mobile phase to make exactly 100 mL. Stir the solu-tion for 10 minutes, centrifuge a part of this solution for 5 minutes at 3000 rpm, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 40 mg of flopropione for assay (previously determine the water <2.48> in the same manner as Flopropione), add 70 mL of the mobile phase, and dissolve by sonicating for 10 minutes. Add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid chromatography <2.01> accord-ing to the following conditions, and determine the peak areas, AT and AS, of flopropione in each solution. phoric acid (114:86:1). Flow rate: Adjust so that the retention time of flopropione is about 3 minutes. System suitability— System performance: Dissolve 50 mg of flopropione in 50 mL of the mobile phase. To 20 mL of the solution add 25 mL of a solution prepared by dissolving 25 mg of ethyl par-ahydroxybenzoate in 30 mL of acetonitrile and add water to make 50 mL, and then add the mobile phase to make 50 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, Flopropione and ethyl par-ahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of flopropione is not more than 1.0z.
Fluconazole Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of fluconazole (C13H12F2N6O: 306.27). Method of preparation Prepare as directed under Cap-sules, with Fluconazole. Identification To an amount of powdered contents of Fluconazole Capsules, equivalent to 25 mg of Fluconazole, add 0.01 mol/L hydrochloric acid-methanol TS to make 100 mL, shake for 30 minutes, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 259 nm and 263 nm and between 265 nm and 269 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To the total amount of the content of 1 capsule of Fluconazole Capsules add the mobile phase to make exactly 100 mL. Disperse the particles by sonicating, stir for 30 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 50 mg of fluconazole (C13H12F2N6O), and use this solution as the sample solution.
To an amount of powdered contents of Fluconazole Capsules, equivalent to 25 mg of Fluconazole, add 0.01 mol/L hydrochloric acid-methanol TS to make 100 mL, shake for 30 minutes, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 259 nm and 263 nm and between 265 nm and 269 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To the total amount of the content of 1 capsule of Fluconazole Capsules add the mobile phase to make exactly 100 mL. Disperse the particles by sonicating, stir for 30 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 50 mg of fluconazole (C13H12F2N6O), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of fluconazole (C13H12F2N6O) ＝ MS × AT/AS × V?/V × 1/5 MS: Amount (mg) of fluconazole for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rates in 90 minutes of 50-mg capsule and 100-mg capsule are not less than 80z and not less than 70z, respec-tively. Start the test with 1 capsule of Fluconazole Capsules, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 28 mg of fluconazole (C13H12F2N6O), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of fluconazole for assay, pre-viously dried at 1059C for 4 hours, and dissolve in the mo-bile phase to make exactly 50 mL. Pipet 5 mL of this solu-tion, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of fluconazole in each solution. ‌1020 Dissolution rate (z) with respect to the labeled amount of fluconazole (C13H12F2N6O) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of fluconazole for assay taken C: Labeled amount (mg) of fluconazole (C13H12F2N6O) in 1 capsule Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fluconazole are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fluconazole is not more than 1.0z.
Take out the contents from not less than 20 Fluconazole Capsules, weigh accurately, and powder, if necessary. Weigh accurately a quantity of the contents, equivalent to about 50 mg of fluconazole (C13H12F2N6O), and add the mobile phase to make exactly 100 mL. Disperse the particles by sonicating, stir for 30 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add the mobile phase to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of fluconazole for assay, previously dried at 1059C for 4 hours, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of fluconazole in each solution. Amount (mg) of fluconazole (C13H12F2N6O) ＝ MS × AT/AS × 2 MS: Amount (mg) of fluconazole for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 261 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. area of fluconazole is not more than 1.0z.
Fluconazole Injection is an aqueous injection. It contains not less than 95.
Fluconazole Injection occurs as a clear and colorless liquid.
(1) Take a volume of Fluconazole Injec-tion, equivalent to 0.1 g of Fluconazole, and evaporate to dryness on a water bath. To the residue add 10 mL of dilute hydrochloric acid, shake, and filter. Add 1 mL of Reinecke salt TS to the filtrate: a light red precipitate is produced. (2) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 259 nm and 263 nm, and between 264 nm and 268 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 0.75 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Fluconazole Injection, equivalent to 10 mg of fluconazole (C13H12F2N6O), add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of fluconazole for assay, previously dried at 1059C for 4 hours, dissolve in a so-lution of sodium chloride (9 in 1000) to make exactly 50 mL. Pipet 10 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 261 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Mobile phase: Dissolve 0.82 g of anhydrous sodium ace-tate in 1000 mL of water, and adjust to pH 5.0 with acetic acid (100). To 700 mL of this solution add 200 mL of metha- Amount (mg) of fluconazole (C ＝ MS × AT/AS × 1/5 13H12F2N6O) nol and 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of fluconazole is about 4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fluconazole are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak MS: Amount (mg) of fluconazole for assay taken
Fludiazepam Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of fludiazepam (C16H12ClFN2O: 302.73). Method of preparation Prepare as directed under Tablets, with Fludiazepam. Identification To a quantity of powdered Fludiazepam Tablets, equivalent to 2 mg of Fludiazepam, add 40 mL of methanol, shake for 20 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 315 nm and 319 nm. Therefore, to 5 mL of the filtrate add methanol to make 50 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 229 nm and 233 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Fludiazepam Tablets add 2V/25 mL of water, disintegrate the fine particles by sonicating, add 3V/25 mL of acetonitrile, and shake for 10 minutes. Add a mixture of acetonitrile and water (3:2) to make exactly V mL so that each mL contains about 5 mg of fludiazepam (C16H12ClFN2O), centrifuge, and use the supernatant liquid as the sample solution.
To a quantity of powdered Fludiazepam Tablets, equivalent to 2 mg of Fludiazepam, add 40 mL of methanol, shake for 20 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 315 nm and 319 nm. Therefore, to 5 mL of the filtrate add methanol to make 50 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 229 nm and 233 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Fludiazepam Tablets add 2V/25 mL of water, disintegrate the fine particles by sonicating, add 3V/25 mL of acetonitrile, and shake for 10 minutes. Add a mixture of acetonitrile and water (3:2) to make exactly V mL so that each mL contains about 5 mg of fludiazepam (C16H12ClFN2O), centrifuge, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of fludiazepam (C16H12ClFN2O) ＝ MS × AT/AS × V/5000 MS: Amount (mg) of fludiazepam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 ‌JP XVIII Official Monographs / Fludrocortisone Acetate 1023 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Fludiazepam Tablets is not less than 80z. Start the test with 1 tablet of Fludiazepam Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.28 mg of fludiazepam (C16H12ClFN2O), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of fludiazepam for assay, previously dried at 609C for 3 hours under reduced pressure, and dis-solve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Then, pipet 2 mL of this solution, and add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of fludiazepam in each solution. Dissolution rate (z) with respect to the labeled amount Amount (mg) of fludiazepam (C16H12ClFN2O) ＝ MS × AT/AS × 1/100 MS: Amount (mg) of fludiazepam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and water (3:2). Flow rate: Adjust so that the retention time of fludiazep- am is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fludiazepam are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times of fludiazepam (C 16H 12ClFN2O) with 20 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of fludiazepam for assay taken C: Labeled amount (mg) of fludiazepam (C16H12ClFN2O) in 1 tablet Operating conditions— Detector, column temperature and flow rate: Proceed as directed in the operating conditions in the Assay. Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Mobile phase: A mixture of water and acetonitrile (1:1). System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fludiazepam are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fludiazepam is not more than 2.0z.
Weigh accurately the mass of not less than 20 tablets of Fludiazepam Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.25 mg of fludiazepam (C16H12ClFN2O), add 4 mL of water, disperse the fine particles by sonicating, add 6 mL of acetonitrile, and shake for 10 minutes. To this solution add a mixture of acetonitrile and water (3:2) to make exactly 50 mL, centri-fuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of fludiazepam for assay, previously dried at 609C for 3 hours under reduced pressure, dissolve in a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Then, pipet 5 mL of this solution, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of fludiazepam in each solution. area of fludiazepam is not more than 1.0z.
Flutoprazepam Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of flutoprazepam (C19H16ClFN2O: 342.79). Method of preparation Prepare as directed under Tablets, with Flutoprazepam. Identification To a quantity of powdered Flutoprazepam Tablets, equivalent to 10 mg of Flutoprazepam, add 20 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000), shake well, and add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 100 mL. Centrifuge this solution, to 10 mL of the supernatant liquid add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 240 nm and 244 nm, between 279 nm and 285 nm, and between 369 nm and 375 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Flutoprazepam Tablets add 60 mL of the mobile phase, shake for 15 minutes to disintegrate, disperse the particle by sonicating, and add the mobile phase to make ‌exactly V mL so that each mL contains about 20 mg of flutoprazepam (C19H16ClFN2O).
To a quantity of powdered Flutoprazepam Tablets, equivalent to 10 mg of Flutoprazepam, add 20 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000), shake well, and add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 100 mL. Centrifuge this solution, to 10 mL of the supernatant liquid add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 240 nm and 244 nm, between 279 nm and 285 nm, and between 369 nm and 375 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Flutoprazepam Tablets add 60 mL of the mobile phase, shake for 15 minutes to disintegrate, disperse the particle by sonicating, and add the mobile phase to make ‌exactly V mL so that each mL contains about 20 mg of flutoprazepam (C19H16ClFN2O). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Proceed as directed in the Assay. Amount (mg) of flutoprazepam (C19H16ClFN2O) ＝ MS × AT/AS × V/1000 MS: Amount (mg) of flutoprazepam for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Flutoprazepam Tablets is not less than 70z. Start the test with 1 tablet of Flutoprazepam Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.2 mg of flutoprazepam (C19H16ClFN2O), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of flutoprazepam for assay, previously dried at 1059C for 2 hours, and dissolve in metha-nol to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed un-der Liquid Chromatography <2.01> according to the follow-ing conditions, and determine the peak areas, AT and AS, of flutoprazepam in each solution. Dissolution rate (z) with respect to the labeled amount of flutoprazepam (C19H16ClFN2O) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of flutoprazepam for assay taken C: Labeled amount (mg) of flutoprazepam (C19H16ClFN2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of flutoprazepam are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flutoprazepam is not more than 1.0z.
Weigh accurately not less than 20 Flutoprazepam Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 2 mg of flutoprazepam (C19H16ClFN2O), add 60 mL of the mobile phase, shake for Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of flutoprazepam in each solution. Amount (mg) of flutoprazepam (C19H16ClFN2O) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of flutoprazepam for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and water (3:1). Flow rate: Adjust so that the retention time of flutoprazepam is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of flutoprazepam are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of flutoprazepam is not more than 1.0z.
Fluvoxamine Maleate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of fluvoxamine maleate (C15H21F3N2O2. C4H4O4: 434.41). Method of preparation Prepare as directed under Tablets, with Fluvoxamine Maleate. Identification Powder Fluvoxamine Maleate Tablets. To a portion of the powder, equivalent to 0.1 g of Fluvoxamine Maleate, add 50 mL of water, shake, then allow to stand, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 0.5 mL of the fil-trate add 50 mL of water, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 243 nm and 247 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Fluvoxamine Maleate Tablets add 4 mL of water, disintegrate the tablet by sonicating, add a mixture of methanol for liquid chromatography and water (7:3) to make exactly 50 mL, and filter. Pipet V mL of the filtrate, equivalent to about 6 mg of fluvoxamine maleate (C15H21F3N2O2.C4H4O4), add exactly 2 mL of the internal standard solution, then add a mixture of methanol for liquid chromatography and water (7:3) to make 50 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) ＝ MS × QT/QS × 6/V MS: Amount (mg) of Fluvoxamine Maleate RS taken, cal-culated on the dried basis Internal standard solution—A solution of diphenylamine in methanol for liquid chromatography (3 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Fluvoxamine Maleate Tablets is not less than 80z. Start the test with 1 tablet of Fluvoxamine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg of fluvoxamine male-ate (C15H21F3N2O2.
Powder Fluvoxamine Maleate Tablets. To a portion of the powder, equivalent to 0.1 g of Fluvoxamine Maleate, add 50 mL of water, shake, then allow to stand, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 0.5 mL of the fil-trate add 50 mL of water, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 243 nm and 247 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Fluvoxamine Maleate Tablets add 4 mL of water, disintegrate the tablet by sonicating, add a mixture of methanol for liquid chromatography and water (7:3) to make exactly 50 mL, and filter. Pipet V mL of the filtrate, equivalent to about 6 mg of fluvoxamine maleate (C15H21F3N2O2.C4H4O4), add exactly 2 mL of the internal standard solution, then add a mixture of methanol for liquid chromatography and water (7:3) to make 50 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) ＝ MS × QT/QS × 6/V MS: Amount (mg) of Fluvoxamine Maleate RS taken, cal-culated on the dried basis Internal standard solution—A solution of diphenylamine in methanol for liquid chromatography (3 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Fluvoxamine Maleate Tablets is not less than 80z. Start the test with 1 tablet of Fluvoxamine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg of fluvoxamine male-ate (C15H21F3N2O2.C4H4O4), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Fluvoxamine Maleate RS (separately determine the loss on drying <2.41> under the same condition as Fluvoxamine Maleate), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 245 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Fluvoxamine Maleate RS taken, cal-culated on the dried basis C: Labeled amount (mg) of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) in 1 tablet
To 10 Fluvoxamine Maleate Tablets add 20 mL of water, disintegrate the tablets by sonicating, then add a mix-ture of methanol for liquid chromatography and water (7:3) to make exactly 250 mL, and filter. Pipet V mL of the fil-trate, equivalent to about 6 mg of fluvoxamine maleate (C15H21F3N2O2.C4H4O4), add exactly 2 mL of the internal standard solution, then add a mixture of methanol for liquid chromatography and water (7:3) to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of Fluvoxamine Maleate RS (separately determine the loss on drying <2.41> under the same condition as Fluvoxamine Maleate), and dissolve in a mixture of meth-anol for liquid chromatography and water (7:3) to make ex-actly 25 mL. Pipet 3 mL of this solution, add exactly 2 mL of the internal standard solution, then add a mixture of methanol for liquid chromatography and water (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of fluvoxamine to that of the internal standard. Amount (mg) of fluvoxamine maleate (C15H21F3N2O2.C4H4O4) in 1 tablet ＝ MS × QT/QS × 3/V MS: Amount (mg) of Fluvoxamine Maleate RS taken, cal-culated on the dried basis Internal standard solution—A solution of diphenylamine in methanol for liquid chromatography (3 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Fluvoxamine Maleate. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, fluvoxamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of fluvoxamine to that of the internal standard is not more than 1.0z.
Folic Acid Injection is an aqueous injection. It contains not less than 95.
Folic Acid Injection is a yellow to orange-yel-low, clear liquid. MS: Amount (mg) of p-Aminobenzoyl Glutamic Acid for Purity RS taken MT: Amount (mg) of Folic Acid taken, calculated on the anhydrous basis pH: 8.0 – 11.0
(1) To a volume of Folic Acid Injection, equivalent to 1.5 mg of Folic Acid, add dilute sodium hy-droxide TS to make 100 mL. Proceed as directed in the Iden- ‌1038 tification (2) under Folic Acid, using this solution as the sample solution. Determine the absorption spectrum of the sample so-lution obtained in (1) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 255 nm and 257 nm, between 281 nm and 285 nm and between 361 nm and 369 nm. Separately, determine the maximal ab-sorbances of the sample solution, A1 and A2, between 255 nm and 257 nm and between 361 nm and 369 nm, respec-tively: the ratio of A1/A2 is between 2.80 and 3.00. Folic Acid Injection responds to Qualitative Tests <1.09> (1) for sodium salt. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Folic Acid Injec-tion, equivalent to about 50 mg of folic acid (C19H19N7O6) add dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Folic Acid RS (separately, determine the water <2.48> in the same manner as Folic Acid), dissolve in dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the standard solu-tion. Proceed with exactly 30 mL each of the sample solution and standard solution as directed in the Assay under Folic Acid. is between 2.80 and 3.00. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Folic Acid Tablets add 50 mL of dilute so-dium hydroxide TS, shake frequently, and filter. Wash the residue with dilute sodium hydroxide TS, combine the fil-trate and the washings, then add dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the sample stock solution. Pipet 30 mL of the sample stock solution, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 60 mL of this solution, add 0.5 g of zinc powder, shake frequently, allow to stand for 20 minutes, and filter the solution through a dried filter paper. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 15 mg of folic acid (C19H19N7O6), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Folic Acid RS (separately determine the water <2.48> in the same manner as Folic Acid), and dissolve in dilute sodium hydroxide TS to make exactly 100 mL. Pipet 30 mL of this solutions, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 60 mL of this solution, add 0.5 g of zinc powder, shake frequently, allow to stand for 20 minutes, and filter the solu-tion through a dried filter paper. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Pipet 4 mL each of the sample solution and standard solution, add 1 mL of water, 1 mL of dilute hydro-chloric acid and 1 mL of sodium nitrite solution (1 in 1000) to them, mix, and allow to stand for 2 minutes. To these so- Amount (mg) of folic acid (C ＝ MS × (AT － AC)/AS 19H19N7O6) lutions add 1 mL of a solution of ammonium amidosulfate (1 in 200), shake, and allow them to stand for 2 minutes. To these solutions add 1 mL of a solution of N,N-diethyl-N?-1- MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis
Folic Acid Tablets contain not less than 90.0z and not more than 115.0z of the labeled amount of folic acid (C19H19N7O6: 441.40). Method of preparation Prepare as directed under Tablets, with Folic Acid. Identification (1) Take a quantity of powdered Folic Acid Tablets, equivalent to 1.5 mg of Folic Acid, add 100 mL of dilute sodium hydroxide TS, shake, and filter. Discard the first 10 mL of the filtrate, use the subsequent filtrate as the sample solution, and proceed as directed in the Identification (2) under Folic Acid. (2) Determine the absorption spectrum of the filtrate ob-tained in (1) as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 255 nm and 257 nm, between 281 nm and 285 nm and between 361 nm and 369 nm. Separately, determine the maximal absorbances of the filtrate, A1 and A2, between 255 nm and 257 nm and between 361 nm and 369 nm, respectively: the ratio of A1/A2 naphthylethylenediamine oxalate (1 in 1000), shake, allow to stand for 10 minutes, and add water to make exactly 20 mL. Separately, pipet 30 mL of the sample stock solution, add 20 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Pipet V mL of this solution, and add water to make exactly V? mL so that each mL contains about 15 mg of folic acid (C19H19N7O6).
(1) Take a quantity of powdered Folic Acid Tablets, equivalent to 1.5 mg of Folic Acid, add 100 mL of dilute sodium hydroxide TS, shake, and filter. Discard the first 10 mL of the filtrate, use the subsequent filtrate as the sample solution, and proceed as directed in the Identification (2) under Folic Acid. (2) Determine the absorption spectrum of the filtrate ob-tained in (1) as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 255 nm and 257 nm, between 281 nm and 285 nm and between 361 nm and 369 nm. Separately, determine the maximal absorbances of the filtrate, A1 and A2, between 255 nm and 257 nm and between 361 nm and 369 nm, respectively: the ratio of A1/A2 naphthylethylenediamine oxalate (1 in 1000), shake, allow to stand for 10 minutes, and add water to make exactly 20 mL. Separately, pipet 30 mL of the sample stock solution, add 20 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Pipet V mL of this solution, and add water to make exactly V? mL so that each mL contains about 15 mg of folic acid (C19H19N7O6). With exactly 4 mL of this solu-tion perform the same procedure described above for ob-taining the sample solution, and use the solution so obtained as the blank solution. Determine the absorbances at 550 nm, AT, AS and AC, of the solutions obtained from the sample solution and standard solution, and the blank solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a control solution obtained with 4 mL of water in the same manner as described above. Amount (mg) of folic acid (C19H19N7O6) ＝ MS × (AT － AC)/AS × V?/V × 1/10 MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Folic Acid Tablets is not less than 75z. Start the test with 1 tablet of Folic Acid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con- tains about 5.6 mg of folic acid (C H N O ), and use this 19 19 7 6 ‌JP XVIII Official Monographs / Formalin Water 1039‌ solution as the sample solution. Separately, weigh accurately about 20 mg of Folic Acid RS (separately determine the water <2.48> in the same manner as Folic Acid), and dissolve in the 2nd fluid for dissolution test to make exactly 100 mL. Pipet 2.5 mL of this solution, add the 2nd fluid for dissolu-tion test to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 280 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of folic acid (C19H19N7O6) ＝ MS × AT/AS × V?/V × 1/C × 45/2 MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of folic acid (C19H19N7O6) in 1 tablet
Weigh accurately and powder not less than 20 Folic Acid Tablets. Weigh accurately a portion of the powder, equivalent to about 50 mg of folic acid (C19H19N7O6). Add 50 mL of dilute sodium hydroxide TS, shake frequently, then filter into a 100-mL volumetric flask, and wash with dilute sodium hydroxide TS. To the combined filtrate and washings add dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of Folic Acid RS (separately, determine the water <2.48> in the same manner as Folic Acid), dissolve in dilute sodium hydroxide TS to make exactly 100 mL, and use this solution as the standard solution. Proceed with exactly 30 mL each of the sample so-lution and standard solution as directed in the Assay under Folic Acid. Amount (mg) of folic acid (C19H19N7O6) ＝ MS × (AT － AC)/AS MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis
シロップ用ホスホマイシンカルシウム
Fosfomycin Calcium for Syrup is a preparation for syrups which is suspended before use. It contains not less than 90.
(1) To an amount of Fosfomycin Calcium for Syrup, equivalent to 40 mg (potency) of Fosfomycin Cal-cium Hydrate, add 10 mL of warm water, shake for 10 to 20 minutes, and collect the insoluble substances by filtration. Dissolve the substances in 3 mL of a solution of perchloric acid (1 in 4), add 1 mL of 0.1 mol/L sodium periodate solu-tion, and warm in a water bath at 609C for 30 minutes. After cooling, add 50 mL of water, neutralize the solution with a saturated solution of sodium hydrogen carbonate, and add 1 mL of potassium iodide TS: the solution does not show a red color. To an amount of Fosfomycin Calcium for Syrup, equivalent to 40 mg (potency) of Fosfomycin Calcium Hy-drate, add 10 mL of warm water, shake for 10 to 20 minutes, and collect the insoluble substances by filtration. Dissolve the substances in 3 mL of a solution of perchloric acid (1 in 4), add 2 mL of 0.1 mol/L sodium periodate solution, and heat in a water bath for 10 minutes. After cooling, add 1 mL of hexaammonium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, and allow to stand for 30 minutes: the solution shows a blue color. To an amount of Fosfomycin Calcium for Syrup, equivalent to 40 mg (potency) of Fosfomycin Calcium Hy-drate, add 10 mL of warm water, shake for 10 to 20 minutes, and collect the insoluble substances by filtration. Dissolve the substances in 25 mL of water: the solution responds to Qualitative Tests <1.09> (3) for calcium salt. Loss on drying <2.41> Not more than 3.0z (2 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Uniformity of dosage units <6.02> Fosfomycin Calcium for Syrup in single-dose packages meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Fosfomycin Calcium for Syrup is not less than 80z. Start the test with an accurately weighed amount of Fos-fomycin Calcium for Syrup, equivalent to about 0.5 g (po-tency) of Fosfomycin Calcium Hydrate, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Fosfomycin Phenethylammonium RS, equivalent to about 28 mg (potency), dissolve in water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of fosfomycin in each solution. Dissolution rate (z) with respect to the labeled amount of fosfomycin (C3H7O4P) ＝ MS/MT × AT/AS × 1/C × 1800 MS: Amount [mg (potency)] of Fosfomycin Phenethylam-monium RS taken MT: Amount (g) of Fosfomycin Calcium for Syrup taken C: Labeled amount [mg (potency)] of fosfomycin (C3H7O4P) in 1 g Operating conditions— Detector: A conductivity detector. Column: A polyetheretherketone column 4.6 mm in inside diameter and 7.5 cm in length, packed with quaternary am-monium group introducing hydrophilic vinyl polymer gel for liquid chromatography (6 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 10.5 g of citric acid monohydrate in water to make 1000 mL. To 800 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of fosfomycin is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fosfomycin are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fosfomycin is not more than 2.0z.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, agar media for seed and base layer, and standard solutions—Proceed as directed in the Assay under Fosfomycin Calcium Hydrate. Sample solutions—Weigh accurately an amount of Fosfomycin Calcium for Syrup, equivalent to about 0.1 g (potency) of Fosfomycin Calcium Hydrate, dissolve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 200 mL. Take exactly a suitable amount of this solution, add exactly 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample so-lution and the low concentration sample solution, respec-tively.
注射用ホスホマイシンナトリウム
Fosfomycin Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Fosfomycin Sodium for Injection occurs as a white crystalline powder.
(1) Dissolve about 0.1 g of Fosfomycin So-dium for Injection in 3 mL of a solution of perchloric acid (1 in 4), add 1 mL of 0.1 mol/L sodium periodate solution, and heat in a water bath at 609C for 30 minutes. After cooling, add 50 mL of water, neutralize with saturated sodium hydro-gen carbonate solution, and add 1 mL of potassium iodide TS; the solution does not reveal a red color, while the blank solution reveals a red color. To 2 mL of a solution of Fosfomycin Sodium for In-jection (1 in 250) add 1 mL of perchloric acid and 2 mL of 0.1 mol/L sodium periodate solution, and heat in a water bath for 10 minutes. After cooling, add 1 mL of hexaammo-nium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphtol-4-sulfonic acid TS, and allow to stand for 30 minutes: a blue color develops. Dissolve an amount of Fosfomycin Sodium for Injec-tion, equivalent to 0.1 g (potency) of Fosfomycin Sodium, in 50 mL of water. Perform the test with this solution as di-rected in the Identification (3) under Fosfomycin Sodium. pH <2.54> The pH of a solution prepared by dissolving an amount of Fosfomycin Sodium for Injection, equivalent to 1.0 g (potency) of Fosfomycin Sodium, in 20 mL of water is between 6.5 and 8.5.
Clarity and color of solution—Dissolve an amount of Fosfomycin Sodium for Injection, equivalent to 1.0 g (potency) of Fosfomycin Sodium, in 10 mL of water: the so-
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, seeded agar layer, and standard solutions—Proceed as directed in the Assay under Fosfomycin Sodium. Sample solutions—Weigh accurately the mass of the contents of not less than 10 Fosfomycin Sodium for Injec-tion. Weigh accurately an amount of the content, equivalent to about 20 mg (potency) of Fosfomycin Sodium, and dis-solve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 50 mL. Take exactly a suitable amount of this solu-tion, add 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively. 3 7 4 Method of preparation Prepare as directed under Injec-tions, with Fosfomycin Sodium. Description Fosfomycin Sodium for Injection occurs as a white crystalline powder. Identification (1) Dissolve about 0.1 g of Fosfomycin So-dium for Injection in 3 mL of a solution of perchloric acid (1 in 4), add 1 mL of 0.1 mol/L sodium periodate solution, and heat in a water bath at 609C for 30 minutes. After cooling, add 50 mL of water, neutralize with saturated sodium hydro-gen carbonate solution, and add 1 mL of potassium iodide TS; the solution does not reveal a red color, while the blank solution reveals a red color. To 2 mL of a solution of Fosfomycin Sodium for In-jection (1 in 250) add 1 mL of perchloric acid and 2 mL of 0.1 mol/L sodium periodate solution, and heat in a water bath for 10 minutes. After cooling, add 1 mL of hexaammo-nium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphtol-4-sulfonic acid TS, and allow to stand for 30 minutes: a blue color develops. Dissolve an amount of Fosfomycin Sodium for Injec-tion, equivalent to 0.1 g (potency) of Fosfomycin Sodium, in 50 mL of water. Perform the test with this solution as di-rected in the Identification (3) under Fosfomycin Sodium. pH <2.54> The pH of a solution prepared by dissolving an amount of Fosfomycin Sodium for Injection, equivalent to 1.0 g (potency) of Fosfomycin Sodium, in 20 mL of water is between 6.5 and 8.5. Purity Clarity and color of solution—Dissolve an amount of Fosfomycin Sodium for Injection, equivalent to 1.0 g (potency) of Fosfomycin Sodium, in 10 mL of water: the so-
Fructose Injection is an aqueous injection. It contains not less than 95.
Fructose Injection is a colorless to pale yellow, clear liquid. It has a sweet taste.
(1) Take a volume of Fructose Injection, equivalent to 1 g of Fructose, dilute with water or concen-trate on a water bath to 20 mL, if necessary, and use this so-lution as the sample solution. Add 2 to 3 drops of the sample solution to 5 mL of boiling Fehling's TS: a red precipitate is produced. (2) To 10 mL of the sample solution obtained in (1) add 0.1 g of resorcinol and 1 mL of hydrochloric acid, and warm in a water bath for 3 minutes: a red color develops. pH <2.54> 3.0 – 6.5 In the case where the labeled concen-tration of the injection exceeds 5z, dilute to 5z with water before the test.
(1) Heavy metals <1.07>—Take a volume of Fruc-tose Injection, equivalent to 5.0 g of Fructose, and evaporate on a water bath to dryness. With the residue, proceed ac-cording to Method 2, and perform the test. Prepare the con-trol solution with 2.0 mL of Standard Lead Solution. (2) Arsenic <1.11>—Take a volume of Fructose Injec- tion, equivalent to 1.5 g of Fructose, dilute with water or concentrate on a water bath to 5 mL, if necessary, and add 5 mL of dilute sulfuric acid and 1 mL of bromine TS. Proceed as directed in the purity (8) under Fructose. Residue on ignition <2.44> Measure exactly a volume of Fructose Injection, equivalent to 2 g of Fructose, evaporate on a water bath to dryness, and perform the test: the residue weighs not more than 2 mg. ‌JP XVIII Official Monographs / Fudosteine 1047 Bacterial endotoxins <4.01> Less than 0.5 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Fructose Injection, equivalent to about 4 g of fructose (C6H12O6), add 0.2 mL of ammonia TS, dilute with water to make exactly 100 mL, shake well, and after allowing to stand for 30 minutes, deter- mine the optical rotation, aD, in a 100-mm cell at 20 ± 19C as directed under Optical Rotation Determination <2.49>. Amount (mg) of fructose (C6H12O6) ＝ |aD| × 1087.0
Fudosteine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of fudosteine (C6H13NO3S: 179.24). Method of preparation Prepare as directed under Tablets, with Fudosteine. Identification Powder Fudostine Tablets. To a portion of the powder, equivalent to 88 mg of Fudosteine, add 10 mL of a mixture of water and methanol (1:1), shake, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 90 mg of fudosteine for assay in 10 mL of a mixture of water and methanol (1:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2.5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 809C for 5 minutes: the princi-pal spot obtained from the sample solution and the spot draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 55.
Powder Fudostine Tablets. To a portion of the powder, equivalent to 88 mg of Fudosteine, add 10 mL of a mixture of water and methanol (1:1), shake, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 90 mg of fudosteine for assay in 10 mL of a mixture of water and methanol (1:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2.5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (3:2:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of ninhydrin in acetone (1 in 50) on the plate, and heat at 809C for 5 minutes: the princi-pal spot obtained from the sample solution and the spot draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 55.6 mg of fudosteine (C6H13NO3S), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of fudosteine for assay, previ-ously dried at 1059C for 3 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of fudosteine in each solution. Dissolution rate (z) with respect to the labeled amount of fudosteine (C6H13NO3S) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of fudosteine for assay taken C: Labeled amount (mg) of fudosteine (C6H13NO3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fudosteine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fudosteine is not more than 2.0z.
Weigh accurately the mass of not less than 20 Fudosteine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.5 g of fudosteine (C6H13NO3S), add 70 mL of the mobile phase, shake vigor-ously for 15 minutes, add the mobile phase to make exactly 100 mL, and centrifuge. Pipet 10 mL of the supernatant liq-uid, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of fudosteine for assay, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of fudosteine to that of the internal standard. from the standard solution show a red-purple color and have Amount (mg) of fudosteine (C H NO S) the same Rf value. ＝ MS × QT/QS × 10 6 13 3 Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Fudosteine Tablets is not less than 85z. Start the test with 1 tablet of Fudosteine Tablets, with- MS: Amount (mg) of fudosteine for assay taken Internal standard solution—A solution of L-methionine in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). ‌JP XVIII Official Monographs / Furosemide 1049 Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A solution of sodium 1-hexanesulfonate in diluted phosphoric acid (1 in 1000) (1 in 1250). Flow rate: Adjust so that the retention time of fudosteine is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, fudosteine and the internal standard are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of fudosteine to that of the internal standard is not more than 1.0z.
Furosemide Injection is an aqueous injection. It contains not less than 95.
Furosemide Injection is a colorless, clear liq-uid.
(1) To a volume of Furosemide Injection, equivalent to 2.5 mg of Furosemide, add 10 mL of 2 mol/L hydrochloric acid TS, heat under a reflux condenser on a water bath for 15 minutes. After cooling, render the solution slightly acid with 18 mL of sodium hydroxide TS: the solu-tion responds to Qualitative Tests <1.09> for primary aro-matic amines. The color of the solution is red to red-purple. (2) To a volume of Furosemide Injection, equivalent to 20 mg of Furosemide, add water to make 100 mL. To 2 mL of this solution add 0.01 mol/L sodium hydroxide TS to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 227 nm and 231 nm, between 269 nm and 273 nm, and between 330 nm and 336 nm. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law.
Pipet a volume of Furosemide Injection, equivalent to 40 mg of Furosemide, add 30 mL of acetone, shake well, and add acetone to make exactly 50 mL. Centrifuge this so-lution, to 1.0 mL of the supernatant liquid add 3.0 mL of water, cool in a ice bath, add 3.0 mL of dilute hydrochloric acid and 0.15 mL of sodium nitrite TS, shake, and allow to stand for 1 minute. To this solution add 1.0 mL of ammo-nium amidosulfate TS, shake well, allow to stand for 3 minutes, add 1.0 mL of N,N-diethyl-N?-1-naphtylethyl-enediamine oxalate TS, shake well, and allow to stand for 5 minutes. Determine the absorbance of this solution at 530 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution, prepared in the same manner with 1.0 mL of acetone, as the blank: the absorbance is not more than 0.10. Bacterial endotoxins <4.01> Less than 1.25 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Furosemide Injection, equivalent to about 20 mg of Furosemide (C12H11ClN2O5S), add water to make exactly 100 mL. Pipet 3 mL of this solution, add 0.01 mol/L sodium hydrochloride TS to make exactly 100 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 20 mg of Furosemide RS, previously dried at 1059C for 4 hours, add 0.01 mol/L sodium hydroxide TS to make exactly 100 mL. Pipet 3 mL of this solution, add 0.01 mol/L sodium hydrox-ide TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 271 nm. Amount (mg) of furosemide (C12H11ClN2O5S) ＝ MS × AT/AS MS: Amount (mg) of Furosemide RS taken
Furosemide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of furosemide (C12H11ClN2O5S: 330.74). Method of preparation Prepare as directed under Tablets, with Furosemide. Identification (1) Shake well a quantity of powdered Furosemide Tablets, equivalent to 0.2 g of Furosemide, with 40 mL of acetone, and filter. To 0.5 mL of the filtrate add 10 mL of 2 mol/L hydrochloric acid TS, and heat under a reflux condenser on a water bath for 15 minutes. After cool-ing, add 18 mL of sodium hydroxide TS to make the solu-tion slightly acidic: the solution responds to Qualitative Tests <1.09> for primary aromatic amines, producing a red to red- purple color. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be- ‌JP XVIII Official Monographs / Fursultiamine Hydrochloride 1051 tween 227 nm and 231 nm, between 269 nm and 273 nm, and between 330 nm and 336 nm. Purity To a quantity of powdered Furosemide Tablets, equivalent to 40 mg of Furosemide, add about 30 mL of ace-tone, shake well, and add acetone to make exactly 50 mL. Centrifuge the solution, add 3.0 mL of water to 1.0 mL of the supernatant liquid, cool in ice, add 3.0 mL of dilute hy-drochloric acid and 0.15 mL of sodium nitrite TS, shake, and allow to stand for 1 minute. Add 1.0 mL of ammonium amidosulfate TS, shake well, allow to stand for 3 minutes, add 1.0 mL of N,N-diethyl-N?-1-naphthylethylenediamine oxalate TS, shake well, and allow to stand for 5 minutes. Perform the test with this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>, using a solution pre-pared in the same manner with 1.0 mL of acetone as the blank: the absorbance at 530 nm is not more than 0.10. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Furosemide Tablets add a suitable amount of 0.05 mol/L sodium hydroxide TS, shake to disintegrate, then add 0.05 mol/L sodium hydroxide TS to make exactly V mL so that each mL contains about 0.
(1) Shake well a quantity of powdered Furosemide Tablets, equivalent to 0.2 g of Furosemide, with 40 mL of acetone, and filter. To 0.5 mL of the filtrate add 10 mL of 2 mol/L hydrochloric acid TS, and heat under a reflux condenser on a water bath for 15 minutes. After cool-ing, add 18 mL of sodium hydroxide TS to make the solu-tion slightly acidic: the solution responds to Qualitative Tests <1.09> for primary aromatic amines, producing a red to red- purple color. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be- ‌JP XVIII Official Monographs / Fursultiamine Hydrochloride 1051 tween 227 nm and 231 nm, between 269 nm and 273 nm, and between 330 nm and 336 nm.
To a quantity of powdered Furosemide Tablets, equivalent to 40 mg of Furosemide, add about 30 mL of ace-tone, shake well, and add acetone to make exactly 50 mL. Centrifuge the solution, add 3.0 mL of water to 1.0 mL of the supernatant liquid, cool in ice, add 3.0 mL of dilute hy-drochloric acid and 0.15 mL of sodium nitrite TS, shake, and allow to stand for 1 minute. Add 1.0 mL of ammonium amidosulfate TS, shake well, allow to stand for 3 minutes, add 1.0 mL of N,N-diethyl-N?-1-naphthylethylenediamine oxalate TS, shake well, and allow to stand for 5 minutes. Perform the test with this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>, using a solution pre-pared in the same manner with 1.0 mL of acetone as the blank: the absorbance at 530 nm is not more than 0.10. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Furosemide Tablets add a suitable amount of 0.05 mol/L sodium hydroxide TS, shake to disintegrate, then add 0.05 mol/L sodium hydroxide TS to make exactly V mL so that each mL contains about 0.4 mg of furosemide (C12H11ClN2O5S). Filter the solution, discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 0.05 mol/L sodium hydroxide TS to make exactly 100 mL, and use this solution as the sample solution. Proceed as di-rected in the Assay. Amount (mg) of furosemide (C12H11ClN2O5S) ＝ MS × AT/AS × V/50 MS: Amount (mg) of Furosemide RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rates of a 20-mg tablet in 15 minutes and a 40-mg tablet in 30 minutes are not less than 80z. Start the test with 1 tablet of Furosemide Tablets, with-draw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 10 mg of furosemide (C12H11ClN2O5S), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Furosemide RS, previously dried at 1059C for 4 hours, and dissolve in 5 mL of methanol, and add the dissolution me-dium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 277 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of furosemide (C12H11ClN2O5S) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Furosemide RS taken C: Labeled amount (mg) of furosemide (C12H11ClN2O5S) in 1 tablet
Weigh accurately the mass of not less than 20 Furosemide Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 40 mg of furosemide (C12H11ClN2O5S), add about 70 mL of 0.05 mol/L sodium hydroxide TS, shake well, and add 0.05 mol/L sodium hy-droxide TS to make exactly 100 mL. Filter, discard the first 10 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 0.05 mol/L sodium hydroxide TS to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Furosemide RS, previously dried at 1059C for 4 hours, and dissolve in 0.05 mol/L sodium hydroxide TS to make exactly 50 mL. Pipet 2 mL of this solution, add 0.05 mol/L sodium hydroxide TS to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 271 nm as directed under the Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of furosemide (C12H11ClN2O5S) ＝ MS × AT/AS × 2 MS: Amount (mg) of Furosemide RS taken
Gallium (67Ga) Citrate Injection is a clear, col-orless or light red liquid. Gati‰oxacin Hydrate
Gati‰oxacin Ophthalmic Solution is an aqueous ophthalmic preparation. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) It contains not less than 95.
Gati‰oxacin Ophthalmic Solution is a clear, pale yellow liquid. Identiˆcation To a volume of Gati‰oxacin Ophthalmic So-lution, equivalent to 6 mg of gati‰oxacin (C19H22FN3O4), add diluted sodium hydroxide TS (1 in 10) to make 30 mL. To 1 mL of this solution add diluted sodium hydroxide TS (1 in 10) to make 20 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm, between 287 nm and 291 nm, and between 336 nm and 340 nm. Osmotic pressure ratio Being speciˆed separately when the drug is granted approval based on the Law. pH Being speciˆed separately when the drug is granted approval based on the Law.
Related substance—To a volume of Gati‰oxacin Ophthalmic Solution, equivalent to 6 mg of gati‰oxacin (C19H22FN3O4), add the diluting solution to make 30 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the diluting solution to make ex-actly 100 mL. Pipet 2 mL of this solution, add the diluting solution to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak area of the related sub-stance A, having the relative retention time of about 1.2 to gati‰oxacin, obtained from the sample solution is not larger than 2 times the peak area of gati‰oxacin from the standard solution, and the area of the peak other than gati‰oxacin and the peak mentioned above from the sample solution is not larger than the peak area of gati‰oxacin from the stand-ard solution. Furthermore, the total area of the peaks other than gati‰oxacin from the sample solution is not larger than 3 times the peak area of gati‰oxacin from the standard solu-tion. Diluting solution: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (4:1). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 325 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of diluted triethylamine (1 in 100) and acetonitrile (22:3), adjusted to pH 4.3 with phos-phoric acid. Mobile phase B: A mixture of diluted triethylamine (1 in 0 – 15 100 0 15 – 30 100 → 0 0 → 100 30 – 40 0 100 Flow rate: 0.9 mL per minute (the retention time of gati‰oxacin is about 16 minutes). Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the diluting solution to make exactly 10 mL. Conˆrm that the peak area of gati‰oxacin obtained with 40 mL of this solution is equivalent to 40 to 60z of that with 40 mL of the standard solution. System performance: Dissolve 20 mg of methyl 4-aminobenzoate in 100 mL of the diluting solution. To 5 mL of this solution and 1 mL of the sample solution add the diluting solution to make 100 mL. When the procedure is run with 40 mL of this solution under the above operating conditions, gati‰oxacin and methyl 4-aminobenzoate are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of gati‰oxacin is not more than 3.0z. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Pipet a volume of Gati‰oxacin Ophthalmic Solution, equivalent to about 6 mg of gati‰oxacin (C19H22FN3O4), and add the diluting solution to make ex-actly 30 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, add the diluting solution to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Gati‰oxacin RS (separately determine the water <2.48> in the same manner as Gati‰oxacin Hydrate), and dissolve in the diluting solution to make exactly 100 mL. Pipet 2 mL of this solution, add ex-actly 3 mL of the internal standard solution, add the diluting solution to make 20 mL, and use this solution as the stand-ard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of gati‰oxacin to that of the internal standard. Amount (mg) of gati‰oxacin (C19H22FN3O4) ＝ MS × QT/QS × 3/10 MS: Amount (mg) of Gati‰oxacin RS taken, calculated on the anhydrous basis Internal standard solution—A solution of methyl 4-aminobenzoate in the diluting solution (1 in 10,000). Diluting solution: A mixture of diluted phosphoric acid (1 ‌1058 in 1000) and acetonitrile (4:1). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and triethylamine (81:18:1), adjusted to pH 4.5 with phosphoric acid. Flow rate: Adjust so that the retention time of gati‰oxacin is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, gati‰oxacin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of gati‰oxacin to that of the internal standard is not more than 1.0z.
Gentamicin Sulfate Injection is an aqueous injec-tion. It contains not less than 90.
Gentamicin Sulfate Injection is a clear and col-orless liquid. Identiˆcation To a volume of Gentamicin Sulfate Injec-tion, equivalent to 40 mg (potency) of Gentamicin Sulfate, add water to make 10 mL, and use this solution as the sam-ple solution. Separately, dissolve an amount of Gentamicin Sulfate RS, equivalent to 20 mg (potency), in 5 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with the lower layer of a mixture of chloroform, ammonia solution (28) and methanol (2:1:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 10 minutes: three principal spots obtained from the sample solution are the same with the corresponding spots from the standard so-lution in color tone and the Rf value, respectively. Osmotic pressure ratio Being speciˆed separately when the drug is granted approval based on the Law. pH <2.54> 4.0 – 6.0 Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, agar media for base and seed layer, agar medium for transferring test organisms, and standard solutions—Proceed as directed in the Assay under Gentami-cin Sulfate. Sample solutions—Pipet a volume of Gentamicin Sulfate Injection, equivalent to about 40 mg (potency) of Gentamicin Sulfate, add 0.1 mol/L phosphate buŠer solu-tion (pH 8.0) to make exactly 200 mL. Pipet a suitable volume of this solution, add 0.1 mol/L phosphate buŠer so-lution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and the low concen-tration sample solution, respectively.
Gentamicin Sulfate Ointment contains not less than 90.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, agar media for base and seed layer, agar medium for transferring test organisms, and standard solutions—Proceed as directed in the Assay under Gentami-cin Sulfate. Sample solutions—Weigh accurately an amount of Gentamicin Sulfate Ointment, equivalent to about 1 mg (po-tency) of Gentamicin Sulfate, transfer to a separator, add 50 mL of diethyl ether, and shake until the solution becomes uniform. Add 25 mL of 0.1 mol/L phosphate buŠer solution (pH 8.0), shake, and collect the water layer. Repeat the same procedure with 25 mL of 0.1 mol/L phosphate buŠer solu-tion (pH 8.0), and combine the water layers. To this solution add 0.1 mol/L phosphate buŠer solution (pH 8.0) to make exactly 100 mL. Pipet a suitable volume of this solution, add 0.1 mol/L phosphate buŠer solution (pH 8.0) to make solu-tions so that each mL contains 4 mg (potency) and 1 mg (po-tency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Gentamicin Sulfate Ophthalmic Solution is an aque-ous ophthalmic preparation. It contains not less than 90.
Gentamicin Sulfate Ophthalmic Solution is a clear, colorless or pale yellow liquid.
To a volume of Gentamicin Sulfate Ophthal-mic Solution, equivalent to 10 mg (potency) of Gentamicin Sulfate, add water to make 5 mL, and use this solution as the sample solution. Separately, dissolve an amount of Gentami-cin Sulfate RS, equivalent to 10 mg (potency), in 5 mL of water, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with the lower layer of a mixture of chloroform, ammonia solution (28) and methanol (2:1:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly 0.2z ninhydrin-water saturated 1-butanol TS on the plate, and heat the plate at 1009C for 5 minutes: three principal spots obtained from the sample so-lution are the same with the corresponding spots from the standard solution in color tone and the Rf value, respec-tively. pH <2.54> 5.5 – 7.5 Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, agar media for seed and base layer, agar medium for transferring test organism, and standard solutions—Proceed as directed in the Assay under Gentami-cin Sulfate. Sample solutions—Pipet a volume of Gentamicin Sulfate Ophthalmic Solution, equivalent to about 12 mg (potency) of Gentamicin Sulfate, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make a solution so that each mL contains about 1 mg (potency). Pipet a suitable volume of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
Gentian and Sodium Bicarbonate Powder oc-curs as a light yellow-brown powder, and has a bitter taste.
(1) To 2 g of Gentian and Sodium Bicar-bonate Powder add 10 mL of water, stir, and filter: the fil-trate responds to the Qualitative Tests <1.09> (1) for bicar-bonate. (2) To 1.5 g of Gentian and Sodium Bicarbonate Powder add 10 mL of methanol, shake for 5 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 1 mg of gentiopicroside for thin-layer chromatography in 1 mL of ‌JP XVIII Crude Drugs and Related Drugs / Ginger 2009‌‌ methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the several spots obtained from the sample solution and a spot from the standard solution show the same color tone and the same Rf value.
Glimepiride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of glimepiride (C24H34N4O5S: 490.62). Method of preparation Prepare as directed under Tablets, with Glimepiride. Identification To a quantity of powdered Glimepiride Tablets, equivalent to 20 mg of Glimepiride, add 40 mL of acetonitrile, shake for 15 minutes, and centrifuge. Evaporate the supernatant liquid on a water bath under reduced pres- solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.3 to glimepiride, obtained from the sample solution is not larger than 2.6 times the peak area of glimepiride from the stand-ard solution, the area of the peak other than glimepiride and the peak mentioned above from the sample solution is not larger than 3/10 times the peak area of glimepiride from the standard solution, and the total area of the peaks other than glimepiride and the peak mentioned above from the sample solution is not larger than the peak area of glimepiride from the standard solution, and the total area of the peaks other than glimepiride from the sample solution is not larger than 3 times the peak area of glimepiride from the standard solu-tion. Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay. Flow rate: Adjust so that the retention time of glimepiride is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of glimepiride. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of glimepiride obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of glimepiride are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Glimepiride Tablets add V/10 mL of water, disintegrate, add V/2 mL of a mixture of acetonitrile for liq-uid chromatography and water (4:1), and shake. To this so-lution add exactly V/5 mL of the internal standard solution, add a mixture of acetonitrile for liquid chromatography and water (4:1) to make V mL so that each mL contains about JP XVIII Official Monographs / Glimepiride Tablets 1071 100 mg of glimepiride (C24H34N4O5S), and centrifuge.
To a quantity of powdered Glimepiride Tablets, equivalent to 20 mg of Glimepiride, add 40 mL of acetonitrile, shake for 15 minutes, and centrifuge. Evaporate the supernatant liquid on a water bath under reduced pres- solutions by the automatic integration method: the area of the peak, having the relative retention time of about 0.3 to glimepiride, obtained from the sample solution is not larger than 2.6 times the peak area of glimepiride from the stand-ard solution, the area of the peak other than glimepiride and the peak mentioned above from the sample solution is not larger than 3/10 times the peak area of glimepiride from the standard solution, and the total area of the peaks other than glimepiride and the peak mentioned above from the sample solution is not larger than the peak area of glimepiride from the standard solution, and the total area of the peaks other than glimepiride from the sample solution is not larger than 3 times the peak area of glimepiride from the standard solu-tion. Operating conditions— Detector, column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay. Flow rate: Adjust so that the retention time of glimepiride is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of glimepiride. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of glimepiride obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of glimepiride are not less than 6000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Glimepiride Tablets add V/10 mL of water, disintegrate, add V/2 mL of a mixture of acetonitrile for liq-uid chromatography and water (4:1), and shake. To this so-lution add exactly V/5 mL of the internal standard solution, add a mixture of acetonitrile for liquid chromatography and water (4:1) to make V mL so that each mL contains about JP XVIII Official Monographs / Glimepiride Tablets 1071 100 mg of glimepiride (C24H34N4O5S), and centrifuge. To 2.5 mL of the supernatant liquid add a mixture of acetonitrile for liquid chromatography and water (4:1) to make 5 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Glimepiride RS (separately determine the water <2.48> in the same manner as Glimepi-ride), and dissolve in a mixture of acetonitrile for liquid chromatography and water (4:1) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal standard solution, add a mixture of acetonitrile for liquid chromatography and water (4:1) to make 20 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay. Amount (mg) of glimepiride (C24H34N4O5S) ＝ MS × QT/QS × V/200 MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of acetonitrile for liquid chroma-tography and water (4:1) (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citric acid buffer solu-tion (pH 7.5) as the dissolution medium, the dissolution rate in 15 minutes of 0.5-mg and 1-mg tablets is not less than 75z, and that in 30 minutes of 3-mg tablet is not less than 70z. Start the test with 1 tablet of Glimepiride Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 0.56 mg of glimepiride (C24H34N4O5S), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Glimepiride RS (separately determine the water <2.48> in the same manner as Glimepiride), and dissolve in acetonitrile for liquid chroma-tography to make exactly 100 mL. Pipet 2 mL of this solu-tion, add 8 mL of acetonitrile for liquid chromatography, and add the dissolution medium to make exactly 200 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glimepiride in each solution. Dissolution rate (z) with respect to the labeled amount of glimepiride (C24H34N4O5S) ＝ MS × AT/AS × V?/V × 1/C × 9/4 MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of glimepiride (C24H34N4O5S) in 1 tablet Operating conditions— Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above opera-tions conditions, the relative standard deviation of the peak area of glimepiride is not more than 1.5z.
Weigh accurately the mass of not less than 20 Glimepiride Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 3 mg of glimepiride (C24H34N4O5S), add 3 mL of water, and shake with 30 mL of a mixture of acetonitrile for liquid chromatography and water (4:1). Add exactly 6 mL of the internal standard solu-tion, and add a mixture of acetonitrile for liquid chroma-tography and water (4:1) to make 50 mL, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Glimepiride RS, (sepa-rately, determine the water <2.48> in the same manner as Glimepiride), dissolve in a mixture of acetonitrile for liquid chromatography and water (4:1) to make exactly 100 mL. Pipet 15 mL of this solution, add exactly 6 mL of the inter-nal standard solution, add a mixture of acetonitrile for liq-uid chromatography and water (4:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of glimepiride to that of the internal standard. Amount (mg) of glimepiride (C24H34N4O5S) ＝ MS × QT/QS × 3/20 MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of acetonitrile for liquid chroma-tography and water (4:1) (1 in 1000). Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 228 nm). Column: A stainless steel column 4 mm in inside diameter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.5 g of sodium dihydrogen phos-phate dihydrate in 500 mL of water, add 500 mL of aceto-nitrile for liquid chromatography, and adjust to pH 3.5 with diluted phosphoric acid (1 in 5). Flow rate: Adjust so that the retention time of glimepiride is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and glimepiride are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of glimepiride to that of the internal standard is not more than 1.0z. ‌1072 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 35 100 → 53 0 → 47 35 – 45 53 → 0 47 → 100
Glucose Injection is an aqueous injection. It contains not less than 95.
Glucose Injection is a clear, colorless liquid. It has a sweet taste. It occurs as a colorless to pale yellow, clear liquid when its labeled concentration exceeds 40z.
Measure a volume of Glucose Injection, equivalent to 0.1 g of glucose (C6H12O6), and, if necessary, add water or evaporate on a water bath to make 2 mL. Add ‌JP XVIII Official Monographs / L-Glutamic Acid 1077 2 to 3 drops of the solution to 5 mL of boiling Fehling's TS: a red precipitate is produced. pH <2.54> 3.5 – 6.5 In the case where the labeled concen-tration of the injection exceeds 5z, dilute to 5z with water before the test.
5-Hydroxymethylfurfural and related substances—Pipet a volume of Glucose Injection, equivalent to 2.5 g of glucose (C6H12O6), and add water to make exactly 100 mL. Determine the absorbance of this solution at 284 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it is not more than 0.80. Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure accurately a volume of Glucose Injection, equivalent to about 4 g of glucose (C6H12O6), and add 0.2 mL of ammonia TS and water to make exactly 100 mL. Shake the solution well, allow to stand for 30 minutes, and determine the optical rotation, aD, at 20 ± 19C in a 100-mm cell as directed under Optical Rotation Determination <2.49>. Amount (mg) of glucose (C6H12O6) ＝ aD × 1895.4
Glycerin and Potash Solution is a clear, color-less liquid, having an aromatic odor. The pH of a solution of Glycerin and Potash Solution (1 in 5) is about 12. Specific gravity d 20: about 1.02
(1) A solution of Glycerin and Potash So-lution (1 in 2) is alkaline (potassium hydroxide). Place 10 mL of a solution of Glycerin and Potash So-lution (1 in 10) in a glass-stoppered test tube, add 2 mL of sodium hydroxide TS and 1 mL of copper (II) sulfate TS, and shake: a blue color is produced (glycerin). Glycerin and Potash Solution responds to Qualitative Tests <1.09> for potassium salt.
Powder Haloperidol Fine Granules. To a portion of the powder, equivalent to 6 mg of Haloperidol, add 70 mL of 2-propanol, and heat to boiling on a water bath while shaking. After cooling, add 2-propanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add 2 mL of 0.1 mol/L hydrochloric acid TS and 2-propanol to make 20 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 219 nm and 223 nm and between 243 nm and 247 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Haloperidol Fine Granules is not less than 70z. Start the test with an accurately weighed amount of Haloperidol Fine Granules, equivalent to about 3 mg of haloperidol (C21H23ClFNO2), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 17 mg of haloperidol for assay, previously dried in vacuum at 609C for 3 hours using phosphorus (V) oxide as a desiccant, and dissolve in methanol to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of haloperidol in each solution. ‌1092 Dissolution rate (z) with respect to the labeled amount of haloperidol (C21H23ClFNO2) ＝ MS/MT × AT/AS × 1/C × 18 MS: Amount (mg) of haloperidol for assay taken MT: Amount (g) of Haloperidol Fine Granules taken C: Labeled amount (mg) of haloperidol (C21H23ClFNO2) in 1 g Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as detected in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 245 nm). System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of haloperidol are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of haloperidol is not more than 2.0z.
Powder Haloperidol Fine Granules. Weigh accu-rately a portion of the powder, equivalent to about 10 mg of haloperidol (C21H23ClFNO2), add 10 mL of water, disperse the particle by sonicating, add exactly 20 mL of the internal standard solution, sonicate, extract for 30 minutes with oc-casional shaking, and add the mobile phase to make 100 mL. Centrifuge after shaking for more 30 minutes, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of haloperidol for assay, previously dried in vacuum at 609C for 3 hours on phosphorus (V) oxide, and dissolve in methanol to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 20 mL of the inter-nal standard solution and the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of haloperidol to that of the in-ternal standard. Amount (mg) of haloperidol (C21H23ClFNO2) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of haloperidol for assay taken Internal standard solution—A solution of diphenyl in meth-anol (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.95 g of trisodium citrate dihy-drate in 900 mL of water, adjust to pH 3.5 with dilute hy-drochloric acid, and add water to make 1000 mL. To 250 mL of this solution add 750 mL of methanol and 1.0 g of sodium lauryl sulfate, and mix to dissolve. Flow rate: Adjust so that the retention time of haloperidol is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, haloperidol and diphenyl are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of haloperidol to that of the internal standard is not more than 1.0z.
Haloperidol Injection is an aqueous injection. It contains not less than 95.
Haloperidol Injection occurs as a colorless to pale yellow, clear liquid.
To a volume of Haloperidol Injection, equivalent to 5 mg of Haloperidol, add 2-propanol to make 100 mL. To 5 mL of this solution add 2 mL of 0.1 mol/L hy-drochloric acid TS and 2-propanol to make 20 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 219 nm and 223 nm and between 243 nm and 247 nm. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 60 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Haloperidol Injection, equivalent to about 10 mg of haloperidol (C21H23ClFNO2), add the mo-bile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of haloperidol for assay, previously dried in vacuum at 609C using phosphorus (V) oxide as a desiccant for 3 hours, dissolve in methanol to make exactly 25 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of haloperidol in each solution. Amount (mg) of haloperidol (C21H23ClFNO2) ＝ MS × AT/AS × 2/5 ‌JP XVIII Official Monographs / Haloperidol Tablets 1093 MS: Amount (mg) of haloperidol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.95 g of trisodium citrate dihy-drate in 900 mL of water, adjust to pH 3.5 with dilute hydro-chloric acid, and add water to make 1000 mL. To 250 mL of this solution add 750 mL of methanol, and add 1.0 g of so-dium lauryl sulfate to dissolve. Flow rate: Adjust so that the retention time of haloperidol is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of haloperidol are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of haloperidol is not more than 1.0z.
Containers—Hermetic containers. Colored containers may be used.
To powdered Haloperidol Tablets, equiva-lent to 6 mg of Haloperidol, add 70 mL of 2-propanol, and heat on a water bath until to boiling while shaking. After cooling, add 2-propanol to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add 2 mL of 0.1 mol/L hy-drochloric acid TS and 2-propanol to make 20 mL. Deter-mine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 219 nm and 223 nm and between 243 nm and 247 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Haloperidol Tablets add 5 mL of the mobile phase, disperse the particle by sonicating, add 30 mL of the mobile phase, sonicate, and extract for 30 minutes with oc-casional shaking. Shake for more 30 minutes, and add the mobile phase to make exactly 50 mL. Centrifuge the solu-tion, pipet V mL of the supernatant liquid, equivalent to about 0.3 mg of haloperidol (C21H23ClFNO2), add exactly 2 mL of the internal standard solution and the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of haloperidol for assay, previously dried in vacuum at 609C for 3 hours on phosphorus (V) oxide, and dissolve in the mobile phase to make exactly 100 mL. Pipet 15 mL of this solution, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 2 mL of the internal standard solution and the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according the following con-ditions, and calculate the ratios, QT and QS, of the peak area of haloperidol to that of the internal standard. Amount (mg) of haloperidol (C21H23ClFNO2) ＝ MS × QT/QS × 1/V × 3/4 MS: Amount (mg) of haloperidol for assay taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 6700). Operating conditions— Proceed as detected in the operating condition in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, haloperidol and diphenyl are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of haloperidol to that of the internal standard is not more than 1.0z. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately, and powder not less than 20 Haloperidol Tablets. Weigh accurately a portion of the powder, equivalent to about 10 mg of haloperidol (C21H23ClFNO2), add 10 mL of water, disperse the particle by sonicating, add exactly 20 mL of the internal standard so-lution, sonicate, extract for 30 minutes with occasional shak-ing, and add the mobile phase to make 100 mL. Centrifuge after shaking for more 30 minutes, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of haloperidol for assay, previously dried in vacuum at 609C for 3 hours on phosphorus (V) oxide, and dissolve in methanol to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 20 mL of the internal standard so-lution and the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of haloperidol to that of the internal standard. Amount (mg) of haloperidol (C21H23ClFNO2) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of haloperidol for assay taken Internal standard solution—A solution of diphenyl in meth-anol (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. ‌1094 Mobile phase: Dissolve 2.95 g of trisodium citrate dihy-drate in 900 mL of water, adjust to pH 3.5 with dilute hy-drochloric acid, and add water to make 1000 mL. To 250 mL of this solution add 750 mL of methanol and 1.0 g of sodium lauryl sulfate, and mix to dissolve. Flow rate: Adjust so that the retention time of haloperidol is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, haloperidol and diphenyl are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of haloperidol to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage—Light-resistant for the tablets without coating.
Heparin Sodium Injection is an aqueous injection. It contains not less than 90z and not more than 110z of the labeled heparin Units.
Heparin Sodium Injection is a clear, colorless to light yellow liquid. pH <2.54> 5.5 – 8.0
Barium—Measure exactly a volume of Heparin So-dium Injection, equivalent to 3000 Units of Heparin So-dium, add water to make 3.0 mL and use this solution as the sample solution. To 1.0 mL of the sample solution add 3 drops of dilute sulfuric acid, and allow to stand for 10 minutes: no turbidity is produced. Bacterial endotoxins <4.01> Less than 0.0030 EU/unit. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Proceed as directed in the Assay under Heparin Sodium, replacing (vii) Heparin sample solutions and (ix) Calculations with the following. (vii) Heparin sample solutions: Take exactly an ap-propriate amount of Heparin Sodium Injection, dilute ex-actly with buffer solution so that each mL contains 0.1 Heparin Units, and use this solution as the sample solution. Make heparin sample solutions T1, T2, T3 and T4 respectively by adding the sample solution to buffer solution as directed in the following table. Heparin sample solution Buffer solution (mL) Sample solution (mL) No. Heparin concentration (Unit/mL) T1 0.005 T2 0.010 T3 0.015 T4 0.020 (ix) Calculations: When the regression expression, y ＝ Ic ＋ AXs ＋ BXt, is obtained using y as log of the absorbance values, Xs as the concentration of the heparin standard solutions and Xt as the concentration of the heparin sample solutions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Injection by the following formula. Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Injection ＝ 0.1 × R × V/a V: Total volume (mL) of the sample solution prepared as containing 0.1 Heparin Units (anti-factor IIa activity) per mL a: Amount (mL) of Heparin Sodium Injection taken for the sample solution However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?Xs ＋ B?Xt ＋ D, where D is a constant term showing the difference between the inter-cepts assumed from the measurement of the blank and the two lines, is not in the range of between －0.2 and 0.2, ana-lyze by excluding the measurements of the blank. The criteria for the test suitability are followed as directed in the Assay under Heparin Sodium. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the potency ratio becomes about 1 using the obtained potency as reference.
Heparin Sodium Lock Solution is a preparation used to prevent blood coagulation in intravenous in-dwelling routes. It contains not less than 90z and not more than 110z of the labeled Heparin Units.
Heparin Sodium Lock Solution is a clear, col-orless to light yellow liquid. Osmotic pressure ratio: 0.9 – 1.1 ‌JP XVIII Official Monographs / Heparin Sodium Solution for Dialysis 1105 pH <2.54> 5.5 – 8.0 Bacterial endotoxins <4.01> Less than 0.0030 EU/unit. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Proceed as directed in the Assay under Heparin So-dium, replacing (vii) Heparin sample solutions and (ix) Cal-culations with the following. (vii) Heparin sample solutions: Pipet a suitable volume of Heparin Sodium Lock Solution, dilute exactly with the buffer solution so that each mL contains 0.1 Heparin Units, and use this solution as the sample solution. Prepare heparin sample solutions T1, T2, T3 and T4 respectively by adding the sample solution to the buffer solution as directed in the fol-lowing table. Heparin sample solution Buffer solution (mL) Sample solution (mL) No. Heparin concentration (Unit/mL) T1 0.005 T2 0.010 T3 0.015 T4 0.020 (ix) Calculations: When the regression expression, y ＝ Ic s t ＋ AX ＋ BX , is obtained using y as log of the absorbance values, xs as the concentration of the heparin standard solu-tions and xt as the concentration of the heparin sample solu-tions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution B: Slope of regression expression of the heparin sample solution Calculate Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Lock Solution by the following formula. Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Lock Solution ＝ 0.1 × R × V/a V: Total volume (mL) of the sample solution prepared as containing 0.1 Heparin Units (anti-factor IIa activity) per mL a: Amount (mL) of Heparin Sodium Lock Solution taken s t However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?X ＋ B?X ＋ D, where D is a constant term showing the difference between the inter-cepts assumed from the measurement of the blank and the two lines, is not in the range of between －0.2 and 0.2, ana-lyze by excluding the measurements of the blank. The criteria for the test suitability are followed as directed in the Assay under Heparin Sodium. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the potency ratio becomes about 1 using the obtained potency as reference.
Heparin Sodium Solution for Dialysis is a prepara-tion used to prevent coagulation of perfused blood during hemodialysis. It contains not less than 90z and not more than 110z of the labeled Heparin Unit.
Heparin Sodium Solution for Dialysis is a clear, colorless to light yellow liquid. Osmotic pressure ratio: 0.9 – 1.1 pH <2.54> 5.5 – 8.0 Bacterial endotoxins <4.01> Less than 0.0030 EU/unit. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Proceed as directed in the Assay under Heparin So-dium, replacing (vii) Heparin sample solutions and (ix) Cal-culations with the following. (vii) Heparin sample solutions: Pipet a suitable volume of Heparin Sodium Solution for Dialysis, dilute exactly with the buffer solution so that each mL contains 0.1 Heparin Units, and use this solution as the sample solution. Prepare heparin sample solutions T1, T2, T3 and T4 respectively by adding the sample solution to the buffer solution as directed in the following table. Heparin sample solution Buffer solution (mL) Sample solution (mL) No. Heparin concentration (Unit/mL) T1 0.005 T2 0.010 T3 0.015 T4 0.020 (ix) Calculations: When the regression expression, y ＝ Ic s t ＋ AX ＋ BX , is obtained using y as log of the absorbance values, xs as the concentration of the heparin standard solu-tions and xt as the concentration of the heparin sample solu-tions, the potency ratio R is B/A. Ic: Common intercept A: Slope of regression expression of the heparin standard solution ‌1106‌ B: Slope of regression expression of the heparin sample solution Calculate Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Solution for Dialysis by the following formula. Heparin Units (anti-factor IIa activity) in 1 mL of Heparin Sodium Solution for Dialysis ＝ 0.1 × R × V/a V: Total volume (mL) of the sample solution prepared as containing 0.1 Heparin Units (anti-factor IIa activity) per mL a: Amount (mL) of Heparin Sodium Solution for Dialysis taken s t However, when a 90z confidence interval of D of the regression expression y ＝ I?c ＋ A?X ＋ B?X ＋ D, where D is a constant term showing the difference between the inter-cepts assumed from the measurement of the blank and the two lines, is not in the range of between －0.2 and 0.2, ana-lyze by excluding the measurements of the blank. The criteria for the test suitability are followed as directed in the Assay under Heparin Sodium. When these criteria are not satisfied, repeat the test after changing the dilution rate so that the potency ratio becomes about 1 using the obtained potency as reference.
Hydralazine Hydrochloride for Injection is a prepa-ration for injection which is dissolved before use. It contains not less than 99.
Hydralazine Hydrochloride for Injection oc-curs as a white to pale yellow powder or mass. It is odorless, and has a bitter taste.
Determine the absorption spectrum of a so-lution of Hydralazine Hydrochloride for Injection (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 238 nm and 242 nm, between 258 nm and 262 nm, between 301 nm and 305 nm, and between 313 nm and 317 nm. pH <2.54> Dissolve 1.0 g of Hydralazine Hydrochloride for Injection in 50 mL of water: the pH of this solution is be-tween 3.5 and 4.5. Bacterial endotoxins <4.01> Less than 5.0 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. (T: 106.0z) Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem- ‌1110‌ brane filtration method: it meets the requirement.
Weigh accurately the contents of not less than 10 samples of Hydralazine Hydrochloride for Injection. Weigh accurately about 0.15 g of the contents, transfer it to a glass-stoppered flask, dissolve in 25 mL of water, add 25 mL of hydrochloric acid, cool to room temperature, and proceed as directed in the Assay under Hydralazine Hydrochloride. Each mL of 0.05 mol/L potassium iodate VS ＝ 9.832 mg of C8H8N4.HCl
Hydralazine Hydrochloride Powder contains not less than 95.
Weigh a portion of Hydralazine Hydrochlo-ride Powder, equivalent to 25 mg of Hydralazine Hydrochlo-ride, add 100 mL of water, shake well, and filter, if neces-sary. Add water to 2 mL of this solution to make 50 mL and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm, between 258 nm and 262 nm, between 301 nm and 305 nm, and between 313 nm and 317 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Hydralazine Hydrochloride Powder is not less than 85z. Start the test with an accurately weighed amount of Hydralazine Hydrochloride Powder, equivalent to about 50 mg of hydralazine hydrochloride (C8H8N4.HCl), withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 5 mL of the first filtrate, pipet 4 mL of the subsequent fil-trate, add water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of hydralazine hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make ex-actly 50 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, at 260 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of hydralazine hydrochloride (C8H8N4.HCl) ＝ MS/MT × AT/AS × 1/C × 180 MS: Amount (mg) of hydralazine hydrochloride for assay taken MT: Amount (g) of the Hydralazine Hydrochloride Pow-der taken C: Labeled amount (mg) of hydralazine hydrochloride (C8H8N4.HCl) in 1 g
Weigh accurately a portion of Hydralazine Hydro- chloride Powder, equivalent to about 0.15 g of hydralazine hydrochloride (C8H8N4.HCl), transfer it to a glass-stoppered flask, add 25 mL of water, shake well, add 25 mL of hydro-chloric acid, cool to room temperature, and proceed as di-rected in the Assay under Hydralazine Hydrochloride. Each mL of 0.05 mol/L potassium iodate VS ＝ 9.832 mg of C8H8N4.HCl
Hydralazine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of hydralazine hydrochloride (C8H8N4.HCl: 196.64). Method of preparation Prepare as directed under Tablets, with Hydralazine Hydrochloride. Identification Weigh a quantity of powdered Hydralazine Hydrochloride Tablets, equivalent to 25 mg of Hydralazine Hydrochloride, add 100 mL of water, mix well, and filter if necessary. To 2 mL of this solution add water to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm, be- tween 258 nm and 262 nm, between 301 nm and 305 nm and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Hydralazine Hydrochloride Tablets add 25 mL of 0.1 mol/L hydrochloric acid TS, disperse the tablet into a small particles by sonicating, then shake well, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and centrifuge. Pipet V mL of the supernatant liquid, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 10 mg of hydralazine hydrochloride (C8H8N4.
Weigh a quantity of powdered Hydralazine Hydrochloride Tablets, equivalent to 25 mg of Hydralazine Hydrochloride, add 100 mL of water, mix well, and filter if necessary. To 2 mL of this solution add water to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 238 nm and 242 nm, be- tween 258 nm and 262 nm, between 301 nm and 305 nm and between 313 nm and 317 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Hydralazine Hydrochloride Tablets add 25 mL of 0.1 mol/L hydrochloric acid TS, disperse the tablet into a small particles by sonicating, then shake well, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL, and centrifuge. Pipet V mL of the supernatant liquid, add 0.1 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 10 mg of hydralazine hydrochloride (C8H8N4.HCl), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of hydralazine hy-drochloride for assay, previously dried at 1059C for 3 hours, dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 2 mL of this solution, add 0.1 mol/L hydro-chloric acid TS to make exactly 100 mL, and use this solu-tion as the standard solution. Determine the absorbances at 260 nm, AT1 and AS1, and at 350 nm, AT2 and AS2, of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>. Amount (mg) of hydralazine hydrochloride (C8H8N4.HCl) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/50 MS: Amount (mg) of hydralazine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Hydralazine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Hydralazine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. ‌JP XVIII Official Monographs / Dilute Hydrochloric Acid 1111‌ Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of hydralazine hydro-chloride (C8H8N4.HCl), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of hydralazine hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 260 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of hydralazine hydrochloride (C8H8N4.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of hydralazine hydrochloride for assay taken C: Labeled amount (mg) of hydralazine hydrochloride (C8H8N4.HCl) in 1 tablet
Weigh accurately not less than 20 Hydralazine Hy-drochloride Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 0.15 g of hydralazine hydrochloride (C8H8N4.HCl), transfer it to a glass-stoppered flask, and proceed as directed in the Assay under Hydrala-zine Hydrochloride. Each mL of 0.05 mol/L potassium iodate VS ＝ 9.832 mg of C8H8N4.HCl
Hydrocortisone and Diphenhydramine Ointment
ヒドロコルチゾン・ジフェンヒドラミン軟膏
Hydrocortisone and Diphenhydramine Oint-ment is white to pale yellow in color.
(1) To 1 g of Hydrocortisone and Diphen-hydramine Ointment add 10 mL of ethanol (95), heat on a water bath for 5 minutes with occasional shaking, cool, and filter. Take 5 mL of the filtrate, distill off the ethanol, and to the residue add 2 mL of sulfuric acid: the solution shows a yellow-green fluorescence immediately and the color of the solution gradually changes through yellow to yellow-brown. Add carefully 10 mL of water to this solution: the color changes to yellow with green fluorescence, and a light yel-low, flocculent precipitate is formed (hydrocortisone ace-tate). To 1 mL of the filtrate obtained in (1) add 5 mL of potassium hydrogen phthalate buffer solution (pH 4.6) and 2 mL of bromophenol blue TS, and add further 5 mL of chloroform. Shake well, and allow to stand: a yellow color develops in the chloroform layer (diphenhydramine). To 0.2 g of Hydrocortisone and Diphenhydramine Ointment add 0.5 mL of methanol, warm, and shake. After cooling, separate the methanol layer, and use this layer as the sample solution. Dissolve 10 mg each of hydrocortisone acetate and diphenhydramine in 10 mL each of methanol, and use these solutions as standard solutions (1) and (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with ‰uorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of ethyl acetate and diethyl ether (4:1) to a distance of about 5 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): two spots obtained from the sample solution show the same Rf value as the corresponding spots from standard solutions (1) and (2).
Hydrophilic Cream is white in color. It has a slight, characteristic odor.
イブプロフェンピコノールクリーム
Ibuprofen Piconol Cream contains not less than 95.
To an amount of Ibuprofen Piconol Cream, equivalent to 50 mg of Ibuprofen Piconol, add 10 mL of methanol, warm in a water bath, mix well, filter after cool-ing, and use the filtrate as the sample solution. Separately, dissolve 50 mg of ibuprofen piconol in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (15:5:1) to a distance of about 13 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. pH Being specified separately when the drug is granted ap-proval based on the Law.
Weigh accurately an amount of Ibuprofen Piconol Cream, equivalent to about 15 mg of ibuprofen piconol (C19H23NO2), add 10 mL of tetrahydrofuran for liquid chro-matography, shake vigorously, and add exactly 10 mL of the internal standard solution. Then, add methanol to make 30 mL, shake vigorously, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 0.15 g of ibuprofen piconol for assay (separately determine the water <2.48> in the same manner as Ibuprofen Piconol), and dissolve in tetrahydrofuran for liquid chromatography to make exactly 100 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution, add methanol to make 30 mL, and use this solution as the standard solu-tion. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of ibuprofen piconol to that of the internal standard. Amount (mg) of ibuprofen piconol (C19H23NO2) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of ibuprofen piconol for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of triphenylmethane in methanol (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- ‌JP XVIII Official Monographs / Ichthammol 1135‌ ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ibuprofen piconol to that of the internal standard. 409C. Mobile phase: A mixture of methanol and acetic acid-sodi-um acetate buffer solution (pH 4.0) (3:1). Amount (mg) of ibuprofen piconol (C ＝ MS × QT/QS × 1/10 19H23NO2) Flow rate: Adjust so that the retention time of ibuprofen piconol is about 6.5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, ibuprofen piconol and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ibuprofen piconol to that of the internal stand-ard is not more than 1.0z.
Ibuprofen Piconol Ointment contains not less than 95.
To an amount of Ibuprofen Piconol Oint-ment, equivalent to 50 mg of Ibuprofen Piconol, add 10 mL of methanol, warm at 609C in a water bath, mix well, and filter after cooling. Use the filtrate as the sample solution. Separately, dissolve 50 mg of ibuprofen piconol in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of hexane, ethyl acetate and acetic acid (100) (15:5:1) to a distance of about 13 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value.
Weigh accurately an amount of Ibuprofen Piconol Ointment, equivalent to about 15 mg of ibuprofen piconol (C19H23NO2), add 10 mL of tetrahydrofuran for liquid chro-matography, shake vigorously, and add exactly 10 mL of the internal standard solution. Then, add methanol to make 30 mL, shake vigorously, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 0.15 g of ibuprofen piconol for assay (separately determine the water <2.48> in the same manner as Ibuprofen Piconol), and dissolve in tetrahydrofuran for liquid chromatography to make exactly 100 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution, add methanol to make exactly 30 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma- peak area of ibuprofen piconol to that of the internal stand-ard is not more than 1.0z.
Idarubicin Hydrochloride for Injection is a prepara-tion for injection, which is dissolved before use. It contains not less than 90.
Idarubicin Hydrochloride for Injection occurs as yellow-red masses.
(1) Dissolve an amount of Idarubicin Hy-drochloride for Injection, equivalent to 2 mg (potency) of Idarubicin Hydrochloride, in 5 mL of sodium hydroxide TS: the solution shows a blue-purple color. (2) Dissolve an amount of Idarubicin Hydrochloride for Injection, equivalent to 1 mg (potency) of Idarubicin Hydro-chloride, in 1 mL of water, and add methanol to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 250 nm and 254 nm, between 285 nm and 289 nm, between 480 nm and 484 nm, and between 510 nm and 520 nm. pH <2.54> The pH of a solution prepared by dissolving an amount of Idarubicin Hydrochloride for Injection, equiva-lent to 5 mg (potency) of Idarubicin Hydrochloride, in 5 mL of water is between 5.0 and 7.0. ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Idarubicin Hydrochloride RS taken
Clarity and color of solution—Dissolve an amount of Idarubicin Hydrochloride for Injection, equivalent to 5 mg (potency) of Idarubicin Hydrochloride, in 5 mL of water: the solution is clear and yellow-red. ‌1138 Water <2.48> Weigh accurately the mass of 1 Idarubicin Hydrochloride for Injection, add 5 mL of methanol for water determination using a syringe, dissolve with thorough shaking, and perform the test with 4 mL of this solution as directed in the Volumetric titration (direct titration). Use 4 mL of methanol for water determination as the blank. De-termine the mass of the content from the difference between the mass of 1 Idarubicin Hydrochloride for Injection ob-tained above and the mass of its bottle and rubber stopper, which are weighed accurately after washing with water then with ethanol (95), drying at 1059C for 1 hour and allowing to cool to room temperature in a desiccator (not more than 4.0z). Bacterial endotoxins <4.01> Less than 8.9 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 Idarubicin Hydrochloride for Injection add the mo-bile phase prepared without addition of sodium lauryl sul-fate to make exactly V mL so that each mL contains 0.2 mg (potency) of idarubicin hydrochloride (C26H27NO9.HCl), and use this solution as the sample solution. Separately, weigh accurately an amount of Idarubicin Hydrochloride RS, equivalent to about 10 mg (potency), dissolve in the mobile phase without sodium lauryl sulfate to make exactly 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Idarubicin Hydrochloride. Amount [mg (potency)] of idarubicin hydrochloride (C26H27NO9.HCl) ＝ MS × AT/AS × V/50 MS: Amount [mg (potency)] of Idarubicin Hydrochloride RS taken Foreign insoluble matter <6.06> Perform the test according to the Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately an amount of Idarubicin Hydro-chloride and Idarubicin Hydrochloride RS, equivalent to about 10 mg (potency), dissolve each in the mobile phase prepared without addition of sodium lauryl sulfate to make exactly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of idarubicin in each solution. Amount [ mg (potency)] of idarubicin hydrochloride (C26H27NO9.HCl) Method of preparation Prepare as directed under Injec-tions, with Idarubicin Hydrochloride. Description Idarubicin Hydrochloride for Injection occurs as yellow-red masses. Identification (1) Dissolve an amount of Idarubicin Hy-drochloride for Injection, equivalent to 2 mg (potency) of Idarubicin Hydrochloride, in 5 mL of sodium hydroxide TS: the solution shows a blue-purple color. (2) Dissolve an amount of Idarubicin Hydrochloride for Injection, equivalent to 1 mg (potency) of Idarubicin Hydro-chloride, in 1 mL of water, and add methanol to make 100 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 250 nm and 254 nm, between 285 nm and 289 nm, between 480 nm and 484 nm, and between 510 nm and 520 nm. pH <2.54> The pH of a solution prepared by dissolving an amount of Idarubicin Hydrochloride for Injection, equiva-lent to 5 mg (potency) of Idarubicin Hydrochloride, in 5 mL of water is between 5.0 and 7.0. ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Idarubicin Hydrochloride RS taken Purity Clarity and color of solution—Dissolve an amount of Idarubicin Hydrochloride for Injection, equivalent to 5 mg (potency) of Idarubicin Hydrochloride, in 5 mL of water: the solution is clear and yellow-red. ‌1138 Water <2.48> Weigh accurately the mass of 1 Idarubicin Hydrochloride for Injection, add 5 mL of methanol for water determination using a syringe, dissolve with thorough shaking, and perform the test with 4 mL of this solution as directed in the Volumetric titration (direct titration). Use 4 mL of methanol for water determination as the blank. De-termine the mass of the content from the difference between the mass of 1 Idarubicin Hydrochloride for Injection ob-tained above and the mass of its bottle and rubber stopper, which are weighed accurately after washing with water then with ethanol (95), drying at 1059C for 1 hour and allowing to cool to room temperature in a desiccator (not more than 4.0z). Bacterial endotoxins <4.01> Less than 8.9 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 Idarubicin Hydrochloride for Injection add the mo-bile phase prepared without addition of sodium lauryl sul-fate to make exactly V mL so that each mL contains 0.2 mg (potency) of idarubicin hydrochloride (C26H27NO9.HCl), and use this solution as the sample solution. Separately, weigh accurately an amount of Idarubicin Hydrochloride RS, equivalent to about 10 mg (potency), dissolve in the mobile phase without sodium lauryl sulfate to make exactly 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Idarubicin Hydrochloride. Amount [mg (potency)] of idarubicin hydrochloride (C26H27NO9.HCl) ＝ MS × AT/AS × V/50 MS: Amount [mg (potency)] of Idarubicin Hydrochloride RS taken Foreign insoluble matter <6.06> Perform the test according to the Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Assay Weigh accurately the mass of the contents of not less than 10 Idarubicin Hydrochloride for Injection. Weigh accu-rately an amount of the content, equivalent to about 5 mg (potency), dissolve in the mobile phase prepared without ad-dition of sodium lauryl sulfate to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Idarubicin Hydrochloride RS, equivalent to about 10 mg (potency), dissolve in the mobile phase without sodium lauryl sulfate to make exactly 50 mL, and use this solution as the standard solution. Proceed as di-rected in the Assay under Idarubicin Hydrochloride. Amount [mg (potency)] of idarubicin hydrochloride (C26H27NO9.HCl) ＝ MS × AT/AS × 1/2 MS: Amount [mg (potency)] of Idarubicin Hydrochloride RS taken
Idoxuridine Ophthalmic Solution contains not less than 90.
Idoxuridine Ophthalmic Solution is a clear, colorless liquid.
(1) To a volume of Idoxuridine Opthalmic Solution, equivalent to 5 mg of Idoxuridine, add 5 mL of diphenylamine-acetic acid TS, and heat for 20 minutes: a light blue color develops. Place a volume of Idoxuridine Ophthalmic Solution, equivalent to 5 mg of Idoxuridine, in a porcelain crucible, add 0.1 g of anhydrous sodium carbonate, heat slowly, evaporate to dryness and ignite until the residue is inciner-ated. Dissolve the residue in 5 mL of water, acidify with hy-drochloric acid, and add 2 to 3 drops of sodium nitrite TS: a yellow-brown color develops. Then add 2 to 3 drops of starch TS: a deep blue color develops. To a volume of Idoxuridine Ophthalmic Solution, equivalent to 2 mg of Idoxuridine, add 0.01 mol/L sodium hydroxide TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 277 nm and 281 nm. pH <2.54> 4.5 – 7.0
5-Iodouracil and 2?-deoxyuridine—To a volume of Idoxuridine Ophthalmic Solution, equivalent to 4.0 mg of Idoxuridine, add water to make exactly 5 mL, and use this solution as the sample solution. Separately, dissolve 12.0 mg of 5-iodouracil for liquid chromatography and 4.0 mg of 2?-deoxyuridine for liquid chromatography in water to make exactly 200 mL. Measure exactly 5 mL of this solution, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas of 5-io-douracil and 2?-deoxyuridine: the peak areas of 5-iodouracil and 2?-deoxyuridine of the sample solution are not larger than the peak areas of 5-iodouracil and 2?-deoxyuridine of the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (24:1). Flow rate: Adjust so that the retention time of 2?-deox- yuridine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating condi-tions, 2?-deoxyuridine and 5-iodouracil are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of 2?-deoxyuridine is not more than 1.0z. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Idoxuridine Oph-thalmic Solution, equivalent to 3 mg of idoxuridine (C9H11IN2O5), add exactly 2 mL of the internal standard so-lution, then add water to make 10 mL, and use this solution as the sample solution. Separately weigh accurately about 10 mg of Idoxuridine RS, previously dried at 609C for 3 hours, dissolve in water to make exactly 10 mL. Measure exactly 3 mL of this solution, add exactly 2 mL of the internal stand-ard solution, then add water to make 10 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of idoxuridine to that of the internal standard, respectively. Amount (mg) of idoxuridine (C9H11IN2O5) ＝ MS × QT/QS × 3/10 MS: Amount (mg) of Idoxuridine RS taken Internal standard solution—A solution of sulfathiazole in the mobile phase (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌1140‌ Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (87:13). Flow rate: Adjust so that the retention time of idoxuridine is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, idoxuridine and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of idoxuridine to that of the internal stand-ard is not more than 1.0z.
Containers—Tight containers. Storage—Light-resistant, in a cold place, and avoid freez-
Ifenprodil Tartrate Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of ifenprodil tartrate [(C H NO ) .C H O : D Optical rotation [a]20: ＋11 – ＋159(1 g calculated on the anhydrous basis, ethanol (95), 20 mL, 100 mm). 800.98]. 21 27 2 2 4 6 6 Melting point: about 1489C (with decomposition). Identification (1) Determine the absorption spectrum of a solution of Ifenprodil Tartrate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ifen-prodil Tartrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.4 g of Ifenprodil Tartrate in 40 mL of water by warming. After cooling, add 0.5 mL of ammonia TS to this solution, extract with two 40-mL portions of chlo- Method of preparation Prepare as directed under Gran-ules, with Ifenprodil Tartrate. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ifenprodil Tartrate Fine Gran-ules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Ifenprodil Tartrate Fine Granules, add 10 mL of water and a suitable amount of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly V mL so that each mL contains about 0.
(1) Determine the absorption spectrum of a solution of Ifenprodil Tartrate in methanol (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ifen-prodil Tartrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 0.4 g of Ifenprodil Tartrate in 40 mL of water by warming. After cooling, add 0.5 mL of ammonia TS to this solution, extract with two 40-mL portions of chlo- Method of preparation Prepare as directed under Gran-ules, with Ifenprodil Tartrate. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Ifenprodil Tartrate Fine Gran-ules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Ifenprodil Tartrate Fine Granules, add 10 mL of water and a suitable amount of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly V mL so that each mL contains about 0.1 mg of ifenprodil tartrate ‌JP XVIII Official Monographs / Ifenprodil Tartrate Tablets 1141 [(C21H27NO2)2.C4H6O6]. Filter through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of ifenprodil tartrate [(C21H27NO2)2.C4H6O6] ＝ MS × AT/AS × V/200 MS: Amount (mg) of ifenprodil tartrate for assay taken, calculated on the anhydrous basis Dissolution Being specified separately when the drug is granted approval based on the Law.
Powder Ifenprodil Tartrate Fine Granules, and weigh accurately a portion of the powder, equivalent to about 10 mg of ifenprodil tartrate [(C21H27NO2)2.C4H6O6], add 5 mL of water and a suitable amount of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly 100 mL. Filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of ifenprodil tartrate for assay (separately determine the water <2.48> in the same manner as Ifenprodil Tartrate), add 10 mL of water and a mixture of ethanol (99.5) and water (3:1) to make exactly 200 mL, and use this solution as the standard solution. Perform Ifenprodil Tartrate Tablets
Ifenprodil Tartrate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of ifenprodil tartrate [(C21H27NO2)2.C4H6O6: 800.98]. Method of preparation Prepare as directed under Tablets, with Ifenprodil Tartrate. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ifenprodil Tartrate Tablets, add V/20 mL of water, and shake until the tablet is completely dis-integrated. Then, add 7V/10 mL of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly V mL so that each mL contains about 0.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ifenprodil Tartrate Tablets, add V/20 mL of water, and shake until the tablet is completely dis-integrated. Then, add 7V/10 mL of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly V mL so that each mL contains about 0.1 mg of ifenprodil tartrate [(C H NO ) .C H O ]. Filter through a membrane filter the test with exactly 20 mL each of the sample solution and 21 27 2 2 4 6 6 standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ifenprodil in each solution. with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of ifenprodil tartrate [(C H NO ) .C H O ] Amount (mg) of ifenprodil tartrate [(C21H27NO2)2.C4H6O6] ＝ MS × AT/AS × 1/2 21 27 2 2 4 6 6 ＝ MS × AT/AS × V/200 MS: Amount (mg) of ifenprodil tartrate for assay taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 224 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.8 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 6.5 with potas-sium hydroxide TS, and add water to make 1000 mL. To 420 mL of this solution add 320 mL of methanol for liquid chromatography and 260 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of ifenprodil is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ifenprodil are not less than 3000 and MS: Amount (mg) of ifenprodil tartrate for assay taken, calculated on the anhydrous basis Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Ifen-prodil Tartrate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of ifen-prodil tartrate [(C21H27NO2)2C4H6O6], add 5 mL of water and a suitable amount of a mixture of ethanol (99.5) and water (3:1), shake thoroughly, and add a mixture of ethanol (99.5) and water (3:1) to make exactly 100 mL. Filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of ifenprodil tartrate for assay (separately determine the water <2.48> in the same manner as Ifenprodil Tartrate), add 10 mL of water and a mixture of ethanol (99.5) and water (3:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ifenprodil tartrate in each solution. not more than 2.0, respectively. Amount (mg) of ifenprodil tartrate [(C H NO ) .C H O ] System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- ＝ MS × AT/AS × 1/2 21 27 2 2 4 6 6 ing conditions, the relative standard deviation of the peak area of ifenprodil is not more than 1.0z.
Imidapril Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of Imidapril Hydrochloride (C20H27N3O6.HCl: 441.91). Method of preparation Prepare as directed under Tablets, with Imidapril Hydrochloride. Identification Weigh accurately an amount of powdered Imidapril Hydrochloride Tablets, equivalent to 25 mg of Imidapril Hydrochloride, add 5 mL of ethanol (99.5), shake well, filter, and use the filtrate as the sample solution. Sepa-rately, dissolve 25 mg of imidapril hydrochloride in 5 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, ethyl acetate, water, ethanol (99.5) and acetic acid (100) (16:16:7:2:2) to a distance of about 13 cm, and air-dry the plate. Examine un-der ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. retention time of imidapril, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add diluted methanol (2 in 5) to make exactly 20 mL. Confirm that the peak area of imidapril obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of imidapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imidapril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Imidapril Hydrochloride Tablets add 2V/5 mL of water, shake vigorously for 10 minutes, add diluted methanol (2 in 3) to make exactly V mL so that each mL contains about 0.
Weigh accurately an amount of powdered Imidapril Hydrochloride Tablets, equivalent to 25 mg of Imidapril Hydrochloride, add 5 mL of ethanol (99.5), shake well, filter, and use the filtrate as the sample solution. Sepa-rately, dissolve 25 mg of imidapril hydrochloride in 5 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. De-velop the plate with a mixture of 1-butanol, ethyl acetate, water, ethanol (99.5) and acetic acid (100) (16:16:7:2:2) to a distance of about 13 cm, and air-dry the plate. Examine un-der ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. retention time of imidapril, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add diluted methanol (2 in 5) to make exactly 20 mL. Confirm that the peak area of imidapril obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of imidapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imidapril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Imidapril Hydrochloride Tablets add 2V/5 mL of water, shake vigorously for 10 minutes, add diluted methanol (2 in 3) to make exactly V mL so that each mL contains about 0.1 mg of imidapril hydrochloride (C20H27N3O6.HCl), filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 10 mg of imida-pril for assay, previously dried at 1059C for 3 hours, dissolve in diluted methanol (2 in 5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of imidapril in each solution.
Related substances—To a quantity of powdered Imidapril Hydrochloride Tablets, equivalent to 25 mg of Imidapril Hydrochloride, add 40 mL of diluted methanol (2 Amount (mg) of imidapril hydrochloride (C ＝ MS × AT/AS × V/100 20H27N3O6.HCl) in 5), shake vigorously for 10 minutes, add diluted ethanol (2 in 5) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Pipet 1 mL of the sample solution, add diluted methanol (2 in 5) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of the peak having the relative retention time of about 0.45 to imidapril, obtained from the sample solution, is not larger than the peak area of imidapril from the standard solution, the area of the peak having the relative retention time of about 0.8 to imidapril from the sample solution is not larger than 7/10 times the peak area of imidapril from the standard solution, and the area of each peak other than imidapril and the peaks mentioned above from the sample solution is not larger than 3/10 times the peak area of imidapril from the standard solution. Furthermore, the total area of the peaks other than imidapril from the sample solution is not larger than 1.5 times the peak area of imidapril from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the MS: Amount (mg) of imidapril hydrochloride for assay taken Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of imidapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imidapril is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Imidapril Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Imidapril Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.8 mg of imidapril hydrochlo-ride (C20H27N3O6.HCl), and use this solution as the sample ‌1144 solution. Separately, weigh accurately about 28 mg of imida-pril hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of imidapril in each solution. Dissolution rate (z) with respect to the labeled amount of imidapril hydrochloride (C20H27N3O6.HCl) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of imidapril hydrochloride for assay taken C: Labeled amount (mg) of imidapril hydrochloride (C20H27N3O6.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of imidapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of imidapril is not more than 2.0z.
Weigh accurately not less than 20 Imidapril Hydro-chloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of imidapril hydro-chloride (C20H27N3O6.HCl), add 30 mL of diluted methanol (2 in 5) and exactly 5 mL of the internal standard solution, shake vigorously for 10 minutes, add diluted methanol (2 in 5) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add diluted methanol (2 in 5) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of imidapril hydrochloride for assay, previously dried at 1059C for 3 hours, dissolve in exactly 5 mL of the internal standard solution, add diluted methanol (2 in 5) to make 50 mL. Pipet 5 mL of this solution, add diluted methanol (2 in 5) to make 20 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of imidapril to that of the internal standard. 409C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust the pH to 2.7 with phosphoric acid. To 600 mL of this solution add 400 mL of methanol. Flow rate: Adjust so that the retention time of imidapril is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, imidapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of imidapril to that of the internal standard is not more than 1.0z.
Imipenem and Cilastatin Sodium for Injection
注射用イミペネム･シラスタチンナトリウム
Imipenem and Cilastatin Sodium for Injection is a preparation for injection which is dissolved or sus-pended before use. It contains not less than 93.
Imipenem and Cilastatin Sodium for Injection occurs as a white to light yellow-white powder.
(1) To 1 mL of a solution of Imipenem and Cilastatin Sodium for Injection (1 in 100) add 1 mL of nin-hydrin TS, heat in a water bath for 5 minutes: a purple color appears (cilastatin). (2) To 2 mL of a solution of Imipenem and Cilastatin ‌Sodium for Injection (1 in 1000) add 0.1 mol/L 3-(N-mor-pholino)propanesulfonic acid buffer solution (pH 7.0) to make 50 mL, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 296 nm and 300 nm (imipenem). pH <2.54> The pH of a solution prepared by dissolving an amount of Imipenem and Cilastatin Sodium for Injection, equivalent to 0.5 g (potency) of Imipenem Hydrate, in 100 7.0) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, ATI and ASI of imipenem, and those, ATC and ASC of cilastatin in each solution. Amount [mg (potency)] of imipenem (C12H17N3O4S) ＝ MSI × ATI/ASI mL of isotonic sodium chloride solution is between 6.5 and 8.0. The pH of the Injection intended for intramuscular use is between 6.0 and 7.5. Amount (mg) of cilastatin (C16 ＝ MSC × ATC/ASC × 0.955 H26N2O5S)
Clarity and color of solution—Dissolve an amount of Imipenem and Cilastatin Sodium for Injection, equivalent to 0.5 g (potency) of Imipenem Hydrate, in 100 mL of iso-tonic sodium chloride solution: the solution is clear and col-orless or pale yellow. Loss on drying <2.41> Not more than 3.0z (1 g, in vacu-um, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.25 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test (T: 104.0z). Dissolve the total amount of the content of 1 Imipenem and Cilastatin Sodium for Injection in isotonic sodium chloride solution to make exactly 100 mL. Measure exactly V mL of this solution, equivalent to about 25 mg (potency) of Imipenem Hydrate, add 0.1 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the sample solution. Proceed hereafter as directed in the Assay. Amount [mg (potency)] of imipenem (C12H17N3O4S) ＝ MSI × ATI/ASI × 100/V Amount (mg) of cilastatin (C16H26N2O5S) ＝ MSC × ATC/ASC × 100/V × 0.955 MSI: Amount [mg (potency)] of Imipenem RS taken MSC: Amount (mg) of cilastatin ammonium for assay taken, calculated on anhydrous and ethanol-free basis Foreign insoluble matter <6.06> Perform the test according to Method 2: It meets the requirement. Insoluble particulate matter <6.07> Perform the test ac-cording to the Method 1: the Injection which is dissolved before use meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Imipenem and Cilastatin Sodium for Injections. Weigh accurately an amount of the content, equivalent to 1 Imipenem and Cilastatin Sodium for Injection, dissolve in isotonic sodium chloride solution to make exactly 100 mL. Measure exactly an amount of this solution, equivalent to about 25 mg (potency) of imipenem, add 0.1 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 50 mL, and use this as the sample solution. Separately, weigh accurately an amount of Imipenem RS, equivalent to about 25 mg (potency), and weigh accurately about 25 mg of cilastatin ammonium for assay, dissolve in MSI: Amount [mg (potency)]of Imipenem RS taken MSC: Amount (mg) of cilastatin ammonium for assay taken, calculated on anhydrous and ethanol-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 20 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 0.836 g of 3-(N-morpholino)pro-panesulfonic acid, 1.0 g of sodium 1-hexane sulfonate and 50 mg of disodium dihydrogen ethylenediamine tetraacetate dihydrate in 800 mL of water, adjust to pH 7.0 with 0.1 mol/L sodium hydrate TS, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of imipenem is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, imipenem and cilastatin are eluted in this order with the resolution between these peaks being not less than 2.0, and the symmetry factors of the peak of imipenem and cilastatin are not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the peak area of imipenem and cilastatin are not more than 2.0z, re-spectively.
Imipramine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of imipramine hydrochloride (C19H24N2.HCl: 316.87). Method of preparation Prepare as directed under Tablets, with Imipramine Hydrochloride. Identification (1) Weigh a quantity of powdered Imipra-mine Hydrochloride Tablets, equivalent to 0.25 g of Imipra-mine Hydrochloride, add 25 mL of chloroform, shake thor-oughly, and filter. Evaporate the filtrate on a water bath, and proceed with the residue as directed in the Identification under Imipramine Hydrochloride. Dissolve an amount of the residue obtained in (1), equivalent to 5 mg of Imipramine Hydrochloride, in 250 mL of 0.01 mol/L hydrochloric acid TS, and determine the ab-sorption spectrum as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 249 nm and 253 nm, and a shoulder between 270 nm and 280 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Imipramine Hydrochloride Tablets add ex-actly 40 mL of 0.01 mol/L hydrochloric acid TS, disperse the tablet into a small particles by sonicating, then shake well. Centrifuge the solution, pipet V mL of the supernatant liquid, add water to make exactly V? mL so that each mL contains about 20 mg of imipramine hydrochloride (C19H24N2.
(1) Weigh a quantity of powdered Imipra-mine Hydrochloride Tablets, equivalent to 0.25 g of Imipra-mine Hydrochloride, add 25 mL of chloroform, shake thor-oughly, and filter. Evaporate the filtrate on a water bath, and proceed with the residue as directed in the Identification under Imipramine Hydrochloride. Dissolve an amount of the residue obtained in (1), equivalent to 5 mg of Imipramine Hydrochloride, in 250 mL of 0.01 mol/L hydrochloric acid TS, and determine the ab-sorption spectrum as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 249 nm and 253 nm, and a shoulder between 270 nm and 280 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Imipramine Hydrochloride Tablets add ex-actly 40 mL of 0.01 mol/L hydrochloric acid TS, disperse the tablet into a small particles by sonicating, then shake well. Centrifuge the solution, pipet V mL of the supernatant liquid, add water to make exactly V? mL so that each mL contains about 20 mg of imipramine hydrochloride (C19H24N2.HCl), and use this solution as the sample solu-tion. Separately, weigh accurately about 25 mg of Imipra-mine Hydrochloride RS, previously dried at 1059C for 2 hours, dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances at 251 nm, AT1 and AS1, and at 330 nm, AT2 and AS2, of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of imipramine hydrochloride (C19H24N2.HCl) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 4/125 MS: Amount (mg) of Imipramine Hydrochloride RS taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 60 minutes of Imipramine Hydrochloride Tablets is not less than 75z. ‌1148 Start the test with 1 tablet of Imipramine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL of the filtrate contains about 10 mg of imipramine hydrochloride (C19H24N2.HCl), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Imipramine Hydrochloride RS, previously dried at 1059C for 2 hours, dissolve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and the standard solution at 250 nm as directed under Ultra-violet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount Indapamide
Indapamide Tablets contain not less than 93.0z and not more than 103.0z of the labeled amount of indapamide (C16H16ClN3O3S: 365.83). Method of preparation Prepare as directed under Tablets, with Indapamide. Identification To an amount of powdered Indapamide Tablets, equivalent to 10 mg of Indapamide, add 5 mL of ethyl acetate, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 10 mg of Indapamide RS in 5 mL of ethyl acetate, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, cyclohexane and acetic acid (100) (100:80:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show a blue-purple color and the same Rf value. Amount (mg) of indapamide (C ＝ MS × QT/QS 16H16ClN3O3S) Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. MS: Amount (mg) of Indapamide RS taken, calculated on the dried basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in a mixture of water and ethanol (99.5) (1:1) (3 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 287 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000), acetonitrile and methanol (6:3:1). Flow rate: Adjust so that the retention time of indapamide is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, indapamide and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of indapamide is not more than 1.0z. Containers and storage Containers—Tight containers. Storage—Light-resistant. To 1 tablet of Indapamide Tablets add exactly V/10 mL of the internal standard solution, and add a mixture of water and ethanol (99.5) (1:1) to make V mL so that each mL con-tains about 0.1 mg of indapamide (C16H16ClN3O3S), shake to disintegrate, sonicate for 10 minutes, shake again for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Proceed as directed in the Assay. Amount (mg) of indapamide (C16H16ClN3O3S) ＝ MS × QT/QS × V/200 MS: Amount (mg) of Indapamide RS taken, calculated on the dried basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in a mixture of water and ethanol (99.5) (1:1) (3 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 45 minutes of 1-mg tablet and in 90 minutes of 2-mg tablet are not less than 70z, respectively. Start the test with 1 tablet of Indapamide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.
To an amount of powdered Indapamide Tablets, equivalent to 10 mg of Indapamide, add 5 mL of ethyl acetate, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 10 mg of Indapamide RS in 5 mL of ethyl acetate, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, cyclohexane and acetic acid (100) (100:80:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show a blue-purple color and the same Rf value. Amount (mg) of indapamide (C ＝ MS × QT/QS 16H16ClN3O3S) Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. MS: Amount (mg) of Indapamide RS taken, calculated on the dried basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in a mixture of water and ethanol (99.5) (1:1) (3 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 287 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000), acetonitrile and methanol (6:3:1). Flow rate: Adjust so that the retention time of indapamide is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, indapamide and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of indapamide is not more than 1.0z.
To 20 Indapamide Tablets add 80 mL of a mixture of water and ethanol (99.5) (1:1), shake well to disintegrate, and sonicate for 10 minutes. Shake the solution for 10 minutes, and add a mixture of water and ethanol (99.5) (1:1) to make exactly 100 mL. Pipet a volume of indapamide (C16 H16ClN3O3S), equivalent to about 2 mg, and add exactly 2 mL of the internal standard solution and a mixture of water and ethanol (99.5) (1:1) to make 20 mL. Centrifuge this solu-tion, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Indapamide RS (separately, determine the loss on drying <2.41> under the same condition as Indapamide), and dissolve in a mixture of water and ethanol (99.5) (1:1) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 2 mL of the internal standard solution and a mixture of water and ethanol (99.5) (1:1) to make 20 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Indapa-mide. Amount (mg) of indapamide (C16H16ClN3O3S) ＝ MS × QT/QS × 1/10 MS: Amount [mg (potency)] of Indapamide RS taken, cal-culated on the dried basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in a mixture of water and ethanol (99.5) (1:1) (3 in 1000).
Indigocarmine Injection is an aqueous injection. It contains not less than 95.
Indigocarmine Injection is a dark blue liquid. pH: 3.0 – 5.0
(1) To a volume of Indigocarmine Injec-tion, equivalent to 20 mg of Indigocarmine, add 1 mL of nitric acid: the dark blue color of the liquid disappears, and a yellow-brown color develops. To a volume of Indigocarmine Injection, equivalent to 20 mg of Indigocarmine, add 1 mL of bromine TS: the dark blue color disappears, and a yellow-brown color de-velops. To a volume of Indigocarmine Injection, equivalent to 20 mg of Indigocarmine, add 1 mL of chlorine TS: the Each mL of 0.1 mol/L titanium (III) chloride VS ＝ 23.32 mg of C16H8N2Na2O8S2
Indium (111In) Chloride Injection is an aqueous in-jection. It contains indium-111 (111In) in the form of indium chloride.
Indium (111In) Chloride Injection is a clear, colorless liquid. Indometacin
Indometacin Capsules contain not less than 90.0z and not more than 110.0z of the labeled amount of minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 28 mg of indometacin (C19H16ClNO4), and use this solution as the sample solution.
Powder the contents of Indometacin Cap-sules. To a quantity of the powder, equivalent to 0.1 g of Indometacin, add 20 mL of chloroform, shake well, and centrifuge. Filter the supernatant liquid, and evaporate the filtrate to dryness. After cooling, dissolve the residue in 20 mL of methanol. To 10 mL of this solution add methanol to trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of indometacin (C19H16ClNO4) ＝ MS × AT/AS × 1/C × 90 MS: Amount (mg) of Indometacin RS taken C: Labeled amount (mg) of indometacin (C19H16ClNO4) in 1 capsule ‌1154
Weigh accurately the contents of not less than 20 Indometacin Capsules. Powder the combined contents, and weigh accurately a portion of the powder, equivalent to about 50 mg of indometacin (C19H16ClNO4). Dissolve in 40 mL of methanol, and add methanol to make exactly 50 mL. Filter this solution, discarding the first 10-mL portion of the filtrate. Pipet 5 mL of the subsequent filtrate, add exactly 3 Indometacin Suppositories
二相性イソフェンインスリン ヒト(遺伝子組換え)水性懸濁注射液
Biphasic Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension is a white aqueous suspension. When allowed to stand, it separates into a white precipitate and colorless supernatant liquid, and the precipitate returns to the suspension state on gentle shak-ing. When it is examined microscopically, the precipitate most-ly consists of fine, oblong crystals of 1 to 30 mm in major axis, and does not contain amorphous substances or large aggregates.
Adjust Biphasic Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension to pH between 2.5 and 3.0 with dilute hydrochloric acid: the precipitate dissolves, and the solution is clear and colorless. ‌pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Desamido substance—Perform the test with 20 mL of the sample solution obtained in the Assay (1) as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of the Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Soluble Insulin Human Being specified separately when the drug is granted approval based on the Law.
(1) Insulin human—Pipet 10 mL of gently shaken Biphasic Isophane Insulin Human (Genetical Recombina-tion) Injectable Aqueous Suspension, and add exactly 40 mL of 6 mol/L hydrochloric acid TS. Pipet 2 mL of this solu-tion, add 0.01 mol/L hydrochloric acid TS to make exactly 5 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay under Insulin Human (Ge-netical Recombination). peak, having the relative retention time of about 1.3 to insu-lin human, is not more than 1.5z. Operating conditions— Proceed as directed in the operating conditions in the Assay (1). Amount (Insulin Unit) of insulin human (C257H in 1 mL ＝ (MS × F )/D × (ATI ＋ ATD)/(ASI ＋ ASD) × 1.004 × 5/2 N65O 77S6) System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). Test for required detectability: Pipet 1 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make ex-actly 50 mL. Confirm that the peak area of insulin human obtained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that with 20 mL of the sample solution. System repeatability: Dissolve Insulin Human RS in 0.01 mol/L hydrochloric acid TS so that each mL contains about 4 Insulin Units. When the test is repeated 6 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of insulin human is not more than 2.0z. (2) High-molecular mass protein—Take a suitable volume of gently shaken Biphasic Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension, add 4 mL of 6 mol/L hydrochloric acid TS for each mL of the suspension, and mix until the solution becomes clear. Perform the test with 100 mL of this solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amount of them by the area percentage method: the total amount of the peaks other than insulin human is not more than 2.0z. Operating conditions— Detector, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Insulin Human (Genetical Recombination). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the completion of the elution of insulin human. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) under Insulin Human (Genetical Recombination). Test for required detectability: Pipet 1 mL of the sample solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of insulin human MS: Amount (mg) of Insulin Human RS taken F: Labeled unit (Insulin Unit /mg) of Insulin Human RS D: Volume (mL) of 0.01 mol/L hydrochloric acid TS used to dissolve Insulin Human RS (2) Zinc—Pipet a volume of gently shaken Biphasic Isophane Insulin Human (Genetical Recombination) Injecta-ble Aqueous Suspension, equivalent to 300 Insulin Units, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. If necessary, further add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sam-ple solution. Separately, pipet a suitable volume of Standard Zinc Solution for Atomic Absorption Spectroscopy, dilute with 0.01 mol/L hydrochloric acid TS to make three solu-tions containing 0.20 mg, 0.60 mg and 1.20 mg of zinc (Zn: 65.38) in each mL, respectively, and use these solutions as the standard solutions. Perform the test with the sample so-lution and standard solutions as directed under Atomic Ab-sorption Spectroscopy <2.23> according to the following con-ditions, using 0.01 mol/L hydrochloric acid TS as the blank, and calculate the content of zinc in the sample solution by using the calibration curve obtained from the absorbances of the standard solutions. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm.
過テクネチウム酸ナトリウム (99mTc) 注射液
Sodium Pertechnetate (99mTc) Injection is a clear, colorless liquid. Dibasic Sodium Phosphate Hydrate
Insulin Glargine (Genetical Recombination) Injection
インスリン グラルギン（遺伝子組換え）注射液
Insulin Glargine (Genetical Recombination) Injection occurs as a clear, colorless liquid.
(1) Insulin Glargine (Genetical Recombina-tion) Injection forms a precipitate when adjusted to pH 5.7 – 6.5 with dilute sodium hydroxide TS, and the precipitate dis-appears when adjusted to pH 3.5 – 4.5 with 0.1 mol/L hy-drochloric acid TS. (2) Perform the test with 5 mL each of the sample solu- Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Zinc content Being specified separately when the drug is granted approval based on the Law.
(1) Related substances—Keep the sample solution at 2 – 89C. Perform the test with 5 mL of the sample solution obtained in the Assay as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than insulin glargine is not more than 0.5z, and the total amount of the peaks other than insulin glargine is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Insulin Glargine (Genetical Recombination). Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL. Confirm that the peak area of insulin glargine obtained with 5 mL of this solution is equivalent to 5 to 15z of that with 5 mL of the solution for system suitabil-ity test. System performance: When the procedure is run with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of insulin glargine are not less than 20,000 and not more than 1.8, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the relative standard deviation of the peak area of insulin glargine is not more than 2.0z. (2) High-molecular mass proteins—To a suitable amount of Insulin Glargine (Genetical Recombination) Injection add water so that each mL contains 40 Insulin Units, and use this solution as the sample solution. Then, proceed as directed in the Purity (2) under Insulin Glargine (Genetical Recombina-tion). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. MS: Amount (mg) of insulin glargine in 1 mL of the stand-ard solution d: Dilution factor of the sample solution 0.0364: Mass (mg) of insulin glargine equivalent to 1 Insu-lin Unit
To a suitable amount of Insulin Glargine (Genetical Recombination) Injection add exactly water so that each mL contains 40 Insulin Units, and use this solution as the sample solution. Then, proceed as directed in the Assay under Insu-lin Glargine (Genetical Recombination). tion and the standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the conditions described in the Assay: the retention times of the principal peaks obtained from the sample solution and Amount (Insulin Unit) of insulin glargine (C in 1 mL ＝ MS × AT/AS × d × 1/0.0364 H404 N72 O78 S6) standard solution are the same. pH Being specified separately when the drug is granted ap-proval based on the Law. Purity (1) Related substances—Keep the sample solution at 2 – 89C. Perform the test with 5 mL of the sample solution obtained in the Assay as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than insulin glargine is not more than 0.5z, and the total amount of the peaks other than insulin glargine is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Insulin Glargine (Genetical Recombination). Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make ex-actly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL. Confirm that the peak area of insulin glargine obtained with 5 mL of this solution is equivalent to 5 to 15z of that with 5 mL of the solution for system suitabil-ity test. System performance: When the procedure is run with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of insulin glargine are not less than 20,000 and not more than 1.8, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution obtained in the Assay under the above operating conditions, the relative standard deviation of the peak area of insulin glargine is not more than 2.0z. (2) High-molecular mass proteins—To a suitable amount of Insulin Glargine (Genetical Recombination) Injection add water so that each mL contains 40 Insulin Units, and use this solution as the sample solution. Then, proceed as directed in the Purity (2) under Insulin Glargine (Genetical Recombina-tion). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. MS: Amount (mg) of insulin glargine in 1 mL of the stand-ard solution d: Dilution factor of the sample solution 0.0364: Mass (mg) of insulin glargine equivalent to 1 Insu-lin Unit
Insulin Human (Genetical Recombination) Injection
インスリン ヒト（遺伝子組換え）注射液
Insulin Human (Genetical Recombination) Injection is an aqueous injection. It contains not less than 95.
Insulin Human (Genetical Recombination) In-jection occurs as a clear, colorless liquid, and slightly a fine precipitate may be observable upon storage.
Insulin Human (Genetical Recombination) Injection forms a precipitate when adjusted to pH 5.3 – 5.5 by addition of dilute hydrochloric acid, and the precipitate disappears when adjusted to pH 2.5 – 3.5 by further addition of the acid. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted approval based on the Law.
(1) Desamido substance—Perform the test with 20 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate their amounts by the area percentage method: the amount of the peak, having the relative retention time of about 1.3 to human insulin, is not more than 1.5z. Operating conditions— Proceed as directed in the operating conditions in the Assay under Insulin Human (Genetical Recombination). System suitability— System performance: Proceed as directed in the system suitability in the Assay under Insulin Human (Genetical Recombination). Test for required detectability: Pipet 1 mL of the sample solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of human insulin obtained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that with 20 mL of the sample solution. System repeatability: Dissolve Insulin Human RS in 0.01 mol/L hydrochloric acid TS so that each mL contains about 4 Insulin Units. When the test is repeated 6 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of human insulin is not more than 2.0z. (2) High-molecular mass proteins—For each mL of Insu-lin Human (Genetical Recombination) Injection add 4 mL of 6 mol/L hydrochloric acid TS, and use this solution as the sample solution. Perform the test with 100 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the total amount of the peaks other than human insulin is not more than 2.0z. Operating conditions— Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Insulin Human (Genetical Recombination). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the completion of the elution of human insulin. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) under Insulin Human (Genetical Recombination). Test for required detectability: Pipet 1 mL of the sample solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of human insulin obtained with 100 mL of this solution is equivalent to 1.4 to 2.6z of that with 100 mL of the sample solution. Bacterial endotoxins <4.01> Less than 0.80 EU/Insulin Unit. Apply to the preparations intended for intravenous administration. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. ‌Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Zinc content To an exact volume of Insulin Human (Genetical Recombination) Injection, equivalent to 300 Insu-lin Units, add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. If necessary, further dilute with 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample solution. Separately, take exactly a suitable amount of Standard Zinc Solution for Atomic Absorption Spectrophotometry, dilute with 0.01 mol/L hy-drochloric acid TS to make three solutions containing 0.20 mg, 0.60 mg and 1.20 mg of zinc (Zn: 65.38) in each mL, re-spectively, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solu-tions as directed under Atomic Absorption Spectrophotome-try <2.23> according to the following conditions, using the 0.01 mol/L hydrochloric acid TS as the blank, and calculate the amount of zinc in the sample solution by using a calibra-tion curve obtained from the absorbances of the standard so-lutions: 10 – 40 mg of zinc (Zn: 65.38) per 100 Insulin Units. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm.
To exactly 10 mL of Insulin Human (Genetical Recombination) Injection add exactly 40 mL of 6 mol/L hy-drochloric acid TS. Pipet 2 mL of this solution, add 0.01 mol/L hydrochloric acid TS to make exactly 5 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay under Insulin Human (Genetical Recom-bination). Amount (Insulin Unit) of human insulin (C257H383N65O77S6) in 1 mL ＝ MS × F/D × (AT1＋ATD)/(AS1＋ASD) × 1.004 × 5/2 MS: Amount (mg) of Insulin Human RS taken F: Labeled unit (Insulin Unit/mg) of Insulin Human RS D: Volume (mL) of 0.01 mol/L hydrochloric acid TS used to dissolve Insulin Human RS
Iodinated (131I) Human Serum Albumin Injection
Iodinated (131I) Human Serum Albumin Injec- ‌1170‌ tion is a clear, colorless or light yellow liquid. Iodine
Iohexol Injection is an aqueous injection. It contains not less than 95.
Iohexol Injection is a clear and colorless liq-uid.
To a volume of Iohexol Injection, equivalent to 0.65 g of Iohexol, add water to make 500 mL. To 1 mL of this solution add water to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 243 nm and 247 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Aromatic primary amine—Conduct this proce-dure using light-resistant vessels. To a volume of Iohexol In-jection, equivalent to 0.20 g of Iohexol add 15 mL of water, cool in ice for 5 minutes, add 1.5 mL of 6 mol/L hydrochlo-ric acid TS and 1 mL of solution of sodium nitrite (1 in 50), prepared before use, shake, and cool in ice for 4 minutes. Then, proceed as directed in the Purity (2) under Iohexol: the absorbance of a solution so obtained is not more than 0.23. (2) Iodine and iodide—To a volume of Iohexol Injec-tion, equivalent to 1.0 g of Iohexol, add 4 mL of water and 1 mL of dilute sulfuric acid, and allow to stand for 10 minutes while occasional shaking. Then, proceed as directed in the Purity (4) under Iohexol: the absorbance of a chloroform layer so obtained is not more than 0.14. Bacterial endotoxins <4.01> Less than 0.47 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Iohexol Injec-tion, equivalent to about 1.5 g of iohexol (C19H26I3N3O9), add water to make exactly 25 mL. Pipet 10 mL of this solu-tion, add 25 mL of a solution of sodium hydroxide (1 in 20) and 0.5 g of zinc powder, and boil under a reflux condenser for 30 minutes. After cooling, wash down the inside of the condenser with 20 mL of water, and filter. Then, proceed as directed in the Assay under Iohexol. Each mL of 0.1 mol/L silver nitrate VS ＝ 27.37 mg of C19H26I3N3O9
Containers—Hermetic containers. Colored containers and plastic containers for aqueous injec-tions may be used.
Iopamidol Injection is an aqueous injection. It contains not less than 95.
Iopamidol Injection occurs as a clear, colorless or faint yellow, liquid, having slight viscosity. It is gradually colored to faint yellow by light.
(1) To a volume of Iopamidol Injection, equivalent to 0.3 g of Iopamidol, add 0.2 mL of sulfuric acid, and mix. When heat the solution over a flame, the color of the solution changes from colorless to purplish brown, and a purple gas is evolved. (2) To a volume of Iopamidol Injection, equivalent to 0.6 g of Iopamidol, add water to make 100 mL, and use this solution as the sample solution. Separately, dissolve 60 mg of iopamidol for assay in 10 mL of water, and use this solu-tion as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 4 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 2-propanol, 2-butanone and ammonia solution (28) (2:2:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the Rf value of the principal spot obtained from the sample solution is the same as that from the standard solution. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Primary aromatic amines—To a volume of Iopamidol Injection, equivalent to 0.18 g of Iopamidol, add 6 mL of water and mix. Add 1 mL of a solution of sodium nitrite (1 in 50) and 12 mL of 2 mol/L hydrochloric acid TS, shake the solution and allow to stand for 2 minutes. Add 1 mL of a solution of ammonium amidosulfate (1 in 10), JP XVIII Official Monographs / Iotalamic Acid 1179 ‌shake well, and allow to stand for 1 minute. Add 1 mL of naphthylethylenediamine TS and water to make exactly 50 mL. Determine the absorbance of this solution at 495 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution prepared in the same manner as the blank: the absorbance is not more than 0.18. Iodine—Take a volume of Iopamidol Injection, equivalent to 2.0 g of Iopamidol, and add 2 mL of 1 mol/L sulfuric acid TS and 1 mL of toluene. Then shake well and allow to stand: the toluene layer is colorless. Free iodine ion—To exactly 10 mL of Iopamidol In-jection add a suitable amount of water, and adjust the pH to about 4.5 with diluted 0.25 mol/L sulfuric acid TS (1 in 10). Titrate <2.50> with 0.001 mol/L silver nitrate VS (potentio-metric titration): the amount of iodine ion contained in Iopamidol Injection is not more than 40 mg per mL. Each mL of 0.001 mol/L silver nitrate VS ＝ 0.1269 mg of I Bacterial endotoxins <4.01> Less than 1.5 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To exactly 1 mL of Iopamidol Injection add water to make exactly 200 mL. Take exactly V mL of this solution, add water to make exactly V? mL so that each mL contains about 80 mg of iopamidol (C17H22I3N3O8), and use this solu-tion as the sample solution. Separately, weigh accurately about 20 mg of iopamidol for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 10 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of iopamidol in each so-lution. Amount (mg) of iopamidol (C17H22I3N3O8) ＝ MS × AT/AS × V?/V × 4/5 MS: Amount (mg) of iopamidol for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Water. Mobile phase B: A mixture of water and methanol (3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 6 6 – 18 92 → 65 8 → 35 18 – 30 65 → 8 35 → 92 30 – 34 Flow rate: 1.5 mL per minute. System suitability— System performance: Dissolve 1 mg each of iopamidol for assay and N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylamino-2,4,6-triiodoisophthalamide in water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, N,N?-bis[2-hydroxy-1-(hydroxymethyl)ethyl]-5-hydroxyacetylamino-2,4,6-triiodoisophthalamide and iopamidol are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of iopamidol is not more than 1.0z.
Ipecac Syrup is a yellow-brown, viscous liquid. It has a sweet taste and a bitter aftertaste.
Take 2 mL of Ipecac Syrup into an evaporat-ing dish, mix with 1 mL of hydrochloric acid, and add small pieces of chlorinated lime: circumference of it turns orange.
Ethanol—Take exactly 5 mL of Ipecac Syrup, add exactly 5 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, pipet 5 mL of ethanol (99.5), and add water to make exactly 100 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the rate of peak height of ethanol to that of the internal standard, QT and QS: QT is not larger than QS. Internal standard solution—A solution of acetonitrile (1 in 20). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass-column about 3 mm in inside diameter and about 1.5 m in length, packed with ethylvinylbenzene-divinylbenzene porous co-polymer for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature of between 1059C and 1159C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethanol is 5 to 10 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, ethanol and the internal standard are eluted in this order with the resolution between these peaks being not less than 1.5. Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 103 CFU/mL and 102 CFU/mL, respectively. Escherichia coli, Salmonella, Pseudomonas aeruginosa and Staphylococcus aureus are not observed.
Take exactly 5 mL of Ipecac Syrup, add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL, and use the so-lution as the sample solution. Separately, weigh accurately about 10 mg of emetine hydrochloride for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide, 509C) for 5 hours, dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, ATE and ATC, of emetine and cephaeline with the sample solution, and the peak area, ASE, of emetine with the standard solution. Amount (mg) of total alkaloids (emetine and cephaeline) ＝ MS ×{ATE ＋ (ATC × 0.971)}/ASE × 1/2 × 0.868 MS: Amount (mg) of emetine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: Dissolve 2.0 g of sodium l-heptane sul-fonate in 500 mL of water, adjust the pH to 4.0 with acetic acid (100), and add 500 mL of methanol. Flow rate: Adjust so that the retention time of emetine is about 14 minutes. System suitability— System performance: Dissolve 1 mg each of emetine hy-drochloride for assay and cephaeline hydrobromide in 10 mL of 0.01 mol/L hydrochloric acid TS. When the procedure is run with 10 mL of this solution under the above operating conditions, cephaeline and emetine are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌JP XVIII Crude Drugs and Related Drugs / Powdered Japanese Angelica Root 2039‌‌ ing conditions, the relative standard deviation of the peak area of emetine is not more than 1.5z.
To a quantity of powdered Ipriflavone Tablets, equivalent to 11 mg of Ipriflavone, add 100 mL of methanol, shake vigorously for 10 minutes, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 247 nm and 251 nm, and between 297 nm and 301 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Ipriflavone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 30 mg of ipriflavone (C18H16O3), add 30 mL of acetonitrile, shake vigorously for 15 minutes, add acetonitrile to make exactly 50 mL, and cen-trifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and add acetonitrile to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Ipriflavone RS, previously dried at 1059C for 2 hours, and dissolve in aceto-nitrile to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and add acetonitrile to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Ipriflavone. Amount (mg) of ipriflavone (C18H16O3) ＝ MS × QT/QS MS: Amount (mg) of Ipriflavone RS taken Internal standard solution—A solution of di-n-butyl phtha-late in acetonitrile (1 in 100).
Irbesartan and Amlodipine Besilate Tablets
イルベサルタン･アムロジピンベシル酸塩錠
Irbesartan and Amlodipine Besilate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of irbesartan (C25H28N6O: 428.53) and amlodipine besilate (C20H25ClN2O5.C6H6O3S: 567.05). Method of preparation Prepare as directed under Tablets, with Irbesartan and Amlodipine Besilate. Identification (1) Perform the test with 5 mL each of the sample solution and standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> accord- ing to the following conditions: the retention time of the gram from the standard solution, and both absorption spec-tra of these peaks exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 237 nm, spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (2). Uniformity of dosage unit <6.02> (1) Irbesartan—Per-form the Mass variation test, or the Content uniformity test according to the following method: it meets the requirement. To 1 tablet of Irbesartan and Amlodipine Besilate Tablets add 4 mL of 0.02 mol/L phosphate buffer solution (pH 3.0), and sonicate. Add 16 mL of methanol, shake vigorously until the tablet is disintegrated completely, and add the mo-bile phase to make exactly 100 mL. Pipet V mL of this solu-tion, add the mobile phase to make exactly V? mL so that each mL contains about 1 mg of irbesartan (C25H28N6O), and filter through a membrane filter with a pore size not exceeding 0.
(1) Perform the test with 5 mL each of the sample solution and standard solution obtained in the Assay as directed under Liquid Chromatography <2.01> accord- ing to the following conditions: the retention time of the gram from the standard solution, and both absorption spec-tra of these peaks exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 237 nm, spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (2). Uniformity of dosage unit <6.02> (1) Irbesartan—Per-form the Mass variation test, or the Content uniformity test according to the following method: it meets the requirement. To 1 tablet of Irbesartan and Amlodipine Besilate Tablets add 4 mL of 0.02 mol/L phosphate buffer solution (pH 3.0), and sonicate. Add 16 mL of methanol, shake vigorously until the tablet is disintegrated completely, and add the mo-bile phase to make exactly 100 mL. Pipet V mL of this solu-tion, add the mobile phase to make exactly V? mL so that each mL contains about 1 mg of irbesartan (C25H28N6O), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed under the Assay (1). Amount (mg) of irbesartan (C25H28N6O) ＝ MS × AT/AS × V?/V × 2 MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis Amlodipine besilate—Perform the test according to the following method: it meets the requirement of the Con-tent uniformity test. To 1 tablet of Irbesartan and Amlodipine Besilate Tablets add 4 mL of 0.02 mol/L phosphate buffer solution (pH 3.0), and sonicate. Add 16 mL of methanol, shake vigorously until the tablet is disintegrated completely, and add the mo-bile phase to make exactly 100 mL. Pipet V mL of this solu-tion, add the mobile phase to make exactly V? mL so that each mL contains about 69 mg of amlodipine besilate (C20H25 ClN2O5.C6H6O3S), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Then, proceed as directed under the Assay (2). Amount (mg) of amlodipine besilate peak of irbesartan in the chromatogram from the sample so- lution is the same with that of the principal peak in the chro- (C20H25 ClN2O5.C6H6O3S) matogram from the standard solution, and both absorption ＝ MS × AT/AS × V?/V × 1/5 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Dissolution <6.10> (1) Irbesartan—When the test is per-formed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Ir-besartan and Amlodipine Besilate Tablets is not less than 70z. Start the test with 1 tablet of Irbesartan and Amlodipine Besilate Tablets, withdraw not less than 15 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add the mobile phase to make exactly V? mL so that each mL contains about 0.11 mg of irbesartan (C25H28N6O). Pipet 2 mL of this solu-tion, add exactly 2 mL of the mobile phase, and use this so-lution as the sample solution. Separately, weigh accurately about 20 mg of irbesartan for assay (separately determine the water <2.48> in the same manner as Irbesartan), dissolve in the mobile phase to make exactly 25 mL, and use this so-lution as the irbesartan standard stock solution. Pipet 7 mL of the irbesartan standard stock solution, and add the mo-bile phase to make exactly 50 mL. Pipet 5 mL of this solu-tion, add exactly 5 mL of the dissolution medium, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of irbesartan in each solution. Dissolution rate (z) with respect to the labeled amount of irbesartan (C25H28N6O) ＝ MS × AT/AS × V?/V × 1/C × 504 MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of irbesartan (C25H28N6O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: To 7 mL of the irbesartan standard stock solution and 5 mL of the amlodipine besilate standard stock solution obtained in (2) add the mobile phase to make 50 mL. To 5 mL of this solution add 5 mL of the dissolution medium. When the procedure is run with 10 mL of this solu-tion under the above operating conditions, amlodipine and irbesartan are eluted in this order with the resolution be-tween these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irbesartan is not more than 2.0z. (2) Amlodipine besilate—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolu-tion medium, the dissolution rate in 30 minutes of Irbesartan and Amlodipine Besilate Tablets is not less than 75z. Start the test with 1 tablet of Irbesartan and Amlodipine Besilate Tablets, withdraw not less than 15 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add the mobile phase to make exactly V? mL so that each mL contains about 7.7 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S). Pipet 2 mL of this solution, add exactly 2 mL of the mobile phase, and use this solution as the sample solution. Sepa-rately, weigh accurately about 26 mg of Amlodipine Besilate RS (separately determine the water <2.48> in the same man-ner as Amlodipine Besilate), and dissolve in the mobile phase to make exactly 50 mL. Pipet 15 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the amlodipine besilate standard stock solution. Pipet 5 mL of the amlodipine besilate standard stock solution, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the dissolution medium, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amlodipine in each solution. Dissolution rate (z) with respect to the labeled amount of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × AT/AS × V?/V × 1/C × 27 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: To 7 mL of the irbesartan standard stock solution obtained in (1) and 5 mL of the amlodipine besilate standard stock solution add the mobile phase to make 50 mL. To 5 mL of this solution add 5 mL of the dis-solution medium. When the procedure is run with 10 mL of this solution under the above operating conditions, amlodi-pine and irbesartan are eluted in this order with the resolu-tion between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of amlodipine is not more than 2.0z.
(1) Irbesartan—To 10 tablets of Irbesartan and Amlodipine Besilate Tablets add 20 mL of 0.02 mol/L phos-phate buffer solution (pH 3.0), and sonicate. Add 120 mL of methanol, shake vigorously until the tablets are disintegrated completely, and add the mobile phase to make exactly 200 mL. Pipet V mL of this solution, add the mobile phase to make exactly V? mL so that each mL contains about 1 mg of irbesartan (C25H28N6O), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Separately, weigh accurately about 50 mg of ir-besartan for assay (separately determine the water <2.48> in the same manner as Irbesartan), dissolve in methanol to make exactly 25 mL, and use this solution as the irbesartan standard stock solution. Pipet 10 mL of the irbesartan stand-ard stock solution, add 2 mL of methanol, add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and stand-ard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of irbesartan in each solution. Amount (mg) of irbesartan (C25H28N6O) in 1 tablet ＝ MS × AT/AS × V?/V × 2/5 ‌JP XVIII Official Monographs / Irinotecan Hydrochloride Hydrate 1187 MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 75 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (2.2 mm in particle di-ameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and 0.02 mol/L phosphate buffer solution (pH 3.0) (3:2). Flow rate: Adjust so that the retention time of irbesartan is about 3 minutes. System suitability— System performance: To 10 mL of the irbesartan standard stock solution and 2 mL of the amlodipine besilate standard stock solution obtained in (2) add 0.02 mol/L phosphate buffer solution (pH 3.0) to make 20 mL. When the proce-dure is run with 5 mL of this solution under the above operat-ing conditions, amlodipine and irbesartan are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of irbesartan is not more than 1.0z. (2) Amlodipine besilate—To 10 tablets of Irbesartan and Amlodipine Besilate Tablets add 20 mL of 0.02 mol/L phos-phate buffer solution (pH 3.0), and sonicate. Add 120 mL of methanol, shake vigorously until the tablets are disintegrated completely, and add the mobile phase to make exactly 200 mL. Pipet V mL of this solution, add the mobile phase to make exactly V? mL so that each mL contains about 69 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 35 mg of Amlodipine Besilate RS (separate-ly determine the water <2.48> in the same manner as Amlodi-pine Besilate), dissolve in methanol to make exactly 50 mL, and use this solution as the amlodipine besilate standard stock solution. Pipet 2 mL of the amlodipine besilate stand-ard stock solution, add 10 mL of methanol, add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and stand-ard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of amlodipine in each solution. Amount (mg) of amlodipine besilate (C H ClN O .C H O S) in 1 tablet order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of amlodipine is not more than 1.0z.
Irbesartan Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of ir- Tablets, equivalent to about 25 mg of Irbesartan, add 2 mL of acetone, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Evaporate the filtrate to dryness, and determine the infrared absorption spectrum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tions at the wave numbers of about 1733 cm－1, 1617 cm－1, 1435 cm－1 and 758 cm－1. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Irbesartan Tablets add 1.5 mL of water, shake vigorously to disintegrate, and add 15 mL of metha-nol. Shake vigorously for 15 minutes, add methanol to make exactly 20 mL, and centrifuge. Pipet V mL of the superna-tant liquid, equivalent to about 20 mg of irbesartan (C25H28N6O), and add a mixture of water and acetonitrile for liquid chromatography (3:2) to make exactly 20 mL. Pipet 2.5 mL of this solution, add a mixture of water and aceto-nitrile for liquid chromatography (3:2) to make exactly 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of irbesartan (C25H28N6O) ＝ MS × AT/AS × 16/V MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of 50-mg and 100-mg tablets is not less than 85z, respectively, and that in 60 minutes of a 200-mg tablet is not less than 70z. Start the test with 1 tablet of Irbesartan Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 3 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg of irbesartan (C25H28N6O), and use this solution as the sample solution.
To a quantity of powdered Irbesartan natant liquid, equivalent to about 20 mg of irbesartan (C25H28N6O), and add a mixture of water and acetonitrile for liquid chromatography (3:2) to make exactly 20 mL. Pipet JP XVIII Official Monographs / Irbesartan and Amlodipine Besilate Tablets 1185 ‌2.5 mL of this solution, add a mixture of water and aceto-nitrile for liquid chromatography (3:2) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of irbesartan for assay (separately determine the water <2.48> in the same manner as Irbesartan), and dissolve in methanol to make exactly 10 mL. Pipet 2.5 mL of this solution, add a mixture of water and acetonitrile for liquid chromatography (3:2) to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of irbesartan in each solution. spectra of these peaks exhibit similar intensities of absorp-tion at the same wavelengths. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 237 nm, spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). (2) Perform the test with 5 mL each of the sample solu-tion and standard solution obtained in the Assay (2) as di-rected under Liquid Chromatography <2.01> according to Amount (mg) of irbesartan (C ＝ MS × AT/AS × 16/V 25H 28N6O) in 1 tablet the following conditions: the retention time of the peak of amlodipine in the chromatogram from the sample solution is the same with that of the principal peak in the chromato- MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 5.5 mL of phosphoric acid add 950 mL of water, adjust to pH 3.0 with triethylamine, and add water to make 1000 mL. To 3 volume of this solution add 2 volume of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of irbesartan is about 13 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of irbesartan are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irbesartan is not more than 1.0z.
To 10 Irbesartan Tablets add 15 mL of water, shake vigorously to disintegrate, and add 150 mL of methanol. besartan (C25 H28 N6O: 428.53). Shake vigorously for 15 minutes, add methanol to make exactly 200 mL, and centrifuge. Pipet V mL of the super- Method of preparation Prepare as directed under Tablets, with Irbesartan. Identification To a quantity of powdered Irbesartan natant liquid, equivalent to about 20 mg of irbesartan (C25H28N6O), and add a mixture of water and acetonitrile for liquid chromatography (3:2) to make exactly 20 mL. Pipet JP XVIII Official Monographs / Irbesartan and Amlodipine Besilate Tablets 1185 ‌2.5 mL of this solution, add a mixture of water and aceto-nitrile for liquid chromatography (3:2) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of irbesartan for assay (separately determine the water <2.48> in the same manner as Irbesartan), and dissolve in methanol to make exactly 10 mL. Pipet 2.5 mL of this solution, add a mixture of water and acetonitrile for liquid chromatography (3:2) to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of irbesartan in each solution. spectra of these peaks exhibit similar intensities of absorp-tion at the same wavelengths. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 237 nm, spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). (2) Perform the test with 5 mL each of the sample solu-tion and standard solution obtained in the Assay (2) as di-rected under Liquid Chromatography <2.01> according to Amount (mg) of irbesartan (C ＝ MS × AT/AS × 16/V 25H 28N6O) in 1 tablet the following conditions: the retention time of the peak of amlodipine in the chromatogram from the sample solution is the same with that of the principal peak in the chromato- MS: Amount (mg) of irbesartan for assay taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 5.5 mL of phosphoric acid add 950 mL of water, adjust to pH 3.0 with triethylamine, and add water to make 1000 mL. To 3 volume of this solution add 2 volume of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of irbesartan is about 13 minutes. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of irbesartan are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irbesartan is not more than 1.0z.
Irinotecan Hydrochloride Injection is an aqueous injection. It contains not less than 95.
Irinotecan Hydrochloride Injection is a clear and pale yellow liquid. It is gradually decomposed by light.
To a volume of Irinotecan Hydrochloride Injection, equivalent to 20 mg of Irinotecan Hydrochloride Hydrate, add water to make 10 mL. To 1 mL of this solution add water to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 253 nm and 257 nm, between 354 nm and 358 nm, and between 368 nm and 372 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—To a volume of Irinotecan Hy-drochloride Injection, equivalent to 40 mg of Irinotecan Hy-drochloride Hydrate, add a mixture of diluted 0.1 mol/L po-tassium dihydrogen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) and 1 mL of 1 mol/L hydrochloric acid TS to make 20 mL, and use this solution as the sample solu-tion. Pipet 1 mL of this solution, add a mixture of diluted 0.1 mol/L potassium dihydrogen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of related substance IA, having the relative retention time of about 0.3 to irinotecan, obtained from the sample solution is not larger than 1/2 times the peak area of irinotecan from the standard solution, the areas of related substances A and B, having the relative retention time of about 0.8, from the sample solution are not larger than 3/10 times the peak area of irinotecan from the standard solution, the area of related substance IB, having the relative retention time of about 1.3, from the sample solution is not larger than 1/3 times the peak area of irinotecan from the standard solution, the areas of related substances C and D, having the relative retention time of about 1.6, and the related substance IC, having the relative retention time of about 2.2, from the sample solu-tion are not larger than 1/5 times the peak area of irinotecan from the standard solution, and the area of the peak other than irinotecan and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of irinotecan from the standard solution. Furthermore, the total area of the peaks other than irinotecan from the sample solution is not larger than 1.5 times the peak area of irinote-can from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Irinotecan Hydrochloride Hydrate. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add a mixture of diluted 0.1 mol/L potassium di-hydrogen phosphate TS (1 in 10), methanol and acetonitrile (6:4:3) to make exactly 20 mL. Confirm that the peak area of irinotecan obtained with 25 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 25 mL of the standard solu-tion. System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of irinotecan are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of irinotecan is not more than 2.0z. Bacterial endotoxins <4.01> Less than 1.8 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Irinotecan Hydrochloride Injec-tion, equivalent to about 20 mg of irinotecan hydrochloride hydrate (C33H38N4O6.HCl.3H2O), and add a mixture of methanol and acetic acid-sodium acetate buffer solution (pH 4.0) (11:9) to make exactly 50 mL. Pipet 10 mL of this solu-tion, add the internal standard solution to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of irinotecan hydrochloride hydrate for assay (separately determine the water <2.48> in the same manner as Irinotecan Hydrochloride Hydrate), dis-solve in a mixture of methanol and acetic acid-sodium ace-tate buffer solution (pH 4.0) (11:9) to make exactly 50 mL. Pipet 10 mL of this solution, add the internal standard solu-tion to make exactly 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of irinotecan to that of the internal standard. Amount (mg) of irinotecan hydrochloride hydrate (C33H38N4O6.HCl.3H2O) ＝ MS × QT/QS × 1.087 MS: Amount (mg) of irinotecan hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—Dissolve 33.3 mg of propyl par-ahydroxybenzoate in a mixture of methanol and acetic acid-sodium acetate buffer solution (pH 4.0) (11:9) to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.01 g of sodium 1-heptanesul- ‌1190 fonate in a mixture of methanol and acetic acid-sodium ace-tate buffer solution (pH 4.0) (11:9) to make 1000 mL. Flow rate: Adjust so that the retention time of irinotecan is about 7 minutes. System Suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, irinotecan and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of irinotecan to that of the internal standard is not more than 1.0z.
イルソグラジンマレイン酸塩細粒
Irsogladine Maleate Fine Granules contain not less than 93.0z and not more than 107.0z of the labeled amount of irsogladine maleate (C9H7Cl2N5.C4H4O4: 372.16). Method of preparation Prepare as directed under Gran-ules, with Irsogladine Maleate. Identification To a quantity of powdered Irsogladine Male-ate Fine Granules, equivalent to 2 mg of Irsogladine Male-ate, add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 2 mg of irsogladine maleate in 5 mL of meth-anol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of petroleum ether, acetone and ace-tic acid (100) (12:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave- length: 254 nm): the spot from the sample solution has the same Rf value as the spot from the standard solution. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Irsogladine Maleate Fine Gran-ules in single-dose packages meet the requirement of the Content uniformity test. Take out the total contents of 1 package of Irsogladine Maleate Fine Granules, add 2 mL of water, add 2 mL metha-nol per mg of irsogladine maleate (C9H7Cl2N5.C4H4O4), sonicate for 10 minutes with occasional shaking, and add water to make exactly V mL so that each mL contains about 40 mg of irsogladine maleate (C9H7Cl2N5.
To a quantity of powdered Irsogladine Male-ate Fine Granules, equivalent to 2 mg of Irsogladine Male-ate, add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 2 mg of irsogladine maleate in 5 mL of meth-anol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of petroleum ether, acetone and ace-tic acid (100) (12:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave- length: 254 nm): the spot from the sample solution has the same Rf value as the spot from the standard solution. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Irsogladine Maleate Fine Gran-ules in single-dose packages meet the requirement of the Content uniformity test. Take out the total contents of 1 package of Irsogladine Maleate Fine Granules, add 2 mL of water, add 2 mL metha-nol per mg of irsogladine maleate (C9H7Cl2N5.C4H4O4), sonicate for 10 minutes with occasional shaking, and add water to make exactly V mL so that each mL contains about 40 mg of irsogladine maleate (C9H7Cl2N5.C4H4O4). Centri-fuge this solution, pipet 1 mL of the supernatant liquid, and add water to make exactly 20 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, discard the first 10 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately about 20 mg of irsogladine maleate for assay, previ-ously dried at 1059C for 4 hours, dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and de-termine the absorbances, AT and AS, at 210 nm. Amount (mg) of irsogladine maleate (C9H7Cl2N5.C4H4O4) ＝ MS × AT/AS × V/500 MS: Amount (mg) of irsogladine maleate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Irsogladine Maleate Fine Granules is not less than 70z. Start the test with an accurately weighed amount of Irso-gladine Maleate Fine Granules, equivalent to about 4 mg of irsogladine maleate (C9H7Cl2N5.C4H4O4), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of irsogladine maleate for assay, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solution, and add water to make ex-actly 20 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank, and determine the absorbances, AT and AS, at 210 nm. Dissolution rate (z) with respect to the labeled amount of irsogladine maleate (C9H7Cl2N5.C4H4O4) ＝ MS/MT × AT/AS × 1/C × 9 MS: Amount (mg) of irsogladine maleate for assay taken MT: Amount (g) of Irsogladine Maleate Fine Granules taken C: Labeled amount (mg) of irsogladine maleate (C9H7Cl2N5.C4H4O4) in 1 g
Weigh accurately an amount of powdered Irsogla-dine Maleate Fine Granules, equivalent to about 5 mg of irsogladine maleate (C9H7Cl2N5.C4H4O4), add exactly 5 mL of the internal standard solution, shake until it is dispersed, ‌1192 and add 5 mL of water. To the solution add 25 mL of ethyl-ene glycol, sonicate for 10 minutes with occasional shaking, and add ethylene glycol to make 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, discard the first 10 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately about 25 mg of irsogladine maleate for assay, previ-ously dried at 1059C for 4 hours, and dissolve in ethylene glycol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add 5 mL of water and ethylene glycol to make 50 mL, and use this so-lution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of irsogladine to that of the internal standard. Amount (mg) of irsogladine maleate (C9H7Cl2N5.C4H4O4) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of irsogladine maleate for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in methanol (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (750:250:3). Flow rate: Adjust so that the retention time of irsogladine test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of petroleum ether, acetone and acetic acid (100) (12:4:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot from the sample solution has the same Rf value as the spot from the standard solution. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Irsogladine Maleate Tablets add 2 mL of water, add 2 mL of methanol per mg of irsogladine maleate (C9H7Cl2N5.C4H4O4), sonicate for 10 minutes with occa-sional shaking, add water to make exactly V mL so that each mL contains about 40 mg of irsogladine maleate (C9H7Cl2N5.C4H4O4). Centrifuge this solution, pipet 1 mL of the supernatant liquid, and add water to make exactly 20 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 20 mg of irsogla-dine maleate for assay, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solution, and add water to make exactly 20 mL. Pipet 2 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and determine the absor- bances, AT and AS, at 210 nm. Amount (mg) of irsogladine maleate (C H Cl N .C H O ) is about 9 minutes. System suitability— ＝ MS × AT/AS × V/500 9 7 2 5 4 4 4 System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, irsogladine and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of irsogladine to that of the internal standard is not more than 1.0z.
Irsogladine Maleate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of irsogladine maleate (C H Cl N .C H O : MS: Amount (mg) of irsogladine maleate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Irsogladine Maleate Tablets is not less than 80z. Start the test with 1 tablet of Irsogladine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.
To a quantity of powdered Irsogladine Male-ate Tablets, equivalent 2 mg of Irsogladine Maleate, add 5 mL of methanol, shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 2 mg of irsogladine maleate in 5 mL of methanol, and use this solution as the standard solution. Perform the <2.24>, using water as the blank, and determine the absor-bances, AT and AS, at 210 nm. Dissolution rate (z) with respect to the labeled amount of irsogladine maleate (C9H7Cl2N5.C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of irsogladine maleate for assay taken C: Labeled amount (mg) of irsogladine maleate (C9H7Cl2N5.C4H4O4) in 1 tablet ‌JP XVIII Official Monographs / Isepamicin Sulfate 1193
Weigh accurately the mass of not less than 20 Irso-gladine Maleate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 5 mg of irsogla-dine maleate (C9H7Cl2N5.C4H4O4), add exactly 5 mL of the internal standard solution, shake until it is dispersed, and add 5 mL of water. To this solution add 25 mL of ethylene glycol, sonicate for 10 minutes with occasional shaking, and add ethylene glycol to make 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, discard the first 10 mL of the filtrate, and use the subse-quent filtrate as the sample solution. Separately, weigh accu-rately about 25 mg of irsogladine maleate for assay, previ-ously dried at 1059C for 4 hours, and dissolve in ethylene glycol to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add 5 mL of water and ethylene glycol to make 50 mL, and use this so-lution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of irsogladine to that of the internal standard. Amount (mg) of irsogladine maleate (C H Cl N .C H O ) Isepamicin Sulfate
Isepamicin Sulfate Injection is an aqueous injection. It contains not less than 90.
Isepamicin Sulfate Injection is a clear, color-less liquid.
To a volume of Isepamicin Sulfate Injection, equivalent to 20 mg (potency) of Isepamicin Sulfate, add water to make 10 mL, and use this solution as the sample so-lution. Separately, dissolve an amount of Isepamicin Sulfate RS, equivalent to 20 mg (potency) in 10 mL of water, and use this solution as the standard solution. Proceed with these solutions as directed in the Identification (2) under Isepami-cin Sulfate. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> 5.5 – 7.5.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area of the sample solution by the automatic integration method and calculate the amounts of their peaks by the area percentage method: the amount of isoserine, having the relative retention time of about 0.3 to isepamicin, is not more than 2.0z, and the amount of gen-tamicin B, having the relative retention time of about 1.3, is not more than 4.0z. For the peak area of gentamicin B, multiply the correction factor, 1.11. ‌JP XVIII Official Monographs / Isoflurane 1195 Operating conditions— Apparatus, detector, column, column temperature, reac-tion coil, mobile phase, reaction reagent, reaction tempera-ture, flow rate of mobile phase, and flow rate of reaction reagent: Proceed as directed in the operating conditions in the Assay under Isepamicin Sulfate. Time span of measurement: About 2 times as long as the retention time of isepamicin. System suitability— System performance and system repeatability: Proceed as directed in the Assay under Isepamicin Sulfate. Test for required detectability: To 1 mL of the sample so-lution add water to make 10 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solu-tion for system suitability test, and add water to make ex-actly 10 mL. Confirm that the peak area of isepamicin ob-tained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suitability test. Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Isepamicin Sulfate Injection, equivalent to about 0.2 g (potency) of Isepamicin Sulfate, add water to make exactly 100 mL. Pipet 10 mL of this solu-tion, add water to make exactly 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Isepamicin Sulfate RS, equivalent to about 20 mg (potency), dissolve in water to make exactly 100 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Isepamicin Sulfate. Amount [mg (potency)] of isepamicin (C22H43N5O12) ＝ MS × AT/AS × 10 MS: Amount [mg (potency)] of Isepamicin Sulfate RS taken
Isoniazid Injection is an aqueous injection. It contains not less than 95.
Isoniazid Injection occurs as a clear, colorless liquid. pH: 6.5 – 7.5
To a volume of Isoniazid Injection, equiva-lent to 20 mg of Isoniazid, and add water to make 200 mL. To 5 mL of the solution add 1 mL of 0.1 mol/L hydrochloric acid TS and water to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 264 nm and 268 nm. Bacterial endotoxins <4.01> Less than 0.50 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Isoniazid Injec-tion, equivalent to about 50 mg of isoniazid (C6H7N3O), add water to make exactly 100 mL. Pipet 5 mL of the solution, add exactly 5 mL of the internal standard solution and the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of isoniazid for assay, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solu-tion and the mobile phase to make 50 mL, and use this solu-tion as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of isoniazid to that of the internal standard. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of isoniazid to that of the internal standard is not more than 1.3z.
Isoniazid Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of iso-niazid (C6H7N3O: 137.14). Method of preparation Prepare as directed under Tablets, with Isoniazid. Identification Take a quantity of powdered Isoniazid Tablets, equivalent to 20 mg of Isoniazid, add 200 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of 0.1 mol/L hydrochloric acid TS and water to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 264 nm and 268 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isoniazid Tablets add water to make exactly V mL so that each mL contains about 0.
Take a quantity of powdered Isoniazid Tablets, equivalent to 20 mg of Isoniazid, add 200 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of 0.1 mol/L hydrochloric acid TS and water to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 264 nm and 268 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isoniazid Tablets add water to make exactly V mL so that each mL contains about 0.5 mg of isoniazid (C6H7N3O), and shake well to disintegrate. Filter this solu-tion, discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of isoniazid (C6H7N3O) ＝ M S × QT/QS Amount (mg) of isoniazid (C6H7N3O) ＝ MS × AT/AS × V/100 MS: Amount (mg) of isoniazid for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave- MS: Amount (mg) of isoniazid for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Isoniazid Tablets is not less than 75z. ‌1202 Start the test with 1 tablet of Isoniazid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size of not more than 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 50 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 0.1 g of isoniazid for assay, previously dried at 1059C for 2 hours, dissolve in water to make exactly 100 mL, then pipet 5 mL of this solution, add water to make exactly 50 mL, and then pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 267 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of isoniazid (C6H7N3O) ＝ MS × AT/AS × 1/C × 90 MS: Amount (mg) of isoniazid for assay taken C: Labeled amount (mg) of isoniazid (C6H7N3O) in 1 tablet
Weigh accurately and powder not less than 20 Isoniazid Tablets. Weigh accurately a quantity of the pow- with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isoniazid is not more than 1.0z.
Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension
イソフェンインスリン ヒト(遺伝子組換え)水性懸濁注射液
Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension is an aqueous suspen-sion for injection. It contains not less than 95.
Isophane Insulin Human (Genetical Recombi-nation) Injectable Aqueous Suspension is a white aqueous suspension. When allowed to stand, it separates into a white precipitate and colorless supernatant liquid, and the precipi-tate returns to the suspension state on gentle shaking. When it is examined microscopically, the precipitate most-ly consists of fine, oblong crystals of 1 to 30 mm in major axis, and does not contain amorphous substances or large aggregates.
Adjust Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension to pH be-tween 2.5 and 3.0 with dilute hydrochloric acid: the precipi-tate dissolves, and the solution is clear and colorless. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Desamido substance—Perform the test with 20 mL of the sample solution obtained in the Assay (1) as di-rected under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of the peak, having the relative retention time of about 1.3 to insu-lin human, is not more than 1.5z. Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: Proceed as directed in the system suitability in the Assay (1). Test for required detectability: Pipet 1 mL of the sample solution, add 0.01 mol/L hydrochloric acid TS to make ex-actly 50 mL. Confirm that the peak area of insulin human obtained with 20 mL of this solution is equivalent to 1.4 to 2.6z of that with 20 mL of the sample solution. System repeatability: Dissolve Insulin Human RS in 0.01 mol/L hydrochloric acid TS so that each mL contains about 4 Insulin Units. When the test is repeated 6 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of insulin human is not more than 2.0z. Dissolved insulin human—Centrifuge Isophane Insu-lin Human (Genetical Recombination) Injectable Aqueous Suspension, and use the supernatant liquid as the sample so-lution. Separately, dissolve exactly Insulin Human RS in 0.01 mol/L hydrochloric acid TS to make a solution so that each mL contains about 1.0 Insulin Units, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of insulin human by the automatic integration method, and calculate the amount of dissolved insulin human by the following equation: not more than 0.5 Insulin Units per mL. Amount (mg) of dissolved insulin human (Insulin Unit /mL) ＝ (MS × F )/D × AT/AS MS: Amount (mg) of Insulin Human RS taken ‌JP XVIII Official Monographs / Biphasic Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension 1159 F: Labeled unit (Insulin Unit /mg) of Insulin Human RS D: Volume (mL) of 0.01 mol/L hydrochloric acid TS used to dissolve Insulin Human RS Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: When the procedure is run with 20 mL of insulin human desamido substance-containing TS under the above operating conditions, insulin human and in-sulin human desamido substance are eluted in this order with the resolution between these peaks being not less than 2.0, and the symmetry factor of the peak of insulin human is not more than 1.6. System repeatability: When the test is repeated 4 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of insulin human is not more than 6.0z. High-molecular mass protein—Take a suitable volume of gently shaken Isophane Insulin Human (Genetical Recombination) Injectable Aqueous Suspension, add 4 mL of 6 mol/L hydrochloric acid TS for each mL of the suspen-sion, and mix until the solution becomes clear. Perform the test with 100 mL of this solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area per-centage method: the total amount of the peaks other than in-sulin human is not more than 2.5z. Operating conditions— Detector, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Insulin Human (Genetical Recombination). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with hydrophilic silica gel for liquid chromatography. Time span of measurement: From the retention time cor-responding to the exclusion volume of the size-exclusion column to the completion of the elution of insulin human. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) under Insulin Human (Genetical Recombination). Test for required detectability: Pipet 1 mL of the sample solution, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Confirm that the peak area of insulin human obtained with 100 mL of this solution is equivalent to 1.4 to 2.6z of that with 100 mL of the sample solution. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Insulin human—Pipet 10 mL of gently shaken Isophane Insulin Human (Genetical Recombination) Injecta-ble Aqueous Suspension, and add exactly 40 mL of 6 mol/L hydrochloric acid TS. Pipet 2 mL of this solution, add 0.01 mol/L hydrochloric acid TS to make exactly 5 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay under Insulin Human (Genetical Recom-bination). Amount (Insulin Unit) of insulin human (C257H383N65O77S6) in 1 mL ＝ (MS × F )/D × (ATI ＋ ATD)/(ASI ＋ ASD) × 1.004 × 5/2 MS: Amount (mg) of Insulin Human RS taken F: Labeled unit (Insulin Unit /mg) of Insulin Human RS D: Volume (mL) of 0.01 mol/L hydrochloric acid TS used to dissolve Insulin Human RS (2) Zinc—Pipet a volume of gently shaken Isophane In-sulin Human (Genetical Recombination) Injectable Aqueous Suspension, equivalent to 300 Insulin Units, and add 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. If nec-essary, further add 0.01 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the sample so-lution. Separately, pipet a suitable volume of Standard Zinc Solution for Atomic Absorption Spectroscopy, dilute with 0.01 mol/L hydrochloric acid TS to make three solutions containing 0.20 mg, 0.60 mg and 1.20 mg of zinc (Zn: 65.38) in each mL, respectively, and use these solutions as the stand-ard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectroscopy <2.23> according to the following conditions, using 0.01 mol/L hydrochloric acid TS as the blank, and cal-culate the content of zinc in the sample solution by using the calibration curve obtained from the absorbances of the standard solutions. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Zinc hollow cathode lamp. Wavelength: 213.9 nm.
Isosorbide Dinitrate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of isosorbide dinitrate (C6H8N2O8: 236.14). Method of preparation Prepare as directed under Tablets, with Isosorbide Dinitrate. Identification Weigh a quantity of powdered Isosorbide Dinitrate Tablets, equivalent to 0.1 g of Isosorbide Dinitrate, add 50 mL of diethyl ether, shake well, and filter. Measure 5 mL of the filtrate, evaporate to dryness cautiously, add 1 mL of water to the residue, and dissolve by adding 2 mL of sul-furic acid cautiously. After cooling, superimpose 3 mL of iron (II) sulfate TS, and allow to stand for 5 to 10 minutes: a brown ring is produced at the zone of contact. Purity Nitrate—Weigh accurately a quantity of powdered Isosorbide Dinitrate Tablets, equivalent to 50 mg of Isosor-bide Dinitrate, transfer to a separator, add 30 mL of tolu-ene, shake thoroughly, extract with three 20-mL portions of water, and proceed as directed in Purity (3) under Isosorbide Dinitrate. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isosorbide Dinitrate Tablets add 1 mL of ‌1206 water, and shake to disintegrate. To this solution add a mix-ture of water and methanol (1:1) to make exactly V mL so that each mL contains about 0.
Weigh a quantity of powdered Isosorbide Dinitrate Tablets, equivalent to 0.1 g of Isosorbide Dinitrate, add 50 mL of diethyl ether, shake well, and filter. Measure 5 mL of the filtrate, evaporate to dryness cautiously, add 1 mL of water to the residue, and dissolve by adding 2 mL of sul-furic acid cautiously. After cooling, superimpose 3 mL of iron (II) sulfate TS, and allow to stand for 5 to 10 minutes: a brown ring is produced at the zone of contact.
Nitrate—Weigh accurately a quantity of powdered Isosorbide Dinitrate Tablets, equivalent to 50 mg of Isosor-bide Dinitrate, transfer to a separator, add 30 mL of tolu-ene, shake thoroughly, extract with three 20-mL portions of water, and proceed as directed in Purity (3) under Isosorbide Dinitrate. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isosorbide Dinitrate Tablets add 1 mL of ‌1206 water, and shake to disintegrate. To this solution add a mix-ture of water and methanol (1:1) to make exactly V mL so that each mL contains about 0.1 mg of isosorbide dinitrate (C6H8N2O8), and shake for 10 minutes. Centrifuge this solu-tion, and use the supernatant liquid as the sample solution. Proceed as directed in the Assay. Amount (mg) of isosorbide dinitrate (C6H8N2O8) ＝ MS × AT/AS × V × 1/500 MS: Amount (mg) of isosorbide dinitrate for assay taken, calculated on the anhydrous basis Disintegration <6.09> It meets the requirement. For Sublingual Tablets, the time limit of the test is 2 minutes, and omit the use of the disk.
Weigh accurately the mass of not less than 20 tablets of Isosorbide Dinitrate Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 5 mg of isosorbide dinitrate (C6H8N2O8), add a mixture of water and methanol (1:1) to make exactly 50 mL, and shake for 10 minutes. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of isosorbide dinitrate for assay (separately, de-termine the water <2.48> in the same manner as Isosorbide Dinitrate), dissolve in a mixture of water and methanol (1:1) to make exactly 100 mL. Pipet 10 mL of this solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of isosorbide dinitrate in each so-lution. Amount (mg) of isosorbide dinitrate (C H N O ) Isosorbide Mononitrate 70%/Lactose 30%
Isosorbide Mononitrate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of isosorbide mononitrate (C6H9NO6: 191.14). Method of preparation Prepare as directed under Tablets, with Isosorbide Mononitrate 70z/Lactose 30z. Identification Shake well a portion of powdered Isosorbide Mononitrate Tablets, equivalent to 50 mg of isosorbide mononitrate (C6H9NO6), with 5 mL of acetone, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 10 mg of isosorbide mononitrate for assay in 1 mL of acetone, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (2:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly a solution of potassium permanganate in potassium hydroxide TS (1 in 50), and allow to stand for about 50 minutes: the principal spot obtained with the sample solution and the spot from the standard solution are yellow, and their Rf values are the same. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isosorbide Mononitrate Tablets add 30 mL of water, allow standing to disintegrate the tablet, and dis-perse the fine particles by sonicating. Add exactly V/10 mL of the internal standard solution, and add water to make V mL so that each mL contains about 0.
Shake well a portion of powdered Isosorbide Mononitrate Tablets, equivalent to 50 mg of isosorbide mononitrate (C6H9NO6), with 5 mL of acetone, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 10 mg of isosorbide mononitrate for assay in 1 mL of acetone, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and hexane (2:1) to a distance of about 15 cm, and air-dry the plate. Spray evenly a solution of potassium permanganate in potassium hydroxide TS (1 in 50), and allow to stand for about 50 minutes: the principal spot obtained with the sample solution and the spot from the standard solution are yellow, and their Rf values are the same. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Isosorbide Mononitrate Tablets add 30 mL of water, allow standing to disintegrate the tablet, and dis-perse the fine particles by sonicating. Add exactly V/10 mL of the internal standard solution, and add water to make V mL so that each mL contains about 0.2 mg of isosorbide mononitrate (C6H9NO6). Centrifuge this solution, filter the supernatant liquid through a membrane filter with a pore MS: Amount (mg) of isosorbide mononitrate for assay taken Internal standard solution—A solution of benzyl alcohol (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Isosorbide Mononitrate Tablets is not less than 85z. Start the test with 1 tablet of Isosorbide Mononitrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of isosorbide mononi-trate (C6H9NO6), and use this solution as the sample solu-tion. Separately, weigh accurately about 22 mg of isosorbide mononitrate for assay, previously dried in vacuum (silica gel) for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of isosorbide mononitrate in each solution. Dissolution rate (z) with respect to the labeled amount of isosorbide mononitrate (C6H9NO6) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of isosorbide mononitrate for assay taken C: Labeled amount (mg) of isosorbide mononitrate (C6H9NO6) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of isosorbide mononitrate are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isosorbide mononitrate is not more than 2.0z.
Weigh accurately the mass of not less than 20 Isosorbide Mononitrate Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 20 mg of isosorbide mononitrate (C6H9NO6), add 30 mL of water, and disperse the fine particles by sonicating. Add exactly 10 mL of the internal standard solution and water to make 50 mL. JP XVIII Official Monographs / Isoxsuprine Hydrochloride 1209 Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding ‌0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of isosorbide mononitrate for assay, previously dried, and dissolve in 30 mL of water, add exactly 10 mL of the internal standard solution, then, add water to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of isosorbide mononitrate to that of the internal standard. Amount (mg) of isosorbide mononitrate (C6H9NO6) ＝ MS × QT/QS MS: Amount (mg) of isosorbide mononitrate for assay taken Internal standard solution—A solution of benzyl alcohol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and methanol (4:1). Flow rate: Adjust so that the retention time of isosorbide mononitrate is about 4.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, isosorbide mononitrate and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of isosorbide mononitrate to that of the inter-nal standard is not more than 1.0z.
Isotonic Sodium Chloride Solution is an aqueous in-jection. It contains not less than 0.
Isotonic Sodium Chloride Solution is a clear, colorless liquid. It has a slightly saline taste.
Isotonic Sodium Chloride Solution responds to Qualitative Tests <1.09> for sodium salt and for chloride. pH <2.54> 4.5 – 8.0
(1) Heavy metals <1.07>—Concentrate 100 mL of Isotonic Sodium Chloride Solution to about 40 mL on a water bath, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution and 2 mL of dilute acetic acid, and add water to make 50 mL (not more than 0.3 ppm). (2) Arsenic <1.11>—Prepare the test solution with 20 mL of Isotonic Sodium Chloride Solution, and perform the test (not more than 0.1 ppm). Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly 20 mL of Isotonic Sodium Chloride Solution, add 30 mL of water, and titrate <2.50> with 0.1 mol/L silver nitrate VS with vigorous shaking (indicator: 3 drops of fluorescein sodium TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl
Isoxsuprine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of isoxsuprine hydrochloride (C18H23NO3.HCl: 337.84). Method of preparation Prepare as directed under Tablets, with Isoxsuprine Hydrochloride. Identification To a quantity of powdered Isoxsuprine Hy-drochloride Tablets, equivalent to 10 mg of Isoxsuprine Hy-drochloride, add 150 mL of water, shake, and then add water to make 200 mL. Centrifuge this solution, filter the su-pernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of filtrate, and determine the absorption spectrum of the subsequent fil-trate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 267 nm and 271 nm, and between 272 nm and 276 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Add methanol to 1 tablet of Isoxsuprine Hydrochloride Tablets, and shake to disintegrate. Add methanol to make exactly V mL so that each mL contains about 0.
To a quantity of powdered Isoxsuprine Hy-drochloride Tablets, equivalent to 10 mg of Isoxsuprine Hy-drochloride, add 150 mL of water, shake, and then add water to make 200 mL. Centrifuge this solution, filter the su-pernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of filtrate, and determine the absorption spectrum of the subsequent fil-trate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 267 nm and 271 nm, and between 272 nm and 276 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Add methanol to 1 tablet of Isoxsuprine Hydrochloride Tablets, and shake to disintegrate. Add methanol to make exactly V mL so that each mL contains about 0.4 mg of isoxsuprine hydrochloride (C18H23NO3.HCl). Centrifuge this solution, and use the supernatant liquid as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of isoxsuprine hydrochloride (C18H23NO3.HCl) solution. Separately, weigh accurately about 28 mg of isox-suprine hydrochloride for assay, previously dried at 1059C for 1 hour, and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of isoxsuprine in each solution. Dissolution rate (z) with respect to the labeled amount of isoxsuprine hydrochloride (C18H23NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of isoxsuprine hydrochloride for assay taken C: Labeled amount (mg) of isoxsuprine hydrochloride (C18H23NO3.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of isoxsuprine are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isoxsuprine is not more than 2.0z.
Weigh accurately not less than 20 Isoxsuprine Hy-drochloride Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 40 mg of isoxsuprine hydrochloride (C18H23NO3.HCl), add 60 mL of methanol, shake for 20 minutes, and then add methanol to make ex-actly 100 mL. Centrifuge a portion of this solution, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of isoxsuprine hy-drochloride for assay, previously dried at 1059C for 1 hour, and dissolve in methanol to make exactly 100 mL. Filter through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of isoxsuprine in each solution. ＝ MS × AT/AS × V × 1/100 ‌JP XVIII Official Monographs / Itraconazole 1211 Amount (mg) of isoxsuprine hydrochloride (C18H23NO3.HCl) ＝ MS × AT/AS MS: Amount (mg) of isoxsuprine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 269 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.3 g of diammonium hydrogen phosphate and 3.2 g of sodium 1-pentane sulfonate in water to make 1000 mL, and adjust to pH 2.5 with phosphoric acid. To 600 mL of this solution add 400 mL of methanol. Flow rate: Adjust so that the retention time of isoxsuprine is about 9 minutes. System suitability— System performance: To exactly 1 mL of the standard so-lution add the mobile phase to make exactly 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of isoxsuprine are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of isoxsuprine is not more than 1.0z.
Josamycin Tablets contain not less than 90.0z and not more than 110.0z of the labeled potency of josamycin (C42H69NO15: 827.99). Method of preparation Prepare as directed under Tablets, with Josamycin. Identification To a quantity of powdered Josamycin Tablets, equivalent to 10 mg (potency) of Josamycin, add 100 mL of methanol, shake vigorously, and centrifuge. To 5 mL of the supernatant liquid, add methanol to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 229 nm and 233 nm. Loss on drying <2.41> Not more than 5.0z (0.5 g, in vacu-um, 609C, 3 hours). Uniformity of dosage units <6.02>—Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Josamycin Tablets, add 5 mL of water, and shake vigorously to disintegrate the tablet. Add metha-nol and sonicate to disperse the particles, add methanol to make exactly V mL so that each mL contains about 2 mg (potency) of Josamycin, and centrifuge.
To a quantity of powdered Josamycin Tablets, equivalent to 10 mg (potency) of Josamycin, add 100 mL of methanol, shake vigorously, and centrifuge. To 5 mL of the supernatant liquid, add methanol to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 229 nm and 233 nm. Loss on drying <2.41> Not more than 5.0z (0.5 g, in vacu-um, 609C, 3 hours). Uniformity of dosage units <6.02>—Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take 1 tablet of Josamycin Tablets, add 5 mL of water, and shake vigorously to disintegrate the tablet. Add metha-nol and sonicate to disperse the particles, add methanol to make exactly V mL so that each mL contains about 2 mg (potency) of Josamycin, and centrifuge. Pipet 3 mL of the supernatant liquid, and add methanol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make ex-actly 50 mL, and use this solution as the sample solution. Separately, accurately weigh about 50 mg (potency) of Josamycin RS, dissolve in 5 mL of water and methanol to make exactly 25 mL. Pipet 3 mL of this solution, and add methanol to make exactly 100 mL. Pipet 10 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 231 nm as directed under Ultraviolet-visible Spectrophotometry X <2.24>. However, — in the formula for calculation of accep- tance value is the result of the assay. Amount [mg (potency)] of josamycin (C42H69NO15) ＝ MS × AT/AS × V/25 MS: Amount [mg (potency)] of Josamycin RS taken Disintegration <6.09> Perform the test using the disk: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, and standard solu-tions—Proceed as directed in the Assay under Josamycin. Sample solutions—Weigh accurately the mass of not less than 20 Josamycin Tablets and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.3 g (po-tency) of Josamycin, add 50 mL of methanol, shake vigor-ously, and add water to make exactly 1000 mL. Take exactly an appropriate amount of this solution, add water to prepare solutions containing 30 mg (potency) and 7.5 mg (potency) per mL, and use these solutions as the high and the low con-centration sample solutions, respectively.
Kainic Acid and Santonin Powder contains not less than 9.
Kainic Acid and Santonin Powder occurs as a white powder.
(1) Shake 1 g of Kainic Acid and Santonin Powder with 10 mL of chloroform, and filter [use the residue for the test (2)]. Distil off the chloroform of the fil-trate, and dissolve the residue in 2 mL of potassium hydrox-ide-ethanol TS: a red color is produced (santonin). (2) Shake the residue obtained in (1) with 20 mL of warm water, filter, and to 1 mL of the filtrate add 10 mL of water and 1 mL of ninhydrin-L-ascorbic acid TS. Warm in a water bath between 609C and 709C for 5 minutes: a yellow color is produced (kainic acid).
(1) Clarity and color of solution—Dissolve 0.10 g of Kainic Acid Hydrate in 10 mL of water: the solution is clear and colorless. (2) Chloride <1.03>—Take 0.5 g of Kainic Acid Hydrate in a platinum crucible, dissolve in 5 mL of sodium carbonate TS, and evaporate on a water bath to dryness. Heat the cru-cible slowly at first, and then ignite until the sample is almost incinerated. After cooling, add 12 mL of dilute nitric acid to the residue, dissolve by warming, and filter. Wash the residue with 15 mL of water, combine the washings and the filtrate, and add water to make 50 mL. Perform the test using this solution as the test solution. Control solution: Add 5 mL of sodium carbonate TS to 0.30 mL of 0.01 mol/L hydrochloric acid VS, and proceed as Kainic Acid and Santonin Powder contains not less than 9.0z and not more than 11.0z of santonin (C15H18O3: 246.30), and not less than 1.80z and not more than 2.20z of kainic acid hydrate (C10H15NO4.H2O: 231.25). Method of preparation Santonin 100 g Kainic Acid Hydrate 20 g Starch, Lactose Hydrate or their mixture a sufficient quantity To make 1000 g Prepare as directed under Powders, with the above ingre- ‌1216 dients. Description Kainic Acid and Santonin Powder occurs as a white powder. Identification (1) Shake 1 g of Kainic Acid and Santonin Powder with 10 mL of chloroform, and filter [use the residue for the test (2)]. Distil off the chloroform of the fil-trate, and dissolve the residue in 2 mL of potassium hydrox-ide-ethanol TS: a red color is produced (santonin). (2) Shake the residue obtained in (1) with 20 mL of warm water, filter, and to 1 mL of the filtrate add 10 mL of water and 1 mL of ninhydrin-L-ascorbic acid TS. Warm in a water bath between 609C and 709C for 5 minutes: a yellow color is produced (kainic acid).
(1) Santonin—Weigh accurately about 0.25 g of Kainic Acid and Santonin Powder and about 25 mg of santo-nin for assay, add 20 mL each of ethanol (95), shake thor-oughly for 5 minutes, and filter. Wash the residue with three 10-mL portions of ethanol (95), and filter. Combine the fil-trate and the washings, and add ethanol (95) to make exactly 50 mL. Pipet 2 mL each of these solutions, add ethanol (95) to make exactly 100 mL, and use these solutions as the sam-ple solution and the standard solution, respectively. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 240 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of santonin (C15H18O3) ＝ MS × AT/AS MS: Amount (mg) of santonin for assay taken (2) Kainic acid—Weigh accurately about 1.25 g of Kainic Acid and Santonin Powder, add 20 mL of diluted pyridine (1 in 10), shake thoroughly for 5 minutes, and filter. Wash the residue with three 10-mL portions of diluted pyridine (1 in 10), and filter. Combine the filtrate and the washings, and add diluted pyridine (1 in 10) to make exactly 50 mL. Pipet 2 mL of this solution, add diluted pyridine (1 in 10) to make exactly 25 mL, and use this solution as the sample so-lution. Separately, dissolve about 25 mg of kainic acid hy-drate for assay, previously dried at 1059C for 4 hours and accurately weighed, in diluted pyridine (1 in 10) to make ex-actly 50 mL. Pipet 2 mL of this solution, add diluted pyri-dine (1 in 10) to make exactly 25 mL, and use this solution as the standard solution. Pipet 2 mL each of the sample solu-tion and standard solution, add 2 mL of ninhydrin-L-ascor-bic acid TS, and heat on a water bath for 30 minutes. After cooling immediately, shake vigorously for 2 minutes, add water to make exactly 20 mL, and allow to stand for 15 minutes. Determine the absorbances, AT and AS, of these so-lutions at 425 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using the solution prepared in the same manner with 2 mL of diluted pyridine (1 in 10) instead of the sample solution as the blank. Amount (mg) of kainic acid hydrate (C10H15NO4.H2O) ＝ MS × AT/AS × 1.085 MS: Amount (mg) of kainic acid hydrate for assay taken
Ketoconazole Cream contains not less than 95.
To a quantity of Ketoconazole Cream, equivalent to 0.1 g of Ketoconazole, add 20 mL of 2-propanol, shake for 20 minutes, centrifuge, and use the su-pernatant liquid as the sample solution. Separately, dissolve 25 mg of ketoconazole in 5 mL of 2-propanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, methanol, water and ammonia so-lution (28) (40:40:25:2:1) to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution.
Weigh accurately an amount of Ketoconazole Cream, equivalent to about 25 mg of ketoconazole (C26H28Cl2N4O4), dissolve in methanol to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 4 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of ketoconazole for assay, previously dried at 1059C for 4 hours, and dissolve in metha-nol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 4 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of ketocona-zole to that of the internal standard. Amount (mg) of ketoconazole (C26H28Cl2N4O4) ＝ MS × QT/QS ‌JP XVIII Official Monographs / Ketoconazole Solution 1223 MS: Amount (mg) of ketoconazole for assay taken ‌Internal standard solution—A solution of xanthone in meth-anol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To ammonium acetate solution (1 in 200) add acetic acid (100) to adjust the pH to 5.0. To 250 mL of this solution add 750 mL of methanol. Flow rate: Adjust so that the retention time of ketocona-zole is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ketoconazole are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ketoconazole to that of the internal stand-ard is not more than 1.0z.
Ketoconazole Solution is a liquid for external use. Ketoconazole Solution contains not less than 95.
Ketoconazole Solution is a clear liquid.
To a volume of Ketoconazole Solution, equivalent to 10 mg of Ketoconazole, add methanol to make 10 mL, and use this solution as the sample solution. Sepa-rately, dissolve 10 mg of ketoconazole in 10 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo- ‌1224 rescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane, methanol, water and ammonia solution (28) (40:40:30:2:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. pH Being specified separately when the drug is granted ap-proval based on the Law.
To an exact amount of Ketoconazole Solution, equivalent to about 10 mg of ketoconazole (C26H28Cl2N4O4), add exactly 5 mL of the internal standard solution, and add 15 mL of methanol. To 1 mL of this solution add methanol to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of ketoconazole for assay, previously dried at 1059C for 4 hours, and dis-solve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solu-tion, add methanol to make 20 mL. Take 1 mL of this solu-tion, add methanol to make 25 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ketoconazole to that of the internal standard. Amount (mg) of ketoconazole (C26H28Cl2N4O4) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of ketoconazole for assay taken Internal standard solution—A solution of bifonazole in methanol (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of diisopropyla-mine in methanol (1 in 500), ammonium acetate solution (1 in 200) and acetic acid (100) (1800:600:1). Flow rate: Adjust so that the retention time of ketocona-zole is about 11 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, ketoconazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ketoconazole to that of the internal stand-ard is not more than 1.0z.
L-Arginine Hydrochloride Injection is an aqueous injection. It contains not less than 9.
L-Arginine Hydrochloride Injection is a clear, colorless liquid.
(1) To 5 mL of a solution of L-Arginine Hydrochloride Injection (1 in 100) add 1 mL of ninhydrin TS, and heat for 3 minutes: a blue-purple color develops. (2) To 5 mL of a solution of L-Arginine Hydrochloride Injection (1 in 10) add 2 mL of sodium hydroxide TS and 1 to 2 drops of a solution of 1-naphthol in ethanol (95) (1 in 1000), allow to stand for 5 minutes, and add 1 to 2 drops of sodium hypochlorite TS: a red-orange color develops. pH <2.54> 5.0 – 6.0 Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet 20 mL of L-Arginine Hydrochloride Injection, add 7.5 mol/L hydrochloric acid TS to make exactly 100 mL, and determine the optical rotation aD as directed under Optical Rotation Determination <2.49> at 20 ± 19C in a 100-mm cell. Amount (mg) of L-arginine hydrochloride (C6H14N4O2.HCl) ＝ aD × 4444
L-Carbocisteine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of L-carbocisteine (C5H9NO4S: 179.19). Method of Preparation Prepare as directed under Tablets, with L-Carbocisteine. Identification Powder L-Carbocisteine Tablets. To a por-tion of the powder, equivalent to 0.18 g of L-Carbocisteine, add 50 mL of water, stir for 10 minutes, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and heat in a water bath for 3 minutes: a purple color develops. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 250-mg tablet and in 30 minutes of 500-mg ‌JP XVIII Official Monographs / Carbon Dioxide 627 tablet are not less than 80z and not less than 85z, respec-tively. Start the test with 1 tablet of L-Carbocisteine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 0.
Powder L-Carbocisteine Tablets. To a por-tion of the powder, equivalent to 0.18 g of L-Carbocisteine, add 50 mL of water, stir for 10 minutes, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and heat in a water bath for 3 minutes: a purple color develops. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 250-mg tablet and in 30 minutes of 500-mg ‌JP XVIII Official Monographs / Carbon Dioxide 627 tablet are not less than 80z and not less than 85z, respec-tively. Start the test with 1 tablet of L-Carbocisteine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 0.14 mg of L-carbocisteine (C5H9NO4S), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of L-carbocisteine for assay, previously dried at 1059C for 2 hours, and dis-solve in the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following condition, and determine the peak areas, AT and AS, of L-carbocisteine in each solution. Dissolution rate (z) with respect to the labeled amount of L-carbocisteine (C H NO S) length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Diluted trifluoroacetic acid (1 in 1000). Flow rate: Adjust so that the retention time of L-carbo- cisteine is about 2 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, L-carbocisteine and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of L-carbocisteine to that of the internal standard is not more than 1.0z. 5 9 4 ＝ MS × AT/AS × V?/V × 1/C × 450 MS: Amount (mg) of L-carbocisteine for assay taken C: Labeled amount (mg) of L-carbocisteine (C5H9NO4S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of L-carbocisteine are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of L-carbocisteine is not more than 1.0z.
To 10 L-Carbocisteine Tablets add 220 mL of 0.5 mol/L hydrochloric acid TS, stir for 30 minutes, add 0.5 mol/L hydrochloric acid TS to make exactly 250 mL, and stir additionally for 30 minutes. Filter this solution, discard the first 20 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add (V—50)/25 mL of 0.5 mol/L hydrochloric acid TS, then add exactly V/25 mL of the internal standard solu-tion, add water to make V mL so that each mL contains about 0.4 mg of L-carbocisteine (C5H9NO4S), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of L-carbocisteine for assay, previously dried at 1059C for 2 hours, add 2 mL of 0.5 mol/L hydrochloric acid TS, and exactly 2 mL of the internal standard solution. Then add water to dissolve to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of L-carbocisteine to that of the internal standard. Amount (mg) of L-carbocisteine (C H NO S) in 1 tablet
イソロイシン・ロイシン・バリン顆粒
Dissolve an amount of powdered L-Isoleu-cine, L-Leucine and L-Valine Granules, equivalent to about 92 mg of L-Isoleucine, in the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, dis-solve 0.46 g of L-isoleucine, 0.92 g of L-leucine and 0.55 g of L-valine in the mobile phase to make 100 mL. Take 10 mL of this solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and the standard solution as directed under Liquid Chromatography <2.01> according to the following conditions: the retention times of the peak in the chromatograms obtained from the sample solution and the standard solution are the same. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 31.2 g of sodium dihydrogen phos-phate dihydrate in 1000 mL of water, and adjust to pH 2.8 with phosphoric acid. To 970 mL of this solution add 30 mL of acetonitrile. rected in the Assay. Amount (mg) of L-isoleucine (C6H13NO2) ＝ MSa × QTa/QSa × V/50 Amount (mg) of L-leucine (C6H13NO2) ＝ MSb × QTb/QSb × V/50 Amount (mg) of L-valine (C5H11NO2) ＝ MSc × QTc/QSc × V/50 MSa: Amount (mg) of L-isoleucine for assay taken MSb: Amount (mg) of L-leucine for assay taken MSc: Amount (mg) of L-valine for assay taken Internal standard solution—A solution of glycine in 0.1 mol/L hydrochloric acid TS (1 in 20). Disintegration <6.09> It meets the requirement. Carry out the test for 15 minutes.
Powder the total amount of the content of not less than ten packages of L-Isoleucine, L-Leucine and L-Valine Granules. Weigh accurately a portion of the powder, equiva-lent to about 0.95 g of L-isoleucine (C6H13NO2), add exactly 10 mL of the internal standard solution, dissolve in 0.1 mol/L hydrochloric acid TS to make 250 mL. To 2 mL of this solution add 0.02 mol/L hydrochloric acid TS to make 200 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.2 g of L-isoleucine for assay, about 0.4 g of L-leucine for assay and about 0.24 g of L-valine for assay, previously these are dried at 1059C for 3 hours, add exactly 2 mL of the internal standard solution, dissolve in 0.1 mol/L hydrochloric acid TS to make 100 mL. To 2 mL of this solution add 0.02 mol/L hydrochloric acid TS to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Calculate the ratios, QTa, QTb and QTc of the peak area of L-isoleucine, L-leucine and L-valin to that of the internal standard obtained from the sample solution, and the ratios, QSa, QSb and QSc of the peak area of L-isoleucine, L-leucine and L-valin to that of the internal standard from the stand-ard solution. ‌1198 Amount (mg) of L-isoleucine (C6H13NO2) ＝ MSa × QTa/QSa × 5 Amount (mg) of L-leucine (C6H13NO2) ＝ MSb × QTb/QSb × 5 Amount (mg) of L-valine (C5H11NO2) ＝ MSc × QTc/QSc × 5 MSa: Amount (mg) of L-isoleucine for assay taken MSb: Amount (mg) of L-leucine for assay taken MSc: Amount (mg) of L-valine for assay taken Internal standard solution—A solution of glycine in 0.1 mol/L hydrochloric acid TS (1 in 20). Operating conditions— Detector: A visible absorption photometer (wavelength: 570 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 6 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with a sulfonated polystyrene (3 mm in particle diameter) (sodium type). Column temperature: A constant temperature of about 579C. Reaction vessel temperature: A constant temperature of about 1309C. Reaction time: About 1 minute. Mobile phase A Mobile phase B Mobile phase C Mobile phase D Mobile phase E Citric acid 19.80 g monohydrate 22.00 g 12.80 g 6.10 g — Trisodium citrate 6.19 g 7.74 g 13.31 g 26.67 g — Sodium chlo- ride 5.66 g 7.07 g 3.74 g 54.35 g — Sodium hydroxide — — — — 8.00 g Ethanol (99.5) 130 mL 20 mL 4 mL — 100 mL Thiodiglycol 5 mL 5 mL 5 mL — — Benzyl alcohol — — — 5 mL — Lauromacrogol solution (1 in 4) 4 mL 4 mL 4 mL 4 mL 4 mL Mobile phase: After prepare the mobile phases A, B, C, D and E according to the following table, add 0.1 mL caprylic acid to each mobile phase. dihydrate Water a sufficient a sufficient a sufficient a sufficient a sufficient amount amount amount amount amount Total amount 1000 mL 1000 mL 1000 mL 1000 mL 1000 mL Switching of mobile phase: Switch the mobile phases A, B, C, D and E sequentially so that when proceed with 20 mL of the standard solution under the conditions above, the in-ternal standard, valine, isoleucine and leucine are eluted in this order with the resolution between the peaks of isoleucine the standard solution under the above operating conditions, the internal standard, valine, isoleucine and leucine are eluted in this order with the resolution between the peaks of isoleucine and leucine being not less than 1.2. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the ratio of the peak area of isoleucine, leucine and valine to that of the internal standard are not more than 1.0z, respectively.
Labetalol Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled let-visible Spectrophotometry <2.24>. Amount (mg) of labetalol hydrochloride (C19H24N2O3.HCl) ＝ MS × AT/AS × V/40 MS: Amount (mg) of labetalol hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Labetalol Hydrochloride Tablets is not less than 75z. Start the test with 1 tablet of Labetalol Hydrochloride Tablets, withdraw not less than 20 mL of the medium at spe-cified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 50 mg of labetalol hydrochloride (C19H24N2O3.
(1) To a quantity of powdered Labetalol Hydrochloride Tablets equivalent to 5 mg of Labetalol Hy-drochloride, add 100 mL of 0.05 mol/L sulfuric acid TS, shake, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 300 nm and 304 nm. (2) To a quantity of powdered Labetalol Hydrochloride Tablets equivalent to 0.25 g of Labetalol Hydrochloride, add 25 mL of methanol, shake vigorously for 30 minutes, filter, and use the filtrate as the sample solution. Separately, dis-solve 10 mg of labetalol hydrochloride in 1 mL of methanol, and use this solution as the standard solution. Perform the test using these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, 2-propanol, water, and am-monia solution (28) (25:15:8:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Labetalol Hydrochloride Tablets add 5 mL of 0.5 mol/L sulfuric acid TS and 30 mL of water, shake vig-orously for 30 minutes, add water to make exactly 50 mL, and filter. Discard the first 5 mL of the filtrate, pipet 4 mL of the subsequent filtrate, add 0.05 mol/L sulfuric acid TS to make exactly V mL so that each mL contains about 40 mg of labetalol hydrochloride (C19H24N2O3.HCl), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of labetalol hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfu-ric acid TS to make exactly 50 mL. Pipet 5 mL of this solu-tion, add 0.05 mol/L sulfuric acid TS to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 302 nm as directed under Ultravio- mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 302 nm. Dissolution rate (z) with respect to the labeled amount of labetalol hydrochloride (C19H24N2O3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of labetalol hydrochloride for assay taken C: Labeled amount (mg) of labetalol hydrochloride (C19H24N2O3.HCl) in 1 tablet
Weigh accurately not less than 20 Labetalol Hydro-chloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 1 g of labetalol hydrochlo-ride (C19H24N2O3.HCl), add 100 mL of 0.5 mol/L sulfuric acid TS and 600 mL of water, shake vigorously for 30 minutes, add water to make exactly 1000 mL, and filter. Dis-card the first 5 mL of the filtrate, pipet 5 mL of the subse-quent filtrate, and add 0.05 mol/L sulfuric acid TS to make exactly 25 mL. Pipet 5 mL of this solution, add 0.05 mol/L sulfuric acid TS to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of labetalol hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfuric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add 0.05 mol/L sulfuric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 302 nm. Amount (mg) of labetalol hydrochloride (C19H24N2O3.HCl) ＝ MS × AT/AS × 25 MS: Amount (mg) of labetalol hydrochloride for assay taken
Lafutidine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of lafuti-dine (C22H29N3O4S: 431.55). Method of preparation Prepare as directed under Tablets, with Lafutidine. Identification Powder Lafutidine Tablets. To a portion of the powder, equivalent to 10 mg of Lafutidine, add 10 mL of methanol, shake thoroughly, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its an absorption maximum between 271 nm and 275 nm. Purity Related substances—To 10 Lafutidine Tablets add 4V/5 mL of the mobile phase, disintegrate the tablets by sonicating, then shake vigorously for not less than 30 minutes, and add the mobile phase to make V mL so that each mL contains about 1 mg of lafutidine (C22H29N3O4S).
Powder Lafutidine Tablets. To a portion of the powder, equivalent to 10 mg of Lafutidine, add 10 mL of methanol, shake thoroughly, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its an absorption maximum between 271 nm and 275 nm.
Related substances—To 10 Lafutidine Tablets add 4V/5 mL of the mobile phase, disintegrate the tablets by sonicating, then shake vigorously for not less than 30 minutes, and add the mobile phase to make V mL so that each mL contains about 1 mg of lafutidine (C22H29N3O4S). Centrifuge, and use the supernatant liquid as the sample so-lution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, other than lafuti-dine and the peak having the relative retention time of about 0.85 to lafutidine, obtained from the sample solution is not larger than 1/5 times the peak area of lafutidine from the standard solution, and the total area of the peaks, other than lafutidine and the peak having the relative retention time of about 0.85, from the sample solution is not larger than 3/5 times the peak area of lafutidine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 6 times as long as the retention time of lafutidine. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of lafutidine obtained with 5 mL of this solution is equivalent to 3.5 to 6.5z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of lafutidine are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of lafutidine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Lafutidine Tablets add exactly V mL of the internal standard solution so that each mL contains about 2 mg of lafutidine (C22H29N3O4S), disintegrate the tablet by sonicating, then shake vigorously for 30 minutes. Centrifuge this solution, filter the supernatant liquid through a mem-brane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accu-rately about 0.1 g of lafutidine for assay, previously dried under a reduced pressure (not exceeding 0.67 kPa) using phosphorus (V) oxide as desiccant for 4 hours, dissolve in ex-actly 50 mL of the internal standard solution, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of lafutidine (C22H29N3O4S) ＝ MS × QT/QS × V/50 MS: Amount (mg) of lafutidine for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in a mixture of acetonitrile and water (4:1) (3 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Lafutidine Tablets is not less than 75z. Start the test with 1 tablet of Lafutidine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter ‌1238 with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of lafutidine (C22H29N3O4S), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of lafutidine for assay, previously dried under a reduced pressure (not exceeding 0.67 kPa) using phosphorus (V) oxide as desiccant for 4 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolu-tion medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of lafutidine in each solution. Dissolution rate (z) with respect to the labeled amount of lafutidine (C H N O S) Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.87 g of sodium 1-pentanesul-fonate in 1000 mL of diluted phosphoric acid (1 in 1000). To 850 mL of this solution add 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of lafutidine is about 15 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, lafutidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of lafutidine to that of the internal standard is not more than 1.0z. 22 29 3 4 ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of lafutidine for assay taken C: Labeled amount (mg) of lafutidine (C22H29N3O4S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of lafutidine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lafutidine is not more than 2.0z.
To 20 Lafutidine Tablets add 4V/5 mL of the inter-nal standard solution, disintegrate the tablets with the aid of ultrasonic waves, then shake vigorously for 30 minutes. Add the internal standard solution to make exactly V mL so that each mL contains about 2 mg of lafutidine (C22H29N3O4S), centrifuge, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, and use the fil-trate as the sample solution. Separately, weigh accurately about 0.1 g of lafutidine for assay, previously dried under a reduced pressure (not exceeding 0.67 kPa) using phosphorus (V) oxide as desiccant for 4 hours, dissolve in the internal standard solution to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of lafutidine to that of the internal standard. Amount (mg) of lafutidine (C22H29N3O4S) in 1 tablet ＝ MS × QT/QS × V/1000 MS: Amount (mg) of lafutidine for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in a mixture of acetonitrile and water (4:1) (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter).
Lanoconazole Cream contains not less than 95.
Conduct this procedure using light-resistant vessels. Weigh accurately a quantity of Lanoconazole Cream, ‌equivalent to about 15 mg of lanoconazole (C14H10ClN3S2), add 80 mL of methanol, sonicate to disperse, and add ex-actly 10 mL of the internal standard solution. Add methanol to make 100 mL, ˆlter through a membrane ˆlter with a pore size of 0.45 mm if necessary, and use this solution as the sam-ple solution. Separately, weigh accurately about 15 mg of Lanoconazole RS, previously dried at 1059C for 2 hours, dissolve in methanol, and add exactly 10 mL of the internal standard solution. Add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lanoconazole to that of the internal standard.‌ Amount (mg) of lanoconazole (C14H10ClN3S2) ＝ MS × QT/QS MS: Amount (mg) of Lanoconazole RS taken Internal standard solution—A solution of diisopropyl 1,3-dithiolan-2-ylidenemalonate in methanol (1 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Lanoconazole. System suitability— matography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, toluene, methanol and am-monia solution (28) (400:400:20:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Assay Conduct this procedure using light-resistant vessels. Pipet a volume of Lanoconazole Cutaneous Solution, equivalent to about 50 mg of lanoconazole (C14H10ClN3S2), and add methanol to make exactly 50 mL. Pipet 15 mL of this solution, add exactly 10 mL of the internal standard so-lution, add methanol to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 15 mg of Lanoconazole RS, previously dried at 1059C for 2 hours, dissolve in methanol, and add exactly 10 mL of the internal standard solution. Add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lanoconazole to that of the internal standard. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lanoconazole and the internal standard are eluted in Amount (mg) of lanoconazole (C ＝ MS × QT/QS × 10/3 14H10ClN3S2) this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lanoconazole to that of the internal stand-ard is not more than 1.0z.
Lanoconazole Cutaneous Solution is a liquid for ex-ternal use. It contains not less than 95.
Lanoconazole Ointment contains not less than 93.
Conduct this procedure using light-resistant vessels. Weigh accurately a quantity of Lanoconazole Ointment, equivalent to about 15 mg of lanoconazole (C14H10ClN3S2), add 20 mL of tetrahydrofuran, sonicate to disperse, and add exactly 10 mL of the internal standard solution. Add metha-nol to make 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 15 mg of Lanoconazole RS, previously dried at 1059C for 2 hours, dissolve in methanol, and add exactly 10 mL of the internal standard solution. Add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lanoconazole to that of the internal standard. Lansoprazole
Lansoprazole Delayed-release Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of lansoprazole (C16H14F3N3O2S: 369.36). Method of preparation Prepare as directed under Cap-sules, with Lansoprazole. Identification Take out the contents of Lansoprazole Delayed-release Capsules, and powder. To a portion of the powder, equivalent to 5 mg of Lansoprazole, add 5 mL of methanol, shake thoroughly, and centrifuge. To 0.1 mL of the supernatant liquid add 10 mL of methanol, and deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 282 nm and 286 nm. ‌JP XVIII Official Monographs / Lansoprazole Delayed-release Orally Disintegrating Tablets 1243 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Lansoprazole Delayed-release Capsules, add 3V/10 mL of dilute sodium hydroxide TS, and sonicate with occasional stirring to disintegrate the contents completely. Add acetonitrile to make exactly V mL so that each mL contains about 0.
Take out the contents of Lansoprazole Delayed-release Capsules, and powder. To a portion of the powder, equivalent to 5 mg of Lansoprazole, add 5 mL of methanol, shake thoroughly, and centrifuge. To 0.1 mL of the supernatant liquid add 10 mL of methanol, and deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 282 nm and 286 nm. ‌JP XVIII Official Monographs / Lansoprazole Delayed-release Orally Disintegrating Tablets 1243 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Lansoprazole Delayed-release Capsules, add 3V/10 mL of dilute sodium hydroxide TS, and sonicate with occasional stirring to disintegrate the contents completely. Add acetonitrile to make exactly V mL so that each mL contains about 0.15 mg of lansoprazole (C16H14F3N3O2S). Centrifuge this solution, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, pipet 4 mL of the subsequent filtrate, add a mix-ture of acetonitrile and dilute sodium hydroxide TS (7:3) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Lan-soprazole RS (separately determine the water <2.48> in the same manner as Lansoprazole), and dissolve in 60 mL of dilute sodium hydroxide TS, and add acetonitrile to make exactly 200 mL. Pipet 4 mL of this solution, add a mixture of acetonitrile and dilute sodium hydroxide TS (7:3) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 294 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of lansoprazole (C16H14F3N3O2S) ＝ MS × AT/AS × V/200 MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Dissolution Being specified separately when the drug is granted approval based on the Law.
Take out the contents of not less than 20 capsules of Lansoprazole Delayed-release Capsules. Weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 0.3 g of lansopra-zole (C16H14F3N3O2S), add 60 mL of dilute sodium hydrox-ide TS, sonicate, and shake thoroughly. To this solution add 20 mL of acetonitrile and exactly 20 mL of the internal standard solution, shake thoroughly, and centrifuge. To 1 mL of the supernatant liquid add diluting solution to make 30 mL, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 5 mL of the first filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 30 mg of Lansopra-zole RS (separately determine the water <2.48> in the same manner as Lansoprazole), dissolve in 6 mL of dilute sodium hydroxide TS and 2 mL of acetonitrile, and add exactly 2 mL of the internal standard solution. To 1 mL of this solu-tion add diluting solution to make 30 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of lansoprazole to that of the internal stand-ard. Operating conditions— Proceed as directed in the operating conditions in the Assay under Lansoprazole. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lansoprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lansoprazole to that of the internal standard is not more than 1.0z.
Lansoprazole Delayed-release Orally Disintegrating Tablets
ランソプラゾール腸溶性口腔内崩壊錠
Lansoprazole Delayed-release Orally Disintegrating Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of lansoprazole (C16H14F3N3O2S: 369.36). Method of preparation Prepare as directed under Tablets, with Lansoprazole. Identification Powder 10 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets. To a portion of the powder, equivalent to 5 mg of Lansoprazole, add 5 mL of methanol, shake thoroughly, and centrifuge. To 0.1 mL of the supernatant liquid add 10 mL of methanol, and deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 282 nm and 286 nm. Purity Related substances—Keep the sample solution and standard solution at not exceeding 59C, and use them within 12 hours. Powder not less than 10 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets. To a portion of the powder, equivalent to 25 mg of Lansoprazole, add 10 mL of a mixture of dilute sodium hydroxide TS and metha-nol (3:1), sonicate, shake thoroughly, and centrifuge. To 2 mL of the supernatant liquid add diluting solution to make 20 mL, filter through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add diluting solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having a relative Amount (mg) of lansoprazole (C ＝ MS × QT/QS × 10 16H14F3N3O2S) retention time of about 1.1 to lansoprazole, obtained from the sample solution is not larger than 2/5 times the peak area of lansoprazole from the standard solution, and the area of MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 4?-ethoxya-cetophenone in acetonitrile (3 in 400). Diluting solution: A mixture of water, acetonitrile and triethylamine (60:40:1), adjusted to pH 11.0 with phosphoric acid. the peak other than lansoprazole and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of lansoprazole from the standard solution. Furthermore, the total area of the peaks other than lan-soprazole from the sample solution is not larger than 1.6 times the peak area of lansoprazole from the standard solu-tion. Diluting solution: A mixture of acetonitrile, water and triethylamine (160:40:1), adjusted to pH 11.0 with phos-phoric acid. To 100 mL of this solution add 900 mL of water. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, and time span of measurement: Proceed as directed in the operating conditions in the Purity (4) under Lansoprazole. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Dissolution Being specified separately when the drug is granted approval based on the Law. Assay Weigh accurately the mass of not less than 20 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.3 g of lansoprazole (C16H14F3N3O2S), add 60 mL of dilute sodium hydroxide TS, sonicate, and shake thoroughly. To this solution add 20 mL of acetonitrile and exactly 20 mL of the internal standard solution, shake thoroughly, and centrifuge. To 1 mL of the supernatant liquid add diluting solution to make 30 mL, and Time after injection 0 – 30 90 → 20 10 → 80 30 – 40 of sample (min) Mobile phase A (volz) Mobile phase B (volz) filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 30 mg of Lansoprazole RS (separately determine the water <2.48> in the same manner as Lansoprazole), dissolve in 6 mL of dilute sodium hydroxide Flow rate: About 0.8 mL per minute (the retention time of lansoprazole is about 24 minutes). System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add diluting solution to make exactly 20 mL. Confirm that the peak area of lansoprazole obtained with 40 mL of this solution is equivalent to 4 to 6z of that with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of lansoprazole are not less than 150,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lansoprazole is not more than 3.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Lansoprazole Delayed-release Orally Disintegrating Tablets add 3V/10 mL of dilute sodium hy-droxide TS, and sonicate with occasional stirring to disinte-grate the tablet completely. Add acetonitrile to make exactly V mL so that each mL contains about 0.
Powder 10 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets. To a portion of the powder, equivalent to 5 mg of Lansoprazole, add 5 mL of methanol, shake thoroughly, and centrifuge. To 0.1 mL of the supernatant liquid add 10 mL of methanol, and deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 282 nm and 286 nm.
Related substances—Keep the sample solution and standard solution at not exceeding 59C, and use them within 12 hours. Powder not less than 10 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets. To a portion of the powder, equivalent to 25 mg of Lansoprazole, add 10 mL of a mixture of dilute sodium hydroxide TS and metha-nol (3:1), sonicate, shake thoroughly, and centrifuge. To 2 mL of the supernatant liquid add diluting solution to make 20 mL, filter through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Pipet 1 mL of the sample solution, add diluting solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak, having a relative Amount (mg) of lansoprazole (C ＝ MS × QT/QS × 10 16H14F3N3O2S) retention time of about 1.1 to lansoprazole, obtained from the sample solution is not larger than 2/5 times the peak area of lansoprazole from the standard solution, and the area of MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 4?-ethoxya-cetophenone in acetonitrile (3 in 400). Diluting solution: A mixture of water, acetonitrile and triethylamine (60:40:1), adjusted to pH 11.0 with phosphoric acid. the peak other than lansoprazole and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of lansoprazole from the standard solution. Furthermore, the total area of the peaks other than lan-soprazole from the sample solution is not larger than 1.6 times the peak area of lansoprazole from the standard solu-tion. Diluting solution: A mixture of acetonitrile, water and triethylamine (160:40:1), adjusted to pH 11.0 with phos-phoric acid. To 100 mL of this solution add 900 mL of water. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, and time span of measurement: Proceed as directed in the operating conditions in the Purity (4) under Lansoprazole. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 tablets of Lansoprazole Delayed-release Orally Disintegrating Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.3 g of lansoprazole (C16H14F3N3O2S), add 60 mL of dilute sodium hydroxide TS, sonicate, and shake thoroughly. To this solution add 20 mL of acetonitrile and exactly 20 mL of the internal standard solution, shake thoroughly, and centrifuge. To 1 mL of the supernatant liquid add diluting solution to make 30 mL, and Time after injection 0 – 30 90 → 20 10 → 80 30 – 40 of sample (min) Mobile phase A (volz) Mobile phase B (volz) filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 30 mg of Lansoprazole RS (separately determine the water <2.48> in the same manner as Lansoprazole), dissolve in 6 mL of dilute sodium hydroxide Flow rate: About 0.8 mL per minute (the retention time of lansoprazole is about 24 minutes). System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add diluting solution to make exactly 20 mL. Confirm that the peak area of lansoprazole obtained with 40 mL of this solution is equivalent to 4 to 6z of that with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of lansoprazole are not less than 150,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lansoprazole is not more than 3.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Lansoprazole Delayed-release Orally Disintegrating Tablets add 3V/10 mL of dilute sodium hy-droxide TS, and sonicate with occasional stirring to disinte-grate the tablet completely. Add acetonitrile to make exactly V mL so that each mL contains about 0.15 mg of lansopra-zole (C16H14F3N3O2S). Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the fil-trate, pipet 4 mL of the subsequent filtrate, add a mixture of acetonitrile and dilute sodium hydroxide TS (7:3) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Lansoprazole RS (separately determine the water <2.48> in the same man-ner as Lansoprazole), and dissolve in 60 mL of dilute sodium hydroxide TS, and add acetonitrile to make exactly 200 mL. Pipet 4 mL of this solution, add a mixture of acetonitrile and dilute sodium hydroxide TS (7:3) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 294 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of lansoprazole (C16H14F3N3O2S) ＝ MS × AT/AS × V/200 MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Disintegration Being specified separately when the drug is granted approval based on the Law. TS and 2 mL of acetonitrile, and add exactly 2 mL of the internal standard solution. To 1 mL of this solution add diluting solution to make 30 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of lansoprazole to that of the internal standard. Amount (mg) of lansoprazole (C16H14F3N3O2S) ＝ MS × QT/QS × 10 MS: Amount (mg) of Lansoprazole RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 4?-ethoxya-cetophenone in acetonitrile (3 in 400). Diluting solution: A mixture of water, acetonitrile and triethylamine (60:40:1), adjusted to pH 11.0 with phosphoric acid. Operating conditions— Proceed as directed in the operating conditions in the Assay under Lansoprazole. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lansoprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lansoprazole to that of the internal standard is not more than 1.0z.
Levallorphan Tartrate Injection is an aqueous injec-tion. It contains not less than 93.
Levallorphan Tartrate Injection is a clear, col-orless liquid. pH: 3.0 – 4.5
Take an exact volume of Levallorphan Tar-trate Injection, equivalent to 3 mg of Levallorphan Tartrate, add 5 mL of water and 2 drops of dilute hydrochloric acid, and wash with five 15-mL portions of diethyl ether by a vigorous shaking. Take the water layer, evaporate the diethyl ether remained by warming on a water bath, and after cool-ing, add 0.01 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it ‌JP XVIII Official Monographs / Levodopa 1255 exhibits a maximum between 277 nm and 281 nm. Bacterial endotoxins <4.01> Less than 150 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take exactly a volume of Levallorphan Tartrate In-jection, equivalent to about 2 mg of levallorphan tartrate (C19H25NO.C4H6O6), add exactly 10 mL of the internal standard solution, and use this solution as the sample solu-tion. Separately, weigh accurately about 0.1 g of levallor-phan tartrate for assay, previously dried at 809C for 4 hours on phosphorus (V) oxide under reduced pressure, and dis-solve in water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solu-tion, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of levallorphan to that of the internal standard: Amount (mg) of levallorphan tartrate (C19H25NO.C4H6O6) ＝ MS × QT/QS × 1/50 MS: Amount (mg) of levallorphan tartrate for assay taken Internal standard solution—Dissolve 0.04 g of isobutyl par-ahydroxybenzoate in 10 mL of ethanol (95), add water to make 100 mL, and to 10 mL of this solution add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 300 mL of this solution add 200 mL of acetonitrile. Flow rate: Adjust so that the retention time of levallor-phan is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and levallorphan are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of levallorphan to that of the internal stand-ard is not more than 1.0z.
Levofloxacin Fine Granules contain not less than 93.0z and not more than 107.0z of the labeled amount of levofloxacin (C18H20FN3O4: 361.37). Method of preparation Prepare as directed under Gran-ules, with Levofloxacin Hydrate. Identification To an amount of Levofloxacin Fine Gran-ules, equivalent to 50 mg of levofloxacin (C18H20FN3O4), add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 50 mL, and stir for 20 minutes. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, dis-card the first 10 mL of the filtrate, and to 1 mL of the subse-quent filtrate add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 225 nm and 229 nm and between 292 nm and 296 nm, and a shoulder be-tween 321 nm and 331 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Levofloxacin Fine Granules add diluted 3 mol/L hydrochlo-ric acid TS (1 in 100) to make exactly V mL so that each mL contains about 1 mg of levofloxacin (C18H20FN3O4), and stir for 20 minutes.
To an amount of Levofloxacin Fine Gran-ules, equivalent to 50 mg of levofloxacin (C18H20FN3O4), add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 50 mL, and stir for 20 minutes. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, dis-card the first 10 mL of the filtrate, and to 1 mL of the subse-quent filtrate add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 225 nm and 229 nm and between 292 nm and 296 nm, and a shoulder be-tween 321 nm and 331 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Levofloxacin Fine Granules add diluted 3 mol/L hydrochlo-ric acid TS (1 in 100) to make exactly V mL so that each mL contains about 1 mg of levofloxacin (C18H20FN3O4), and stir for 20 minutes. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 1 mL of the subsequent fil-trate, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 25 mg of levoflox- Amount (mg) of levofloxacin (C18H20FN3O4) ＝ MS × AT/AS × V/25 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Levofloxacin Fine Granules is not less than 70z. Start the test with an accurately weighed amount of Levofloxacin Fine Granules, equivalent to about 0.1 g of levofloxacin (C18H20FN3O4), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 28 mg of levoflox-acin hydrate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Determine the absorbances, AT and AS, at 289 nm of the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of levofloxacin (C18H20FN3O4) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis MT: Amount (g) of Levofloxacin Fine Granules taken C: Labeled amount (mg) of levofloxacin (C18H20FN3O4) in 1 g
Weigh accurately an amount of Levofloxacin Fine Granules, powder if necessary, equivalent to about 50 mg of levofloxacin (C18H20FN3O4), add diluted 3 mol/L hydrochlo-ric acid TS (1 in 100) to make exactly 50 mL, stir for 20 minutes, and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of levofloxacin hy-drate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in diluted 3 mol/L hydrochloric acid TS (1 in 100) to make ex-actly 50 mL. Pipet 5 mL of this solution, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of levofloxacin in each solution. acin hydrate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in diluted 3 mol/L hydrochloric acid TS (1 in 100) to make Amount (mg) of levofloxacin (C ＝ MS × AT/AS 18H20FN3O4) exactly 50 mL. Pipet 2 mL of this solution, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 327 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ‌1258 ‌gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 1.00 g of copper (II) sulfate penta-hydrate, 1.41 g of L-valine and 6.17g of ammonium acetate in 800 mL of water, and add 200 mL of methanol. Flow rate: Adjust so that the retention time of levofloxa-cin is about 20 minutes. System suitability— System performance: Dissolve 10 mg of ofloxacin in 20 mL of diluted 3 mol/L hydrochloric acid TS (1 in 100). To 1 weigh accurately about 50 mg of levofloxacin hydrate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in diluted 1 mol/L hydrochloric acid TS (3 in 100) to make exactly 50 mL. Pipet 5 mL of this solution, add diluted 1 mol/L hydro-chloric acid TS (3 in 100) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of levofloxacin in each solution. mL of this solution add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating condi- Amount (mg) of levofloxacin (C ＝ MS × AT/AS 18H20FN3O4) tions, levofloxacin and an enantiomer are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of levofloxacin is not more than 1.0z.
Levofloxacin Injection is an aqueous injection. It contains not less than 95.
Levofloxacin Injection is yellow to greenish yellow, clear liquid.
To a volume of Levofloxacin Injection, equivalent to 50 mg of levofloxacin (C18H20FN3O4), add diluted 1 mol/L hydrochloric acid TS (3 in 100) to make 50 mL. To 1 mL of this solution add diluted 1 mol/L hydro-chloric acid TS (3 in 100) to make 100 mL. Determine the ab-sorption spectrum of this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 225 nm and 229 nm and between 292 nm and 296 nm, and a shoulder between 321 nm and 331 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxin <4.01> Less than 0.60 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Levofloxacin Injection, equivalent to about 50 mg of levofloxacin (C18H20FN3O4), add diluted 1 mol/L hydrochloric acid TS (3 in 100) to make exactly 50 mL. Pipet 5 mL of this solution, add diluted 1 mol/L hydrochloric acid TS (3 in 100) to make exactly 100 mL, and use this solution as the sample solution. Separately, MS: Amount (mg) of levofloxacin for assay taken, calcu-lated on the anhydrous basis Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Purity (2) under Levofloxacin Hydrate. Mobile phase: Dissolve 1.00 g of copper (II) sulfate penta-hydrate, 1.41 g of L-valine and 6.17 g of ammonium acetate in 800 mL of water, and add 200 mL of methanol. Flow rate: Adjust so that the retention time of levofloxa-cin is about 20 minutes. System suitability— System performance: Dissolve 10 mg of ofloxacin in 20 mL of diluted 1 mol/L hydrochloric acid TS (3 in 100). To 1 mL of this solution add diluted 1 mol/L hydrochloric acid TS (3 in 100) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, levofloxacin and the enantiomer are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of levofloxacin is not more than 1.0z.
Levofloxacin Ophthalmic Solution is an aqueous ophthalmic preparation. It contains not less than 95.
Levofloxacin Ophthalmic Solution occurs as a clear, pale yellow to yellow liquid.
(1) To a volume of Levofloxacin Ophthal-mic Solution, equivalent to 5 mg of Levofloxacin Hydrate, add 0.01 mol/L hydrochloric acid TS to make 100 mL. To 2 mL of this solution add 0.01 mol/L hydrochloric acid TS to make 20 mL, and use this solution as the sample solution. Determine the absorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 225 nm and 229 nm, and between 292 nm and 296 nm. JP XVIII Official Monographs / Levofloxacin Tablets 1259 ‌(2) To a volume of Levofloxacin Ophthalmic Solution, equivalent to 5 mg of Levofloxacin Hydrate, add a mixture of water and methanol (1:1) to make 5 mL, and use this solu-tion as the sample solution. Separately, dissolve 10 mg of levofloxacin hydrate for assay in 10 mL of a mixture of water and methanol (1:1), and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions: the retention time of the principal peaks in the chro-matogram obtained from the sample solution and the stand-ard solution is the same. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 1.25 g of copper (II) sulfate penta-hydrate, 1.76 g of L-valine and 7.71 g of ammonium acetate in water to make 1000 mL, and add 250 mL of methanol. Flow rate: Adjust so that the retention time of levofloxa-cin is about 22 minutes. System suitability— System performance: Dissolve 10 mg of ofloxacin in 20 mL of a mixture of water and methanol (1:1). To 1 mL of this solution add a mixture of water and methanol (1:1) to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolu-tion between the peak of levofloxacin and the peak having the relative retention time of about 1.2 to levofloxacin is not less than 3. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exact volume of Levofloxacin Ophthalmic Solution, equivalent to about 5 mg of levofloxacin hydrate (C18H20FN3O4.1/2H2O) add exactly 2 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of levofloxacin hydrate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in water to make exactly 50 mL. Pipet 10 mL of this solution, add ex-actly 2 mL of the internal standard solution, then add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of levofloxacin to that of the internal standard. Amount (mg) of levofloxacin hydrate (C18H20FN3O4.1/2H2O) ＝ MS × QT/QS × 1/5 × 1.025 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of naphazoline hy-drochloride in the mobile phase (3 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 13.61 g of potassium dihydrogen phosphate and 0.77 g of ammonium acetate in 900 mL of water, adjust to pH 3.0 with 1 mol/L hydrochloric acid TS, and add water to make 1000 mL. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of levofloxa-cin is about 17 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, levofloxacin and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of levofloxacin to that of the internal standard is not more than 1.0z.
Levofloxacin Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of levofloxacin (C18H20FN3O4: 361.37). Method of preparation Prepare as directed under Tablets, with Levofloxacin Hydrate. Identification To an amount of powdered Levofloxacin Tablets, equivalent to 0.1 g of levofloxacin (C18H20FN3O4), add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL, and stir for 20 minutes. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and to 1 mL of the subsequent filtrate add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 225 nm and 229 nm and between 292 nm and 296 nm, and a shoulder between 321 nm and 331 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Levofloxacin Tablets add about 70 mL of diluted 3 mol/L hydrochloric acid TS (1 in 100), sonicate to disintegrate the tablet, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 100 mL, and stir for 20 minutes. Pipet V mL the solution, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly V? mL so that each mL contains about 50 mg of levofloxacin (C18H20FN3O4), and filter this solution through a membrane filter with a pore size not exceeding 0.
To an amount of powdered Levofloxacin Tablets, equivalent to 0.1 g of levofloxacin (C18H20FN3O4), add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL, and stir for 20 minutes. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and to 1 mL of the subsequent filtrate add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 225 nm and 229 nm and between 292 nm and 296 nm, and a shoulder between 321 nm and 331 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Levofloxacin Tablets add about 70 mL of diluted 3 mol/L hydrochloric acid TS (1 in 100), sonicate to disintegrate the tablet, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 100 mL, and stir for 20 minutes. Pipet V mL the solution, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly V? mL so that each mL contains about 50 mg of levofloxacin (C18H20FN3O4), and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of levofloxacin (C18H20FN3O4) ＝ MS × AT/AS × V?/V × 1/5 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis Dissolution <6.10> (1) For a 100-mg Tablet When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes is not less than 80z. Start the test with 1 tablet of Levofloxacin Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 28 mg of levofloxacin hydrate for assay (separately de-termine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 289 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of levofloxacin hydrate (C H FN O .1/ H O) portion of the powder, equivalent to about 1 g of levofloxa-cin (C18H20FN3O4), add 150 mL of diluted 3 mol/L hydro-chloric acid TS (1 in 100), sonicate for 5 minutes, and add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make ex-actly 200 mL, and stir for 10 minutes. Pipet 2 mL of this so-lution, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 200 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of levofloxacin hydrate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hy-drate), and dissolve in diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 50 mL. Pipet 2 mL of this solu-tion, add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of levofloxacin in each solution. Amount (mg) of levofloxacin (C18H20FN3O4) ＝ MS × AT/AS × 40 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 340 nm). Column: A stainless steel column 4.6 mm in inside diame- 18 20 3 4 2 2 ter and 15 cm in length, packed with octadecylsilanized silica ＝ MS × AT/AS × 18/5 × 1.025 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis (2) For a 250-mg Tablet and 500-mg Tablet When the tests are performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolu-tion test as the dissolution medium, the dissolution rate in 30 minutes is not less than 80z. Start the test with 1 tablet of Levofloxacin Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11.2 mg of levofloxacin (C18H20FN3O4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of levofloxacin hy-drate for assay (separately determine the water <2.48> in the same manner as Levofloxacin Hydrate), and dissolve in the dissolution medium to make exactly 50 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 287 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of levofloxacin (C18H20FN3O4) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of levofloxacin hydrate for assay taken, calculated on the anhydrous basis C: Amount (mg) of levofloxacin (C18H20FN3O4) in 1 g
Accurately weigh the mass of not less than 20 Levofloxacin Tablets, and powder them. Weigh accurately a gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 1.00 g of copper (II) sulfate penta-hydrate, 1.41 g of L-valine and 6.17g of ammonium acetate in 800 mL of water, and add 200 mL of methanol. Flow rate: Adjust so that the retention time of levofloxa-cin is about 20 minutes. System suitability— System performance: Dissolve 10 mg of ofloxacin in 20 mL of diluted 3 mol/L hydrochloric acid TS (1 in 100). To 1 mL of this solution add diluted 3 mol/L hydrochloric acid TS (1 in 100) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, levofloxacin and an enantiomer are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of levofloxacin is not more than 1.0z.
(1) Weigh a quantity of powdered Levo-thyroxine Sodium Tablets, equivalent to 0.5 mg of Levo-thyroxine Sodium Hydrate, add 8 mL of a mixture of water, ethanol (95), hydrochloric acid and sodium hydroxide TS (6:5:2:2), warm in a water bath for 2 minutes, cool, and filter. To the filtrate add 0.1 mL of sodium nitrite TS, and allow to stand in a dark place for 20 minutes. Add 1.5 mL of ammonia solution (28): a yellowish red color develops. (2) To a quantity of powdered Levothyroxine Sodium Tablets, equivalent to 1 mg of Levothyroxine Sodium Hy-drate, add 10 mL of ethanol (95), shake, filter, and use the filtrate as the sample solution. Dissolve 0.01 g of levothyroxine sodium for thin-layer chromatography in 100 mL of ethanol (95), and use this solution as the standard so-lution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of t-butyl alcohol, t-amyl alcohol, water, am-monia solution (28) and 2-butanone (59:32:17:15:7) to a dis-tance of about 12 cm, and air-dry the plate. Spray evenly a solution of 0.3 g of ninhydrin in 100 mL of a mixture of 1-butanol and acetic acid (100) (97:3) on the plate, and heat the plate at 1009C for 3 minutes: the spots obtained from the sample solution and the standard solution show a red-purple color, and has the same Rf value.
Soluble halides—Weigh a quantity of powdered Levothyroxine Sodium Tablets, equivalent to 2.5 mg of Levothyroxine Sodium Hydrate, add 25 mL of water, warm to 409C, shake for 5 minutes, add 3 drops of dilute nitric acid, and filter. To the filtrate add 3 drops of silver nitrate TS, and mix: the solution has no more opalescence than the following control solution. Control solution: To 0.25 mL of 0.01 mol/L hydrochloric acid VS add 25 mL of water and 3 drops of dilute nitric acid, and proceed as directed above. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Place 1 tablet of Levothyroxine Sodium Tablets in a glass-stoppered centrifuge tube, add exactly 10 mL of 0.01 mol/L sodium hydroxide TS, warm at 509C for 15 minutes, and shake vigorously for 20 minutes. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 1 mL of the internal standard solution, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratio of the peak area of levothyroxine to that of the internal standard. Calculate the mean value from the ratios of each peak area of 10 samples: the deviation (z) of the mean value and the ratio of each peak area should be not more than 15z. When the deviation (z) is more than 15z, and 1 sam- ‌JP XVIII Official Monographs / Lidocaine 1263 ple shows not more than 25z, perform another test with 20 samples. Calculate the deviation (z) of the mean value of the 30 samples used in the 2 tests and the ratio of each peak area: there should be not more than 1 sample with the devia-tion more than 15z but not more than 25z, and no sample should deviate by more than 25z. Internal standard solution—A solution of ethinylestradiol in a mixture of acetonitrile and diluted phosphoric acid (1 in 10) (9:1) (3 in 40,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: a constant wavelength between 220 nm and 230 nm). Column: A stainless steel column 4 to 6 mm in inside di-ameter and 10 to 25 cm in length, packed with octadecyl- silanized silica gel (5 mm in particle diameter). Column temperature: A constant temperature at about 259C. Mobile phase: A mixture of methanol, water and phos-phoric acid (1340:660:1). Flow rate: Adjust so that the retention time of levothyroxine is about 9 minutes. Selection of column: To 5 mL of a solution of levothyroxine sodium in 0.01 mol/L sodium hydroxide TS (1 in 200,000) add 1 mL of the internal standard solution. Proceed with 20 mL of this solution under the above operat-ing conditions, and calculate the resolution. Use a column giving elution of levothyroxine and the internal standard in this order with the resolution between these peaks being not less than 2.0. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately about 25 mg of Levothyroxine So-dium Hydrate, and proceed as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 10 mL of so-dium hydroxide solution (1 in 100) and 1 mL of a freshly prepared sodium bisulfate solution (1 in 100) as the absorb-ing liquid, and prepare the test solution. Apply a small amount of water to the upper part of apparatus A, pull out C carefully, and wash C, B and the inner wall of A with 40 mL of water. To the test solution add 1 mL of bromine-acetic acid TS, insert the stopper C, and shake vigorously for 1 minute. Remove the stopper, rinse the stopper, the sample holder and the inner wall of the flask with 40 mL of water, and add 0.5 mL of formic acid. Stopper the flask with C, and shake vigorously for 1 minute again. Remove the stop-per, and rinse the stopper, the sample holder and the inner wall of the flask with 40 mL of water. Bubble the solution with enough nitrogen gas in the flask to remove the oxygen and excess bromine, add 0.5 g of potassium iodide to the so-lution, and dissolve. Add immediately 3 mL of dilute sulfu-ric acid, mix, and allow to stand for 2 minutes. Titrate <2.50> the solution with 0.02 mol/L sodium thiosulfate VS (indica-tor: 3 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.02 mol/L sodium thiosulfate VS ＝ 0.6657 mg of C15H10I4NNaO4
Lidocaine Injection is an aqueous injection. It contains not less than 95.
Lidocaine Injection is a colorless, clear liquid. pH: 5.0 – 7.0
To a volume of Lidocaine Injection, equiva-lent to 20 mg of lidocaine hydrochloride (C14H22N2O.HCl), add 1 mL of sodium hydroxide TS, and extract with 20 mL of hexane. To 10 mL of the hexane extract add 20 mL of 1 mol/L hydrochloric acid TS, and shake vigorously. Deter-mine the absorption spectrum of the water layer as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 261 nm and 265 nm. Bacterial endotoxins <4.01> Less than 1.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require- standard solution under the above operating conditions, lidocaine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of lidocaine to that of the internal standard is not more than 1.0z.
Lincomycin Hydrochloride Injection is an aqueous injection. It contains not less than 93.
Lincomycin Hydrochloride Injection is a clear, colorless liquid.
To a volume of Lincomycin Hydrochloride Injection, equivalent to 30 mg (potency) of Lincomycin Hy-drochloride Hydrate, add 30 mL of water, and use this solu-tion as the sample solution. Separately, dissolve 10 mg (po-tency) of Lincomycin Hydrochloride RS in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Dissolve 150 g of ammonium acetate in 800 mL of water, adjust the pH to 9.6 with ammonia solu-tion (28), and add water to make 1000 mL. To 80 mL of this solution add 40 mL of 2-propanol and 90 mL of ethyl ace-tate, shake, develop the plate with the upper layer of this so-lution to a distance of about 15 cm, and air-dry the plate. Spray evenly a solution of potassium permanganate (1 in 1000) on the plate: the principal spot obtained from the sam-ple solution and the spot from the standard solution show the same Rf value. pH <2.54> 3.5 – 5.5 Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Lincomycin Hydrochloride Injec-tion, equivalent to about 0.3 g (potency) of Lincomycin Hy-drochloride Hydrate, add the mobile phase to make exactly 30 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Lincomy-cin Hydrochloride RS, equivalent to 20 mg (potency), dis-solve in the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay under Lincomycin Hydrochloride Hy-drate. Liothyronine Sodium
(1) To a glass-stoppered centrifuge tube add a portion of finely powdered Liothyronine Sodium Tablets, equivalent to 0.1 mg of Liothyronine Sodium, add 30 mL of dilute sodium hydroxide TS, shake vigorously, and centrifuge. Transfer the supernatant liquid to a separator, add 10 mL of dilute hydrochloric acid, and extract with two 20-mL portions of ethyl acetate. Filter each extract succes-sively through absorbent cotton previously overlaid with 8 g of anhydrous sodium sulfate. Evaporate the filtrate on a water bath to dryness with the aid of a current of nitrogen. Dissolve the residue in 0.5 mL of methanol, and use this so-lution as the sample solution. Separately, dissolve 10 mg of liothyronine sodium for thin-layer chromatography in 50 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of t-butyl alcohol, t-amyl alcohol, water, ammonia solution (28) and 2-butanone (59:32:17:15:7) to a distance of about 12 cm, and air-dry the plate. Spray evenly a solution of 0.3 g of ninhydrin in 100 mL of a mixture of 1-butanol and acetic acid (100) (97:3) on the plate, and dry the plate at 1009C for 3 minutes: the spots obtained from the sample solution and the standard solution show a red-purple color, and has the same Rf value. (2) The colored solution obtained in the Assay is blue in color. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Place 1 tablet of Liothyronine Sodium Tablets in a glass-stoppered centrifuge tube, add exactly 10 mL of 0.01 mol/L sodium hydroxide TS, warm at 509C for 15 minutes, and shake vigorously for 20 minutes. Centrifuge for 5 minutes, and filter the supernatant liquid, if necessary. Pipet a definite volume of this solution, and add a volume of 0.01 mol/L sodium hydroxide TS to prepare a definite volume of a solution containing about 0.5 mg of liothyronine sodium (C15H11I3NNaO4) per mL. Pipet 5 mL of this solution, add exactly 1 mL of the internal standard solution, and use this solution as the sample solution. Perform the test with 200 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratio of the peak area of the liothyronine to that of the internal standard. Calculate the mean value of the ratios of each peak area of 10 samples: the deviation (z) of each ratio of the peak area from the mean value should be not more than 15z. When the deviation (z) is more than 15z, and 1 sample shows not more than 25z, perform another test with 20 samples. Calculate the deviation (z) of each ratio of the peak area from the mean value of the 30 samples used in the two tests: there should be not more than 1 sample with the deviation more than 15z but not more than 25z, and no sample should deviate by more than 25z. Internal standard solution—A solution of propylpara-hydroxybenzoate in a mixture of methanol and diluted phos-phoric acid (1 in 10) (9:1) (1 in 250,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsylanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: Diluted methanol (57 in 100). Flow rate: Adjust so that the retention time of liothyro-nine is about 9 minutes. ‌JP XVIII Official Monographs / Lisinopril Hydrate 1269 System suitability— System performance: To 5 mL of a solution of liothyro-nine sodium in 0.01 mol/L sodium hydroxide TS (1 in 2,000,000) add 1 mL of the internal standard solution, and use this solution as the solution for system suitability test. When the procedure is run with 200 mL of this solution under the above operating conditions, the internal standard and liothyronine are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 200 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the ratios of the peak area of liothyronine to that of the internal standard is not more than 1.0z.
Weigh accurately not less than 20 Liothyronine So-dium Tablets, and finely powder. Place an accurately weighed portion of the powder, equivalent to about 50 mg of liothyronine sodium (C15H11I3NNaO4), in an agate mortar, Lisinopril Hydrate
Lisinopril Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of lisinopril (C21H31N3O5: 405.49). Method of preparation Prepare as directed under Tablets, with Lisinopril Hydrate. Identification To an amount of powdered Lisinopril Tablets, equivalent to 10 mg of lisinopril (C21H31N3O5), add 10 mL of methanol, shake for 20 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 10 mg of lisinopril in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 30 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetonitrile, acetic acid (100), water and ethyl acetate (2:2:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 1209C: the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show a red-purple color and their Rf values are the same. Purity Related substances—Powder not less than 20 Lisinopril Tablets. Take a portion of the powder, equivalent to about 25 mg of lisinopril (C21H31N3O5), add 25 mL of water, shake for 20 minutes, filter, and use the filtrate as the sample solution. Pipet 3 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the peak area of lisinopril diketopiperazine, having the relative retention time of about 2.0 to lisinopril obtained from the sample solution, is not larger than 2/3 times the peak area of lisinopril from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Lisinopril Hydrate. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) under Lisinopril Hydrate. Test for required detectability: To exactly 2.5 mL of the standard solution add water to make exactly 50 mL. Con-firm that the peak area of lisinopril obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lisinopril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Lisinopril Tablets add exactly 5 mL of the internal standard solution per 1 mg of lisinopril (C21H31N3O5), shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Hereafter, proceed as directed in the Assay. Amount (mg) of lisinopril (C21H31N3O5) ＝ MS × QT/QS × C/10 MS: Amount (mg) of lisinopril for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of lisinopril (C21H31N3O5) in 1 tablet Internal standard solution—A solution of anhydrous caffeine (1 in 20,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate ‌JP XVIII Official Monographs / Lithium Carbonate 1271 of a 5-mg tablet in 60 minutes and that of a 10-mg tablet in 90 minutes is not less than 80z, and that of a 20-mg tablet in 90 minutes is not less than 75z. Start the test with 1 tablet of Lisinopril Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.
To an amount of powdered Lisinopril Tablets, equivalent to 10 mg of lisinopril (C21H31N3O5), add 10 mL of methanol, shake for 20 minutes, filter, and use the filtrate as the sample solution. Separately, dissolve 10 mg of lisinopril in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 30 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of acetonitrile, acetic acid (100), water and ethyl acetate (2:2:1:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly ninhydrin TS on the plate, and heat the plate at 1209C: the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show a red-purple color and their Rf values are the same.
Related substances—Powder not less than 20 Lisinopril Tablets. Take a portion of the powder, equivalent to about 25 mg of lisinopril (C21H31N3O5), add 25 mL of water, shake for 20 minutes, filter, and use the filtrate as the sample solution. Pipet 3 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: the peak area of lisinopril diketopiperazine, having the relative retention time of about 2.0 to lisinopril obtained from the sample solution, is not larger than 2/3 times the peak area of lisinopril from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Lisinopril Hydrate. System suitability— System performance: Proceed as directed in the system suitability in the Purity (2) under Lisinopril Hydrate. Test for required detectability: To exactly 2.5 mL of the standard solution add water to make exactly 50 mL. Con-firm that the peak area of lisinopril obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of lisinopril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Lisinopril Tablets add exactly 5 mL of the internal standard solution per 1 mg of lisinopril (C21H31N3O5), shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Hereafter, proceed as directed in the Assay. Amount (mg) of lisinopril (C21H31N3O5) ＝ MS × QT/QS × C/10 MS: Amount (mg) of lisinopril for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of lisinopril (C21H31N3O5) in 1 tablet Internal standard solution—A solution of anhydrous caffeine (1 in 20,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate ‌JP XVIII Official Monographs / Lithium Carbonate 1271 of a 5-mg tablet in 60 minutes and that of a 10-mg tablet in 90 minutes is not less than 80z, and that of a 20-mg tablet in 90 minutes is not less than 75z. Start the test with 1 tablet of Lisinopril Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of lisinopril (C21H31N3O5), and use this solution as the sample solution. Separately, weigh accurately about 15 mg of lisinopril for assay (separately determined the water <2.48> in the same manner as Lisinopril Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with ex-actly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of lisinopril in each solution. Dissolution rate (z) with respect to the labeled amount of lisinopril (C21H31N3O5) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of lisinopril for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of lisinopril (C21H31N3O5) in 1 tablet Operating conditions— Detector, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay. Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust so that the retention time of lisinopril is Internal standard solution—A solution of anhydrous caffeine (1 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 609C. Mobile phase: A mixture of diluted 0.05 mol/L sodium di-hydrogen phosphate TS (1 in 2) and acetonitrile for liquid chromatography (19:1). Flow rate: Adjust so that the retention time of lisinopril is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, lisinopril and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lisinopril to that of the internal standard is not more than 1.0z.
Losartan Potassium and Hydrochlorothiazide Tablets
ロサルタンカリウム・ヒドロクロロチアジド錠
(1) Shake thoroughly a portion of pow-dered Losartan Potassium and Hydrochlorothiazide Tablets, equivalent to 50 mg of Losartan Potassium, with 10 mL of methanol, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 50 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of losartan po-tassium in methanol to make 10 mL. To 5 mL of this solu-tion add methanol to make 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and acetic acid (100) (75:25:1) to a dis-tance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the two spots obtained from the sample solution and the spot from the standard solution show the same Rf value. (2) Shake well a portion of powdered Losartan Potassi- Amount (mg) of losartan potassium (C H ClKN O) in um and Hydrochlorothiazide Tablets, equivalent to 12.5 mg 1 tablet ＝ MS × AT/AS × V/50 22 22 6 of Hydrochlorothiazide, with 10 mL of methanol, and cen-trifuge. To 5 mL of the supernatant liquid add methanol to make 50 mL, and use this solution as the sample solution. MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.5 with phos-phoric acid, and add water to make 1000 mL. To 600 mL of this solution add 400 mL of acetonitrile Flow rate: Adjust so that the retention time of losartan is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry Separately, dissolve 25 mg of hydrochlorothiazide in metha-nol to make 10 mL. To 5 mL of this solution add methanol to make 100 mL, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and acetic acid (100) (75:25:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the two spots obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Losartan potassium—Perform the Mass variation test, or the Content uniformity test according to the follow-ing method: it meets the requirement. To 1 tablet of Losartan Potassium and Hydrochloro-thiazide Tablets add V/2 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), and stir for JP XVIII Official Monographs / Losartan Potassium and Hydrochlorothiazide Tablets 1277 60 minutes to disintegrate the tablet, add sodium dihydrogen phosphate TS (pH 2.5) to make exactly V mL so that each mL contains about 0.5 mg of losartan potassium (C22H22ClKN6O). Pipet 10 mL of this solution, add 45 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 46 mg of Losartan Potassium RS (separately determine the water <2.48> in the same manner as Losartan Potassium), and dissolve in 50 mL of a mixture of acetonitrile and so-dium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the losartan potassium standard stock solution. Pipet 12 mL of the losartan potassium stand-ard stock solution, add 44 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of losartan in each solu-tion. Amount (mg) of losartan potassium (C22H22ClKN6O) ＝ MS × AT/AS × 3V/250 MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.5 with phos-phoric acid, and add water to make 1000 mL. To 900 mL of this solution add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of losartan is about 5 minutes. System suitability— System performance: To 12 mL of the losartan potassium standard stock solution and 4 mL of the hydrochlorothiazide standard stock solution obtained in (2), add 42 mL of a mix-ture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), and add sodium dihydrogen phosphate TS (pH 2.5) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrochlorothiazide and losartan are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of losartan is not more than 1.0z. Hydrochlorothiazide—Perform the test according to the following method: it meets the requirement of the Con-tent uniformity test. To 1 tablet of Losartan Potassium and Hydrochloro-thiazide Tablets add V/2 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), and stir for 60 minutes to disintegrate the tablet, add sodium dihydrogen phosphate TS (pH 2.5) to make exactly V mL so that each mL contains about 0.125 mg of hydrochlorothiazide (C7H8ClN3O4S2). Pipet 10 mL of this solution, add 45 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 35 mg of Hydrochlorothiazide RS (separately determine the loss on drying <2.41> under the same conditions as Hydro-chlorothiazide), and dissolve in 50 mL of a mixture of aceto-nitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the hydrochloro-thiazide standard stock solution. Pipet 4 mL of the hydro-chlorothiazide standard stock solution, add 48 mL of a mix-ture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of hydrochlorothiazide in each solution. Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × AT/AS × V/250 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis Operating conditions— Proceed as directed in the operating conditions in (1). System suitability— System performance: To 12 mL of the losartan potassium standard stock solution obtained in (1) and 4 mL of the hy-drochlorothiazide standard stock solution, add 42 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), and add sodium dihydrogen phosphate TS (pH 2.5) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrochlorothiazide and losartan are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z. Dissolution <6.10> (1) Losartan potassium—When the test is performed at 100 revolutions per minute according to the Basket method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Losartan Potassium and Hydrochlorothiazide Tablets is not less than 85z. Start the test with 1 tablet of Losartan Potassium and Hy-drochlorothiazide Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of losartan potassium (C22H22ClKN6O), and use this solution as the sample solution. Separately, weigh accurately about 46 mg of Losartan Potassium RS (separately determine the water <2.48> in the same manner as Losartan Potassium), and dissolve in water to make exactly 100 mL, and use this solution as the losartan potassium standard stock solution. Pipet 12 mL of the losartan potassium standard stock solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of losartan in each solution. Dissolution rate (z) with respect to the labeled amount of losartan potassium (C22H22ClKN6O) ＝ MS × AT/AS × V?/V × 1/C × 108 MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of losartan potassium (C22H22ClKN6O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units (1). System suitability— System performance: To 12 mL of the losartan potassium standard stock solution and 8 mL of the hydrochlorothiazide standard stock solution obtained in (2), add water to make 100 mL. When the procedure is run with 20 mL of this solu-tion under the above operating conditions, hydrochloro-thiazide and losartan are eluted in this order with the resolu-tion between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of losartan is not more than 1.0z. (2) Hydrochlorothiazide—When the test is performed at 100 revolutions per minute according to the Basket method, using 900 mL of water as the dissolution medium, the disso-lution rate in 45 minutes of Losartan Potassium and Hydro-chlorothiazide Tablets is not less than 80z. Start the test with 1 tablet of Losartan Potassium and Hy-drochlorothiazide Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 13.9 mg Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units (1). System suitability— System performance: To 12 mL of the losartan potassium standard stock solution obtained in (1) and 8 mL of the hy-drochlorothiazide standard stock solution, add water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydro-chlorothiazide and losartan are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z.
(1) Losartan potassium—To 10 Losartan Potassi-um and Hydrochlorothiazide Tablets add 21V/25 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), stir for 60 minutes to disintegrate the tablets, add sodium dihydrogen phosphate TS (pH 2.5) to make ex-actly V mL so that each mL contains about 2 mg of losartan potassium (C22H22ClKN6O), and sonicate for 2 minutes. Pipet 10 mL of this solution, add 10 mL of acetonitrile, and add sodium dihydrogen phosphate TS (pH 2.5) to make ex-actly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 40 mg of Losar-tan Potassium RS (separately determine the water <2.48> in the same manner as Losartan Potassium), and dissolve in 30 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phos-phate TS (pH 2.5) to make exactly 50 mL, and use this solu-tion as the losartan potassium standard stock solution. Pipet 10 mL of the losartan potassium standard stock solution, add 4 mL of a mixture of acetonitrile and sodium dihydro-gen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of losartan in each solution. of hydrochlorothiazide (C7H8ClN3O4S2), and use this solu-tion as the sample solution. Separately, weigh accurately about 35 mg of Hydrochlorothiazide RS (separately deter-mine the loss on drying <2.41> under the same conditions as Amount (mg) of losartan potassium (C in 1 tablet ＝ MS × AT/AS × V/200 22H22ClKN6O) Hydrochlorothiazide), dissolve in 20 mL of methanol, and add water to make exactly 200 mL, and use this solution as the hydrochlorochiazide standard stock solution. Pipet 8 mL of hydrochlorothiazide standard stock solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of hy-drochlorothiazide in each solution. Dissolution rate (z) with respect to the labeled amount of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis C: Labeled amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) in 1 tablet MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Dissolve 1.25 g of potassium dihydrogen phosphate and 1.5 g of anhydrous disodium hydrogen phos-phate in water to make 1000 mL. To 930 mL of this solution add 70 mL of acetonitrile. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. ‌JP XVIII Official Monographs / Loxoprofen Sodium Hydrate 1279 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 12 100 → 92 0 → 8 12 – 28 92 → 38 8 → 62 Flow rate: Adjust so that the retention time of losartan is about 20 minutes. System suitability— System performance: To 25 mL of the losartan potassium standard stock solution and 10 mL of the hydrochloro-thiazide standard stock solution obtained in (2), add sodium dihydrogen phosphate TS (pH 2.5) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrochlorothiazide and losar-tan are eluted in this order, and the number of theoretical plates of the peak of hydrochlorothiazide and the symmetry factor of the peak of losartan are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of losartan is not more than 1.0z. (2) Hydrochlorothiazide—To 10 Losartan Potassium and Hydrochlorothiazide Tablets add 21V/25 mL of a mix-ture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), stir for 60 minutes to disintegrate the tablets, add sodium dihydrogen phosphate TS (pH 2.5) to make exactly V mL so that each mL contains about 0.5 mg of hy-drochlorothiazide (C7H8ClN3O4S2), and sonicate for 2 minutes. Pipet 10 mL of this solution, add 10 mL of aceto-nitrile, and add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Separately, weigh accurately about 25 mg of Hydrochlorothiazide RS (separately determine the loss on drying <2.41> under the same conditions as Hydrochloro-thiazide), and dissolve in a mixture of acetonitrile and so-dium dihydrogen phosphate TS (pH 2.5) (3:2), to make ex-actly 50 mL, and use this solution as the hydrochlorothiazide standard stock solution. Pipet 20 mL of the hydrochloro-thiazide standard stock solution, add 30 mL of a mixture of acetonitrile and sodium dihydrogen phosphate TS (pH 2.5) (3:2), add sodium dihydrogen phosphate TS (pH 2.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of hydro-chlorothiazide in each solution. Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) in 1 tablet ＝ MS × AT/AS × V/500 MS: Amount (mg) of Hydrochlorothiazide RS taken, cal-culated on the dried basis Operating conditions— Proceed as directed in the operating conditions in (1). System suitability— System performance: To 25 mL of the losartan potassium standard stock solution obtained in (1) and 10 mL of the hy-drochlorothiazide standard stock solution, add sodium dihy-drogen phosphate TS (pH 2.5) to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, hydrochlorothiazide and losartan are eluted in this order, and the number of theoretical plates of the peak of hydrochlorothiazide and the symmetry factor of the peak of losartan are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z.
Losartan Potassium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of losartan potassium (C22H22ClKN6O: 461.00). Method of preparation Prepare as directed under Tablets, with Losartan Potassium. Identification To an amount of powdered Losartan Potas-sium Tablets, equivalent to 25 mg of losartan potassium, add 10 mL of methanol, shake well, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 25 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of losartan potassium in 10 mL of methanol. To 5 mL of this solution add methanol to make 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and acetic acid (100) (75:25:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Losartan Potassium Tablets add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly 100 mL, and stir until the tablet is completely disin-tegrated. Pipet 5 mL of this solution, add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly V mL so that each mL contains about 50 mg of losartan po-tassium (C22H22ClKN6O), centrifuge, and use the superna-tant liquid as the sample solution.
To an amount of powdered Losartan Potas-sium Tablets, equivalent to 25 mg of losartan potassium, add 10 mL of methanol, shake well, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 25 mL, and use this solution as the sample solution. Separately, dissolve 25 mg of losartan potassium in 10 mL of methanol. To 5 mL of this solution add methanol to make 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, methanol and acetic acid (100) (75:25:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Losartan Potassium Tablets add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly 100 mL, and stir until the tablet is completely disin-tegrated. Pipet 5 mL of this solution, add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly V mL so that each mL contains about 50 mg of losartan po-tassium (C22H22ClKN6O), centrifuge, and use the superna-tant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of losartan potassium (C22H22ClKN6O) ＝ MS × AT/AS × V/25 MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute for 25-mg and 50-mg tablets and at 75 revo-lutions per minute for 100-mg tablet according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of 25-mg and 50-mg tablets, and in 30 minutes of 100-mg tablet are not less than ‌1276 85z, respectively. Start the test with 1 tablet of Losartan Potassium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of losartan potassium (C22H22ClKN6O), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Losartan Potassium RS (separately determine the water <2.48> in the same manner as Losartan Potassium), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 256 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of losartan potassium (C22H22ClKN6O) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Losartan Potassium RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of losartan potassium (C22H22ClKN6O) in 1 tablet
To 20 Losartan Potassium Tablets add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly 1000 mL, and stir until the tablets are completely dis-integrated. Pipet 5 mL of this solution, add diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly V mL so that each mL contains about 50 mg of losar-tan potassium (C22H22ClKN6O), centrifuge, and use the su-pernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of Losartan Potassium RS (separate-ly determine the water <2.48> in the same manner as Losar-tan Potassium), dissolve in diluted 0.1 mol/L phosphate buffer solution (pH 8.0) (1 in 10) to make exactly 500 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of losartan in each solution. factor of the peak of losartan are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of losartan is not more than 1.0z.
Loxoprofen Sodium Tablets contain not less than 95.0z and not more than 105.0z of the labeled violet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not amount of loxoprofen sodium (C 15H 17NaO3: 268.28). more intense than the spot from the standard solution. Water <2.48> 11.0 – 13.0z (0.2 g, volumetric titration, direct titration). Assay Weigh accurately about 60 mg of Loxoprofen So-dium Hydrate, and dissolve in diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, add ex-actly 10 mL of the internal standard solution, add diluted methanol (3 in 5) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Loxoprofen RS, previously dried in a desiccator (in vacuum, 609C) for 3 hours, and dissolve in diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, proceed in the same manner as directed for the preparation of the sample solution, and use so obtained solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of loxoprofen to that of the internal standard. Method of preparation Prepare as directed under Tablets, with Loxoprofen Sodium Hydrate. Identification To a quantity of powdered Loxoprofen So-dium Tablets, equivalent to 60 mg of loxoprofen sodium (C15H17NaO3), add 20 mL of methanol, shake vigorously for 10 minutes, and centrifuge. To 1 mL of the supernatant liq-uid add methanol to make 20 mL. To 2 mL of this solution add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Loxoprofen Sodium Tablets add exactly V mL of the internal standard solution so that each mL con-tains about 3 mg of loxoprofen sodium (C15H17NaO3). After sonicating for 10 minutes with occasional shaking, centri-fuge the solution. To 2 mL of the supernatant liquid add Amount (mg) of loxoprofen sodium (C H NaO ) diluted methanol (3 in 5) to make 100 mL, and use this solu- ＝ MS × QT/QS × 1.089 15 17 3 tion as the sample solution. Then, proceed as directed in the Assay. MS: Amount (mg) of Loxoprofen RS taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (3 in 5) (7 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water, acetic acid (100) and triethylamine (600:400:1:1). Flow rate: Adjust so that the retention time of loxoprofen is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, loxoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of loxoprofen to that of the internal stand-ard is not more than 1.0z. Containers and storage Containers—Tight containers. Amount (mg) of loxoprofen sodium (C15H17NaO3) ＝ MS × QT/QS × V/10 × 1.089 MS: Amount (mg) of Loxoprofen RS taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (3 in 5) (3 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Loxoprofen Sodium Tablets is not less than 85z. Start the test with 1 tablet of Loxoprofen Sodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add 2nd fluid for dissolution test to make ex-actly V? mL so that each mL contains about 13 mg of lox-oprofen sodium (C15H17NaO3).
L-Lysine Acetate occurs as white, crystals or crystalline powder. It has a characteristic odor and a slightly acid taste. It is very soluble in water, freely soluble in formic acid, oprofen sodium (C15 H17 NaO3), add exactly 20 mL of the in- and practically insoluble in ethanol (99.5). It is deliquescent. ‌ternal standard solution, and shake vigorously for 15 minutes. Centrifuge this solution, and to 2 mL of the super-natant liquid add diluted methanol (3 in 5) to make 100 mL. Use this solution as the sample solution. Separately, weigh accurately about 30 mg of Loxoprofen RS, previously dried in vacuum at 609C for 3 hours, and dissolve in exactly 10 mL of the internal standard solution. To 2 mL of this solution add diluted methanol (3 in 5) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of loxoprofen to that of the internal standard. Amount (mg) of loxoprofen sodium (C15H17NaO3) ＝ MS × QT/QS × 2 × 1.089 MS: Amount (mg) of Loxoprofen RS taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (3 in 5) (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water, acetic acid (100) and triethylamine (600:400:1:1). Flow rate: Adjust so that the retention time of loxoprofen is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, loxoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of loxoprofen to that of the internal standard is not more than 1.0z.
To a quantity of powdered Loxoprofen So-dium Tablets, equivalent to 60 mg of loxoprofen sodium (C15H17NaO3), add 20 mL of methanol, shake vigorously for 10 minutes, and centrifuge. To 1 mL of the supernatant liq-uid add methanol to make 20 mL. To 2 mL of this solution add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Loxoprofen Sodium Tablets add exactly V mL of the internal standard solution so that each mL con-tains about 3 mg of loxoprofen sodium (C15H17NaO3). After sonicating for 10 minutes with occasional shaking, centri-fuge the solution. To 2 mL of the supernatant liquid add Amount (mg) of loxoprofen sodium (C H NaO ) diluted methanol (3 in 5) to make 100 mL, and use this solu- ＝ MS × QT/QS × 1.089 15 17 3 tion as the sample solution. Then, proceed as directed in the Assay. MS: Amount (mg) of Loxoprofen RS taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (3 in 5) (7 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water, acetic acid (100) and triethylamine (600:400:1:1). Flow rate: Adjust so that the retention time of loxoprofen is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, loxoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of loxoprofen to that of the internal stand-ard is not more than 1.0z.
Weigh accurately about 60 mg of Loxoprofen So-dium Hydrate, and dissolve in diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, add ex-actly 10 mL of the internal standard solution, add diluted methanol (3 in 5) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Loxoprofen RS, previously dried in a desiccator (in vacuum, 609C) for 3 hours, and dissolve in diluted methanol (3 in 5) to make exactly 100 mL. Pipet 5 mL of this solution, proceed in the same manner as directed for the preparation of the sample solution, and use so obtained solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of loxoprofen to that of the internal standard. Method of preparation Prepare as directed under Tablets, with Loxoprofen Sodium Hydrate. Identification To a quantity of powdered Loxoprofen So-dium Tablets, equivalent to 60 mg of loxoprofen sodium (C15H17NaO3), add 20 mL of methanol, shake vigorously for 10 minutes, and centrifuge. To 1 mL of the supernatant liq-uid add methanol to make 20 mL. To 2 mL of this solution add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 221 nm and 225 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Loxoprofen Sodium Tablets add exactly V mL of the internal standard solution so that each mL con-tains about 3 mg of loxoprofen sodium (C15H17NaO3). After sonicating for 10 minutes with occasional shaking, centri-fuge the solution. To 2 mL of the supernatant liquid add Amount (mg) of loxoprofen sodium (C H NaO ) diluted methanol (3 in 5) to make 100 mL, and use this solu- ＝ MS × QT/QS × 1.089 15 17 3 tion as the sample solution. Then, proceed as directed in the Assay. MS: Amount (mg) of Loxoprofen RS taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (3 in 5) (7 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water, acetic acid (100) and triethylamine (600:400:1:1). Flow rate: Adjust so that the retention time of loxoprofen is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, loxoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of loxoprofen to that of the internal stand-ard is not more than 1.0z.
Magnesium Sulfate Injection is an aqueous injec-tion. It contains not less than 95.
Magnesium Sulfate Injection is a clear, color-less liquid.
Measure a volume of Magnesium Sulfate In-jection, equivalent to 0.5 g of Magnesium Sulfate Hydrate, and add water to make 20 mL: the solution responds to Qualitative Tests <1.09> for magnesium salt and for sulfate. pH <2.54> 5.5 – 7.0 When the labeled concentration exceeds 5z, prepare a solution of 5z with water, and per-form the test. Bacterial endotoxins <4.01> Less than 0.09 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Magnesium Sulfate Injection, equivalent to about 0.3 g of magnesium sulfate hydrate (MgSO4.7H2O), and add water to make 75 mL. Then add 5 mL of ammonia-ammonium chloride buffer so-lution (pH 10.7), and proceed as directed in the Assay under Magnesium Sulfate Hydrate. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 12.32 mg of MgSO .7H O of Magnesium Sulfate Hydrate according to Method 1, and 4 2 perform the test (not more than 2 ppm). Loss on ignition <2.43> 45.0 – 52.0z (1 g, after drying at 1059C for 2 hours, ignite at 4509C for 3 hours). Assay Weigh accurately about 0.6 g of Magnesium Sulfate Hydrate, previously ignited at 4509C for 3 hours after drying at 1059C for 2 hours, and dissolve in 2 mL of dilute hydro-chloric acid and water to make exactly 100 mL. Pipet 25 mL of this solution, add 50 mL of water and 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate
To a quantity of powdered Manidipine Hydrochloride Tablets, equivalent to 10 mg of Manidipine Hydrochloride, add 5 mL of methanol, shake vigorously, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of Manidipine Hydrochlo-ride RS in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and diethylamine (200:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Manidipine Hydrochloride Tablets, add exactly 1 mL of the internal standard solution per 1 mg of manidipine hydrochloride (C35H38N4O6.2HCl), disintegrate by adding a mixture of water and acetonitrile (1:1) to make V mL so that each mL contains about 0.1 mg of manidipine hydrochloride ‌(C35H38N4O6.2HCl), shake vigorously for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of manidipine hydrochloride (C35H38N4O6.2HCl) ＝ MS × QT/QS × V/250 MS: Amount (mg) of Manidipine Hydrochloride RS taken Internal standard solution—A solution of butyl benzoate in acetonitrile (7 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 45 minutes of Manidipine Hydrochloride Tablets is not less than 75z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Manidipine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of manidipine hydrochloride (C35H38N4O6.2HCl). Pipet 2 mL of this solu-tion, add exactly 2 mL of methanol, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Manidipine Hydrochloride RS, previously dried, dis-solve in a mixture of water and acetonitrile (1:1) to make ex-actly 50 mL. Pipet 1 mL of this solution, and add the disso-lution medium to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of methanol, and use this solu-tion as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of manidipine in each solution. with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of manidipine is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Weigh accurately not less than 20 Manidipine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of manidipine hydrochlo-ride (C35H38N4O6.2HCl), add exactly 10 mL of the internal standard solution, add a mixture of water and acetonitrile (1:1) to make 100 mL, shake vigorously for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 25 mg of Manidipine Hydro-chloride RS, previously dried, and dissolve in the mixture of water and acetonitrile (1:1) to make exactly 50 mL. Pipet 20 mL of this solution, add exactly 10 mL of the internal stand-ard solution, add the mixture of water and acetonitrile (1:1) to make 100 mL, and use this solution as the standard solu-tion. Then, proceed as directed in the Assay under Manidi-pine Hydrochloride. Amount (mg) of manidipine hydrochloride (C35H38N4O6.2HCl) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of Manidipine Hydrochloride RS taken Internal standard solution—A solution of butyl benzoate in acetonitrile (7 in 10,000).
Mecobalamin Tablets contain not less than 92.0z and not more than 108.0z of the labeled amount of MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis Operating conditions— Proceed as directed in the operating conditions in the Assay under Mecobalamin. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mecobalamin are not less than 2000 and 0.8 to 1.1, respectively. System repeatability: When the test is repeated 6 times mecobalamin (C63 H91 CoN13 O14 P: 1344.38). with 10 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak Method of preparation Prepare as directed under Tablets, with Mecobalamin. Identification (1) Conduct this procedure without ex-posure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, add 10 mL of hydrochloric acid-potassium chloride buffer solution (pH 2.0), sonicate, and add hydro-chloric acid-potassium chloride buffer solution (pH 2.0) to make 20 mL. Centrifuge this solution, and filter the superna-tant liquid through a membrane filter with a pore size not exceeding 0.8 mm. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 262 nm and 266 nm, between 303 nm and 307 nm, and between 461 nm and 465 nm. (2) Conduct this procedure without exposure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, area of mecobalamin is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Mecobalamin Tablets is not less than 80z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 tablet of Mecobalamin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 0.
(1) Conduct this procedure without ex-posure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, add 10 mL of hydrochloric acid-potassium chloride buffer solution (pH 2.0), sonicate, and add hydro-chloric acid-potassium chloride buffer solution (pH 2.0) to make 20 mL. Centrifuge this solution, and filter the superna-tant liquid through a membrane filter with a pore size not exceeding 0.8 mm. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 262 nm and 266 nm, between 303 nm and 307 nm, and between 461 nm and 465 nm. (2) Conduct this procedure without exposure to light, using light-resistant vessels. To a quantity of powdered Mecobalamin Tablets, equivalent to 1 mg of Mecobalamin, area of mecobalamin is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Mecobalamin Tablets is not less than 80z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 tablet of Mecobalamin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 0.28 mg of mecobalamin (C63H91CoN13O14P). Use this solution as the sample solution. Separately, weigh accurately about 28 mg of Mecobalamin RS (separately determine the water <2.48> in the same manner as Mecobalamin), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, ‌add water to make exactly 100 mL. Pipet 2 mL of this solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mecobalamin in each solution. Dissolution rate (z) with respect to the labeled amount of mecobalamin (C H CoN O P) Operating conditions— Proceed as directed in the operating conditions in the Assay under Mecobalamin. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mecobalamin are not less than 3000 and 0.8 to 1.1, respectively. 63 91 13 14 System repeatability: When the test is repeated 6 times ＝ MS × AT/AS × V?/V × 1/C × 9/10 MS: Amount (mg) of Mecobalamin RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of mecobalamin (C63H91CoN13O14P) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 264 nm). Column: A stainless steel column of 4.6 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 3.0 of a solution of 6.0 g of L-tartaric acid in 1000 mL of water with a solution of 14.3 g of disodium hydrogen phosphate dodecahydrate in 1000 mL of water. To 630 mL of this solution add 370 mL of metha-nol. Flow rate: Adjust so that the retention time of mecobala-min is about 8 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mecobalamin is not more than 1.0z.
(1) Weigh a quantity of powdered Mefru-side Tablets, equivalent to 0.3 g of Mefruside, shake with 15 mL of heated methanol for 20 minutes, and filter. Add 25 mL of water to the filtrate, and allow to stand while ice-cool-ing for 30 minutes. Filter the white precipitate formed, wash with water, and dry at 1059C for 2 hours: the precipitate melts <2.60> between 1499C and 1529C. C: Labeled amount (mg) of mefruside (C13H19ClN2O5S2) in 1 tablet
Weigh accurately not less than 20 Mefruside Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 65 mg of mefruside (C13H19ClN2O5S2), shake with 70 mL of methanol for 15 minutes, then add methanol to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, take exactly 10 mL of the subsequent filtrate, add methanol to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 65 mg of mefruside for assay, previously dried at 1059C for 2 hours, and dissolve in metha-nol to make exactly 100 mL. Pipet 10 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 285 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. (2) Weigh a quantity of powdered Mefruside Tablets, Amount (mg) of mefruside (C H ClN O S ) equivalent to 0.01 g of Mefruside, shake with 70 mL of methanol strongly for 15 minutes, add methanol to make ＝ MS × AT/AS 13 19 2 5 2 100 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 274 nm and 278 nm, and between 283 nm and 287 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mefruside Tablets add 40 mL of methanol, disintegrate the tablet by sonicating with occasional stirring, then further sonicate for 10 minutes, and add methanol to make exactly V mL of a solution containing about 0.5 mg of mefruside (C13H19ClN2O5S2) per mL. Centrifuge the solu-tion, pipet 5 mL of the supernatant liquid, add methanol to make exactly 20 mL, and use this solution as the sample so-lution. Then, proceed as directed in the Assay. Amount (mg) of mefruside (C13H19ClN2O5S2) ＝ MS × AT/AS × V/125 MS: Amount (mg) of mefruside for assay taken
Meglumine Iotalamate Injection is an aqueous injec-tion. It contains not less than 95.
Meglumine Iotalamate Injection is a clear, col- internal standard solution, add the mobile phase to make 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.4 g of iotalamic acid for assay, previously dried at 1059C for 4 hours, dissolve in 100 mL of water and 1 mL of sodium hydroxide TS, and add water to make exactly 200 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iotalamic acid to that of the internal standard. (2) Amidotorizoic Acid (anhydrous) 597.30 g Sodium Hydroxide 6.29 g Meglumine 159.24 g Amount (mg) of iotalamic acid (C ＝ MS × QT/QS × 10 11H9I3N2O4) Water for Injection or Sterile Water for Injection in Containers a sufficient quantity To make 1000 mL MS: Amount (mg) of iotalamic acid for assay taken Internal standard solution—A solution of L-tryptophan in the mobile phase (3 in 2500). ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 3.9 g of phosphoric acid and 2.8 mL of triethylamine in water to make 2000 mL. To this solu-tion add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of iotalamic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, iotalamic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of iotalamic acid to that of the internal standard is not more than 1.0z.
(1) Primary aromatic amines—To a volume of Meglumine Iotalamate Injection, equivalent to 0.20 g of Iotalamic Acid, add 15 mL of water, shake, add 4 mL of a solution of sodium nitrite (1 in 100) under ice-cooling, and proceed as directed in the Purity (2) under Iotalamic Acid: the absorbance is not more than 0.17. (2) Iodine and iodide—Take a volume of Meglumine Iotalamate Injection, equivalent to 1.5 g of Iotalamic Acid, add 20 mL of water and 5 mL of dilute sulfuric acid, shake well, and filter the precipitate by suction throuth a glass filter (G4). To the filtrate add 5 mL of toluene, and shake vigorously: the toluene layer is colorless. Then add 2 mL of a solution of sodium nitrite (1 in 100), and shake vigorously: the toluene layer has no more color than the following con-trol solution. Control solution: Dissolve 0.25 g of potassium iodide in water to make 1000 mL. To 2.0 mL of this solution add 20 mL of water, 5 mL of dilute sulfuric acid, 5 mL of toluene and 2 mL of a solution of sodium nitrite (1 in 100), and shake vigorously. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Meglumine Iotalamate Injection, equivalent to about 4 g of iotalamic acid (C H I N O ), add water to make exactly 200 mL. 11 9 3 2 4 (2) To make 1000 mL Pipet 2 mL of this solution, add water to make exactly 200 mL. To exactly 5 mL of this solution add exactly 5 mL of the Iotalamic Acid 455 g Meglumine 145 g Water for Injection or Sterile Water for Injection in Containers a sufficient quantity To make 1000 mL Prepare as directed under Injections, with the above ingre-dients (1) or (2). Description Meglumine Iotalamate Injection is a clear, col- internal standard solution, add the mobile phase to make 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.4 g of iotalamic acid for assay, previously dried at 1059C for 4 hours, dissolve in 100 mL of water and 1 mL of sodium hydroxide TS, and add water to make exactly 200 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of iotalamic acid to that of the internal standard. (2) Amidotorizoic Acid (anhydrous) 597.30 g Sodium Hydroxide 6.29 g Meglumine 159.24 g Amount (mg) of iotalamic acid (C ＝ MS × QT/QS × 10 11H9I3N2O4) Water for Injection or Sterile Water for Injection in Containers a sufficient quantity To make 1000 mL MS: Amount (mg) of iotalamic acid for assay taken Internal standard solution—A solution of L-tryptophan in the mobile phase (3 in 2500). ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: Dissolve 3.9 g of phosphoric acid and 2.8 mL of triethylamine in water to make 2000 mL. To this solu-tion add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of iotalamic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, iotalamic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of iotalamic acid to that of the internal standard is not more than 1.0z.
Meglumine Sodium Amidotrizoate Injection
アミドトリゾ酸ナトリウムメグルミン注射液
Meglumine Sodium Amidotrizoate Injection is an aqueous injection. It contains not less than 95.
Meglumine Sodium Amidotrizoate Injection is a clear, colorless to pale yellow, slightly viscous liquid. It gradually changes in color by light.
(1) To a volume of Meglumine Sodium Amidotrizoate Injection, equivalent to 1 g of Amidotrizoic Acid, add 25 mL of water, and add 2.5 mL of dilute hydro-chloric acid with stirring: a white precipitate is produced. Filter the precipitate by suction through a glass filter (G4), wash with two 10-mL portions of water, and dry at 1059C for 1 hour. Proceed with the precipitate so obtained as di-rected in the Identification (2) under Amidotrizoic Acid. To 1 mL of Meglumine Sodium Amidotrizoate Injec-tion add 1 mL of potassium 1,2-naphthoquinone-4-sulfonate TS and 0.2 mL of sodium hydroxide TS: a deep red color de-velops. Meglumine Sodium Amidotrizoate Injection re-sponds to Qualitative Tests <1.09> (1) for sodium salt. Optical rotation <2.49> D Method of preparation (1) a20: －2.91 – －3.369(100 mm). D Method of preparation (2) a20: －3.69 – －4.279(100 mm). pH <2.54> 6.0 – 7.7
(1) Primary aromatic amines—To a volume of Meglumine Sodium Amidotrizoate Injection, equivalent to 0.20 g of Amidotrizoic Acid, add 6 mL of water, mix, add 4 mL of a solution of sodium nitrite (1 in 100) and 10 mL of 1 mol/L hydrochloric acid TS, and shake. Proceed as directed in the Purity (2) under Amidotrizoic Acid: the absorbance is not more than 0.19. (2) Iodine and iodide—To a volume of Meglumine Sodium Amidotrizoate Injection, equivalent to 0.25 g of Amidotrizoic Acid, add water to make 20 mL, add 5 mL of dilute nitric acid, shake well, and filter by suction through a glass filter (G4). Add 5 mL of chloroform to the filtrate, and shake vigorously: no color develops in the chloroform layer. Then add 1 mL of hydrogen peroxide (30), and shake vigor-ously: the chloroform layer has no more color than the fol-lowing control solution. Control solution: Dissolve 0.10 g of potassium iodide in water to make 100 mL. Add 20 mL of water to 0.10 mL of this solution, add 5 mL of dilute nitric acid, 5 mL of chlo-roform and 1 mL of hydrogen peroxide (30), and shake vig-orously. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Meglumine So- ‌dium Amidotrizoate Injection, equivalent to about 0.5 g of amidotrizoic acid (C11H9I3N2O4), add water to make exactly 200 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.25 g of amidotrizoic acid for assay (separately determine the loss on drying <2.41> under the same condition as Amidotrizoic Acid), dissolve in a solution of meglumine (3 in 1000) to make exactly 100 mL, then pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and water to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of amidotrizoic acid to that of the internal standard. Amount (mg) of amidotrizoic acid (C11H9I3N2O4) ＝ MS × QT/QS × 2 MS: Amount (mg) of amidotrizoic acid for assay taken, calculated on the dried basis Internal standard solution—Dissolve 0.06 g of acetrizoic acid in a solution of meglumine (3 in 1000) to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.7 g of tetrabutylammonium di-hydrogen phosphate and 7.0 g of dipotassium hydrogen-phosphate in 750 mL of water, adjust the pH to 7.0 with diluted phosphoric acid (1 in 10), add water to make 800 mL, then add 210 mL of acetonitrile, and mix. Flow rate: Adjust so that the retention time of amidotrizoic acid is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, amidotrizoic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of amidotrizoic acid to that of the internal standard is not more than 1.0z.
Mepivacaine Hydrochloride Injection is an aqueous injection. It contains not less than 95.
Mepivacaine Hydrochloride Injection is a clear, colorless liquid.
To a volume of Mepivacaine Hydrochloride Injection, equivalent to 20 mg of Mepivacaine Hydrochlo-ride, add 1 mL of sodium hydrochloride TS, and extract with 20 mL of hexane. To 8 mL of the hexane extract add 20 mL of 1 mol/L hydrochloric acid TS, shake vigorously, and determine the absorption spectrum of the water layer sepa-rated as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 261 nm and 265 nm, and between 270 nm and 273 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 0.6 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Mepivacaine Hydrochloride Injec-tion, equivalent to about 40 mg of mepivacaine hydrochlo-ride (C15H22N2O.HCl), add exactly 4 mL of the internal standard solution and 0.001 mol/L hydrochloric acid TS to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of mepivacaine hy-drochloride for assay, previously dried at 1059C for 3 hours, dissolve in 0.001 mol/L hydrochloric acid TS, add exactly 4 mL of the internal standard solution and 0.001 mol/L hy-drochloride TS to make 20 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of mepivacaine to that of the internal standard. Amount (mg) of mepivacaine hydrochloride (C15H22N2O.HCl) ＝ MS × QT/QS MS: Amount (mg) of mepivacaine hydrochloride for assay taken Internal standard solution—A solution of benzophenone in methanol (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). ‌Column: A stainless steel column about 4 mm in inside di-ameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diam-eter).‌ Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.88 g of sodium lauryl sulfate in 1000 mL of a mixture of 0.02 mol/L phosphate buffer solu-tion (pH 3.0) and acetonitrile (11:9). Flow rate: Adjust so that the retention time of mepivacaine is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, mepivacaine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of mepivacaine to that of the internal standard is not more than 1.0z.
Mequitazine Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of mequitazine (C20H22N2S: 322.47). Method of preparation Prepare as directed under Tablets, with Mequitazine. Identification Powder Mequitazine Tablets. To a portion of the powder, equivalent to 3 mg of Mequitazine, add 50 mL of ethanol (95), shake thoroughly, and add ethanol (95) to make 100 mL. Centrifuge, if necessary, and filter the su-pernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard 10 mL of the first filtrate, to 4 mL of the subsequent filtrate add ethanol (95) to make 25 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 253 nm and 257 nm and between 301 nm and 311 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mequitazine Tablets add 50 mL of a mix-ture of methanol and water (4:3), and disperse to fine parti-cles by sonicating. Shake this solution thoroughly, and add methanol to make exactly 100 mL. Centrifuge, if necessary, ‌and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 4.
Powder Mequitazine Tablets. To a portion of the powder, equivalent to 3 mg of Mequitazine, add 50 mL of ethanol (95), shake thoroughly, and add ethanol (95) to make 100 mL. Centrifuge, if necessary, and filter the su-pernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard 10 mL of the first filtrate, to 4 mL of the subsequent filtrate add ethanol (95) to make 25 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 253 nm and 257 nm and between 301 nm and 311 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mequitazine Tablets add 50 mL of a mix-ture of methanol and water (4:3), and disperse to fine parti-cles by sonicating. Shake this solution thoroughly, and add methanol to make exactly 100 mL. Centrifuge, if necessary, ‌and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 4.8 mg of mequitazine (C20H22N2S), and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. Storage—Light-resistant. Mercaptopurine Hydrate
Meropenem for Injection is a preparation for injec-tion, which is dissolved before use. It contains not less than 93.
Meropenem for Injection occurs as a white to light yellow crystalline powder.
Determine the infrared absorption spectrum of Meropenem for Injection as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 3410 cm－1, 1750 cm－1, 1655 cm－1, 1583 cm－1 and 1391 cm－1. pH <2.54> Dissolve an amount of Meropenem for Injec-tion, equivalent to 0.25 g (potency) of Meropenem Hydrate, in 5 mL of water: the pH of the solution is between 7.3 and 8.3.
(1) Clarity and color of solution—Dissolve an amount of Meropenem for Injection, equivalent to 1.0 g (po-tency) of Meropenem Hydrate, in 20 mL of water: the solu-tion is clear and is not more intensely colored than the fol-lowing control solution. Control solution: To a mixture of 0.3 mL of Cobalt (II) Chloride CS and 1.2 mL of Iron (III) Chloride CS add 18.5 mL of diluted hydrochloric acid (1 in 40). Amount [mg (potency)] of meropenem (C ＝ MS × QT/QS × 1000 17H25N3O5S) (2) Related substances—Dissolve an amount of Meropenem for Injection, equivalent to 0.10 g (potency) of Meropenem Hydrate, in triethylamine-phosphate buffer so- MS: Amount [mg (potency)] of Meropenem RS taken Internal standard solution—A solution of benzyl alcohol in triethylamine-phosphate buffer solution (pH 5.0) (1 in 300). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of triethylamine-phosphate buffer solution (pH 5.0) and methanol (5:1). Flow rate: Adjust so that the retention time of meropenem is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, meropenem and the internal standard are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating lution (pH 5.0) to make 25 mL, and use this solution as the sample solution. Prepare the sample solution before use. Pipet 1 mL of the sample solution, add triethylamine-phosphate buffer solution (pH 5.0) to make exactly 100 mL. Pipet 5 mL of this solution, add triethylamine-phosphate buffer solution (pH 5.0) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the peak area of ring-opened meropenem and meropenem dimer, respectively having the relative retention time of about 0.5 and about 2.2 to meropenem obtained from the sample solution is not larger than the peak area of meropenem from the standard solution, the area of the peak, other than meropenem and the peaks mentioned above, is not larger than 1/5 times the peak area of meropenem from the standard solution, and the total area of the peaks other than meropenem is not larger than 3 times the peak area of meropenem from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the ‌Purity (3) under Meropenem Hydrate. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add triethylamine-phosphate buffer solution (pH 5.0) to make exactly 25 mL. Confirm that the peak area of meropenem obtained with 10 mL of this solution is equivalent to 16 to 24z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the sample solution, previously allowed to stand at 609C for 30 minutes, under the above operating conditions, the ring-opened meropenem, meropenem and the meropen-em dimer are eluted in this order, and the resolution between the peaks of the ring-opened meropenem and meropenem is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of meropenem is not more than 1.5z. Loss on drying <2.41> 9.5 – 12.0z (0.1 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.12 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Meropenem for Injection. Weigh accu-rately an amount of the contents, equivalent to about 50 mg (potency) of Meropenem Hydrate, dissolve in exactly 10 mL of the internal standard solution, add triethylamine-phosphate buffer solution (pH 5.0) to make 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Meropenem RS, equivalent to about 50 mg (potency), dissolve in exactly 10 mL of the internal stand-ard solution, add triethylamine-phosphate buffer solution (pH 5.0) to make 100 mL, and use this solution as the stand-ard solution. Then, proceed as directed in the Assay under Meropenem Hydrate. Mesalazine
Mesalazine Extended-release Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of mesalazine (C7H7NO3: 153.14). Method of preparation Prepare as directed under Tablets, with Mesalazine. Identification Powder Mesalazine Extended-release Tablets. To a portion of the powder, equivalent to 20 mg of Mesalazine, add 100 mL of diluted phosphoric acid (1 in 1000) and shake vigorously. To 5 mL of this solution add diluted phosphoric acid (1 in 1000) to make 50 mL, filter, and determine the absorbance spectrum of the filtrate as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm, and between 298 nm and 302 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Mesalazine Extended-release Tablets add 6V/25 mL of diluted phosphoric acid (1 in 1000), shake until the tablet is disintegrated, then add 3V/5 mL of methanol, and sonicate for 30 minutes. Add diluted phosphoric acid (1 in 1000) to make exactly V mL so that each mL contains about 1 mg of mesalazine (C7H7NO3), and centrifuge.
Powder Mesalazine Extended-release Tablets. To a portion of the powder, equivalent to 20 mg of Mesalazine, add 100 mL of diluted phosphoric acid (1 in 1000) and shake vigorously. To 5 mL of this solution add diluted phosphoric acid (1 in 1000) to make 50 mL, filter, and determine the absorbance spectrum of the filtrate as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm, and between 298 nm and 302 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Mesalazine Extended-release Tablets add 6V/25 mL of diluted phosphoric acid (1 in 1000), shake until the tablet is disintegrated, then add 3V/5 mL of methanol, and sonicate for 30 minutes. Add diluted phosphoric acid (1 in 1000) to make exactly V mL so that each mL contains about 1 mg of mesalazine (C7H7NO3), and centrifuge. Pipet 8 mL of the supernatant liquid, add exactly 2 mL of the in-ternal standard solution and 13 mL of methanol, then add diluted phosphoric acid (1 in 1000) to make 50 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of mesalazine (C7H7NO3) ＝ MS × QT/QS × V/40 MS: Amount (mg) of mesalazine for assay taken Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 800). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rates in 3 hours, in 6 hours and in 24 hours of Mesalazine Extended-release Tablets are 10 to 40z, 30 to 60z, and not less than 80z, respectively. Start the test with 1 tablet of Mesalazine Extended-release Tablets, withdraw exactly 20 mL of the medium at the speci-fied minutes after starting the test and supply exactly 20 mL of dissolution medium warmed to 37 ± 0.59C immediately after withdrawing of the medium every time. Filter the with-drawn media through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the disso-lution medium to make exactly V? mL so that each mL con- ‌tains about 56 mg of mesalazine (C7H7NO3), and use these solutions as the sample solutions. Separately, weigh accu-rately about 28 mg of mesalazine for assay, previously dried at 1059C for 2 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 25 mL, and use this so-lution as the standard solution. Determine the absorbances, AT(n) and AS, of the sample solutions and standard solution at 330 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. ＋ ∑ Dissolution rate (z) with respect to the labeled amount of mesalazine (C7H7NO3) on the nth medium withdrawing (n ＝ 1, 2, 3) System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of mesalazine to that of the internal standard is not more than 1.0z.
メテノロンエナント酸エステル注射液
Metenolone Enanthate Injection is an oily solution for injection. It contains not less than 90.
Metenolone Enanthate Injection is a clear, pale yellow, oily liquid.
(1) Measure a volume of Metenolone Enanthate Injection, equivalent to 0.1 g of Metenolone Enanthate, add 20 mL of petroleum ether, and extract with three 20-mL portions of diluted acetic acid (100) (5 in 7). Combine the extracts, wash with 20 mL of petroleum ether, add 300 mL of cold water while cooling in an ice bath, and stir sufficiently. Filter the produced precipitate by suction through a glass filter (G4), wash with water until the last washing becomes neutral, and dry in a desiccator (in vacu-um, phosphorus (V) oxide) for 6 hours. With this sample, proceed as directed in the Identification (1) under Meteno-lone Enanthate. (2) Measure a volume of Metenolone Enanthate Injec-tion, equivalent to 10 mg of Metenolone Enanthate, dissolve in 10 mL of chloroform, and use this solution as the sample solution. Separately dissolve 10 mg of metenolone enanthate in 10 mL of chloroform, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with toluene to a distance of about 15 cm, and air-dry the plate. Again develop this plate with a mixture of ethyl acetate and cyclohexane (1:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay To an exactly measured volume of Metenolone Enanthate Injection, equivalent to about 0.1 g of meteno-lone enanthate (C27H42O3), add chloroform to make exactly 100 mL. Pipet 5 mL of this solution, add chloroform to make exactly 50 mL, and use this solution as the sample solution. Weigh accurately about 0.1 g of metenolone enan-thate for assay, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, and prepare the standard solution in the same manner as directed for the preparation of the sample solution. Pipet 3 mL each of the sample solu-tion and standard solution, add exactly 10 mL of isoniazid TS, add methanol to make exactly 20 mL, and allow to stand for 60 minutes. Determine the absorbances, AT and AS, of the solutions from the sample solution and standard solu-tion, respectively, at 384 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution obtained by proceeding with 3 mL of chloroform as the blank. Hydrochloride in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add water to make exactly 50 mL. Pipet 1 mL of this solu-tion, add water to make exactly 10 mL, and use this solution as the standard solution (1). Pipet 5 mL of the standard solu-tion (1), add water to make exactly 10 mL, and use this solu-tion as the standard solution (2). Separately, to 0.10 g of 1-cyanoguanidine add water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution (3). Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1), (2) and (3) on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of 4-methyl-2-pentanone, 2-methoxyethanol, water and acetic acid (100) (30:20:5:3) to a distance of about 10 cm, air-dry the plate, then dry at 1059C for 10 minutes. Amount (mg) of metenolone enanthate (C ＝ MS × AT/AS 27H42O3) Spray evenly sodium pentacyanonitrosylferrate (III)-potas-sium hexacyanoferrate (III) TS on the plate: the spot other than the principal spot obtained from the sample solution is MS: Amount (mg) of metenolone enanthate for assay taken ‌Containers and storage Containers—Hermetic containers. Storage—Light-resistant. Metformin Hydrochloride
Shake an amount of powdered Metformin Hydrochloride Tablets, equivalent to 250 mg of Metformin Hydrochloride, with 25 mL of 2-propanol, and filter. Evaporate the filtrate under reduced pressure in a water bath at 409C, and determine the infrared absorption spectrum of the residue as directed in the potassium chloride disk method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 3370 cm－1, 3160 cm－1, 1627 cm－1, 1569 cm－1 and 1419 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution Being specified separately when the drug is ‌granted approval based on the Law.
Weigh accurately the mass of not less than 20 Met-formin Hydrochloride Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.15 g of metformin hydrochloride (C4H11N5.HCl), add 70 mL of a mixture of water and acetonitrile (3:2), shake for 10 minutes, add the mixture of water and acetonitrile (3:2) to make ex-actly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 3 mL of the subsequent filtrate, add exactly 3 mL of the internal standard solution and the mixture of water and acetonitrile (3:2) to make 50 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 0.15 g of metformin hydrochloride for assay, previ-ously dried at 1059C for 3 hours, and dissolve in the mixture of water and acetonitrile (3:2) to make exactly 100 mL. Pipet 3 mL of this solution, add exactly 3 mL of the internal stand-ard solution and the mixture of water and acetonitrile (3:2) to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of metformin to that of the internal standard. Methamphetamine Hydrochloride
Methotrexate Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of methotrexate (C20H22N8O5: 454.44). Method of preparation Prepare as directed under Cap-sules, with Methotrexate. Identification To an amount of the content of Metho-trexate Capsules, equivalent to 2 mg of Methotrexate, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 10 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 20 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 240 nm and 244 nm and between 304 nm and 308 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To the content of 1 capsule of Methotrexate Capsules add 3V/5 mL of the mobile phase, sonicate for 15 minutes, then shake for 25 minutes, and add the mobile phase to make ex-actly V mL so that each mL contains about 20 mg of methotrexate (C20H22N8O5).
To an amount of the content of Metho-trexate Capsules, equivalent to 2 mg of Methotrexate, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 10 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 20 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 240 nm and 244 nm and between 304 nm and 308 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To the content of 1 capsule of Methotrexate Capsules add 3V/5 mL of the mobile phase, sonicate for 15 minutes, then shake for 25 minutes, and add the mobile phase to make ex-actly V mL so that each mL contains about 20 mg of methotrexate (C20H22N8O5). Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 2 mL of the inter-nal standard solution, then add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Methotrexate RS (separate-ly determine the water <2.48> in the same manner as Methotrexate), and dissolve in the mobile phase to make ex-actly 100 mL. Pipet 10 mL of this solution, and add the mo-bile phase to make exactly 50 mL. Pipet 2 mL of this solu-tion, add exactly 2 mL of the internal standard solution, then add the mobile phase to make 20 mL, and use this solu-tion as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of methotrexate to that of the internal stand-ard. (89:11). Flow rate: Adjust so that the retention time of metho-trexate is about 8 minutes. Amount (mg) of methotrexate (C ＝ MS × QT/QS × V/500 20H22N8O5) System suitability— System performance: Dissolve 10 mg each of Metho-trexate and folic acid in 100 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, folic acid and methotrexate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methotrexate is not more than 1.0z.
Accurately weigh the mass of not less than 20 Methotrexate Capsules, take out all of the content, and ac-curately weigh the mass of the empty capsules. Powder the content, weigh accurately a portion of the powder, equiva-lent to about 10 mg of methotrexate (C20H22N8O5), add 60 mL of the mobile phase, shake for 25 minutes, and add the mobile phase to make exactly 100 mL. Centrifuge this solu-tion, pipet 2 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, then add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Methotrexate RS (separately determine the water <2.48> in the same man-ner as Methotrexate), and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add ex-actly 2 mL of the internal standard solution, then add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01>, and calculate the ratios, QT and QS, of the peak area of methotrexate to that of the internal standard. Amount (mg) of methotrexate (C20H22N8O5) ＝ MS × QT/QS MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis Internal standard solution—A solution of 4-nitrophenol in methanol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 302 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 250 mL of 0.2 mol/L potassium dihy-drogen phosphate TS add 28.5 mL of 0.2 mol/L sodium hy-droxide TS and water to make 1000 mL. To 890 mL of this solution add 110 mL of acetonitrile. Flow rate: Adjust so that the retention time of metho-trexate is about 6 minutes. System suitability— System performance: Dissolve 10 mg each of methotrexate and folic acid in 100 mL of the mobile phase. To 2 mL of this solution add the mobile phase to make 20 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, folic acid and methotrexate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of methotrexate to that of the internal stand-ard is not more than 1.0z.
Methotrexate for Injection is a preparation for in-jection which is dissolved before use. It contains not less than 95.
Methotrexate for Injection occurs as a light yellow to reddish yellow crystalline powder or mass.
To 1 mL of a solution of Methotrexate for Injection (1 in 400) add 0.1 mol/L hydrochloric acid TS to make 250 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 241 nm and 245 nm, and between 305 nm and 309 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Water Being specified separately when the drug is granted approval based on the Law. Bacterial endotoxins <4.01> Less than 0.1 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: Being specified separately when the drug is granted approval based on the Law.). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay Dissolve the contents of 20 containers of Methotrexate for Injection in the mobile phase, wash the containers with the mobile phase, combine the solution of the content and washings, and add the mobile phase to make exactly 1000 mL. Pipet V mL of this solution, add the mo-bile phase to make exactly V? mL so that each mL contains about 0.1 mg of methotrexate (C20H22N8O5), and use this so-lution as the sample solution. Separately, weigh accurately about 10 mg of Methotrexate RS (separately determine the water <2.48> in the same manner as Methotrexate), add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of methotrexate in each solution. tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 241 nm and 245 nm and between 305 nm and 309 nm. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Methotrexate Tablets add the mobile phase, stir, and add the mobile phase to make exactly V mL so that each mL contains about 0.1 mg of methotrexate (C20H22N8O5). Centrifuge this solution, and use the superna-tant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of methotrexate (C20H22N8O5) ＝ MS × AT/AS × V/250 MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate Amount (mg) of methotrexate (C H N O ) in 1 container in 45 minutes of Methotrexate Tablets is not less than 85z. of Methotrexate for Injection ＝ MS × AT/AS × V?/V × 1/2 22 8 5 Start the test with 1 tablet of Methotrexate Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis Operating conditions— Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Methotrexate. Column: A stainless steel column of 4.6 mm in inside di-ameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). System suitability— System performance: Dissolve 10 mg each of methotrexate and folic acid in 100 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, folic acid and methotrexate are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methotrexate is not more than 1.0z.
Methotrexate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of methotrexate (C20H22N8O5: 454.44). (This monograph is applied to only 2.5-mg tablets.) Method of preparation Prepare as directed under Tablets, with Methotrexate. Identification To a quantity of powdered Methotrexate Tablets, equivalent to 2.5 mg of Methotrexate, add 100 mL of diluted hydrochloric acid (1 in 100), shake, and filter or centrifuge. Determine the absorption spectrum of this solu- filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.
To a quantity of powdered Methotrexate Tablets, equivalent to 2.5 mg of Methotrexate, add 100 mL of diluted hydrochloric acid (1 in 100), shake, and filter or centrifuge. Determine the absorption spectrum of this solu- filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.8 mg of methotrexate (C20H22N8O5), and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Methotrexate RS (separately determine the water <2.48> in the same manner as Methotrexate), and dissolve in the mobile phase to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of methotrexate in each solution. Dissolution rate (z) with respect to the labeled amount of methotrexate (C20H22N8O5) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of Methotrexate RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of methotrexate (C20H22N8O5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 302 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 250 mL of 0.2 mol/L of potassium dihy-drogen phosphate TS add 29 mL of 0.2 mol/L sodium hy-droxide TS and water to make 1000 mL. To 890 mL of this solution add 110 mL of acetonitrile. Flow rate: Adjust so that the retention time of methotrexate is about 4 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of methotrexate are not less than 3000 and ‌not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methotrexate is not more than 1.0z.
Weigh accurately the mass of not less than 20 Methotrexate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of methotrexate (C20H22N8O5), add 50 mL of the mobile phase, shake, and add the mobile phase to make exactly 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of Methotrexate RS (separately determine the water <2.48> in the same manner as Methotrexate), dissolve in the mobile phase to make exactly 250 mL, and use this solution Methoxsalen
Methyldopa Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of methyldopa (C10H13NO4: 211.21). Method of preparation Prepare as directed under Tablets, with Methyldopa Hydrate. Identification (1) To a quantity of powdered Methyldopa Tablets, equivalent to 0.1 g of Methyldopa Hydrate, add 10 mL of water, and heat in a water bath for 5 minutes with oc-casional shaking. After cooling, centrifuge for 5 minutes at 2000 rotations per minute, apply 1 drop of the supernatant solution to a filter paper, and dry with warm air. Place 1 drop of ninhydrin TS over the spot, and heat for 5 minutes at 1009C: a purple color develops. To 0.5 mL of the supernatant liquid obtained in (1) add 2 mL of 0.05 mol/L sulfuric acid TS, 2 mL of iron (II) tartrate TS and 4 drops of ammonia TS, and shake well: a deep purple color develops. To 0.7 mL of the supernatant liquid obtained in (1) add 0.1 mol/L hydrochloric acid TS to make 20 mL. To 10 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 277 nm and 283 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Methyldopa Tablets add 50 mL of 0.05 mol/L sulfuric acid TS, shake for 15 minutes, then add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate equivalent to about 5 mg of methyldopa (C10H13NO4), add exactly 5 mL of iron (II) tartrate TS, then Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Methyldopa Tablets is not less than 75z. Start the test with 1 tablet of Methyldopa Tablets, with-draw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 25 mg of methyldopa (C10H13NO4), and use this solution as the sample solution.
(1) To a quantity of powdered Methyldopa Tablets, equivalent to 0.1 g of Methyldopa Hydrate, add 10 mL of water, and heat in a water bath for 5 minutes with oc-casional shaking. After cooling, centrifuge for 5 minutes at 2000 rotations per minute, apply 1 drop of the supernatant solution to a filter paper, and dry with warm air. Place 1 drop of ninhydrin TS over the spot, and heat for 5 minutes at 1009C: a purple color develops. To 0.5 mL of the supernatant liquid obtained in (1) add 2 mL of 0.05 mol/L sulfuric acid TS, 2 mL of iron (II) tartrate TS and 4 drops of ammonia TS, and shake well: a deep purple color develops. To 0.7 mL of the supernatant liquid obtained in (1) add 0.1 mol/L hydrochloric acid TS to make 20 mL. To 10 mL of this solution add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 277 nm and 283 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Methyldopa Tablets add 50 mL of 0.05 mol/L sulfuric acid TS, shake for 15 minutes, then add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate equivalent to about 5 mg of methyldopa (C10H13NO4), add exactly 5 mL of iron (II) tartrate TS, then Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Methyldopa Tablets is not less than 75z. Start the test with 1 tablet of Methyldopa Tablets, with-draw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 25 mg of methyldopa (C10H13NO4), and use this solution as the sample solution. Separately, weigh accu-rately about 56 mg of methyldopa for assay (separately de-ternine the loss on drying <2.41> at 1259C for 2 hours), and dissolve in water to make exactly 200 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and the standard solution at 280 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of methyldopa (C10H13NO4) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of methyldopa for assay taken, calcu-lated on the dried basis C: Labeled amount (mg) of methyldopa (C10H13NO4) in 1 tablet
Weigh accurately and powder not less than 20 Methyldopa Tablets. Weigh accurately a portion of the pow-der, equivalent to about 0.1 g of methyldopa (C10H13NO4), add 50 mL of 0.05 mol/L sulfuric acid TS, shake thoroughly for 15 minutes, add 0.05 mol/L sulfuric acid TS to make ex-actly 100 mL, and filter through a dry filter paper. Discard the first 20 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 0.11 g of Methyldopa RS (separately determine the loss on drying <2.41> at 1259C for 2 hours), dissolve in 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and use this solu-tion as the standard solution. Pipet 5 mL each of the sample solution and standard solution, add exactly 5 mL of iron (II) tartrate TS, and add ammonia-ammonium acetate buffer so-lution (pH 8.5) to make exactly 100 mL. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of 0.05 mol/L sulfuric acid TS in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 520 nm, respectively. add ammonia-ammonium acetate buffer solution (pH 8.5) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.11 g of Methyldopa RS (separately determine the loss on drying <2.41> at 1259C for 2 hours), and dissolve in 0.05 mol/L sulfuric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of iron (II) tartrate TS, then add ammonia-ammonium acetate buffer solution (pH 8.5) to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances at 520 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of methyldopa (C10H13NO4) ＝ MS × AT/AS × 5/V MS: Amount (mg) of Methyldopa RS taken, calculated on the dried basis Amount (mg) of methyldopa (C10H13NO4) ＝ MS × AT/AS MS: amount (mg) of Methyldopa RS taken, calculated on the dried basis
メチルエルゴメトリンマレイン酸塩錠
Methylergometrine Maleate Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of methylergometrine maleate (C20H25N3O2.C4H4O4: 455.50). Method of preparation Prepare as directed under Tablets, with Methylergometrine maleate. Identification (1) The sample solution obtained in the Assay shows a blue fluorescence. (2) The colored solution obtained in the Assay shows a deep blue color. Determine the absorption spectrum of the colored solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 543 nm and 547 nm and between 620 nm and 630 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Methylergometrine Maleate Tablets to a brown glass-stoppered centrifuge tube, add 10 mL of water, shake for 10 minutes vigorously, and disintegrate the tablet. Add 3 g of sodium chloride and 2 mL of ammonia so-lution (28), add exactly 25 mL of chloroform, and after vigorous shaking for 10 minutes, centrifuge for 5 minutes. Discard the water layer, take the chloroform extracts, add chloroform to make exactly V mL of a solution containing about 5 mg of methylergometrine maleate (C20H25N3O2.C4H4O4) per mL, and use this solution as the sample solution. Separately, weigh accurately about 1.25 mg of Methylergometrine Maleate RS, previously dried in a desiccator ( in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution into a brown glass-stoppered centrifuge tube, and add 3 g of sodium chloride and 2 mL of ammonia solu-tion (28). Add exactly 25 mL of chloroform, shake vigor-ously for 10 minutes, and centrifuge for 5 minutes. Discard the water layer, and use the chloroform layer as the standard solution. Pipet 20 mL each of the sample solution and the standard solution separately into brown glass-stoppered centrifuge tubes, add immediately exactly 10 mL of dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS, and shake for 5 minutes vigorously. Centrifuge these solutions for 5 minutes, take the water layers, and allow them to stand for 1 hour. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and standard solution at 545 nm, respectively. Amount (mg) of methylergometrine maleate (C20H25N3O2.C4H4O4) ＝ MS × AT/AS × V/250 ‌MS: Amount (mg) of Methylergometrine Maleate RS taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Methylergometrine Maleate Tablets is not less than 70z. Start the test with 1 tablet of Methylergometrine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis-card not less than 10 mL of the first filtrate, to exactly V mL of the subsequent filtrate add water to make exactly V? mL so that each mL contains about 0.
(1) The sample solution obtained in the Assay shows a blue fluorescence. (2) The colored solution obtained in the Assay shows a deep blue color. Determine the absorption spectrum of the colored solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 543 nm and 547 nm and between 620 nm and 630 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Methylergometrine Maleate Tablets to a brown glass-stoppered centrifuge tube, add 10 mL of water, shake for 10 minutes vigorously, and disintegrate the tablet. Add 3 g of sodium chloride and 2 mL of ammonia so-lution (28), add exactly 25 mL of chloroform, and after vigorous shaking for 10 minutes, centrifuge for 5 minutes. Discard the water layer, take the chloroform extracts, add chloroform to make exactly V mL of a solution containing about 5 mg of methylergometrine maleate (C20H25N3O2.C4H4O4) per mL, and use this solution as the sample solution. Separately, weigh accurately about 1.25 mg of Methylergometrine Maleate RS, previously dried in a desiccator ( in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution into a brown glass-stoppered centrifuge tube, and add 3 g of sodium chloride and 2 mL of ammonia solu-tion (28). Add exactly 25 mL of chloroform, shake vigor-ously for 10 minutes, and centrifuge for 5 minutes. Discard the water layer, and use the chloroform layer as the standard solution. Pipet 20 mL each of the sample solution and the standard solution separately into brown glass-stoppered centrifuge tubes, add immediately exactly 10 mL of dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS, and shake for 5 minutes vigorously. Centrifuge these solutions for 5 minutes, take the water layers, and allow them to stand for 1 hour. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and standard solution at 545 nm, respectively. Amount (mg) of methylergometrine maleate (C20H25N3O2.C4H4O4) ＝ MS × AT/AS × V/250 ‌MS: Amount (mg) of Methylergometrine Maleate RS taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Methylergometrine Maleate Tablets is not less than 70z. Start the test with 1 tablet of Methylergometrine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis-card not less than 10 mL of the first filtrate, to exactly V mL of the subsequent filtrate add water to make exactly V? mL so that each mL contains about 0.13 mg of methylergome-trine maleate (C20H25N3O2.C4H4O4), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Methylergometrine Maleate RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, then pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine im-mediately the intensities of the fluorescence, FT and FS, of the sample solution and standard solution at 338 nm as the excitation wavelength and at 427 nm as the fluorescence wavelength as directed under Fluorometry <2.22>. Dissolution rate (z) with respect to the labeled amount of methylergometrine maleate (C20H25N3O2.C4H4O4) ＝ MS × FT/FS × V?/V × 1/C × 9/20 sample solution and the standard solution add exactly 25 mL each of dilute 4-dimethylaminobenzaldehyde-iron (III) chlo-ride TS, and after vigorous shaking for 5 minutes, allow to stand for 30 minutes. Draw off the water layer, centrifuge, and allow to stand for 1 hour. Perform the test with these so-lutions as directed under Ultraviolet-visible Spectrophotome-try <2.24>, using dilute 4-dimethylaminobenzaldehyde-iron (III) chloride TS as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solu-tion and the standard solution at 545 nm, respectively. Amount (mg) of methylergometrine maleate (C20H25N3O2.C4H4O4) ＝ MS × AT/AS × 3/100 MS: Amount (mg) of Methylergometrine Maleate RS taken
Methyltestosterone Tablets contain not less than 90.0z and not more than 110.0z of the labeled amount of methyltestosterone (C20H30O2: 302.45). Method of preparation Prepare as directed under Tablets, with Methyltestosterone. Identification To a portion of powdered Methyltestoster-one Tablets, equivalent to 10 mg of Methyltestosterone, add 50 mL of acetone, shake for 30 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 10 mL of acetone, and use this solution as the sample solution. Separately, dissolve 10 mg of Methyltestosterone RS in 10 mL of acetone, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (9:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly dilute sulfu-ric acid on the plate, and heat the plate at 1109C for 10 minutes: the spot obtained from the sample solution and the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Methyltestosterone Tablets add 5 mL of water to disintegrate, add 50 mL of methanol, and shake for 30 minutes. Add methanol to make exactly 100 mL, and cen-trifuge. Measure exactly V mL of the supernatant liquid, add methanol to make exactly V? mL of a solution containing about 10 mg of methyltestosterone (C20H30O2) per ml, and use this solution as the sample solution. Separately, weigh Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 1 g of polysor-bate 80 in water to make 5 L as the dissolution medium, the dissolution rate in 30 minutes of a 10-mg tablet is not less than 75z and that in 60 minutes of a 25-mg tablet is not less than 70z. Start the test with 1 tablet of Methyltestosterone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of methyltestosterone (C20H30O2), and use this solution as the sample solution.
To a portion of powdered Methyltestoster-one Tablets, equivalent to 10 mg of Methyltestosterone, add 50 mL of acetone, shake for 30 minutes, and filter. Evaporate the filtrate to dryness, dissolve the residue in 10 mL of acetone, and use this solution as the sample solution. Separately, dissolve 10 mg of Methyltestosterone RS in 10 mL of acetone, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform and ethanol (95) (9:1) to a distance of about 12 cm, and air-dry the plate. Spray evenly dilute sulfu-ric acid on the plate, and heat the plate at 1109C for 10 minutes: the spot obtained from the sample solution and the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Methyltestosterone Tablets add 5 mL of water to disintegrate, add 50 mL of methanol, and shake for 30 minutes. Add methanol to make exactly 100 mL, and cen-trifuge. Measure exactly V mL of the supernatant liquid, add methanol to make exactly V? mL of a solution containing about 10 mg of methyltestosterone (C20H30O2) per ml, and use this solution as the sample solution. Separately, weigh Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 1 g of polysor-bate 80 in water to make 5 L as the dissolution medium, the dissolution rate in 30 minutes of a 10-mg tablet is not less than 75z and that in 60 minutes of a 25-mg tablet is not less than 70z. Start the test with 1 tablet of Methyltestosterone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of methyltestosterone (C20H30O2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Methyltesto-sterone RS, previously dried in vacuum using phosphorus (V) oxide as a desiccant for 10 hours, and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 249 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of methyltestosterone (C20H30O2) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Methyltestosterone RS taken C: Labeled amount (mg) of methyltestosterone (C20H30O2) in 1 tablet
Weigh accurately the mass of not less than 20 Methyltestosterone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg of methyl-testosterone (C20H30O2), add about 70 mL of methanol, shake for 30 minutes, and add methanol to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 5 mL of the in-ternal standard solution and methanol to make 50 mL, filter through a membrane filter (not exceeding 0.45 mm in pore size), and use the filtrate as the sample solution. Separately, weigh accurately about 20 mg of Methyltestosterone RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 10 hours, dissolve in methanol to make exactly 200 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of methyltestosterone to that of the internal standard. accurately about 10 mg of Methyltestosterone RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 10 hours, and dissolve in 5 mL of water and 50 mL of meth- Amount (mg) of methyltestosterone (C ＝ MS × QT/QS × 5/4 20H30O2) anol, then add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and the standard solution at the wavelength of maximum absorption at about 241 nm, respectively, as directed under Ultraviolet-visible Spectrophotometry <2.25>. Amount (mg) of methyltestosterone (C20H30O2) ＝ MS × AT/AS × V?/V × 1/10 MS: Amount (mg) of Methyltestosterone RS taken MS: Amount (mg) of Methyltestosterone RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in methanol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 241 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. ‌Mobile phase: A mixture of acetonitrile and water (11:9). Flow rate: Adjust so that the retention time of methyl- testosterone is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and methyltestosterone are eluted in this order with the resolution between these peaks being not less than 9. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of methyltestosterone to that of the internal standard is not more than 1.0z.
Metoprolol Tartrate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of metoprolol tartrate [(C15H25NO3)2.C4H6O6: 684.81]. Method of preparation Prepare as directed under Tablets, with Metoprolol Tartrate. Identification To an amount of powdered Metoprolol Tar-trate Tablets, equivalent to 10 mg of Metoprolol Tartrate, add 100 mL of ethanol (95), shake for 15 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 274 nm and 278 nm and between 281 nm and 285 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Metoprolol Tartrate Tablets add 1 mL of in 30 minutes of Metoprolol Tartrate Tablets is not less than 80z. Start the test with 1 tablet of Metoprolol Tartrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of metoprolol tartrate [(C15H25NO3)2.
To an amount of powdered Metoprolol Tar-trate Tablets, equivalent to 10 mg of Metoprolol Tartrate, add 100 mL of ethanol (95), shake for 15 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 274 nm and 278 nm and between 281 nm and 285 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Metoprolol Tartrate Tablets add 1 mL of in 30 minutes of Metoprolol Tartrate Tablets is not less than 80z. Start the test with 1 tablet of Metoprolol Tartrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of metoprolol tartrate [(C15H25NO3)2.C 4H6O6], and use this solution as the sample solution. Sepa-rately, weigh accurately about 56 mg of metoprolol tartrate for assay, previously dried in vacuum at 609C for 4 hours, and dissolve in water to make exactly 200 mL. Pipet 8 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of metoprolol in each solution. Dissolution rate (z) with respect to the labeled amount of metoprolol tartrate [(C15H25NO3)2.C4H6O6] ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of metoprolol tartrate for assay taken C: Labeled amount (mg) of metoprolol tartrate [(C15H25NO3)2.C4H6O6] in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of metoprolol are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of metoprolol is not more than 2.0z.
Weigh accurately the mass of not less than 20 Metoprolol Tartrate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.12 g of metoprolol tartrate [(C H NO ) .C H O ], add 60 mL of a water for every 10 mg of Metoprolol Tartrate, shake for 20 15 25 3 2 4 6 6 minutes, then add 75 mL of ethanol (95), shake for 15 minutes, add ethanol (95) to make exactly 100 mL, and cen-trifuge. Pipet V mL of the supernatant liquid, add ethanol (95) to make exactly V? so that each mL contains about 0.1 mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1) and exactly 10 mL of the internal standard solution, shake for 15 minutes, and add the mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1) to make 100 mL. mg of metoprolol tartrate [(C15H25 NO3)2.C4H6O6], and use Centrifuge, and use the supernatant liquid as the sample so- lution. Separately, weigh accurately about 0.12 g of meto- this solution as the sample solution. Separately, weigh accu-rately about 50 mg of metoprolol tartrate for assay, previ-ously dried in vacuum at 609C for 4 hours, dissolve in 5 mL of water, and add ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, add ethanol (95) to make ex-actly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 276 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using ethanol (95) as the blank. Amount (mg) of metoprolol tartrate [(C15H25NO3)2.C4H6O6] ＝ MS × AT/AS × V?/V × 1/5 MS: Amount (mg) of metoprolol tartrate for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate prolol tartrate for assay, previously dried in vacuum at 609C for 4 hours, dissolve in 60 mL of the mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1), add exactly 10 mL of the internal standard solution, then add the mix-ture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1) to make 100 mL, and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of metoprolol to that of the internal standard. Amount (mg) of metoprolol tartrate [(C15H25NO3)2.C4H6O6] ＝ MS × QT/QS MS: Amount (mg) of metoprolol tartrate for assay taken Internal standard solution—A solution of ethyl parahy- ‌droxybenzoate in the mixture of ethanol (99.5) and 1 mol/L hydrochloric acid TS (100:1) (1 in 500).‌ Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 14.0 g of sodium perchlorate monohydrate in 1000 mL of water, and adjust to pH 3.2 with diluted perchloric acid (17 in 2000). To 750 mL of this solution add 250 mL of acetonitrile. Flow rate: Adjust so that the retention time of metoprolol is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, metoprolol and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of metoprolol to that of the internal standard is not more than 1.0z.
Metronidazole Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of metronidazole (C6H9N3O3: 171.15). Method of preparation Prepare as directed under Tablets, with Metronidazole. Identification (1) To an amount of powdered Metronida-zole Tablets, equivalent to 0.1 g of Metronidazole, add 100 mL of 0.1 mol/L hydrochloric acid TS, and allow to stand for 30 minutes with occasional stirring. Then, shake vigor-ously, and centrifuge a part of this solution. To 1 mL of the supernatant liquid add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 275 nm and 279 nm. (2) Shake vigorously a quantity of powdered Metronida-zole Tablets, equivalent to 0.20 g of Metronidazole, with 20 mL of acetone for 10 minutes, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 0.10 g of metronidazole in 10 mL of acetone, and use this so- lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate immediately with a mixture of acetone, water and ethyl acetate (8:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf um on silica gel for 24 hours, dissolve in the mixture of water and methanol (4:1) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of metronidazole in each solution. Amount (mg) of metronidazole (C H N O ) ‌value of the principal spots obtained from the sample solu-tion and the standard solution is the same. ＝ MS × AT/AS × 10 6 9 3 3 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Metronidazole Tablets add 25 mL of a mix-ture of water and methanol (1:1), shake vigorously for 25 minutes, and add the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add a mixture of water and methanol (4:1) to make exactly 100 mL. Filter the solution through a membrane filter with pore size of 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Hereinafter, proceed as directed in the Assay. Amount (mg) of metronidazole (C H N O ) MS: Amount (mg) of metronidazole for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 320 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (4:1). Flow rate: Adjust so that the retention time of metronida- zole is about 5 minutes. System suitability— ＝ MS × AT/AS × 10 6 9 3 3 System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- MS: Amount (mg) of metronidazole for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Metronidazole Tablets is not less than 70z. Start the test with 1 tablet of Metronidazole Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of metronidazole (C6H9N3O3), and use this solution as the sample solution.
(1) To an amount of powdered Metronida-zole Tablets, equivalent to 0.1 g of Metronidazole, add 100 mL of 0.1 mol/L hydrochloric acid TS, and allow to stand for 30 minutes with occasional stirring. Then, shake vigor-ously, and centrifuge a part of this solution. To 1 mL of the supernatant liquid add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 275 nm and 279 nm. (2) Shake vigorously a quantity of powdered Metronida-zole Tablets, equivalent to 0.20 g of Metronidazole, with 20 mL of acetone for 10 minutes, centrifuge, and use the super-natant liquid as the sample solution. Separately, dissolve 0.10 g of metronidazole in 10 mL of acetone, and use this so- lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography, develop the plate immediately with a mixture of acetone, water and ethyl acetate (8:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the Rf um on silica gel for 24 hours, dissolve in the mixture of water and methanol (4:1) to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of metronidazole in each solution. Amount (mg) of metronidazole (C H N O ) ‌value of the principal spots obtained from the sample solu-tion and the standard solution is the same. ＝ MS × AT/AS × 10 6 9 3 3 Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Metronidazole Tablets add 25 mL of a mix-ture of water and methanol (1:1), shake vigorously for 25 minutes, and add the mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add a mixture of water and methanol (4:1) to make exactly 100 mL. Filter the solution through a membrane filter with pore size of 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Hereinafter, proceed as directed in the Assay. Amount (mg) of metronidazole (C H N O ) MS: Amount (mg) of metronidazole for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 320 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and methanol (4:1). Flow rate: Adjust so that the retention time of metronida- zole is about 5 minutes. System suitability— ＝ MS × AT/AS × 10 6 9 3 3 System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- MS: Amount (mg) of metronidazole for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Metronidazole Tablets is not less than 70z. Start the test with 1 tablet of Metronidazole Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of metronidazole (C6H9N3O3), and use this solution as the sample solution. Separately, weigh accu-rately about 22 mg of metronidazole for assay, previously dried in vacuum with silica gel for 24 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 320 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of metronidazole (C6H9N3O3) ditions, the number of theoretical plates and the symmetry factor of the peak of metronidazole are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of metronidazole is not more than 1.0z.
To a quantity of powdered Miglitol Tablets, equivalent to 0.1 g of Miglitol, add 50 mL of a mixture of acetonitrile and water (9:1), shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 50 mg of miglitol in a mixture of acetonitrile and water (9:1) to make 25 mL, and use this solution as the standard solution. Perform the test with these solutions as directed solution, and use the supernatant liquid as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of miglitol (C8H17NO5) ＝ MS × AT/AS × V/50 MS: Amount (mg) of Miglitol RS taken, calculated on the dried basis Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Miglitol Tablets is not less than 85z. Start the test with 1 tablet of Miglitol Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 28 mg of miglitol (C8H17NO5), and use this solu-tion as the sample solution. Separately, weigh accurately about 56 mg of Miglitol RS (separately determine the loss on drying <2.41> under the same conditions as Miglitol), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 200 mL and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of miglitol in each solution. Dissolution rate (z) with respect to the labeled amount of miglitol (C8H17NO5) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Miglitol RS taken, calculated on the dried basis C: Labeled amount (mg) of miglitol (C8H17NO5) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of miglitol are not less than 1000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of miglitol is not more than 2.0z.
Weigh accurately the mass of not less than 20 Miglitol Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of miglitol (C8H17NO5), add 50 mL of a mixture of acetonitrile for liq-uid chromatography and water (4:1), shake, and add a mix-ture of acetonitrile for liquid chromatography and water (4:1) to make exactly 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 50 mg of Miglitol RS (separately de-termine the loss on drying <2.41> under the same conditions as Miglitol), and dissolve in a mixture of acetonitrile for liq-uid chromatography and water (4:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of miglitol in each solution. and in chloroform, and slightly soluble in diethyl ether. The pH of a solution of 1.0 g of Migrenin in 10 mL of water is between 3.0 and 4.0. It is affected by moisture and light. Identification (1) To 5 mL of a solution of Migrenin (1 in 100) add 2 drops of sodium nitrite TS and 1 mL of dilute sul-furic acid: a deep green color develops. To 5 mL of a solution of Migrenin (1 in 50) add 1 drop of hydrochloric acid and 0.2 mL of formaldehyde solu-tion, heat in a water bath for 30 minutes, add an excess of ammonia TS, and filter. Acidify the filtrate with hydrochlo-ric acid, shake with 3 mL of chloroform, and separate the chloroform layer. Evaporate the chloroform solution on a water bath, add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid to the residue, and evaporate on a water bath to dryness: the residue shows a yellow-red color. Invert the residue over a vessel containing 2 to 3 drops of ammonia TS: a red-purple color develops, disappearing on Amount (mg) of miglitol (C8H 17NO5) ＝ MS × AT/AS × 2 the addition of 2 to 3 drops of sodium hydroxide TS. A solution of Migrenin (1 in 10) responds to Qualita- ‌MS: Amount (mg) of Miglitol RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with pentaethylenehexaami-nated polyvinyl alcohol polymer beads for liquid chromatog-raphy (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.6 g of potassium dihydrogen phosphate and 0.28 g of anhydrous disodium hydrogen phosphate in water to make 1000 mL. To 200 mL of this so-lution add 800 mL of acetonitrile for liquid chromatogra-phy. Flow rate: Adjust so that the retention time of miglitol is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of miglitol are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of miglitol is not more than 1.0z.
Minocycline Hydrochloride for Injection is a prepa-ration for injection, which is dissolved before use. It contains not less than 90.
Minocycline Hydrochloride for Injection oc-curs as a yellow to yellow-brown, powder or flakes.
Dissolve 4 mg of Minocycline Hydrochloride for Injection in 250 mL of a solution of hydrochloric acid in methanol (19 in 20,000). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 221 nm and 225 nm, between 261 nm and 265 nm, and between 354 nm and 358 nm. pH <2.54> The pH of a solution, prepared by dissolving an amount of Minocycline Hydrochloride for Injection, equiva-lent to 0.1 g (potency) of Minocycline Hydrochloride, in 10 mL of water is 2.0 to 3.5.
Related substances—Conduct this procedure rapidly after the preparation of the sample solution. Take an amount of Minocycline Hydrochloride for Injection, equiva-lent to 0.1 g (potency) of Minocycline Hydrochloride, dis-solve in the mobile phase to make 100 mL. To 25 mL of this solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method. Calculate the amounts of each peak by the area percentage method: the amount of epiminocycline, having the relative retention time of about 0.83 to minocycline, is not more than 6.0z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Minocycline Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of minocycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Minocycline Hydrochloride. Test for required detectability: Pipet 2 mL of the standard solution obtained in the Assay, add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for sys-tem suitability test, add the mobile phase to make exactly 100 mL. Confirm that the peak area of minocycline obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Weigh accurately the mass of the content of one container of Minocycline Hydrochloride for Injection, dissolve in exactly 2 mL of methanol for water determina-tion, and perform the test with exactly 1 mL of this solution as directed in the Volumetric titration (back titration): not more than 3.0z. Bacterial endotoxins <4.01> Less than 1.25 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Minocycline Hydrochloride for Injec- ‌tion. Weigh accurately an amount of the contents, equiva-lent to about 0.1 g (potency) of Minocycline Hydrochloride, dissolve in the mobile phase to make exactly 100 mL. Pipet 25 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately an amount of Minocycline Hydro-chloride RS, equivalent to about 25 mg (potency), dissolve in the mobile phase to make exactly 50 mL, and use this solu-tion as the standard solution. Then, proceed as directed in the Assay under Minocycline Hydrochloride. Amount [mg (potency)] of minocycline (C23H27N3O7) ＝ MS × AT/AS × 4 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken
Minocycline Hydrochloride Granules contain not less than 90.0z and not more than 110.0z of the labeled potency of minocycline (C23H27N3O7: 457.48). Method of preparation Prepare as directed under Gran-ules, with Minocycline Hydrochloride. Identiˆcation To a quantity of Minocycline Hydrochloride Granules, equivalent to 10 mg (potency) of Minocycline Hy- the relative retention time of about 0.83 to minocycline, is not more than 4.0z. Operating conditions— Detector, column, column temperature, mobile phase, and ‰ow rate: Proceed as directed in the operating conditions in the Assay under Minocycline Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of minocycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Minocycline Hydrochloride. Test for required detectability: To 2 mL of the standard solution obtained in the Assay add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suita-bility test, and add the mobile phase to make exactly 100 mL. Conˆrm that the peak area of minocycline obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Not more than 2.0z (4 g of powdered Minocycline Hydrochloride Granules, volumetric titration, back titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Minocycline Hydrochloride Granules in single-dose packages meet the requirement of the Content uniformity test. To the total content of 1 package of Minocycline Hydro-chloride Granules add water to disintegrate, shake thor-oughly, add water to make exactly 100 mL, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard the ˆrst 10 mL of the ˆltrate, pipet V mL of the subsequent ˆltrate, add water to make exactly V? mL so that each mL contains about 20 mg (potency) of minocy-cline (C23H27N3O7), and use this solution as the sample solution.
Related substances—Conduct this procedure within 30 minutes after the preparation of the sample solution. To a quantity of Minocycline Hydrochloride Granules, equivalent MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Minocycline Hydrochloride Granules is not less than 85z. ‌Start the test with an accurately weighed amount of Minocycline Hydrochloride Granules, equivalent to about 20 mg (potency) of Minocycline Hydrochloride, withdraw not less than 20 mL of the medium at the speciˆed minute after starting the test, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the ˆrst ˆltrate, and use the subsequent ˆltrate as the sam-ple solution. Separately, weigh accurately an amount of Minocycline Hydrochloride RS, equivalent to about 22 mg (potency), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 348 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of minocycline (C23H27N3O7) ＝ MS/MT × AT/AS × 1/C × 90 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken MT: Amount (g) of Minocycline Hydrochloride Granules taken C: Labeled amount [mg (potency)] of minocycline (C23H27N3O7) in 1 g
Weigh accurately an amount of powdered Minocy-cline Hydrochloride Granules, equivalent to about 50 mg (potency) of Minocycline Hydrochloride, add the mobile phase, shake vigorously, and add the mobile phase to make exactly 100 mL. Centrifuge this solution, and use the super-natant liquid as the sample solution. Separately, weigh accu-rately an amount of Minocycline Hydrochloride RS, equiva-lent to about 25 mg (potency), dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Minocycline Hydrochloride. Amount [mg (potency)] of minocycline (C23H27N3O7) ＝ MS × AT/AS × 2 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken
Minocycline Hydrochloride Tablets contain not less than 90.0z and not more than 110.0z of the labeled potency of Minocycline (C23H27N3O7: 457.48). Method of preparation Prepare as directed under Tablets, with Minocycline Hydrochloride. Identification To a quantity of powdered Minocycline Hy-drochloride Tablets, equivalent to 10 mg (potency) of Mino-cycline Hydrochloride, add 625 mL of a solution of hydro-chloric acid in methanol (19 in 20,000), shake well, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 221 nm and 225 nm, between 261 nm and 265 nm, and between 354 nm and 358 nm. Purity Related substances—Conduct this procedure rapidly after preparation of the sample solution. Powder not less than 5 Minocycline Hydrochloride Tablets. Weigh accurately a portion of the powder, equivalent to 50 mg (potency) of Minocycline Hydrochloride, add 60 mL of the mobile phase, shake vigorously, and add the mobile phase to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method. Calcu-late the amounts of these peaks by the area percentage method: the amount of the peak of epiminocycline, having the relative retention time of about 0.83 to minocycline, is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Minocycline Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of minocycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Minocycline Hydrochloride. Test for required detectability: To 2 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of minocycline obtained with 20 mL of this solu-tion is equivalent to 3.5 to 6.5z of that with 20 mL of the so-lution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Not more than 12.0z (0.5 g of powdered Minocycline Hydrochloride Tablets, volumetric titration, back titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Minocycline Hydrochloride Tablets add 60 mL of the mobile phase, sonicate for 15 minutes, and add the mobile phase to make exactly V mL so that each mL con-tains about 0.5 mg (potency) of Minocycline Hydrochloride. Centrifuge this solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of minocycline (C23H27N3O7) ＝ MS × AT/AS × V/50 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Minocycline Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Minocycline Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 9 mg (potency) of Minocy-cline Hydrochloride, and use this solution as the sample so-lution.
To a quantity of powdered Minocycline Hy-drochloride Tablets, equivalent to 10 mg (potency) of Mino-cycline Hydrochloride, add 625 mL of a solution of hydro-chloric acid in methanol (19 in 20,000), shake well, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 221 nm and 225 nm, between 261 nm and 265 nm, and between 354 nm and 358 nm.
Related substances—Conduct this procedure rapidly after preparation of the sample solution. Powder not less than 5 Minocycline Hydrochloride Tablets. Weigh accurately a portion of the powder, equivalent to 50 mg (potency) of Minocycline Hydrochloride, add 60 mL of the mobile phase, shake vigorously, and add the mobile phase to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method. Calcu-late the amounts of these peaks by the area percentage method: the amount of the peak of epiminocycline, having the relative retention time of about 0.83 to minocycline, is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Minocycline Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of minocycline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Minocycline Hydrochloride. Test for required detectability: To 2 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, and add the mobile phase to make exactly 100 mL. Confirm that the peak area of minocycline obtained with 20 mL of this solu-tion is equivalent to 3.5 to 6.5z of that with 20 mL of the so-lution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of minocycline is not more than 2.0z. Water <2.48> Not more than 12.0z (0.5 g of powdered Minocycline Hydrochloride Tablets, volumetric titration, back titration). Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Minocycline Hydrochloride Tablets add 60 mL of the mobile phase, sonicate for 15 minutes, and add the mobile phase to make exactly V mL so that each mL con-tains about 0.5 mg (potency) of Minocycline Hydrochloride. Centrifuge this solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount [mg (potency)] of minocycline (C23H27N3O7) ＝ MS × AT/AS × V/50 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Minocycline Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Minocycline Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 9 mg (potency) of Minocy-cline Hydrochloride, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Minocy-cline Hydrochloride RS, equivalent to about 30 mg (po-tency), and dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 348 nm. Dissolution rate (z) with respect to the labeled amount of minocycline (C23H27N3O7) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken C: Labeled amount [mg (potency)] of minocycline (C23H27N3O7) in 1 tablet
To a number of Minocycline Hydrochloride Tablets, equivalent to about 1 g (potency) of Minocycline Hydrochlo-ride, add 120 mL of the mobile phase, sonicate for 15 minutes, and add the mobile phase to make exactly 200 mL. Centrifuge this solution, pipet 5 mL of the supernatant liq-uid, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Minocycline Hydrochloride RS, equiva-lent to about 25 mg (potency), dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under ‌Minocycline Hydrochloride. Amount [mg (potency)] of minocycline (C H N O ) dryness on a water bath, moisten the residue with 3 drops of hydrochloric acid, add 10 mL of boiling water, and heat for ＝ MS × AT/AS × 40 23 27 3 7 2 minutes. Sonicate this solution, add 1 drop of phenol-phthalein TS, drop ammonia TS until a slight red color de- MS: Amount [mg (potency)] of Minocycline Hydrochlo-ride RS taken
To 5 mL of the sample solution obtained in the Purity, add a mixture of water and acetonitrile (2:1) to make 10 mL, and use this solution as the sample solution. Separately, to 50 mg of mitiglinide calcium hydrate add the mixture of water and acetonitrile (2:1), dissolve by sonicat-ing while occasional shaking, add the mixture of water and acetonitrile (2:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following System performance: Proceed as directed in the system suitability in the Purity.
Related substances—Take not less than 10 tablets of Mitiglinide Calcium Tablets, and powder. Weigh a portion of the powder, equivalent to 50 mg of Mitiglinide Calcium Hydrate, add 35 mL of a mixture of water and acetonitrile (2:1), sonicate whiles occasional shaking, add the mixture of water and acetonitrile (2:1) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 2 mL of the sample solution, add the mixture of water and acetonitrile (2:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of both solu-tions by the automatic integration method: the area of the peak, having the relative retention time of about 0.2 to mitiglinide, obtained from the sample solution is not larger than 1/4 times the peak area of mitiglinide from the stand-ard solution, and the area of peak other than mitiglinide and the peak mentioned above from the sample solution is not larger than 1/8 times the peak area of mitiglinide from the standard solution. In addition, the total area of the peaks other than mitiglinide from the sample solution is not larger than 1/2 times the peak area of mitiglinide from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with palmitamide propyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Adjust to pH 2.0 of a mixture of water, acetonitrile for liquid chromatography and n-amyl alcohol (66:33:1) with phosphoric acid. Flow rate: Adjust so that the retention time of mitiglinide is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of mitiglinide, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2.5 mL of the stand-ard solution, add the mixture of water and acetonitrile (2:1) to make exactly 50 mL. Confirm that the peak area of mitiglinide obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mitiglinide are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitiglinide is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mitiglinide Calcium Tablets add a mixture of water and acetonitrile (2:1), add exactly V/10 mL of the internal standard solution, sonicate while occasional shak-ing, then add the mixture of water and acetonitrile (2:1) to make V mL so that each mL contains about 0.1 mg of mitiglinide calcium hydrate (C H CaN O .2H O), and to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of mitiglinide in each solution. Dissolution rate (z) with respect to the labeled amount of mitiglinide calcium hydrate (C38H48CaN2O6.2H2O) ＝ MS × AT/AS × V?/V × 1/C × 18 × 1.054 MS: Amount (mg) of Mitiglinide Calcium RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of mitiglinide calcium hydrate (C38H48CaN2O6.2H2O) in 1 tablet. 38 48 2 6 2 filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 50 mg of Mitiglinide Calcium RS (separately determine the water <2.48> in the same man-ner as Mitiglinide Calcium Hydrate), add the mixture of water and acetonitrile (2:1), dissolve by sonicating while occasional shaking, and add the mixture of water and aceto-nitrile (2:1) to make exactly 100 mL. Pipet 20 mL of this so-lution, add exactly 10 mL of the internal standard solution, add the mixture of water and acetonitrile (2:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of mitiglinide to that of the internal standard. Operating conditions— Proceed as directed in the operating conditions in the Assay under Mitiglinide Calcium Hydrate. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mitiglinide are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitiglinide is not more than 1.5z.
Weigh accurately the mass of not less than 20 tablets of Mitiglinide Calcium Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 10 mg of mitiglinide calcium hydrate (C H CaN O .2H O), add a 38 48 2 6 2 Amount (mg) of mitiglinide calcium hydrate (C38H48CaN2O6.2H2O) ＝ MS × QT/QS × V/500 × 1.054 MS: Amount (mg) of Mitiglinide Calcium RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of 2-nitrophenol in acetonitrile (1 in 5000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Mitiglinide Calcium Hydrate. System suitability— Proceed as directed in the operating conditions in the Assay. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Mitiglinide Calcium Tablets is not less than 85z. Start the test with 1 tablet of Mitiglinide Calcium Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 1 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mixture of water and acetonitrile (2:1) to make exactly V? mL so that each mL contains about 5.6 mg of mitiglinide calcium hydrate (C38H48CaN2O6.2H2O), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Mitiglinide Calcium RS (separate-ly determine the water <2.48> in the same manner as Mitiglinide Calcium Hydrate), add the mixture of water and acetonitrile (2:1), dissolve by sonicating while occasional shaking, and add the mixture of water and acetonitrile (2:1) mixture of water and acetonitrile (2:1), add exactly 10 mL of the internal standard solution, sonicate while occasional shaking, then add the mixture of water and acetonitrile (2:1) to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 50 mg of Mitiglinide Calcium RS (separately determine the water <2.48> in the same manner as Mitiglinide Calcium Hydrate), add the mixture of water and acetonitrile (2:1), dissolve by sonicating while occasional shaking, and add the mixture of water and acetonitrile (2:1) to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 10 mL of the internal stand-ard solution, add the mixture of water and acetonitrile (2:1) to make 100 mL, and use this solution as the standard solu-tion. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of mitiglinide to that of the internal standard. Amount (mg) of mitiglinide calcium hydrate (C38H48CaN2O6.2H2O) ＝ MS × QT/QS × 1/5 × 1.054 MS: Amount (mg) of Mitiglinide Calcium RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of 2-nitrophenol in acetonitrile (1 in 5000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Mitiglinide Calcium Hydrate. System suitability— System performance: When the procedure is run with 5 mL ‌of the standard solution under the above operating condi-tions, the internal standard and mitiglinide are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak of mitiglinide to that of the internal standard is not more than 1.0z.
Mitomycin C for Injection is a preparation for in-jection, which is dissolved before use. It contains not less than 90.
Mitomycin C for Injection occurs as a blue-purple powder.
Dissolve an amount of Mitomycin C for In-jection, equivalent to 2 mg (potency) of Mitomycin C, in 200 mL of water, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 216 nm and 220 nm, and between 362 nm and 366 nm. pH <2.54> The pH of a solution, prepared by dissolving 0.25 g of Mitomycin C for Injection in 20 mL of water, is 5.5 to 8.5. Loss on drying <2.41> Not more than 1.0z (0.4 g, in vacu-um not exceeding 0.67 kPa, phosphorus (V) oxide, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 10 EU/mg (potency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To the content of 1 container of Mitomycin C for Injec-tion add exactly V mL of N,N-dimethylacetamide so that each mL contains about 0.5 mg (potency) of Mitomycin C, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg (potency) of Mitomycin C RS, add N,N-dimethylacetamide to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Mitomycin C. Amount [mg (potency)] of mitomycin C (C15H18N4O5) ＝ MS × AT/AS × V/50 MS: Amount [mg (potency)] of Mitomycin C RS taken Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Mitomycin C for Injection. Weigh ac-curately an amount of the contents, equivalent to about 10 mg (potency) of Mitomycin C, add exactly 20 mL of N,N-dimethylacetamide, shake, centrifuge, and use the superna-tant liquid as the sample solution. Separately, weigh accu-rately an amount of Mitomycin C RS, equivalent to about 25 mg (potency), dissolve in N,N-dimethylacetamide to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Mitomycin C. Amount [mg (potency)] of mitomycin C (C15H18N4O5) ＝ MS × AT/AS × 2/5 MS: Amount [mg (potency)] of Mitomycin C RS taken
To a quantity of powdered Mizoribine Tablets, equivalent to 0.1 g of Mizoribine, add 5 mL of water, shake, filter, and use the filtrate as the sample solu-tion. Separately, dissolve 20 mg of Mizoribine RS in 1 mL of water, and use this solution as the standard solution. Per-form the test with the sample solution and standard solution as directed under Thin-Layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Then de-velop the plate with a mixture of methanol, ammonia solu-tion (28) and 1-propanol (2:1:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the principal spot obtained from the sample solution and the spot from the standard solution show a red-brown color and the same Rf value.
Related substances—To a quantity of powdered Mizoribine Tablets, equivalent to 0.10 g of Mizoribine, add 30 mL of the mobile phase, shake, then add the mobile phase to make 50 mL. Filter the solution through a mem-brane filter with a pore size not exceeding 0.5 mm and use the filtrate as the sample solution. Pipet 2 mL of the sample so-lution, add the mobile phase to make exactly 20 mL. Pipet 1 mL of the solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography ‌<2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.3 to mizoribine, obtained from the sample solution is not larger than the peak area of mizoribine from the standard solution, and the area of the peak other than mizoribine and the peak mentioned above is not larger than 2/5 times the peak area of mizoribine from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Mizoribine. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 3 times as long as the retention time of mizoribine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mobile phase to make exactly 5 mL. Confirm that the peak area of mizoribine obtained with 5 mL of this solution is equivalent to 14 to 26z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of mizoribine are not less than 10,000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of mizoribine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Mizoribine Tablets add 50 mL of water, shake until the tablet is disintegrated, and add water to make exactly 100 mL. Filter the solution, discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 5 mg of mizoribine (C9H13N3O6), and use this so-lution as the sample solution. Then, proceed as directed in the Assay. AT and AS, at 279 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Dissolution rate (z) with respect to the labeled amount of mizoribine (C9H13N3O6) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Mizoribine RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of mizoribine (C9H13N3O6) in 1 tablet
Weigh accurately not less than 20 Mizoribine Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 25 mg of mizoribine (C9H13N3O6), add 50 mL of water and shake, then add water to make exactly 100 mL. Filter the solution, discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 25 mg of Mizoribine RS (separately determine the water <2.48> in the same manner as Mizoribine), and dissolve in water to make exactly 100 mL. Pipet 2 mL of the solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 279 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of mizoribine (C9H13N3O6) ＝ MS × AT/AS MS: Amount (mg) of Mizoribine RS taken, calculated on the anhydrous basis
モンテルカストナトリウムチュアブル錠
Montelukast Sodium Chewable Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of montelukast (C35H36ClNO3S: 586.18). Method of preparation Prepare as directed under Chewa-ble Tablets, with Montelukast Sodium. Identification To an amount of powdered Montelukast So-dium Chewable Tablets, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm and between 357 nm and 361 nm. Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than 1.5 times the peak area of montelukast from the standard solution, the area of related substance B having the relative retention time of about 0.92 from the sample solu-tion is not larger than 3/20 times the peak area of mon-telukast from the standard solution, and the area of the peaks other than montelukast and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of montelukast from the standard solution. Furthermore, the total area of the peaks other than mon-telukast is not larger than 1.8 times the peak area of mon-telukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related sub-stance E), about 1.16 (related substance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correc-tion factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Montelukast Sodium Chewable Tablets add 50 mL of water to disintegrate the tablet, add a suitable amount of methanol, and disperse the fine particles by sonicating. Add methanol to make exactly 200 mL, and centrifuge or filter. Pipet V mL of this solution, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 25 mg of montelukast (C35H36ClNO3S) and use this so-lution as the sample solution.
To an amount of powdered Montelukast So-dium Chewable Tablets, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm and between 357 nm and 361 nm.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than 1.5 times the peak area of montelukast from the standard solution, the area of related substance B having the relative retention time of about 0.92 from the sample solu-tion is not larger than 3/20 times the peak area of mon-telukast from the standard solution, and the area of the peaks other than montelukast and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of montelukast from the standard solution. Furthermore, the total area of the peaks other than mon-telukast is not larger than 1.8 times the peak area of mon-telukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related sub-stance E), about 1.16 (related substance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correc-tion factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Montelukast Sodium Chewable Tablets add 50 mL of water to disintegrate the tablet, add a suitable amount of methanol, and disperse the fine particles by sonicating. Add methanol to make exactly 200 mL, and centrifuge or filter. Pipet V mL of this solution, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 25 mg of montelukast (C35H36ClNO3S) and use this so-lution as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dis-solve in a mixture of methanol and water (3:1) to make ex-actly 200 mL. Pipet 20 mL of this solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Amount (mg) of montelukast (C35H36ClNO3S) ＝ MS × AT/AS × V?/V × 1/5 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 389 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 10 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A solution of trifluoroacetic acid in a mix-ture of water and acetonitrile for liquid chromatography (1:1) (1 in 500). Flow rate: Adjust so that the retention time of mon-telukast is about 2 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of montelukast are not less than 2000 and not more than 1.5, respectively. ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Disintegrate 10 tablets of Montelukast Sodium Chewable Tablets in 150 mL of a mixture of methanol and water (3:1), disperse the fine particles by sonicating, and add a mixture of methanol and water (3:1) to make exactly 200 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, pipet V mL of the subsequent filtrate, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 0.25 mg of montelukast (C35H36ClNO3S), and use this solu-tion as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dis-solve in a mixture of methanol and water (3:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat- Amount (mg) of montelukast (C 35H 36ClNO3S) in 1 tablet ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of sodium lauryl sulfate (1 in 200) as the dissolution medium, the dissolution rate in 20 minutes of Montelukast Sodium Chewable Tablets is not less than 85z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Montelukast Sodium Chewable Tablets, withdraw not less than 15 mL of the medium at the specified minute after starting the test, and centrifuge. Pipet V mL of the supernatant liquid, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of montelukast (C35H36ClNO3S), and use this solution as the sample solution. Separately, weigh accurately about 35 mg of Montelukast Dicyclohexylamine RS, dissolve in meth-anol to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Dissolution rate (z) with respect to the labeled amount of montelukast (C35H36ClNO3S) ＝ MS × AT/AS × V?/V × 1/C × 18 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken C: Labeled amount (mg) of montelukast (C35H36ClNO3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of montelukast are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 50 mL of the standard solution under the above operat- ＝ MS × AT/AS × V?/V × 1/5 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with phenylhexylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 509C. Mobile phase A: A solution of trifluoroacetic acid (1 in 500). Mobile phase B: A mixture of methanol and acetonitrile for liquid chromatography (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 5 48 → 45 52 → 55 5 – 12 12 – 22 45 → 25 55 → 75 22 – 23 Flow rate: 1.5 mL per minute (the retention time of mon-telukast is about 14 minutes). System suitability— System performance: Take 10 mL of the standard solution in a transparent vessel, add 4 mL of hydrogen peroxide (30), and allow to stand under 4000 lx white light for 10 minutes. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution between the peak of related substance B, having a relative retention time of about 0.92 to montelukast and the peak of montelukast is not less than 1.5. And proceed with 20 mL of the standard solution under the above conditions, the number of theoreti-cal plates and the symmetry factor of the peak of mon-telukast are not less than 5000 and not more than 2.5, re-spectively. ‌System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z.
Montelukast Sodium Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of montelukast (C35H36ClNO3S: 586.18). Method of preparation Prepare as directed under Gran-ules, with Montelukast Sodium. Identification To an amount of Montelukast Sodium Granules, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm, and between 357 nm and 361 nm. Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than the peak area of montelukast from the standard solution, and the peak area of related substance B, having the relative retention time of about 0.92, from the sample so-lution is not larger than 3/20 times the peak area of mon-telukast from the standard solution, and the area of the peak other than montelukast and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of montelukast from the standard solution. Further-more, the total area of the peaks other than montelukast from the sample solution is not larger than 1.2 times the peak area of montelukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related substance E), about 1.16 (related sub-stance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correction factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: Montelukast Sodium Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To the total content of 1 package of Montelukast Sodium Gran-ules add 130 mL of methanol, disperse the fine particles by sonicating, and add methanol to make exactly V mL so that each mL contains about 20 mg of montelukast (C35H36ClNO3S).
To an amount of Montelukast Sodium Granules, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm, and between 357 nm and 361 nm.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than the peak area of montelukast from the standard solution, and the peak area of related substance B, having the relative retention time of about 0.92, from the sample so-lution is not larger than 3/20 times the peak area of mon-telukast from the standard solution, and the area of the peak other than montelukast and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of montelukast from the standard solution. Further-more, the total area of the peaks other than montelukast from the sample solution is not larger than 1.2 times the peak area of montelukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related substance E), about 1.16 (related sub-stance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correction factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: Montelukast Sodium Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To the total content of 1 package of Montelukast Sodium Gran-ules add 130 mL of methanol, disperse the fine particles by sonicating, and add methanol to make exactly V mL so that each mL contains about 20 mg of montelukast (C35H36ClNO3S). Centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dissolve in methanol to make exactly 100 mL. Pipet 8 mL of this so-lution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Amount (mg) of montelukast (C35H36ClNO3S) ＝ MS × AT/AS × V/1250 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 389 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 10 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A solution of trifluoroacetic acid in a mix-ture of water and acetonitrile (1:1) (1 in 500). Flow rate: Adjust so that the retention time of mon-telukast is about 2 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of montelukast are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of sodium lauryl sulfate (1 in 200) as the dissolution medium, the dissolution rate in 15 minutes of Montelukast Sodium Granules is not less than 85z. Conduct this procedure using light-resistant vessels. Start the test with an accurately weighed amount of Montelukast Sodium Granules, equivalent to about 4 mg of montelukast (C35H36ClNO3S), withdraw not less than 15 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding ‌0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 27 mg of Montelukast Dicy-clohexylamine RS, and dissolve in methanol to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolution me-dium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Dissolution rate (z) with respect to the labeled amount of montelukast (C35H36ClNO3S) ＝ MS/MT × AT/AS × 1/C × 18 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken MT: Amount (g) of Montelukast Sodium Granules taken C: Labeled amount (mg) of montelukast (C35H36ClNO3S) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of montelukast are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z.
Conduct this procedure using light-resistant vessels. Weigh accurately an amount of Montelukast Sodium Gran-ules, equivalent to about 48 mg of montelukast (C35H36ClNO3S), and add exactly 200 mL of a mixture of methanol and water (3:1). Disperse the fine particles by sonicating, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dissolve in a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Amount (mg) of montelukast (C35H36ClNO3S) ＝ MS × AT/AS × 2 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with phenylhexylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 509C. Mobile phase A: A solution of trifluoroacetic acid (1 in 500). Mobile phase B: A mixture of methanol and acetonitrile (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 5 48 → 45 52 → 55 5 – 12 12 – 22 45 → 25 55 → 75 22 – 23 Flow rate: 1.5 mL per minute (the retention time of mon-telukast is about 14 minutes). System suitability— System performance: Take 10 mL of the standard solution in a transparent vessel, add 4 mL of hydrogen peroxide (30), and allow to stand under 4000 lx white light for 10 minutes. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution between the peak of related substance B, having the relative retention time of about 0.92 to montelukast, and the peak of mon-telukast is not less than 1.5. And proceed with 20 mL of the standard solution under the above operating conditions, the number of the theoretical plates and the symmetry factor of the peak of montelukast are not less than 5000 and not more than 2.5, respectively. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z.
Montelukast Sodium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of montelukast (C35H36ClNO3S: 586.18). Method of preparation Prepare as directed under Tablets, with Montelukast Sodium. Identification To an amount of powdered Montelukast Sodium Tablets, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm and between 357 nm and 361 nm. Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than the peak area of montelukast from the standard solution, the area of related substance B having the relative retention time of about 0.92 from the sample solution is not larger than 3/20 times the peak area of montelukast from the standard solution, and the area of the peaks other than mon-telukast and the peaks mentioned above from the sample so-lution is not larger than 1/10 times the peak area of mon-telukast from the standard solution. Furthermore, the total area of the peaks other than montelukast from the sample solution is not larger than 1.2 times the peak area of mon-telukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related sub-stance E), about 1.16 (related substance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correc-tion factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Montelukast Sodium Tablets add 50 mL of water to disintegrate the tablet, add a suitable amount of methanol, and disperse the fine particles by sonicating. Add methanol to make exactly 200 mL, and centrifuge or filter. Pipet V mL of this solution, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 25 mg of montelukast (C35H36ClNO3S) and use this solution as the sample solution.
To an amount of powdered Montelukast Sodium Tablets, equivalent to 5 mg of montelukast (C35H36ClNO3S), add 500 mL of a mixture of methanol and water (3:1), shake, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, between 325 nm and 329 nm, be- tween 343 nm and 347 nm and between 357 nm and 361 nm.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the total area of the two peaks of related substance A, having the relative retention time of about 0.45 to montelukast, obtained from the sample solution is not larger than the peak area of montelukast from the standard solution, the area of related substance B having the relative retention time of about 0.92 from the sample solution is not larger than 3/20 times the peak area of montelukast from the standard solution, and the area of the peaks other than mon-telukast and the peaks mentioned above from the sample so-lution is not larger than 1/10 times the peak area of mon-telukast from the standard solution. Furthermore, the total area of the peaks other than montelukast from the sample solution is not larger than 1.2 times the peak area of mon-telukast from the standard solution. However, the peaks of the related substances derived from Montelukast Sodium [having the relative retention time of about 1.04 (related sub-stance E), about 1.16 (related substance C), about 1.18 (related substance D), about 1.24 and about 1.55 (related substance F)] are excluded. For the area of the peak, having the relative retention time of about 0.71, multiply the correc-tion factor 0.6. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.5 times as long as the retention time of montelukast, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 10 mL of the stand-ard solution, and add a mixture of methanol and water (3:1) to make exactly 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Montelukast Sodium Tablets add 50 mL of water to disintegrate the tablet, add a suitable amount of methanol, and disperse the fine particles by sonicating. Add methanol to make exactly 200 mL, and centrifuge or filter. Pipet V mL of this solution, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 25 mg of montelukast (C35H36ClNO3S) and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dissolve in a mixture of methanol and water (3:1) to make exactly 200 mL. Pipet 20 mL of this solution, add a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 389 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 10 cm in length, packed with phenylated silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A solution of trifluoroacetic acid in a mix-ture of water and acetonitrile for liquid chromatography (1:1) (1 in 500). Flow rate: Adjust so that the retention time of mon-telukast is about 2 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of montelukast are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of sodium lauryl sulfate (1 in 200) as the dissolution medium, the dissolution rate in 20 minutes of Montelukast Sodium Tablets is not less than 85z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Montelukast Sodium Tablets, with-draw not less than 15 mL of the medium at the specified minute after starting the test, and centrifuge. Pipet V mL of the supernatant liquid, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of montelukast (C35H36ClNO3S), and use this solution as the sample solution. Separately, weigh accurately about 35 mg of Montelukast Dicyclohexylamine RS, and dissolve in methanol to make exactly 100 mL. Pipet 2 mL of this solu-tion, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Dissolution rate (z) with respect to the labeled amount of montelukast (C35H36ClNO3S) ＝ MS × AT/AS × V?/V × 1/C × 18 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken C: Labeled amount (mg) of montelukast (C35H36ClNO3S) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Uniformity of dosage units. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of montelukast are not less than 2000 and not more than 1.5, respectively. Amount (mg) of montelukast (C 35H36ClNO3S) System repeatability: When the test is repeated 5 times with 50 mL of the standard solution under the above operat- ＝ MS × AT/AS × V?/V × 1/5 × 0.764 ‌ing conditions, the relative standard deviation of the peak area of montelukast is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Disintegrate 10 tablets of Montelukast Sodium Tablets in 150 mL of a mixture of methanol and water (3:1), disperse the fine particles by sonicating, and add a mixture of metha-nol and water (3:1) to make exactly 200 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 1 mL of the filtrate, pipet V mL of the subse-quent filtrate, add a mixture of methanol and water (3:1) to make exactly V? mL so that each mL contains about 0.25 mg of montelukast (C35H36ClNO3S), and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Montelukast Dicyclohexylamine RS, and dissolve in a mixture of methanol and water (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of montelukast in each solution. Amount (mg) of montelukast (C35H36ClNO3S) in 1 tablet ＝ MS × AT/AS × V/V? × 1/5 × 0.764 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with phenylhexylsilanized silica gel for liquid chromatography (3 mm in particle diame-ter). Column temperature: A constant temperature of about 509C. Mobile phase A: A solution of trifluoroacetic acid (1 in 500). Mobile phase B: A mixture of methanol and acetonitrile for liquid chromatography (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 5 48 → 45 52 → 55 5 – 12 12 – 22 45 → 25 55 → 75 22 – 23 Flow rate: 1.5 mL per minute (the retention time of mon-telukast is about 14 minutes). System suitability— System performance: Take 10 mL of the standard solution in a transparent vessel, add 4 mL of hydrogen peroxide (30), and allow to stand under 4000 lx white light for 10 minutes. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolution between the peak of related substance B, having a relative retention time of about 0.92 to montelukast and the peak of montelukast is not less than 1.5. And proceed with 20 mL of the standard solution under the above conditions, the number of theoreti-cal plates and the symmetry factor of the peak of mon-telukast are not less than 5000 and not more than 2.5, re-spectively. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 1.0z.
Morphine and Atropine Injection is an aqueous in-jection. It contains not less than 0.
Morphine and Atropine Injection is a clear, colorless liquid. It is gradually colored by light. pH: 2.5 – 5.0
To 2 mL of Morphine and Atropine Injec-tion add 2 mL of ammonia TS, and extract with 10 mL of diethyl ether. Filter the extract with a filter paper, evaporate the filtrate on a water bath to dryness, dissolve the residue in 1 mL of ethanol (99.5), and use this solution as the sample solution. Separately, dissolve 0.1 g of morphine hydrochlo-ride hydrate in 10 mL of water, perform with 2 mL of this solution the same procedure as used for preparation of the sample solution, and use the solution so obtained as the standard solution (1). Separately, dissolve 3 mg of atropine sulfate hydrate in 10 mL of water, perform with 2 mL of this solution the same procedure as used for preparation of the sample solution, and use the solution so obtained as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of methanol and am-monia solution (28) (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS on the plate: the two spots obtained from the sample solution show the same color tone and the same Rf value with either spot of orange color from the standard solution (1) or the stand-ard solution (2) (morphine and atropine). Extractable volume <6.05> It meets the requirement.
(1) Morphine hydrochloride hydrate—Pipet 2 mL of Morphine and Atropine Injection, add exactly 10 mL of the internal standard solution, then add water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride hydrate for assay, add exactly 10 mL of the internal standard solution to dissolve, then add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O) ＝ MS × QT/QS × 1.168 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH with sodium hydroxide TS to 3.0. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 1.0z. (2) Atropine sulfate hydrate—Pipet 2 mL of Morphine and Atropine Injection, add exactly 2 mL of the internal standard solution, and use this solution as the sample solu-tion. Separately, weigh accurately about 15 mg of Atropine Sulfate RS (separately determine the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak areas of atropine to that of the inter- ‌nal standard. Amount (mg) of atropine sulfate hydrate solution of Morphine Sulfate Hydrate in dilute sodium hy-droxide TS (1 in 10,000) as directed under Ultraviolet-visible [(C17H23 NO3)2.H2SO4.H2O] Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 2: both spectra exhibit similar inten- ＝ MS × QT/QS × 1/25 × 1.027 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 12,500). Operating conditions— Column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay (1). Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Flow rate: Adjust so that the retention time of morphine is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the sample solution under the above operating condi-tions, morphine, the internal standard and atropine are eluted in this order, and the resolution between morphine and the internal standard is not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of atropine to that of the internal standard is not more than 1.0z.
Morphine Hydrochloride Injection is an aqueous in-jection. It contains not less than 93.
Morphine Hydrochloride Injection is a clear, colorless or pale yellow-brown liquid. It gradually becomes yellow-brown by light. pH: 2.5 – 5.0
Take a volume of Morphine Hydrochloride Injection, equivalent to 0.04 g of Morphine Hydrochloride Hydrate, add water to make 20 mL, and use this solution as the sample solution. To 5 mL of the sample solution add water to make 100 mL, and determine the absorption spec-trum as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 283 nm and 287 nm. And to 5 mL of the sample solution add dilute sodium hydroxide TS to make 100 mL, and determine the absorp-tion spectrum as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 296 nm and 300 nm. Bacterial endotoxins <4.01> Less than 1.5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take exactly a volume of Morphine Hydrochloride Injection, equivalent to about 80 mg of morphine hydrochlo-ride hydrate (C17H19NO3.HCl.3H2O), and add water to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard solution, add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O) ＝ MS × QT/QS × 4 × 1.168 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 1.0z.
Morphine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O: 375.84). Method of preparation Prepare as directed under Tablets, with Morphine Hydrochloride Hydrate. Identification Weigh a quantity of powdered Morphine Hydrochloride Tablets equivalent to 0.01 g of Morphine Hy-drochloride Hydrate, add 100 mL of water, shake for 10 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. And weigh a quantity of powdered Morphine Hy-drochloride Tablets equivalent to 0.01 g of Morphine Hydro-chloride Hydrate, add 100 mL of dilute sodium hydroxide TS, shake for 10 minutes, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 296 nm and 300 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Morphine Hydrochloride Tablets add ex-actly 1 mL of the internal standard solution per 2 mg of mor-phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), dis-perse the tablet into a small particles by sonicating, then sonicate for 15 minutes with occasional stirring, and add water to make V mL so that each mL contains about 0.
Weigh a quantity of powdered Morphine Hydrochloride Tablets equivalent to 0.01 g of Morphine Hy-drochloride Hydrate, add 100 mL of water, shake for 10 minutes, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 283 nm and 287 nm. And weigh a quantity of powdered Morphine Hy-drochloride Tablets equivalent to 0.01 g of Morphine Hydro-chloride Hydrate, add 100 mL of dilute sodium hydroxide TS, shake for 10 minutes, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 296 nm and 300 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Morphine Hydrochloride Tablets add ex-actly 1 mL of the internal standard solution per 2 mg of mor-phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), dis-perse the tablet into a small particles by sonicating, then sonicate for 15 minutes with occasional stirring, and add water to make V mL so that each mL contains about 0.4 mg of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O). Filter the solution, and use the filtrate as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O) ＝ MS × QT/QS × V/50 × 1.168 sequent filtrate as the sample solution. Separately, weigh ac-curately about 28 mg of morphine hydrochloride hydrate for assay (separately, determine the water <2.48> in the same manner as Morphine Hydrochloride Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solu-tion, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of morphine in each solution. Dissolution rate (z) with respect to the labeled amount of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O) ＝ MS × AT/AS × 1/C × 36 × 1.168 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of morphine hydrochloride hy-drate (C17H19NO3.HCl.3H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of morphine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of morphine is not more than 2.0z.
Take not less than 20 Morphine Hydrochloride Tablets, weigh accurately, and powder. Weigh accurately a quantity of the powder, equivalent to about 20 mg of mor-phine hydrochloride hydrate (C17H19NO3.HCl.3H2O), add exactly 10 mL of the internal standard solution, extract the mixture by sonicating for 10 minutes, and add water to make 50 mL. Filter this solution, and use the filtrate as the sample solution. Separately, weigh accurately about 25 mg of mor-phine hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard solution, add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine hydrochloride hydrate (C17H19NO3.HCl.3H2O) MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Morphine Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Morphine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, and use the sub- ＝ MS × QT/QS × 1.168 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in ‌500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 1.0z.
(1) Powder Mosapride Citrate Powder. To a portion of the powder, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), add 10 mL of dilute acetic acid, shake for 10 minutes, and filter. To 5 mL of the filtrate add 0.3 mL of Dragendorff's TS: an orange precipitate is formed. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 271 nm and 275 nm and between 306 nm and 310 nm.
Related substances—Powder Mosapride Citrate Powder. To a portion of the powder, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), moisten with 1 mL of water, then add 9 mL of methanol, shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 20 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 20 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the two peaks, having the relative retention time of about 0.60 and about 0.85 to mosapride obtained from the sample solution, is not larger than the peak area of mosapride from the standard so-lution, the area of other than mosapride and the peaks men-tioned above from the sample solution is not larger than 2/5 times the peak area of mosapride from the standard solu-tion, and the total area of the peak other than mosapride from the sample solution is not larger than 2 times the peak area of mosapride from the standard solution. Operating conditions— Detector, column, column temperature, mobile phases A and B, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Mosapride Citrate Hy-drate. Flowing of mobile phase: Control the gradient by mixing Uniformity of dosage units <6.02> Perform the test accord- ing to the following method: the powder in single-dose pack-ages meets the requirement of the Content uniformity test. To the total amount of the content of 1 package of Mosapride Citrate Powder add 5 mL of water, and shake. Then, add 20 mL of methanol, shake for 20 minutes, and add methanol to make exactly 50 mL. Centrifuge this solu-tion, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL so that each mL contains about 20 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), and use this so-lution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Mosapride Citrate Powder is not less than 70z. Start the test with an amount of Mosapride Citrate Powder, equivalent to about 2.5 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 30 mg of mosapride citrate hydrate for assay (separately determine the water <2.48> in the same manner as Mosapride Citrate Hydrate), and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, and determine the peak areas, AT and AS, of mosapride in each solution. Dissolution rate (z) with respect to the labeled amount of mosapride citrate (C H ClFN O .C H O ) the mobile phases A and B as directed in the following table. 21 25 3 3 6 8 7 Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) ＝ MS/MT × AT/AS × 1/C × 9 MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis MT: Amount (g) of Mosapride Citrate Powder taken 0 – 40 85 – 45 15 – 55 Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add methanol to make exactly 25 mL. Confirm that the peak area of mosapride obtained with 10 mL of this solution is equivalent to 3.0 to 5.0z of that with 10 mL of the standard solution. C: Labeled amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) in 1 g Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 8.82 g of trisodium citrate dihy-drate in 800 mL of water, adjust to pH 3.3 with dilute hydro-chloric acid, and add water to make 1000 mL. To 240 mL of this solution add 90 mL of methanol and 70 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of mosapride is about 9 minutes. System suitability— ‌System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mosapride are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mosapride is not more than 2.0z.
Powder Mosapride Citrate Powder. Weigh accu-rately a portion of the powder, equivalent to about 10 mg of (C21H25ClFN3O3.C6H8O7), with 1 mL of water. Add 9 mL of methanol, shake for 20 minutes, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of this solution, add methanol to make exactly 20 mL. Pipet 2 mL of the sample solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peaks having the relative retention times of about 0.60 and about 0.85 to mosapride obtained from the sample solu-tion is not larger than the peak area of mosapride from the standard solution, and the area of each peak other than mosapride and these peaks mentioned above from the sam-ple solution is not larger than 2/5 times the peak area of mosapride from the standard solution. Furthermore, the total area of the peaks other than mosapride from the sam-ple solution is not larger than 2 times the peak area of mosapride citrate (C21H25 ClFN3O3.C6H8O7), moisten with 2 mosapride from the standard solution. Operating conditions— mL of water, add 70 mL of methanol, shake for 20 minutes, then add methanol to make exactly 100 mL, and centrifuge. Pipet 10 mL of the supernatant liquid, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 53 mg of mosapride citrate hydrate for assay (separately determine the water <2.48> in the same manner as Mosapride Citrate Hy-drate), and dissolve in methanol to make exactly 100 mL. To Detector, column, column temperature, mobile phases A and B, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Mosapride Citrate Hy-drate. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. mL of this solution add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) absorbances, AT and AS, of the sample solution and the standard solution at 273 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis
Mosapride Citrate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of mosapride citrate (C21H25ClFN3O3.C6H8O7: 614.02). Method of preparation Prepare as directed under Tablets, with Mosapride Citrate Hydrate. Identification (1) To an amount of powdered Mosapride Citrate Tablets, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), add 10 mL of dilute acetic acid, shake for 10 minutes, and filter. To 5 mL of the filtrate add 0.3 mL of Dragendorff's TS: an orange precipitate is formed. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 271 nm and 275 nm, and between 306 nm and 310 nm. Purity Related substances—Powder not less than 20 tablets of Mosapride Citrate Tablets. Moisten a portion of the powder, equivalent to 10 mg of mosapride citrate 0 – 40 85 → 45 15 → 55 Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add methanol to make exactly 25 mL. Confirm that the peak area of mosapride obtained with 10 mL of this solution is equivalent to 3.0 to 5.0z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mosapride are not less than 40,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mosapride is not more than 3.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mosapride Citrate Tablets add 5 mL of water, and shake well to disintegrate. Add 20 mL of metha-nol, shake for 20 minutes, and add methanol to make exactly 50 mL. Centrifuge this solution, pipet V mL of the superna-tant liquid, add methanol to make exactly V? mL so that each mL contains about 20 mg of mosapride citrate (C21H25ClFN3O3.
(1) To an amount of powdered Mosapride Citrate Tablets, equivalent to 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), add 10 mL of dilute acetic acid, shake for 10 minutes, and filter. To 5 mL of the filtrate add 0.3 mL of Dragendorff's TS: an orange precipitate is formed. (2) Determine the absorption spectrum of the sample so-lution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 271 nm and 275 nm, and between 306 nm and 310 nm.
Related substances—Powder not less than 20 tablets of Mosapride Citrate Tablets. Moisten a portion of the powder, equivalent to 10 mg of mosapride citrate 0 – 40 85 → 45 15 → 55 Time span of measurement: For 40 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add methanol to make exactly 25 mL. Confirm that the peak area of mosapride obtained with 10 mL of this solution is equivalent to 3.0 to 5.0z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mosapride are not less than 40,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mosapride is not more than 3.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Mosapride Citrate Tablets add 5 mL of water, and shake well to disintegrate. Add 20 mL of metha-nol, shake for 20 minutes, and add methanol to make exactly 50 mL. Centrifuge this solution, pipet V mL of the superna-tant liquid, add methanol to make exactly V? mL so that each mL contains about 20 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), and use this solution as the sam-ple solution. Proceed as directed in the Assay. Amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 45 minutes of Mosapride Citrate Tablets is not less than 80z.‌ Start the test with 1 tablet of Mosapride Citrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less mosapride citrate hydrate for assay (separately, determine the water <2.48> in the manner as Mosapride Citrate Hy-drate), and dissolve in methanol to make exactly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 273 nm. than 10 mL of the first filtrate, pipet V mL of the subsequent Amount (mg) of mosapride citrate (C H ClFN O .C H O ) filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 2.8 mg of mosapride citrate ＝ MS × AT/AS × 1/5 21 25 3 6 8 7 (C21H25ClFN3O3.C6H8O7), and use this solution as the sam-ple solution. Separately, weigh accurately about 30 mg of mosapride citrate hydrate for assay (separately, determine the water <2.48> in the same manner as Mosapride Citrate Hydrate), and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mosapride in each solution. Dissolution rate (z) with respect to the labeled amount of mosapride citrate (C21H25ClFN3O3.C6H8O7) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of mosapride citrate (C21H25ClFN3O3.C6H8O7) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 8.82 g of trisodium citrate dihy-drate in 800 mL of water, adjust the pH to 3.3 with dilute MS: Amount (mg) of mosapride citrate hydrate for assay taken, calculated on the anhydrous basis
To an amount of Mupirocin Calcium Oint-ment, equivalent to 10 mg (potency) of Mupirocin Calcium Hydrate, add 5 mL of water, and warm on a water bath at 609C for 10 minutes while occasional shaking. After cooling, filter, and to 1 mL of the filtrate add water to make 100 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 220 nm and 224 nm.
Related substances—To an amount of Mupirocin Calcium Ointment, equivalent to 50 mg (potency) of Mupirocin Calcium Hydrate, add 5 mL of diluted tetra-hydrofuran (3 in 4), and shake vigorously. Then, add 5 mL of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0), shake vigorously, filter through a glass wool filter, and use the filtrate as the sample solution. Pipet 2 mL of the sample solution, add a mixture of 0.1 mol/L acetic acid-so-dium acetate buffer solution (pH 4.0) and diluted tetra-hydrofuran (3 in 4) (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the area of the peak other than mupirocin obtained from the sample solution and the peak area of mupirocin from the standard solution by the automatic integration method. Calculate the amount of each related substance using the following equa-tion: the amount of the related substance having the relative retention time of about 0.7 to mupirocin is not more than 4.0z, the amount of the related substance other than that is not more than 1.5z, and the total amount of the related substances is not more than 6.0z. Amount (z) of each related substance ＝ A/(SA ＋ Am) × 100 A: Peak area of each related substance obtained from the sample solution. SA: Total area of the peaks other than mupirocin ob-tained from the sample solution. Am: Amount of 50 times the peak area of mupirocin ob-tained from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Mupirocin Calcium Hydrate. Time span of measurement: About 5 times as long as the retention time of mupirocin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Mupirocin Calcium Hydrate. Test for required detectability: To exactly 1 mL of the standard solution add a mixture of 0.1 mol/L acetic acid- sodium acetate buffer solution (pH 4.0) and diluted tetra-hydrofuran (3 in 4) (1:1) to make exactly 20 mL. Confirm that the peak area of mupirocin obtained with 20 mL of this solution is equivalent to 4 to 6z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mupirocin is not more than 2.0z.
Weigh accurately an amount of Mupirocin Calcium Ointment, equivalent to about 2 mg (potency) of Mupirocin Calcium Hydrate, add exactly 10 mL of diluted tetra-hydrofuran (3 in 4), and shake vigorously. To this solution add exactly 10 mL of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0), shake vigorously, filter through a glass wool filter, and use the filtrate as the sample solution. Separately, weigh accurately an amount of Mupirocin Lithi-um RS, equivalent to about 20 mg (potency), dissolve in a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solu-tion (pH 4.0) and diluted tetrahydrofuran (3 in 4) (1:1) to make exactly 200 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Mupirocin Calcium Hydrate. Amount [mg (potency)] of mupirocin (C26H44O9) ＝ MS × AT/AS × 1/10 MS: Amount [mg (potency)] of Mupirocin Lithium RS taken
Nabumetone Tablets contain not less than 95.0z Flow rate: 1.3 mL per minute. Time span of measurement: About 3 times as long as the retention time of nabumetone, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, and add acetonitrile to make exactly 10 mL. Con-firm that the peak area of nabumetone obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nabumetone is not more than 5.0z. Water <2.48> Not more than 0.2z (1 g, volumetric titra-tion, direct titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 20 mg each of Nabumetone and Nabumetone RS (separately determine the water <2.48> in the same manner as Nabumetone), dissolve them in aceto- and not more than 105.0z of the labeled amount of nabumetone (C15H16O2: 228.29). Method of preparation Prepare as directed under Tablets, with Nabumetone. Identification To a quantity of powdered Nabumetone Tablets, equivalent to 80 mg of Nabumetone, add 50 mL of methanol, shake for 10 minutes and centrifuge the solution. To 1 mL of the supernatant liquid, add methanol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 259 nm and 263 nm, be- tween 268 nm and 272 nm, between 316 nm and 320 nm, and between 330 nm and 334 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 3 g of polysorbate 80 in water to make 100 mL as the dissolution medium, the dissolution rate in 60 minutes of Nabumetone Tablets is not less than 70z. ‌Start the test with 1 tablet of Nabumetone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a solution, prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL, to make exactly V? mL so that each mL contains about 89 mg of nabumetone (C15H16O2), and use this solution as the sample solution.
To a quantity of powdered Nabumetone Tablets, equivalent to 80 mg of Nabumetone, add 50 mL of methanol, shake for 10 minutes and centrifuge the solution. To 1 mL of the supernatant liquid, add methanol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 259 nm and 263 nm, be- tween 268 nm and 272 nm, between 316 nm and 320 nm, and between 330 nm and 334 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 3 g of polysorbate 80 in water to make 100 mL as the dissolution medium, the dissolution rate in 60 minutes of Nabumetone Tablets is not less than 70z. ‌Start the test with 1 tablet of Nabumetone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a solution, prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL, to make exactly V? mL so that each mL contains about 89 mg of nabumetone (C15H16O2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Nabumetone RS (separately determine the water <2.48> in the same manner as Nabumetone), and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 10 mL of this solution, add the dissolution medium to make exactly 25 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 331 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using a solution prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL as the blank. Dissolution rate (z) with respect to the labeled amount of nabumetone (C15H16O2) ＝ MS × AT/AS × V?/V × 1/C × 360 MS: Amount (mg) of Nabumetone RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of nabumetone (C15H16O2) in 1 tablet
Weigh accurately about 20 mg each of Nabumetone and Nabumetone RS (separately determine the water <2.48> in the same manner as Nabumetone), dissolve them in aceto- and not more than 105.0z of the labeled amount of nabumetone (C15H16O2: 228.29). Method of preparation Prepare as directed under Tablets, with Nabumetone. Identification To a quantity of powdered Nabumetone Tablets, equivalent to 80 mg of Nabumetone, add 50 mL of methanol, shake for 10 minutes and centrifuge the solution. To 1 mL of the supernatant liquid, add methanol to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 259 nm and 263 nm, be- tween 268 nm and 272 nm, between 316 nm and 320 nm, and between 330 nm and 334 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 3 g of polysorbate 80 in water to make 100 mL as the dissolution medium, the dissolution rate in 60 minutes of Nabumetone Tablets is not less than 70z. ‌Start the test with 1 tablet of Nabumetone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a solution, prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL, to make exactly V? mL so that each mL contains about 89 mg of nabumetone (C15H16O2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Nabumetone RS (separately determine the water <2.48> in the same manner as Nabumetone), and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 10 mL of this solution, add the dissolution medium to make exactly 25 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 331 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using a solution prepared by adding to 20 mL of ethanol (99.5) the dissolution medium to make 50 mL as the blank. Dissolution rate (z) with respect to the labeled amount of nabumetone (C15H16O2) ＝ MS × AT/AS × V?/V × 1/C × 360 MS: Amount (mg) of Nabumetone RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of nabumetone (C15H16O2) in 1 tablet Assay Weigh accurately the mass of not less than 20 tablets of Nabumetone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.2 g of nabume-tone (C15H16O2), add 10 mL of water and shake, add 40 mL of methanol, shake for 30 minutes, and then add methanol to make exactly 100 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, then add methanol to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Nabumetone RS (separately deter-mine the water <2.48> in the same manner as Nabumetone), dissolve by adding 50 mL of methanol and exactly 20 mL of the internal standard solution, then add methanol to make 200 mL, and use this solution as the standard solution. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nabumetone to that of the internal standard. Amount (mg) of nabumetone (C15H16O2) ＝ MS × QT/QS × 5 MS: Amount (mg) of Nabumetone RS taken, calculated on the anhydrous basis Internal standard solution—Dissolve 0.12 g of 2-ethylhexyl parahydroxybenzoate in methanol to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (550:450:1). Flow rate: Adjust so that the retention time of nabume-tone is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, nabumetone and the internal standard are eluted in this order with the resolution between these peaks being not less than 13. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nabumetone to that of the internal standard is not more than 1.0z.
Naftopidil Orally Disintegrating Tablets
Naftopidil Orally Disintegrating Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of naftopidil (C24H28N2O3: 392.49). Method of preparation Prepare as directed under Tablets, with Naftopidil. Identification Powder Naftopidil Orally Disintegrating Tablets. To a portion of the powder, equivalent to 25 mg of Naftopidil add 100 mL of methanol, shake thoroughly, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 6 mL of the filtrate add methanol to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 281 nm and 285 nm, and between 318 nm and 322 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Naftopidil Orally Disintegrating Tablets add V/10 mL of water, disintegrate and disperse the tablet by sonicating. To this solution add V/2 mL of methanol, shake thoroughly, then add methanol to make exactly V mL so that each mL contains about 0.
Powder Naftopidil Orally Disintegrating Tablets. To a portion of the powder, equivalent to 25 mg of Naftopidil add 100 mL of methanol, shake thoroughly, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 6 mL of the filtrate add methanol to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 281 nm and 285 nm, and between 318 nm and 322 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Naftopidil Orally Disintegrating Tablets add V/10 mL of water, disintegrate and disperse the tablet by sonicating. To this solution add V/2 mL of methanol, shake thoroughly, then add methanol to make exactly V mL so that each mL contains about 0.25 mg of naftopidil (C24H28N2 O3), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 6 mL of the subsequent filtrate, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of naftopidil for assay, previously dried at 1059C for 3 hours, dissolve in methanol to make exactly 100 mL. Pipet 3 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Determine the absorbances, AT and AS, at 283 nm of the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of naftopidil (C24H28N2O3) ＝ MS × AT/AS × V/200 MS: Amount (mg) of naftopidil for assay taken Disintegration Being specified separately when the drug is granted approval based on the Law. Dissolution <6.10> When the test is performed at 50 revolu-tion per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 30 minutes of Naftopidil Orally Disintegrating Tablets is not less than 75z. Start the test with 1 tablet of Naftopidil Orally Disin-tegrating Tablets, withdraw not less than 20 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 28 mg of naftopidil (C24H28N2O3), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of naftopidil for assay, previously dried at 1059C for 3 hours, dissolve in 50 mL of methanol, then add the disso-lution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 283 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution me-dium as the control. Dissolution rate (z) with respect to the labeled amount of naftopidil (C24H28N2O3) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of naftopidil for assay taken C: Labeled amount (mg) of naftopidil (C24H28N2O3) in 1 tablet
Weigh accurately the mass of not less than 20 Naftopidil Orally Disintegrating Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of naftopidil (C H N O ), add 30 mL of meth- 24 28 2 3 ‌anol, shake thoroughly, add diluted 0.1 mol/L potassium di-hydrogen phosphate TS (pH 2.0) (1 in 2) to make exactly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, add a mixture of meth-anol and water (3:2) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of naftopidil for assay, previously dried at 1059C for 3 hours, dissolve in 30 mL of methanol, add diluted 0.1 mol/L potassium dihydrogen phosphate TS (pH 2.0) (1 in 2) to make exactly 50 mL. Pipet 10 mL of this solution, add ex-actly 10 mL of the internal standard solution, add a mixture of methanol and water (3:2) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS of the peak area of naftopidil to that of the internal stand-ard. Amount (mg) of naftopidil (C24H28N2O3) ＝ MS × QT/QS MS: Amount (mg) of naftopidil for assay taken Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of methanol and water (3:2) (3 in 2000). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Naftopidil. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, naftopidil and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of naftopidil to that of the internal standard is not more than 1.0z.
Naftopidil Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of naftopidil (C24H28N2O3: 392.49). Method of preparation Prepare as directed under Tablets, with Naftopidil. Identification Powder Naftopidil Tablets. To a portion of the powder, equivalent to 25 mg of Naftopidil, add 100 mL of methanol, shake thoroughly, and centrifuge, if necessary. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 6 mL of the filtrate add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, and between 318 nm and 322 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Naftopidil Tablets add V/10 mL of water, disintegrate and disperse the tablet with the aid of ultrasonic waves. To this dispersed solution add V/2 mL of methanol, shake thoroughly, add methanol to make exactly V mL so that each mL contains about 0.
Powder Naftopidil Tablets. To a portion of the powder, equivalent to 25 mg of Naftopidil, add 100 mL of methanol, shake thoroughly, and centrifuge, if necessary. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 6 mL of the filtrate add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 281 nm and 285 nm, and between 318 nm and 322 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Naftopidil Tablets add V/10 mL of water, disintegrate and disperse the tablet with the aid of ultrasonic waves. To this dispersed solution add V/2 mL of methanol, shake thoroughly, add methanol to make exactly V mL so that each mL contains about 0.25 mg of naftopidil (C24H28N2O3). Centrifuge this solution, if necessary, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet 6 mL of the subsequent filtrate, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of naftopidil for assay, previously dried at 1059C for 3 hours, dissolve in methanol to make exactly 100 mL. Pipet 3 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 283 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of naftopidil (C24H28N2O3) ＝ MS × AT/AS × V/200 MS: Amount (mg) of naftopidil for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 15 minutes of 25-mg and 50-mg tablet and in 30 minutes of 75-mg tablet is not less than 75z. Start the test with 1 tablet of Naftopidil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add the dissolution medium to make exactly V? mL so that each mL contains about 28 mg of naftopidil (C24H28N2O 3), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of naftopidil for assay, previ-ously dried at 1059C for 3 hours, dissolve in 50 mL of meth-anol, and add the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 283 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the control. Dissolution rate (z) with respect to the labeled amount of naftopidil (C24H28N2O3) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of naftopidil for assay taken C: Labeled amount (mg) of naftopidil (C24H28N2O3) in 1 tablet
Weigh accurately the mass of not less than 20 Naftopidil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of naftopidil (C24H28N2O3), add 30 mL of methanol, shake thoroughly, and add diluted 0.1 mol/L potassium dihydrogen phosphate TS (pH 2.0) (1 in 2) to make exactly 50 mL. Centrifuge this solution, if necessary, filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 10 mL of the filtrate, pipet 10 mL of the subse-quent filtrate, add exactly 10 mL of the internal standard so-lution, add a mixture of methanol and water (3:2) to make ‌100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of naftopidil for assay, previously dried at 1059C for 3 hours, dissolve in 30 mL of methanol, add diluted 0.1 mol/L potassium dihydrogen phosphate TS (pH 2.0) (1 in 2) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add a mixture of methanol and water (3:2) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS of the peak area of TS (1 in 200,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Nali-dixic Acid, previously dried, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Melting point <2.60> 225 – 2319C naftopidil to that of the internal standard. Purity (1) Chloride <1.03>—To 2.0 g of Nalidixic Acid Amount (mg) of naftopidil (C24H28N2O3) ＝ M S × QT/QS add 50 mL of water, warm at 709C for 5 minutes, cool quickly, and filter. To 25 mL of the filtrate add 6 mL of MS: Amount (mg) of naftopidil for assay taken Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of methanol and water (3:2) (3 in 2000). Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Naftopidil. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, naftopidil and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of naftopidil to that of the internal standard is not more than 1.0z.
Naphazoline and Chlorpheniramine Solution
ナファゾリン・クロルフェニラミン液
Naphazoline and Chlorpheniramine Solution is a clear, colorless liquid.
(1) To 20 mL of Naphazoline and Chlor-pheniramine Solution add 2 mL of a solution of potassium ‌hydroxide (7 in 10) and 5 mL of pyridine, and heat at 1009C for 5 minutes: a red color is produced (chlorobutanol). Place 10 mL of Naphazoline and Chlorpheniramine Solution in a glass-stoppered test tube, add 10 mL of ethanol (95), 2 mL of sodium hydroxide TS and 1 mL of a solution of copper (II) chloride dihydrate in ethanol (95) (1 in 10), and shake: a blue color is produced (glycerin). To 20 mL of Naphazoline and Chlorpheniramine Solution add 5 mL of sodium hydroxide TS, extract with 10 mL of diethyl ether, and separate the diethyl ether layer. Take 5 mL of this solution, distil off the solvent, dissolve the residue in 5 mL of methanol, and use this solution as the sample solution. Separately, dissolve 0.01 g each of napha-zoline nitrate and Chlorpheniramine Maleate RS in 10 mL and 5 mL of methanol, respectively, and use these solutions as standard solutions (1) and (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol, acetone and ammonia so-lution (28) (73:15:10:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): two spots obtained from the sample solu-tion exhibit the same Rf values as the spots from standard solutions (1) and (2). Spray evenly Dragendorff's TS on the plate: the spots from standard solutions (1) and (2) and the corresponding spot from the sample solutions reveal an orange color.
Pipet 4 mL of Naphazoline and Chlorpheniramine Solution, add exactly 4 mL of the internal standard solution, then add water to make 10 mL, and use this solution as the sample solution. Weigh accurately about 50 mg of naphazo-line nitrate for assay, dried at 1059C for 2 hours, and about 0.1 g of Chlorpheniramine Maleate RS, dried at 1059C for 3 hours, dissolve in water to make exactly 100 mL. Pipet 4 mL of this solution, add exactly 4 mL of the internal standard solution, then add water to make 10 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solutions as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QTa and QTb, of the peak height of naphazoline and chlorphenira-mine to that of the internal standard of the sample solution, and the ratios, QSa and QSb, of the peak height of naphazo-line and chlorpheniramine to that of the internal standard of the standard solution. Amount (mg) of naphazoline nitrate (C14H14N2.HNO3) ＝ MSa × QTa/QSa × 1/25 Amount (mg) of chlorpheniramine maleate Column temperature: Room temperature. Mobile phase: A mixture of acetonitrile and a solution of sodium laurylsulfate (1 in 500) in diluted phosphoric acid (1 in 1000) (1:1). Flow rate: Adjust so that the retention time of chlor-pheniramine is about 10 minutes. Selection of column: Proceed with 10 mL of the standard solution under the above operating conditions. Use a column giving well-resolved peaks of the internal standard, naphazo-line and chlorpheniramine in this order.
Nartograstim for Injection (Genetical Recombination)
注射用ナルトグラスチム（遺伝子組換え）
Nartograstim for Injection (Genetical Recom-bination) occurs as white, masses or powder.
Dissolve the content of 1 package of Nar-tograstim for Injection (Genetical Recombination) in 1 mL of tris-sodium chloride buffer solution (pH 8.0). To a suita-ble amount of this solution add tris-sodium chloride buffer solution (pH 8.0) so that each mL contains 1 mg of Nar-tograstim (Genetical Recombination), and use this solution as the sample solution. Then, proceed with the sample solu-tion as directed in the Identification (1) under Nartograstim (Genetical Recombination). pH <2.54> Being specified separately when the drug is granted approval based on the Law.
(1) Clarity and color of solution—A solution of Nartograstim for Injection (Genetical Recombination) in water, containing 100 mg of Nartograstim (Genetical Recom-bination) in each mL, is clear and colorless. (2) Lactose conjugate—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not more than 3.0z (50 mg, coulometric titration). Bacterial endotoxins <4.01> Less than 0.62 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2, using 3 mL of water for injection per 1 Nar-tograstim for Injection (Genetical Recombination) to dis-solve the content: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method, using the sample solution prepared by dissolving the sample in water in a concentration to be used for the injection: it meets the requirement. Specific activity Nartograstim for Injection (Genetical Recombination), when perform the assay and the following test, contains not less than 4.0 × 108 units of nartograstim (genetical recombination) per mg nartograstim (genetical recombination). Wash out each content of 10 Nartograstim for Injection (Genetical Recombination) with a suitable amount of po-tency measuring medium for nartograstim test, wash the empty containers with the same medium, combine all wash-ings, and add the same medium to make exactly 50 mL. To an exact amount of this solution add the same medium so that the concentration of nartograstim (genetical recombina-tion) is equivalent to 50z to 150z of that of the standard solution, and use this solution as the sample solution. Sepa-rately, weigh accurately a suitable amount of Nartograstim RS, dissolve in the potency measuring medium for nar-tograstim test so that each mL contains exactly 1.2 × 104 units of nartograstim according to the labeled unit, and use this solution as the standard solution. Then, determine the nartograstim potency (unit) in 1 Nartograstim for Injection (Genetical Recombination) by proceeding as directed in the Assay (2) under Nartograstim (Genetical Recombination), and calculate the ratio against the amount of nartograstim obtained in the Assay. Nartograstim (genetical recombination) potency (unit) in 1 Nartograstim for Injection (Genetical Recombination) ＝ S × mean relative potency of the sample solution × d × 5 S: Concentration (unit/mL) of the standard solution d: Dilution factor for the sample solution 5: Amount (mL) of the medium used to dissolve per 1 sample Relative activity of sample solution ＝ 2a S2b × 1 a: nT1 ＋ (AT1 － AM)/(AT1 － AT2) b: nS1 ＋ (AS1 － AM)/(AS1 － AS2) System suitability— Proceed as directed in the system suitability in the Assay (2) under Nartograstim (Genetical Recombination).
Weigh accurately the mass of each content of not ‌less than 10 Nartograstim for Injection (Genetical Recombi-nation). Weigh accurately an amount of the content, equiva-lent to about 0.25 mg of Nartograstim (Genetical Recombi-nation), dissolve in exactly 5 mL of the mobile phase, and use this solution as the sample solution. Separately, dissolve a suitable amount of Nartograstim RS in the mobile phase so that each mL contains about 50 mg of nartograstim, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of nartograstim in each solution. Amount (mg) of nartograstim (genetical recombination) in 1 Nartograstim for Injection (Genetical Recombination) ＝ MS × AT/AS × M/MT × 5 MS: Amount (mg) of nartograstim in 1 mL of the standard solution M: Mean mass (mg) of each content MT: Amount (mg) of Nartograstim for Injection (Geneti-cal Recombination) taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with porous silica gel for liq-uid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 15.6 g of sodium dihydrogen phos-phate dihydrate and 1.0 g of sodium lauryl sulfate in 700 mL of water, adjust to pH 6.5 with sodium hydroxide TS, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of nar-tograstim is about 16 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nartograstim are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nartograstim is not more than 1.5z.
Nateglinide Tablets contain not less than 96.0z and not more than 104.0z of the labeled amount of nateglinide (C19H27NO3: 317.42). Method of preparation Prepare as directed under Tablets, with Nateglinide. Identification To an amount of powdered Nateglinide Tablets, equivalent to 20 mg of Nateglinide, add 20 mL of methanol, shake, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 246 nm and 250 nm, between 251 nm and 255 nm, between 257 nm and 261 nm and between 262 nm and 266 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Nateglinide Tablets add 10 mL of 0.05 mol/L sodium dihydrogen phosphate TS adjusted to pH 2.5 with phosphoric acid, shake to disintegrate the tablet, and disperse to fine particles by sonicating. Add exactly 3V/50 mL of the internal standard solution, add 3V/5 mL of aceto-nitrile, shake for 10 minutes, and add acetonitrile to make V mL so that each mL contains about 0.
To an amount of powdered Nateglinide Tablets, equivalent to 20 mg of Nateglinide, add 20 mL of methanol, shake, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 246 nm and 250 nm, between 251 nm and 255 nm, between 257 nm and 261 nm and between 262 nm and 266 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Nateglinide Tablets add 10 mL of 0.05 mol/L sodium dihydrogen phosphate TS adjusted to pH 2.5 with phosphoric acid, shake to disintegrate the tablet, and disperse to fine particles by sonicating. Add exactly 3V/50 mL of the internal standard solution, add 3V/5 mL of aceto-nitrile, shake for 10 minutes, and add acetonitrile to make V mL so that each mL contains about 0.6 mg of nateglinide (C 19H27NO3). Filter the solution through a membrane filter with a pore size not exceeding 0.45 mm, and discard the first 5 mL of the filtrate. To 8 mL of the subsequent filtrate add the mobile phase to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Nateglinide RS, previously dried at 1059C for 2 hours, and dissolve in acetonitrile to make exactly 10 mL. Pipet 6 mL of this solution, add exactly 3 mL of the internal stand-ard solution, and add the mobile phase to make 25 mL. To 8 mL of this solution add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01>, and calculate the ratios, QT and QS, of the peak area of nateglinide to that of the internal standard. Column temperature: A constant temperature of about 409C. Mobile phase: Adjust 0.05 mol/L sodium dihydrogen Amount (mg) of nateglinide (C ＝ MS × QT/QS × 3V/250 19H27NO3) phosphate TS to pH 2.5 with phosphoric acid. To 550 mL of this solution add 450 mL of acetonitrile for liquid chroma-tography. Flow rate: Adjust so that the retention time of nateglinide is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and nateglinide are eluted in this order with the resolution between these peaks being not less than 19. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nateglinide to that of the internal standard is not more than 1.0z. MS: Amount (mg) of Nateglinide RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in acetonitrile (1 in 250). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and nateglinide are eluted in this order with the resolution between these peaks being not less than 19. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nateglinide to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of a 30-mg tablet and that in 30 minutes of a 90-mg tablet of Nateglinide Tablets is not less than 75z, respectively. ‌Start the test with 1 tablet of Nateglinide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 33 mg of nateglinide (C19H27NO3), and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Nateglinide RS, previously dried at 1059C for 2 hours, and dissolve in aceto-nitrile to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of nateglinide in each solution. Dissolution rate (z) with respect to the labeled amount of nateglinide (C19H27NO3) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Nateglinide RS taken C: Labeled amount (mg) of nateglinide (C19H27NO3) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nateglinide are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nateglinide is not more than 2.0z.
To 20 Nateglinide Tablets add V/5 mL of 0.05 mol/L sodium dihydrogen phosphate TS adjusted to pH 2.5 with phosphoric acid, shake to disintegrate the tablets, and disperse to fine particles by sonicating. Then, add V/2 mL of acetonitrile and exactly V/10 mL of the internal standard so-lution, shake for 10 minutes, and add acetonitrile to make V mL so that each mL contains about 6 mg of nateglinide (C19 H27NO3). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, to 4 mL of the subsequent filtrate add the mobile phase to make 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 60 mg of Nateglinide RS, previously dried at 1059C for 2 hours, add exactly 1 mL of the internal standard solution, and add acetonitrile to make 10 mL. To 4 mL of this solution add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of nateglinide to that of the internal standard. Amount (mg) of nateglinide (C19H27NO3) in 1 tablet ＝ MS × QT/QS × V/200 MS: Amount (mg) of Nateglinide RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in acetonitrile (3 in 125). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 2.5 of 0.05 mol/L sodium di-hydrogen phosphate TS with phosphoric acid. To 550 mL of this solution add 450 mL of acetonitrile. Flow rate: Adjust so that the retention time of nateglinide is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and nateglinide are eluted in this order with the resolution between these peaks being not less than 19. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nateglinide to that of the internal standard is not more than 1.0z.
ネオスチグミンメチル硫酸塩注射液
Neostigmine Methylsulfate Injection is an aqueous injection. It contains not less than 93.
Neostigmine Methylsulfate Injection is a clear, colorless liquid. It is slowly affected by light. pH: 5.0 – 6.5
Take a volume of Neostigmine Methylsul-fate Injection, equivalent to 5 mg of neostigmine methylsul-fate, add water to make 10 mL if necessary, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 257 nm and 261 nm. Bacterial endotoxins <4.01> Less than 5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. and AS, of neostigmine in each solution. Amount (mg) of neostigmine methylsulfate (C ＝ MS × AT/AS 13H22N2O6S) Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. MS: Amount (mg) of Neostigmine Methylsulfate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 259 nm). Column: A stainless steel column 4.6 mm in inside diame-
Use Neostigmine Methylsulfate Injection as the sample solution. Separately, weigh accurately about 25 mg of Neostigmine Methylsulfate RS, previously dried at 1059C for 3 hours, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Neostigmine Methylsulfate. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about Amount (mg) of neostigmine methylsulfate (C ＝ MS × AT/AS 13H22N2O6S) 259C. Mobile phase: Dissolve 3.12 g of sodium dihydrogenphos-phate dihydrate in 1000 mL of water, adjust to pH 3.0 with phosphoric acid, and add 0.871 g of sodium 1-pentanesul-fonate to dissolve. To 890 mL of this solution add 110 mL of acetonitrile. Flow rate: Adjust so that the retention time of neostig-mine is about 9 minutes. System suitability— System performance: Dissolve 25 mg of Neostigmine Methylsulfate and 4 mg of dimethylaminophenol in 50 mL of the mobile phase. When the procedure is run with 10 mL of this solution under the above operating conditions, dimethylaminophenol and neostigmine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak MS: Amount (mg) of Neostigmine Methylsulfate RS taken
Nicardipine Hydrochloride Injection is an aqueous injection. It contains not less than 93.
Nicardipine Hydrochloride Injection occurs as a clear, pale yellow liquid. It is gradually changed by light.
To a volume of Nicardipine Hydrochloride Injection, equivalent to 1 mg of Nicardipine Hydrochloride, add ethanol (99.5) to make 100 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 235 nm and 239 nm, and between 351 nm and 355 nm. pH <2.54> 3.0 – 4.5 ‌Purity Related substances—Conduct the procedure with-out exposure to light, using light-resistant vessels. To a volume of Nicardipine Hydrochloride Injection, equivalent to 5 mg of Nicardipine Hydrochloride, add the mobile phase to make 10 mL, and use this solution as the sample solution. To exactly 2 mL of the sample solution add the mobile phase to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas of these solutions by the automatic integration method: the areas of the peaks other than nicardipine obtained from the sample solution are not larger than the peak area of nicardipine from the stand-ard solution, and the total of the areas of the peaks other than nicardipine is not larger than 2 times of the peak area of nicardipine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of nicardipine, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of nicardipine obtained with 10 mL of this solution is equivalent to 8 to 12z of that with 10 mL of the standard solution. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of nicardipine is not more than 1.0z. Bacterial endotoxins <4.01> Less than 8.33 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Conduct the procedure without exposure to light, using light-resistant vessels. To an exact volume of Nicar-dipine Hydrochloride Injection, equivalent to about 2 mg of nicardipine hydrochloride (C26H29N3O6.HCl), add exactly 5 mL of the internal standard solution and methanol to make 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 50 mg of nicardipine hydro-chloride for assay, previously dried at 1059C for 2 hours, dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 5 mL of the internal standard solu-tion and methanol to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nicardipine to that of the internal standard. Amount (mg) of nicardipine hydrochloride (C26H29N3O6.HCl) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of nicardipine hydrochloride for assay taken Internal standard solution—A solution of di-n-butyl phtha-late in methanol (1 in 625). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL. To 320 mL of this so-lution add 680 mL of methanol. Flow rate: Adjust so that the retention time of nicardipine is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, nicardipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of nicardipine is not more than 1.0z.
Nicergoline Powder contains not less than 95.
Vigorously shake for 10 minutes a quantity of Nicergoline Powder, equivalent to 10 mg of Nicergoline, with 20 mL of diluted ethanol (4 in 5), and centrifuge for 10 minutes. To 2 mL of the supernatant liquid add ethanol (99.5) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 226 nm and 230 nm, and between 286 nm and 290 nm.
Related substances—Perform the test with 20 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of substances other than nicergoline by the area percentage method: the total amount of them is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of nicergoline, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. ‌Test for required detectability: To 1 mL of the standard solution obtained in the Assay add a mixture of acetonitrile and water (17:3) to make 50 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solu-tion for system suitability test, add the mixture of aceto-nitrile and water (17:3) to make exactly 100 mL. Confirm that the peak area of nicergoline obtained with 20 mL of this solution is equivalent to 3 to 7z of that with 20 mL of the so-lution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of nicergoline is not more than 1.5z. Uniformity of dosage units <6.02> The Nicergoline Powder in single-dose packages meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 15 minutes of Nicergoline Powder is not less than 80z. Start the test with an accurately weighed amount of Nicergoline Powder, equivalent to about 5 mg of nicergoline (C24H26BrN3O3), withdraw not less than about 20 mL of the medium at the specified minute after starting the test, and filter through a laminated polyester fiber. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 50 mg of nicergoline for assay, previously dried in vacuum at 609C for 2 hours, and dissolve in 0.1 mol/L hy-drochloric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, and add the dissolution medium to make ex-actly 100 mL. Pipet 10 mL of this solution, add the dissolu-tion medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances at 225 nm, AT1 and AS1, and at 250 nm, AT2 and AS2, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of nicergoline (C24H26BrN3O3) ＝ MS/MT × (AT1 － AT2)/(AS1 － AS2) × 1/C × 9 MS: Amount (mg) of nicergoline for assay taken MT: Amount (g) of Nicergoline Powder taken C: Labeled amount (mg) of nicergoline (C24H26BrN3O3) in 1 g
Weigh accurately a quantity of Nicergoline Powder, equivalent to about 20 mg of nicergoline (C24H26BrN3O3), add exactly 20 mL of a mixture of acetonitrile and water (17:3), vigorously shake for 10 minutes, centrifuge for 10 minutes, and use the supernatant liquid as the sample solu-tion. Separately, weigh accurately about 20 mg of nicer-goline for assay, previously dried in vacuum at 609C for 2 hours, dissolve in exactly 20 mL of the mixture of aceto-nitrile and water (17:3), and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of nicergoline in each solution. Amount (mg) of nicergoline (C24H26BrN3O3) ＝ MS × AT/AS MS: Amount (mg) of nicergoline for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 288 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust the pH of 0.05 mol/L potassium di-hydrogen phosphate TS to 7.0 with triethylamine. To 350 mL of this solution add 350 mL of methanol and 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of nicergoline is about 25 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nicergoline are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nicergoline is not more than 1.0z.
Take a quantity of powdered Nicergoline Tablets, equivalent to 10 mg of Nicergoline, add 20 mL of ethanol (99.5), shake vigorously for 10 minutes, and filter through a 0.45-mm pore-size membrane filter. To 2 mL of the filtrate add ethanol (99.5) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 226 nm and 230 nm, and between 286 nm and 290 nm.
Related substances—Perform the test with 20 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate the amount of substances other than nicergoline by the area percentage method: the total amount of them is not more than 2.0z. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of nicergoline, beginning after the solvent peak. System suitability— ‌System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 1 mL of the standard solution obtained in the Assay add a mixture of acetonitrile and water (17:3) to make 50 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solu-tion for system suitability test, add the mixture of aceto-nitrile and water (17:3) to make exactly 100 mL. Confirm that the peak area of nicergoline obtained with 20 mL of this solution is equivalent to 3 to 7z of that with 20 mL of the so-lution for system suitability test. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of nicergoline is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Nicergoline Tablets add exactly 25 mL of diluted ethanol (4 in 5), disperse to fine particles by sonicat-ing, and shake for 5 minutes. Centrifuge this solution for 10 minutes, pipet 4 mL of the supernatant liquid, add diluted ethanol (4 in 5) to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of nicergoline for assay, previously dried in vacuum at 609C for 2 hours, and dissolve in exactly 25 mL of diluted ethanol (4 in 5). Pipet 4 mL of this solution, add diluted ethanol (4 in 5) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances at 288 nm, AT1 and AS1, and at 340 nm, AT2 and AS2, of the sample solution and the standard solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>. Amount (mg) of nicergoline (C24H26BrN3O3) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × 1/2 MS: Amount (mg) of nicergoline for assay taken Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Nicer-goline Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of nicergoline (C24H26BrN3O3), add exactly 20 mL of a mixture of aceto-nitrile and water (17:3), vigorously shake for 10 minutes, centrifuge for 10 minutes, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of nicergoline for assay, previously dried in vacuum at 609C for 2 hours, dissolve in exactly 20 mL of the mixture of acetonitrile and water (17:3), and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of nicergoline in each solution. Amount (mg) of nicergoline (C24H26BrN3O3) ＝ MS × AT/AS MS: Amount (mg) of nicergoline for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 288 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust the pH of 0.05 mol/L potassium di-hydrogen phosphate TS to 7.0 with triethylamine. To 350 mL of this solution add 350 mL of methanol and 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of nicergoline is about 25 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nicergoline are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nicergoline is not more than 1.0z.
Weigh accurately not less than 20 Nicomol Tablets and powder. Weigh accurately a portion of the powder, equivalent to about 1 g of nicomol (C34H32N4O9), add 100 mL of 1 mol/L hydrochloric acid TS, shake well, add water to make exactly 500 mL, and filter. Discard the first 50 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 50 mL of 1 mol/L hydrochloric acid TS and water to make ex-actly 250 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of nicomol for assay, previously dried at 1059C for 4 hours, dissolve in 50 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add 20 mL of 1 mol/L hydrochloric acid TS and water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 262 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of nicomol (C34H32N4O9) ＝ MS × AT/AS × 25/2 MS: Amount (mg) of nicomol for assay taken
Nicotinic Acid Injection is an aqueous injection. It contains not less than 95.
Nicotinic Acid Injection is a clear, colorless liquid. pH: 5.0 – 7.0
(1) To a volume of Nicotinic Acid Injec-tion, equivalent to 0.1 g of Nicotinic Acid, add 0.3 mL of dilute hydrochloric acid, and evaporate on a water bath to 2 mL. After cooling, collect the crystals formed, wash with small portions of ice-cold water until the last washing shows no turbidity on the addition of silver nitrate TS, and dry at 1059C for 1 hour: the crystals melt <2.60> between 2349C and 2389C. With the crystals, proceed as directed in the Identification (1) under Nicotinic Acid. (2) Dissolve 0.02 g of the dried crystals obtained in (1) in water to make 1000 mL, and determine the absorption spec-trum as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 261 nm and 263 nm, and a minimum between 235 nm and 239 nm. Sepa-rately, determine the absorbances of this solution, A1 and A2, at each wavelength of maximum and minimum absorp-tion, respectively: the ratio A2/A1 is between 0.35 and 0.39. Bacterial endotoxins <4.01> Less than 3.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. MS: Amount (mg) of Nicotinic Acid RS taken Internal standard solution—A solution of caffeine in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.1 g of sodium 1-octane sulfonate in a mixture of 0.05 mol/L sodium dihydrogenphosphate TS (pH 3.0) and methanol (4:1) to make 1000 mL. Flow rate: Adjust so that the retention time of caffeine is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, nicotinic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of nicotinic acid to that of the internal standard is not more than 1.0z.
Nifedipine Delayed-release Fine Granules
Nifedipine Delayed-release Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of nifedipine (C17H18N2O6: 346.33). Method of preparation Prepare as directed under Gran-ules, with Nifedipine. Identification Conduct this procedure without exposure to light, using light-resistant vessels. Shake for 15 minutes a quantity of powdered Nifedipine Delayed-release Fine Gran-ules, equivalent to 3 mg of Nifedipine, with 100 mL of meth- anol, and filter. Determine the absorption spectrum of the filtrate so obtained as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a broad absorption maximum between 335 nm and 356 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To the total content of 1 package of Nifedipine Delayed-release Fine Granules add 50 mL of a mixture of methanol and water (9:1), sonicate for 15 minutes with occasional shaking, and shake for further 15 minutes. Then, add methanol to make exactly V mL so that each mL contains about 0.
Conduct this procedure without exposure to light, using light-resistant vessels. Shake for 15 minutes a quantity of powdered Nifedipine Delayed-release Fine Gran-ules, equivalent to 3 mg of Nifedipine, with 100 mL of meth- anol, and filter. Determine the absorption spectrum of the filtrate so obtained as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a broad absorption maximum between 335 nm and 356 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To the total content of 1 package of Nifedipine Delayed-release Fine Granules add 50 mL of a mixture of methanol and water (9:1), sonicate for 15 minutes with occasional shaking, and shake for further 15 minutes. Then, add methanol to make exactly V mL so that each mL contains about 0.1 mg of nifedipine (C17H18N2O6). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of nifedipine for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 100 mL, and use this so-lution as the standard solution. Then, proceed as directed in the Assay. Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nifedipine are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nifedipine is not more than 1.0z.
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately a portion of powdered Nifedipine Delayed-release Fine Granules, equiva-lent to about 10 mg of nifedipine (C17H18N2O6), add 50 mL of a mixture of methanol and water (9:1), shake vigorously for 15 minutes, and add methanol to make exactly 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of nifedi-pine for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 100 mL, and use Amount (mg) of nifedipine (C ＝ MS × AT/AS × V/500 17H18N2O6) this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac- MS: Amount (mg) of nifedipine for assay taken Dissolution <6.10> When the tests are performed at 50 revolutions per minute according to the Paddle method, using 900 mL each of 1st and 2nd fluids for dissolution test as the dissolution medium, the dissolution rate in the test using the 1st fluid for dissolution test in 60 minutes is not more than 15z, and that in the test using the 2nd fluid for dissolution test in 30 minutes is not less than 75z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with an accurately weighed amount of Nifedipine Delayed-release Fine Gran-ules, equivalent to about 20 mg of nifedipine (C17H18N2O6), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add the dissolution medium to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of nifedipine for assay, previ-ously dried at 1059C for 2 hours, dissolve in 50 mL of meth-anol, and add the dissolution medium to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01>, and determine the peak areas, AT and AS, of nifedipine in each solution. Dissolution rate (z) with respect to the labeled amount of nifedipine (C H N O ) cording to the following conditions, and determine the peak areas, AT and AS, of nifedipine in each solution. Amount (mg) of nifedipine (C17H18N2O6) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of nifedipine for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 6.1 of a mixture of methanol and diluted 0.05 mol/L disodium hydrogen phosphate TS (1 in 5) (11:9) with phosphoric acid. Flow rate: Adjust so that the retention time of nifedipine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nifedipine are not less than 4000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nifedipine is not more than 1.0z. 17 18 2 6 ＝ MS/MT × AT/AS × 1/C × 72 MS: Amount (mg) of nifedipine for assay taken MT: Amount (g) of Nifedipine Delayed-release Fine Gran-ules taken C: Labeled amount (mg) of nifedipine (C17H18N2O6) in 1 g Operating conditions— Proceed as directed in the operating conditions in the
Conduct this procedure without exposure to light, using light-resistant vessels. Take out the content of Nifedipine Extended-release Capsules, and powder. To an amount of the powder, equivalent to 3 mg of Nifedipine, add 100 mL of methanol, shake for 15 minutes, and centri-fuge. Determine the absorption spectrum of the supernatant liquid so obtained as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a broad absorption maxi-mum between 335 nm and 356 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To the total content of 1 capsule of Nifedipine Extended-release Capsules, add 50 mL of a mix-ture of methanol and water (9:1), sonicate for 15 minutes with occasional shaking, and shake for further 15 minutes. Then, add methanol to make exactly V mL so that each mL contains about 0.1 mg of nifedipine (C17H18N2O6). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of nifedipine for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 100 mL, and use this so-lution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of nifedipine (C17H18N2O6) ＝ MS × AT/AS × V/500 MS: Amount (mg) of nifedipine for assay taken Dissolution Being specified separately when the drug is granted approval based on the Law.
Conduct this procedure without exposure to light, using light-resistant vessels. Take out the contents of not less than 20 Nifedipine Extended-release Capsules, weigh accu-rately the mass of the contents, and powder. Weigh accu-rately a portion of the powder, equivalent to about 10 mg of nifedipine (C17H18N2O6), add 50 mL of a mixture of metha-nol and water (9:1), shake vigorously for 15 minutes, and add methanol to make exactly 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of nifedipine for assay, previ-ously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add metha-nol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of nifedipine in each solution. Amount (mg) of nifedipine (C17H18N2O6) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of nifedipine for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 6.1 of a mixture of methanol and diluted 0.05 mol/L disodium hydrogen phosphate TS (1 in 5) (11:9) with phosphoric acid. Flow rate: Adjust so that the retention time of nifedipine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nifedipine are not less than 4000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nifedipine is not more than 1.0z.
Nifedipine Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of nifedipine (C17H18N2O6: 346.33). Method of preparation Prepare as directed under Gran-ules, with Nifedipine. Identification Conduct this procedure without exposure to light, using light-resistant vessels. Shake for 15 minutes a quantity of powdered Nifedipine Fine Granules, equivalent to 6 mg of Nifedipine, with 200 mL of methanol, and centri-fuge. Determine the absorption spectrum of the supernatant liquid so obtained as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a broad absorption maxi-mum between 335 nm and 356 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To the total content of 1 package of Nifedipine Fine Granules add 50 mL of a mixture of metha-nol and water (9:1), sonicate for 15 minutes with occasional shaking, and shake for further 15 minutes. Then, add meth-anol to make exactly V mL so that each mL contains about 0.
Conduct this procedure without exposure to light, using light-resistant vessels. Shake for 15 minutes a quantity of powdered Nifedipine Fine Granules, equivalent to 6 mg of Nifedipine, with 200 mL of methanol, and centri-fuge. Determine the absorption spectrum of the supernatant liquid so obtained as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a broad absorption maxi-mum between 335 nm and 356 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To the total content of 1 package of Nifedipine Fine Granules add 50 mL of a mixture of metha-nol and water (9:1), sonicate for 15 minutes with occasional shaking, and shake for further 15 minutes. Then, add meth-anol to make exactly V mL so that each mL contains about 0.1 mg of of nifedipine (C17H18N2O6). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of nifedipine for assay, previ-ously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add metha-nol to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of nifedipine (C17H18N2O6) ＝ MS × AT/AS × V/500 MS: Amount (mg) of nifedipine for assay taken ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Nifedipine Fine Granules is not less than 85z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with an accurately weighted amount of Nifedipine Fine Granules, equivalent to about 10 mg of nifedipine (C17H18N2O6), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 28 mg of nifedipine for assay, previously dried at 1059C for 2 hours, dissolve in 50 mL of methanol, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of nifedi-pine in each solution. Dissolution rate (z) with respect to the labeled amount of nifedipine (C17H18N2O6) ＝ MS/MT × AT/AS × 1/C × 36 MS: Amount (mg) of nifedipine for assay taken MT: Amount (g) of Nifedipine Fine Granules taken C: Labeled amount (mg) of nifedipine (C17H18N2O6) in 1 g Operating conditions— Proceed as directed in the operating conditions under the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nifedipine are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nifedipine is not more than 1.0z.
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately a protein of powdered Nifedipine Fine Granules, equivalent to about 10 mg of nifedipine (C17H18N2O6), add 50 mL of a mixture of methanol and water (9:1), shake vigorously for 15 minutes, and add methanol to make exactly 100 mL. Filter this solu-tion through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of nifedipine for assay, previously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solu-tion, add methanol to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of nifedipine in each solution. Amount (mg) of nifedipine (C17H18N2O6) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of nifedipine for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 6.1 of a mixture of methanol and diluted 0.05 mol/L disodium hydrogen phosphate TS (1 in 5) (11:9) with phosphoric acid. Flow rate: Adjust so that the retention time of nifedipine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nifedipine are not less than 4000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nifedipine is not more than 1.0z.
Nilvadipine Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of nil-vadipine (C19H19N3O6: 385.37). Method of preparation Prepare as directed under Tablets, with Nilvadipine. Identification To a quantity of powdered Nilvadipine Tablets, equivalent to 1 mg of Nilvadipine, add 100 mL of ethanol (99.5), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Determine the ab-sorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 239 nm and 243 nm and a maximum hav-ing a broad-ranging absorption between 371 nm and 381 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Nilvadipine Tablets add V mL of a mixture of acetonitrile and water (7:3) so that each mL of the solu-tion contains about 0.
To a quantity of powdered Nilvadipine Tablets, equivalent to 1 mg of Nilvadipine, add 100 mL of ethanol (99.5), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Determine the ab-sorption spectrum of the sample solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 239 nm and 243 nm and a maximum hav-ing a broad-ranging absorption between 371 nm and 381 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Nilvadipine Tablets add V mL of a mixture of acetonitrile and water (7:3) so that each mL of the solu-tion contains about 0.2 mg of nilvadipine (C19H19N3O6), add exactly V mL of the internal standard solution, and disperse the particles by sonicating. Centrifuge for 10 minutes, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Nilvadipine RS, dissolve in the mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 25 mL of the internal standard solution and the mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay. Amount (mg) of nilvadipine (C19H19N3O6) ＝ MS × QT/QS × V/100 MS: Amount (mg) of Nilvadipine RS taken Internal standard solution—A solution of acenaphthene in acetonitrile (1 in 500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Nilvadipine Tablets is not less than 85z. Start the test with 1 tablet of Nilvadipine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet 10 mL of the subse-quent filtrate, add exactly 1 mL of methanol, and use this solution as the sample solution. Separately, weigh accurately an amount of Nilvadipine RS, equivalent to 10 times the labeled amount of Nilvadipine Tablets, and dissolve in meth-anol to make exactly 50 mL. Pipet 5 mL of this solution, and add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add exactly 10 mL of water, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of nilvadipine in each solution. Dissolution rate (z) with respect to the labeled amount of nilvadipine (C19H19N3O6) ＝ MS × AT/AS × 1/C × 9 MS: Amount (mg) of Nilvadipine RS taken C: Labeled amount (mg) of nilvadipine (C19H19N3O6) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of phosphate buffer solution (pH 7.4), methanol and acetonitrile (7:7:6). Flow rate: Adjust so that the retention time of nilvadipine is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nilvadipine are not less than 2000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nilvadipine is not more than 1.5z.
Weigh accurately not less than 20 Nilvadipine Tablets, and powder. Weigh accurately an amount of the powder, equivalent to about 5 mg of nilvadipine (C19H19N3O6), add 10 mL of a mixture of acetonitrile and water (7:3) and exactly 25 mL of the internal standard solu-tion, shake for 15 minutes, and add the mixture of aceto-nitrile and water (7:3) to make 50 mL. Centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Nilvadipine RS, dissolve in the mixture of acetonitrile and water (7:3) to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 25 mL of the internal standard solution and the mixture of acetonitrile and water (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nilvadipine to that of the internal standard. Amount (mg) of nilvadipine (C19H19N3O6) ＝ MS × QT/QS × 1/4 MS: Amount (mg) of Nilvadipine RS taken Internal standard solution—A solution of acenaphthene in acetonitrile (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.5 g of diammonium hydrogen phosphate in 1000 mL of water, add 10 mL of tetrabutylam-monium hydroxide TS, adjust the pH to 7.0 with diluted phosphoric acid (1 in 10), and add 900 mL of acetonitrile. Flow rate: Adjust so that the retention time of nilvadipine is about 12 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, nilvadipine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of nilvadipine to that of the internal standard is not more than 1.0z.
Nitrendipine Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of nitrendipine (C18H20N2O6: 360.36). Method of preparation Prepare as directed under Tablets, with Nitrendipine. Identification Shake a quantity of powdered Nitrendipine Tablets, equivalent to 5 mg of Nitrendipine, with 70 mL of methanol, then add methanol to make 100 mL, and centri-fuge. To 5 mL of the supernatant liquid add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 234 nm and 238 nm, and between 350 nm and 354 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Nitrendipine Tablets add 15 mL of diluted aceto-nitrile (4 in 5), stir until the tablet is completely disintegrat-ed, and further stir for 10 minutes. Add diluted acetonitrile (4 in 5) to make exactly 20 mL, and centrifuge. Pipet V mL of the supernatant liquid, equivalent to about 1 mg of nitren-dipine (C18H20N2O6), add exactly 5 mL of the internal stand-ard solution, then add diluted acetonitrile (4 in 5) to make 25 mL, and use this solution as the sample solution. Proceed as directed in the Assay. standard solution add the mobile phase to make exactly 10 Amount (mg) of nitrendipine (C H N O ) mL. Confirm that the peak area of nitrendipine obtained with 10 mL of this solution is equivalent to 14 to 26z of that ＝ MS × QT/QS × 1/V × 1/5 20 2 6 with 10 mL of the standard solution. System performance: Dissolve 10 mg of Nitrendipine and 3 mg of propyl parahydroxybenzoate in 5 mL of acetonitrile, and add the mobile phase to make 100 mL. When the proce-dure is run with 5 mL of this solution under the above operat-ing conditions, propyl parahydroxybenzoate and nitrendi-pine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitrendipine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.3 g of Nitrendipine, previ-ously dried, dissolve in 60 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 100), add 50 mL of water, and titrate <2.50> with 0.1 mol/L serium (IV) tetraammonium sulfate VS until the red-orange color of the solution vanishes (indi-cator: 3 drops of 1,10-phenanthroline TS). Perform a blank determination in the same manner, and make any necessary correction. MS: Amount (mg) of nitrendipine for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (4 in 5) (1 in 10,000). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of the dissolution medium containing 3 g of polysor-bate 80 in 5 L of water for 5-mg tablet and the dissolution medium containing 3 g of polysorbate 80 in 2000 mL of water for 10-mg tablet, the dissolution rate in 45 minutes of Nitrendipine Tablets is not less than 70z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Nitrendipine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL, add the dis-solution medium to make exactly V? mL so that each mL contains about 5.
Shake a quantity of powdered Nitrendipine Tablets, equivalent to 5 mg of Nitrendipine, with 70 mL of methanol, then add methanol to make 100 mL, and centri-fuge. To 5 mL of the supernatant liquid add methanol to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 234 nm and 238 nm, and between 350 nm and 354 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Nitrendipine Tablets add 15 mL of diluted aceto-nitrile (4 in 5), stir until the tablet is completely disintegrat-ed, and further stir for 10 minutes. Add diluted acetonitrile (4 in 5) to make exactly 20 mL, and centrifuge. Pipet V mL of the supernatant liquid, equivalent to about 1 mg of nitren-dipine (C18H20N2O6), add exactly 5 mL of the internal stand-ard solution, then add diluted acetonitrile (4 in 5) to make 25 mL, and use this solution as the sample solution. Proceed as directed in the Assay. standard solution add the mobile phase to make exactly 10 Amount (mg) of nitrendipine (C H N O ) mL. Confirm that the peak area of nitrendipine obtained with 10 mL of this solution is equivalent to 14 to 26z of that ＝ MS × QT/QS × 1/V × 1/5 20 2 6 with 10 mL of the standard solution. System performance: Dissolve 10 mg of Nitrendipine and 3 mg of propyl parahydroxybenzoate in 5 mL of acetonitrile, and add the mobile phase to make 100 mL. When the proce-dure is run with 5 mL of this solution under the above operat-ing conditions, propyl parahydroxybenzoate and nitrendi-pine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitrendipine is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.3 g of Nitrendipine, previ-ously dried, dissolve in 60 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 100), add 50 mL of water, and titrate <2.50> with 0.1 mol/L serium (IV) tetraammonium sulfate VS until the red-orange color of the solution vanishes (indi-cator: 3 drops of 1,10-phenanthroline TS). Perform a blank determination in the same manner, and make any necessary correction. MS: Amount (mg) of nitrendipine for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (4 in 5) (1 in 10,000). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of the dissolution medium containing 3 g of polysor-bate 80 in 5 L of water for 5-mg tablet and the dissolution medium containing 3 g of polysorbate 80 in 2000 mL of water for 10-mg tablet, the dissolution rate in 45 minutes of Nitrendipine Tablets is not less than 70z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Nitrendipine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet the subsequent V mL, add the dis-solution medium to make exactly V? mL so that each mL contains about 5.6 mg of nitrendipine (C18H20N2O6), and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of nitrendipine for assay, previously dried at 1059C for 2 hours, dissolve in methanol to make ex-actly 100 mL, then pipet 5 mL of this solution, and add the dissolution medium to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine ‌the peak areas, AT and AS, of nitrendipine in each solution. Dissolution rate (z) with respect to the labeled amount of nitrendipine (C18H20N2O6) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of nitrendipine for assay taken C: Labeled amount (mg) of nitrendipine (C18H20N2O6) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 356 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, tetrahydrofuran and acetonitrile (14:6:5). Flow rate: Adjust so that the retention time of nitrendi-pine is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of nitrendipine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitrendipine is not more than 2.0z. Assay Conduct this procedure using light-resistant vessels. To 20 tablets of Nitrendipine Tablets add 150 mL of diluted acetonitrile (4 in 5), stir until the tablets completely disinte-grate, and stir for further 10 minutes. Add diluted aceto-nitrile (4 in 5) to make exactly 200 mL, and centrifuge. Pipet a volume of the supernatant liquid, equivalent to about 2 mg of nitrendipine (C18H20N2O6), add exactly 10 mL of the inter-nal standard solution and diluted acetonitrile (4 in 5) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of nitrendipine for assay, previously dried at 1059C for 2 hours, and dissolve in diluted acetonitrile (4 in 5) to make exactly 200 mL. Pipet 4 mL of this solution, add exactly 10 mL of the internal stand-ard solution and diluted acetonitrile (4 in 5) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of nitrendipine to that of the in-ternal standard. and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, tetrahydrofuran and acetonitrile (14:6:5). Flow rate: Adjust so that the retention time of nitrendi-pine is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and nitrendipine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitrendipine is not more than 1.0z.
(1) Weigh a quantity of powdered Nitroglycerin Tablets, equivalent to 6 mg of nitroglycerin (C3H5N3O9), shake thoroughly with 12 mL of diethyl ether, filter, and use the filtrate as the sample solution. Evaporate 5 mL of the sample solution, dissolve the residue in 1 to 2 drops of sulfuric acid, and add 1 drop of diphenylamine TS: a deep blue color develops. (2) Evaporate 5 mL of the sample solution obtained in (1), add 5 drops of sodium hydroxide TS, heat over a low flame, and concentrate to about 0.1 mL. Cool, heat the residue with 0.02 g of potassium hydrogen sulfate: the odor of acrolein is perceptible.
Free nitrate ion—Transfer an accurately measured quantity of powdered Nitroglycerin Tablets, equivalent to 20 mg of nitroglycerin (C H N O ), to a separator, add 40 mL 30 minutes, and observe the tubes horizontally: the sample solution has no more color than the standard solution. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Transfer 1 tablet of Nitroglycerin Tablets to a glass-stoppered centrifuge tube, and add exactly V mL of acetic acid (100) to provide a solution containing about 30 mg of nitroglycerin (C3H5N3O9) per ml. Shake vigorously for 1 hour, and after disintegrating the tablet, centrifuge, and use the supernatant liquid as the sample solution. When the tablet does not disintegrate during this procedure, transfer 1 tablet of Nitroglycerin Tablets to a glass-stoppered centri-fuge tube, wet the tablet with 0.05 mL of acetic acid (100), and grind down it with a glass rod. While rinsing the glass rod, add acetic acid (100) to make exactly V mL of a solution containing about 30 mg of nitroglycerin (C3H5N3O9) per ml. Shake for 1 hour, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 90 mg of potassium nitrate, previously dried at 1059C for 4 hours, dissolve in 5 mL of water, and add acetic acid (100) to make exactly 100 mL. Pipet 5 mL of the solution, add acetic acid (100) to make exactly 100 mL, and use this solution as the standard solution. Measure exactly 2 mL each of the sample solution and the standard solution, add 2 mL each of salicylic acid TS shake, allow to stand for 15 minutes, and add 10 mL each of water. Render the solution alkaline with about 12 mL of a solution of sodium hydroxide (2 in 5) while cooling in ice, and add water to make exactly 50 mL. Per-form the test with these solutions as directed under Ultravio-let-visible Spectrophotometry <2.24>, using a solution, pre-pared with 2 mL of acetic acid (100) in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 410 nm, respectively. Amount (mg) of nitroglycerin (C3H5N3O9) ＝ MS × AT/AS × V/2000 × 0.749 MS: Amount (mg) of potassium nitrate taken Calculate the average content from the contents of 10 tablets: it meets the requirements of the test when each con-tent deviates from the average content by not more than 25z. When there is 1 tablet showing a deviation exceeding 25z and not exceeding 30z, determine the content of an additional 20 tablets in the same manner. Calculate the 30 deviations from the new average of all 30 tablets: it meets the requirements of the test when 1 tablet may deviate from the average content by between 25z and 30z, but no tablet deviates by more than 30z. Disintegration <6.09> It meets the requirement, provided that the time limit of the test is 2 minutes, and the use of the disks is omitted. 3 5 3 9 of isopropylether and 40 mL of water, shake for 10 minutes, and allow the layers to separate. Collect the aqueous layer, add 40 mL of isopropylether, shake for 10 minutes, collect the aqueous layer, filter, and use the filtrate as the sample solution. Separately, transfer 10 mL of Standard Nitric Acid Solution to a separator, add 30 mL of water and 40 mL of the isopropyl ether layer of the first extraction of the sample solution, shake for 10 minutes, continue the procedure in the same manner as the sample solution, and use the solution so obtained as the standard solution. Transfer 20 mL each of the sample solution and the standard solution to Nessler tubes, respectively, shake well with 30 mL of water and 0.06 g of Griess-Romijin's nitric acid reagent, allow to stand for
Weigh accurately and disintegrate, by soft pressing, not less than 20 Nitroglycerin Tablets. Weigh accurately a portion of the powder, equivalent to about 3.5 mg of nitroglycerin (C3H5N3O9), add exactly 50 mL of acetic acid (100), shake for 1 hour, filter, and use this filtrate as the sample solution. Separately, weigh accurately about 90 mg of potassium nitrate, previously dried at 1059C for 4 hours, dissolve in 5 mL of water, and add acetic acid (100) to make exactly 100 mL. Pipet 10 mL of the solution, add acetic acid (100) to make exactly 100 mL, and use this solution as the standard solution. Measure exactly 2 mL each of the sample solution and the standard solution, to each solution add 2 mL of salicylic acid TS, shake, allow to stand for 15 ‌minutes, and add 10 mL of water. Render the solution alka-line with about 12 mL of a solution of sodium hydroxide (2 in 5) while cooling in ice, and add water to make exactly 50 mL. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solu-tion, prepared with 2 mL of acetic acid (100) in the same manner, as the blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 410 nm, respectively. Amount (mg) of nitroglycerin (C3H5N3O9) ＝ MS × AT/AS × 1/20 × 0.749 MS: Amount (mg) of potassium nitrate taken
Containers—Tight containers. Storage—Light-resistant, and not exceeding 209C.
Nizatidine Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of nizati-dine (C12H21N5O2S2: 331.46). Method of preparation Prepare as directed under Cap-sules, with Nizatidine. Identification Take out the contents of Nizatidine Cap-sules, and powder. To a portion of the powder, equivalent to 50 mg of Nizatidine, add 50 mL of methanol, shake well, and filter. Pipet 1 mL of the filtrate, and add methanol to make 100 mL. Determine the absorption spectrum of the so- MS: Amount (mg) of Nizatidine RS taken Internal standard solution—A solution of phenol in the mo-bile phase (1 in 100). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using a sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Nizatidine Capsules is not less than 80z. Start the test with 1 capsule of Nizatidine Capsules, with-draw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 10 mg of nizatidine (C12H21N5O2S2).
Take out the contents of Nizatidine Cap-sules, and powder. To a portion of the powder, equivalent to 50 mg of Nizatidine, add 50 mL of methanol, shake well, and filter. Pipet 1 mL of the filtrate, and add methanol to make 100 mL. Determine the absorption spectrum of the so- MS: Amount (mg) of Nizatidine RS taken Internal standard solution—A solution of phenol in the mo-bile phase (1 in 100). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using a sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Nizatidine Capsules is not less than 80z. Start the test with 1 capsule of Nizatidine Capsules, with-draw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 10 mg of nizatidine (C12H21N5O2S2). Use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Nizatidine RS, previously dried at 100 9C for 1 hour, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 314 nm. Dissolution rate (z) with respect to the labeled amount of nizatidine (C12H21N5O2S2) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of Nizatidine RS taken C: Labeled amount (mg) of nizatidine (C12H21N5O2S2) in 1 capsule
Take out the contents of not less than 10 Nizatidine Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equiva-lent to about 0.15 g of nizatidine (C12H21N5O2S2), add ex-actly 50 mL of the mobile phase, shake vigorously for 10 minutes, and centrifuge. Pipet 5 mL of the supernatant liq-uid, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 15 mg of Nizatidine RS, previously dried at 1009C for 1 hour, dis-solve in 30 mL of the mobile phase, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of nizatidine to that of the internal standard. lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 239 nm and 244 nm, and between 323 nm and 327 nm. Amount (mg) of nizatidine (C ＝ MS × QT/QS × 10 12H21N5O2S2) Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the contents from 1 capsule of Nizatidine Cap-sules, add the mobile phase to make exactly V mL so that each mL contains about 1.5 mg of nizatidine (C12H21N5O2S 2). After shaking vigorously for 10 minutes, centrifuge. Pipet 10 mL of the supernatant liquid, add exactly 5 mL of the in-ternal standard solution and add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, MS: Amount (mg) of Nizatidine RS taken Internal standard solution—A solution of phenol in the mo-bile phase (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ‌409C.‌ Mobile phase: Dissolve 5.9 g of ammonium acetate in 760 mL of water, add 1 mL of diethylamine, and adjust to pH 7.5 with acetic acid (100). To this solution add 240 mL of methanol. Flow rate: Adjust so that the retention time of nizatidine is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and nizatidine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of nizatidine to that of the internal standard is not more than 1.0z.
Norepinephrine Injection is a clear, colorless liquid. It gradually becomes a pale red color by light and by air. pH: 2.3 – 5.0
Transfer a volume of Noradrenaline Injec-tion, equivalent to 1 mg of Noradrenaline, to each of two test tubes A and B, and add 1 mL of water to each tube. Add 10 mL of potassium hydrogen phthalate buffer solution (pH 3.5) to A, and 10 mL of phosphate buffer solution (pH 6.5) to B. To each of these solutions add 1.0 mL of iodine TS, allow to stand for 5 minutes, and add 2.0 mL of sodium thiosulfate TS: no color or a pale red color develops in test tube A, and a deep red-purple color develops in test tube B.
(1) Arterenone—Measure a volume of Noradrena-line Injection, equivalent to 10 mg of Noradrenaline, add water to make exactly 20 mL, and determine the absorbance of this solution at 310 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>: the absorbance is not more than 0.10. (2) Adrenaline—Measure a volume of Noradrenaline In-jection, equivalent to 5 mg of Noradrenaline, add 1 mL of diluted acetic acid (100) (1 in 2) and water to make exactly 10 mL, and proceed as directed in the Purity (3) under Noradrenaline. Bacterial endotoxins <4.01> Less than 300 EU/mg. ‌Extractable volume <6.05> It meets the requirement.‌ Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Noradrenaline Injection, equiva-lent to about 5 mg of dl-noradrenaline (C8H11NO3), add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Noradrenaline Bitartrate RS, previously dried in a desic-cator (in vacuum, silica gel) for 24 hours, dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Pipet 5 mL each of the sample solution and stand-ard solution, add 0.2 mL each of starch TS, then add iodine TS dropwise with swirling until a persistent blue color is produced. Add 2 mL of iodine TS, and shake. Adjust the pH of the solution to 6.5 with 0.05 mol/L disodium hydro-genphosphate TS, add 10 mL of phosphate buffer solution slightly soluble in water. It is affected by light. Identification (1) To 2 mg of Norethisterone add 2 mL of sulfuric acid: the solution shows a red-brown color and a yel-low-green fluorescence. Add 10 mL of water to this solution cautiously: a yellow color develops and a yellow-brown pre-cipitate is formed. (2) Determine the infrared absorption spectrum of Norethisterone as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: －32 – －379(after drying, 0.25 g, acetone, 25 mL, 100 mm). Melting point <2.60> 203 – 2099C Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (0.5 g). Assay Weigh accurately about 0.2 g of Norethisterone, previously dried, dissolve in 40 mL of tetrahydrofuran, add 10 mL of a solution of silver nitrate (1 in 20), and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS (pH 6.5), and shake. Immediately after allowing to stand for 3 minutes, add sodium thiosulfate TS dropwise until a red- ＝ 29.84 mg of C20 H26O2 purple color develops, then add water to make exactly 50 mL. Determine the absorbances, AT and AS, of the subse-quent solutions of the sample solution and the standard solu-tion at 515 nm within 5 minutes as directed under Ultravio-let-visible Spectrophotometry <2.24>. Amount (mg) of dl-noradrenaline (C8H11NO3) ＝ MS × AT/AS × 0.502 MS: Amount (mg) of Noradrenaline Bitartrate RS taken
ノルゲストレル・エチニルエストラジオール錠
(1) To a quantity of powdered Norgestrel and Ethinylestradiol Tablets, equivalent to 10 mg of Norges-trel add 10 mL of ethyl acetate, shake for 10 minutes, and filter. To 2 mL of the filtrate add 6 mL of sodium hydroxide TS, shake vigorously, and centrifuge. Take 1 mL of the ethyl acetate layer, evaporate on a water bath to dryness, dissolve the residue in 2 mL of ethanol (95), and add 1 mL of sulfuric acid: a red-purple color develops. Examine under ultraviolet light (main wavelength: 365 nm): this solution shows a red-orange fluorescence (norgestrel). Take 1 mL of the filtrate obtained in (1), evaporate on a water bath to dryness, add 1 mL of boric acid-methanol buffer solution to the residue, shake, and cool in ice. Add 1 mL of ice-cold diazo TS, shake, add 1 mL of sodium hy-droxide TS, and shake: a red-orange color develops (ethinylestradiol). Use the filtrate obtained in (1) as the sample solution. Separately, dissolve 10 mg of Norgestrel RS and 1 mg of Ethinylestradiol RS, respectively, in 10 mL of ethyl acetate, and use these solutions as the standard solution (1) and the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatogra-phy. Develop the plate with a mixture of pentane and ethyl acetate (3:2) to a distance of about 7 cm, and air-dry the plate. Spray evenly a solution of p-toluenesulfonic acid monohydrate in ethanol (95) (1 in 5) on the plate, and heat the plate at 1059C for 5 minutes. Examine under ultraviolet light (main wavelength: 365 nm): two spots obtained from the sample solution show the similar color tone and Rf value to each spot from the standard solutions (1) and (2). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Add 2 mL of diluted methanol (7 in 10) to 1 tablet of Nor-gestrel and Ethinylestradiol Tablets, add exactly 2 mL of the internal standard solution, shake for 20 minutes, and centri- ratios, QTa and QTb, of the peak areas of norgestrel and ethinylestradiol to the peak area of the internal standard of the sample solution and also the ratios, QSa and QSb, of the peak areas of norgestrel and ethinylestradiol to the peak area of the internal standard of the standard solution. Then, proceed as directed in the Assay. Amount (mg) of norgestrel (C21H28O2) ＝ MSa × QTa/QSa × 1/100 Amount (mg) of ethinylestradiol (C20H24O2) ＝ MSb × QTb/QSb × 1/100 MSa: Amount (mg) of Norgestrel RS taken MSb: Amount (mg) of Ethinylestradiol RS taken Internal standard solution—A solution of diphenyl in diluted methanol (7 in 10) (1 in 50,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Norgestrel and Ethinylestradiol Tablets is not less than 70z. Start the test with 1 tablet of Norgestrel and Ethinyles-tradiol Tablets, withdraw not less than 50 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet exactly V mL of the subsequent filtrate, equivalent to about 17 mg of norgestrel (C21H28O2) and about 1.7 mg of ethinylestradiol (C20H24O2), transfer into a chromatography column [prepared by packing 0.36 g of octadecylsilanized silica gel for pretreatment (55 to 105 mm in particle diameter) in a tube about 1 cm in inside diameter]. After washing the column with 15 mL of water, elute with 3 mL of methanol, and evaporate the effluent in a water bath to dryness at about 409C with the aid of a current air. Dissolve the residue in exactly 2 mL of diluted methanol (7 in 10), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Norgestrel RS and about 2.5 mg of Ethinylestradiol RS, dissolve in diluted methanol (7 in 10) to make exactly 100 mL, then pipet 3 mL of this solution, add diluted methanol (7 in 10) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, ATa and ATb, of norgestrel and ethinylestradiol from the sample solution, and the peak areas, ASa and ASb, of norgestrel and ethinylestradiol from the standard solution. Dissolution rate (z) with respect to the labeled amount fuge. Filter the supernatant liquid through a membrane filter with a pore size of not more than 0.2 mm, and use this fil- of norgestrel (C 21H 28O2) trate as the sample solution. Separately, weigh accurately ＝ MSa × ATa/ASa × 1/V × 1/Ca × 54 ‌Dissolution rate (z) with respect to the labeled amount of ethinylestradiol (C20H24O2) ＝ MSb × ATb/ASb × 1/V × 1/Cb × 54 MSa: Amount (mg) of Norgestrel RS taken MSb: Amount (mg) of Ethinylestradiol RS taken Ca: Labeled amount (mg) of norgestrel (C21H28O2) in 1 tablet Cb: Labeled amount (mg) of ethinylestradiol (C20H24O2) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Proceed as directed in the system suitability in the Assay.
Weigh accurately not less than 20 Norgestrel and with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ethinylestradiol and norgestrel to that of the internal standard are not more than 1.0z, respectively.
Nortriptyline Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of nortriptyline (C19H21N: 263.38). Method of preparation Prepare as directed under Tablets, with Nortriptyline Hydrochloride. Identiˆcation (1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, using diluted 0.1 mol/L hydrochloric acid TS (1 in 50) as the blank: it exhibits a maximum between 237 nm and 241 nm. (2) To a quantity of powdered Nortriptyline Hydrochlo-ride Tablets, equivalent to 10 mg of nortriptyline (C19H21N), add 10 mL of ethanol (99.5), shake thoroughly, centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, dissolve 11 mg of nortriptyline hydrochloride in 10 mL of ethanol (99.5), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with ‰uorescent indicator for thin-layer chromatog-raphy. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (4:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord- the subsequent ˆltrate, add water to make exactly V? mL so that each mL contains about 11 mg of nortriptyline (C19H21N), and use this solution as the sample solution.
Weigh accurately the mass of not less than 20 tablets of Nortriptyline Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of nortriptyline (C19H21N), add 50 mL of 0.1 mol/L hy-drochloric acid TS, sonicate, and extract for 15 minutes while occasional shaking. Shake for 15 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Centri-fuge this solution, pipet 2 mL of the supernatant liquid, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of nortriptyline hydrochloride for assay, previously dried at 1059C for 2 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 239 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using diluted 0.1 mol/L hydrochloric acid TS (1 in 50) as the blank. ‌Amount (mg) of nortriptyline (C19H21N) ＝ MS × AT/AS × 2 × 0.878 MS: Amount (mg) of nortriptyline hydrochloride for assay taken
Olmesartan Medoxomil Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of olmesartan medoxomil (C29H30N6O6: 558.59). Method of preparation Prepare as directed under Tablets, with Olmesartan Medoxomil. Identification To a quantity of powdered Olmesartan Medoxomil Tablets, equivalent to 20 mg of Olmesartan Medoxomil, add 60 mL of a mixture of acetonitrile and water (3:2), sonicate for 10 minutes, and add a mixture of acetonitrile and water (3:2) to make 100 mL. Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (3:2) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 255 nm and 259 nm. Purity Related substances—To a quantity of powdered Olmesartan Medoxomil Tablets, equivalent to 20 mg of Ol-mesartan Medoxomil, add 20 mL of a mixture of acetonitrile and water (9:1), sonicate for 15 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the areas of the peaks, having the relative retention time of about 0.2 and about 1.6 to olmesartan medoxomil, obtained from the sam-ple solution are not larger than 3/5 times the peak area of ol-mesartan medoxomil from the standard solution, and the area of the peak other than olmesartan medoxomil and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of olmesartan medox-omil from the standard solution. Furthermore, the total area of the peaks other than olmesartan medoxomil from the sample solution is not larger than 1.4 times the peak area of olmesartan medoxomil from the standard solution. For the areas of the peaks, having the relative retention time of about 0.7 and about 3.4 to olmesartan medoxomil, multiply their collection factors, 0.65 and 1.39, respectively. Operating conditions— Detector, column, column temperature, mobile phase, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Olmesar-tan Medoxomil. Time span of measurement: For 45 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (9:1) to make exactly 20 mL. Confirm that the peak area of olmesartan medoxomil obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the stand-ard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of olmesartan medoxomil are not less than 5500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olmesartan medoxomil is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Olmesartan Medoxomil Tablets add 5V/7 mL of a mixture of acetonitrile and water (3:2) and exactly V/10 mL of the internal standard solution. Sonicate for 10 minutes with occasional stirring, and add a mixture of aceto-nitrile and water (3:2) to make V mL so that each mL con-tains about 0.2 mg of olmesartan medoxomil (C29H30N6O6). Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (3:2) to make 25 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of olmesartan medoxomil (C H N O ) with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according ＝ MS × QT/QS × V/200 29 30 6 6 to the following conditions, and calculate the ratios, QT and QS, of the peak area of olmesartan medoxomil to that of the MS: Amount (mg) of Olmesartan Medoxomil RS taken, calculated on the anhydrous and residual solvent-free internal standard. Amount (mg) of olmesartan medoxomil (C H N O ) basis ＝ MS × QT/QS × 1/2 29 30 6 6 Internal standard solution—A solution of isobutyl para-hydroxybenzoate in a mixture of acetonitrile and water (3:2) (1 in 1000). ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rates in 30 minutes of 5-mg, 10-mg and 20-mg tablets are not less than 80z, and that in 30 minutes of 40-mg tablet is not less than 75z. Start the test with 1 tablet of Olmesartan Medoxomil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 6 mg of ol-mesartan medoxomil (C29H30N6O6), and use this solution as the sample solution.
To a quantity of powdered Olmesartan Medoxomil Tablets, equivalent to 20 mg of Olmesartan Medoxomil, add 60 mL of a mixture of acetonitrile and water (3:2), sonicate for 10 minutes, and add a mixture of acetonitrile and water (3:2) to make 100 mL. Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (3:2) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 255 nm and 259 nm.
Related substances—To a quantity of powdered Olmesartan Medoxomil Tablets, equivalent to 20 mg of Ol-mesartan Medoxomil, add 20 mL of a mixture of acetonitrile and water (9:1), sonicate for 15 minutes, centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.5 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and water (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the areas of the peaks, having the relative retention time of about 0.2 and about 1.6 to olmesartan medoxomil, obtained from the sam-ple solution are not larger than 3/5 times the peak area of ol-mesartan medoxomil from the standard solution, and the area of the peak other than olmesartan medoxomil and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of olmesartan medox-omil from the standard solution. Furthermore, the total area of the peaks other than olmesartan medoxomil from the sample solution is not larger than 1.4 times the peak area of olmesartan medoxomil from the standard solution. For the areas of the peaks, having the relative retention time of about 0.7 and about 3.4 to olmesartan medoxomil, multiply their collection factors, 0.65 and 1.39, respectively. Operating conditions— Detector, column, column temperature, mobile phase, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity (2) under Olmesar-tan Medoxomil. Time span of measurement: For 45 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and water (9:1) to make exactly 20 mL. Confirm that the peak area of olmesartan medoxomil obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the stand-ard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of olmesartan medoxomil are not less than 5500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olmesartan medoxomil is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Olmesartan Medoxomil Tablets add 5V/7 mL of a mixture of acetonitrile and water (3:2) and exactly V/10 mL of the internal standard solution. Sonicate for 10 minutes with occasional stirring, and add a mixture of aceto-nitrile and water (3:2) to make V mL so that each mL con-tains about 0.2 mg of olmesartan medoxomil (C29H30N6O6). Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (3:2) to make 25 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of olmesartan medoxomil (C H N O ) with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according ＝ MS × QT/QS × V/200 29 30 6 6 to the following conditions, and calculate the ratios, QT and QS, of the peak area of olmesartan medoxomil to that of the MS: Amount (mg) of Olmesartan Medoxomil RS taken, calculated on the anhydrous and residual solvent-free internal standard. Amount (mg) of olmesartan medoxomil (C H N O ) basis ＝ MS × QT/QS × 1/2 29 30 6 6 Internal standard solution—A solution of isobutyl para-hydroxybenzoate in a mixture of acetonitrile and water (3:2) (1 in 1000). ‌Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rates in 30 minutes of 5-mg, 10-mg and 20-mg tablets are not less than 80z, and that in 30 minutes of 40-mg tablet is not less than 75z. Start the test with 1 tablet of Olmesartan Medoxomil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 6 mg of ol-mesartan medoxomil (C29H30N6O6), and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Olmesartan Medoxomil RS (separately, determine the water <2.48> and the residual solvent in the same manner as Olmesartan Medoxomil), dissolve in 15 mL of ethanol (99.5) by warming at 50 – 609C, and after cooling add ethanol (99.5) to make exactly 20 mL. Pipet 5 mL of this solution, add ethanol (99.5) to make exactly 50 mL. Then, pipet 5 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 257 nm of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24> using the dissolu-tion medium as the control. Dissolution rate (z) with respect to the labeled amount of olmesartan medoxomil (C29H30N6O6) ＝ MS × AT/AS × V?/V × 1/C × 45/4 MS: Amount (mg) of Olmesartan Medoxomil RS taken, calculated on the anhydrous and residual solvent-free basis C: Labeled amount (mg) of olmesartan medoxomil (C29H30N6O6) in 1 tablet
Weigh accurately the mass of not less than 20 Ol-mesartan Medoxomil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of ol-mesartan medoxomil (C29H30N6O6), add 70 mL of a mixture of acetonitrile and water (3:2) and exactly 10 mL of the in-ternal standard solution. Sonicate for 15 minutes with occa-sional stirring, and add a mixture of acetonitrile and water (3:2) to make 100 mL. Centrifuge this solution, to 5 mL of the supernatant liquid add a mixture of acetonitrile and water (3:2) to make 25 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 40 mg of Olmesartan Medoxomil RS (separately determine the water <2.48> and the residual solvent in the same manner as Ol- mesartan Medoxomil), dissolve in 60 mL of a mixture of acetonitrile and water (3:2), add exactly 20 mL of the inter-nal standard solution, then add a mixture of acetonitrile and water (3:2) to make 100 mL. To 5 mL of this solution add a mixture of acetonitrile and water (3:2) to make 50 mL, and use this solution as the standard solution. Perform the test MS: Amount (mg) of Olmesartan Medoxomil RS taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of isobutyl para-hydroxybenzoate in a mixture of acetonitrile and water (3:2) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 250 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.04 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 3.4 with a solution prepared by dissolving 1.73 g of phosphoric acid in water to make 1000 mL. To 330 mL of this solution add 170 mL of acetonitrile. Flow rate: Adjust so that the retention time of olmesartan medoxomil is about 16 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, olmesartan medoxomil and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviations of the ratio of the peak area of olmesartan medoxomil to that of the in-ternal standard is not more than 1.0z.
Olopatadine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of olopatadine hydrochloride (C21H23NO3. HCl: 373.87). Method of preparation Prepare as directed under Tablets, with Olopatadine Hydrochloride. Identification Shake well a quantity of powdered Olopata-dine Hydrochloride Tablets, equivalent to 5 mg of Olopata-dine Hydrochloride, with 100 mL of 0.01 mol/L hydro-chloric acid TS, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 295 nm and 299 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Olopatadine Hydrochloride Tablets add 4V/5 mL of a mixture of 0.05 mol/L phosphate buffer solu-tion (pH 3.5) and acetonitrile (3:2). To this solution add ex-actly V/10 mL of the internal standard solution, shake well, and add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2) to make V mL so that each mL contains about 50 mg of olopatadine hydrochloride (C21H23NO3.
Shake well a quantity of powdered Olopata-dine Hydrochloride Tablets, equivalent to 5 mg of Olopata-dine Hydrochloride, with 100 mL of 0.01 mol/L hydro-chloric acid TS, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 295 nm and 299 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Olopatadine Hydrochloride Tablets add 4V/5 mL of a mixture of 0.05 mol/L phosphate buffer solu-tion (pH 3.5) and acetonitrile (3:2). To this solution add ex-actly V/10 mL of the internal standard solution, shake well, and add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2) to make V mL so that each mL contains about 50 mg of olopatadine hydrochloride (C21H23NO3.HCl). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, and use this fil-trate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of olopatadine hydrochloride (C21H23NO3.HCl) ＝ MS × QT/QS × V/1000 MS: Amount (mg) of olopatadine hydrochloride for assay taken Internal standard solution—A solution of doxepin hydro-chloride in a mixture of 0.05 mol/L phosphate buffer solu tion (pH 3.5) and acetonitrile (3:2) (7 in 20,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Olopatadine Hydrochloride Tablets is not less than 85z. ‌Start the test with 1 tablet of Olopatadine Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.8 mg of olopatadine hydro-chloride (C21H23NO3.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 28 mg of olopatadine hydrochloride for assay, previously dried at 105 9C for 3 hours, dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL. Then pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of olopata-dine in each solution. Dissolution rate (z) with respect to the labeled amount Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of olopatadine to that of the internal standard. Amount (mg) of olopatadine hydrochloride (C21H23NO3.HCl) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of olopatadine hydrochloride for assay taken Internal standard solution—A solution of doxepin hydro-chloride in a mixture of 0.05 mol/L phosphate buffer solu-tion (pH 3.5) and acetonitrile (3:2) (7 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 299 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2.3 g of sodium lauryl sulfate in a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (11:9) to make 1000 mL. Flow rate: Adjust so that the retention time of olopatadine of olopatadine hydrochloride (C 21H 23NO3.HCl) is about 11 minutes. System suitability— ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of olopatadine hydrochloride for assay taken C: Labeled amount (mg) of olopatadine hydrochloride (C21H23NO3.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of olopatadine are not less than 10,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olopatadine is not more than 1.5z.
Weigh accurately the mass of not less than 20 Olopatadine Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 5 mg of olopatadine hydrochloride (C21H23NO3.HCl), add 80 mL of a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2), and add exactly 10 mL of the internal standard solution. Shake well for 10 minutes, add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2) to make 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, and use this filtrate as the sample solution. Sepa-rately, weigh accurately about 50 mg of olopatadine hydro-chloride for assay, previously dried at 1059C for 3 hours, and dissolve in a mixture of 0.05 mol/L phosphate buffer so-lution (pH 3.5) and acetonitrile (3:2) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 3.5) and acetonitrile (3:2) to make 100 mL, and use this solution as the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, olopatadine and the internal standard are eluted in this order with the resolution between these peaks being not less than 13. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of olopatadine is not more than 1.0z.
Omeprazole Delayed-release Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of omeprazole (C17H19N3O3S: 345.42). Method of preparation Prepare as directed under Tablets, with Omeprazole. Identification Powder Omeprazole Delayed-release Tablets. To a portion of the powder, equivalent to 10 mg of Omeprazole, add 10 mL of ethanol (95), shake for 10 minutes, and centrifuge. To 1 mL of the supernatant liquid add phosphate buffer solution (pH 7.4) to make 50 mL. De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 273 nm and 277 nm, and between 299 nm and 303 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Omeprazole Delayed-release Tablets add V/20 mL of a solution of sodium tetraborate decahydrate (19 in 5000), and shake thoroughly to disintegrate the tablet. Then, proceed as directed in the Assay. Amount (mg) of omeprazole (C17H19N3O3S) ＝ MS × QT/QS × V/50 MS: Amount (mg) of omeprazole for assay taken Internal standard solution—A solution of 1,2-dinitroben-zene in ethanol (95) (1 in 400). Dissolution <6.10> When the tests are performed at 50 revolutions per minute according to the Paddle method, using 900 mL each of 1st fluid for dissolution test and 2nd fluid for dissolution test as the dissolution medium, the dis- ‌solution rates of 10-mg tablet and 20-mg tablet in 120 minutes of the test using the 1st fluid for dissolution test are not more than 5z, respectively, and those of 10-mg tablet in 20 minutes and 20-mg tablet in 15 minutes of the test using the 2nd fluid for dissolution test are not less than 85z, re-spectively. Start the test with 1 tablet of Omeprazole Delayed-release Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of omeprazole (C17H19N3O3S), and use this solution as the sam-ple solution.
Powder Omeprazole Delayed-release Tablets. To a portion of the powder, equivalent to 10 mg of Omeprazole, add 10 mL of ethanol (95), shake for 10 minutes, and centrifuge. To 1 mL of the supernatant liquid add phosphate buffer solution (pH 7.4) to make 50 mL. De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 273 nm and 277 nm, and between 299 nm and 303 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Omeprazole Delayed-release Tablets add V/20 mL of a solution of sodium tetraborate decahydrate (19 in 5000), and shake thoroughly to disintegrate the tablet. Then, proceed as directed in the Assay. Amount (mg) of omeprazole (C17H19N3O3S) ＝ MS × QT/QS × V/50 MS: Amount (mg) of omeprazole for assay taken Internal standard solution—A solution of 1,2-dinitroben-zene in ethanol (95) (1 in 400). Dissolution <6.10> When the tests are performed at 50 revolutions per minute according to the Paddle method, using 900 mL each of 1st fluid for dissolution test and 2nd fluid for dissolution test as the dissolution medium, the dis- ‌solution rates of 10-mg tablet and 20-mg tablet in 120 minutes of the test using the 1st fluid for dissolution test are not more than 5z, respectively, and those of 10-mg tablet in 20 minutes and 20-mg tablet in 15 minutes of the test using the 2nd fluid for dissolution test are not less than 85z, re-spectively. Start the test with 1 tablet of Omeprazole Delayed-release Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of omeprazole (C17H19N3O3S), and use this solution as the sam-ple solution. Separately, weigh accurately about 22 mg of omeprazole for assay, previously dried in vacuum at 509C using phosphorus (V) oxide as desiccant for 2 hours, and dis-solve in ethanol (95) to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24> at 323 nm when the test is per-formed using the 1st fluid as the dissolution medium and at 293 nm when the test is performed using the 2nd fluid as the dissolution medium, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of omeprazole (C17H19N3O3S) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of omeprazole for assay taken C: Labeled amount (mg) of omeprazole (C17H19N3O3S) in 1 tablet
To 20 Omeprazole Delayed-release Tablets add V/20 mL of a solution of sodium tetraborate decahydrate (19 in 5000), shake to disintegrate. To this solution add 3V/5 mL of ethanol (95), shake for 15 minutes, then add ethanol (95) to make exactly V mL so that each mL contains about 0.4 mg of omeprazole (C17H19N3O3S), and centrifuge. Pipet 10 mL of the supernatant liquid, add exactly 4 mL of the inter-nal standard solution, add a mixture of ethanol (95) and a solution of sodium tetraborate decahydrate (19 in 5000) (19:1) to make 20 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 20 mg of omepra-zole for assay, previously dried in vacuum at 509C with phosphorus (V) oxide as the desiccant for 2 hours, dissolve in a mixture of ethanol (95) and a solution of sodium tetraborate decahydrate (19 in 5000) (19:1), add exactly 20 mL of the internal standard solution, add a mixture of ethanol (95) and a solution of sodium tetraborate decahy-drate (19 in 5000) (19:1) to make 100 mL, and use this solu-tion as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of omeprazole to that of the internal standard. Amount (mg) of omeprazole (C17H19N3O3S) in tablet ＝ MS × QT/QS × V/1000 MS: Amount (mg) of omeprazole for assay taken Internal standard solution—A solution of 1,2-dinitroben-zene in ethanol (95) (1 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.83 g of disodium hydrogen phos-phate dodecahydrate and 0.21 g of sodium dihydrogen phos-phate dihydrate in water to make 1000 mL, and adjust to pH 7.6 with diluted phosphoric acid (1 in 100). To 290 mL of this solution add 110 mL of acetonitrile. Flow rate: Adjust so that the retention time of omeprazole is about 8 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, omeprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of omeprazole to that of the internal standard is not more than 1.0z.
アヘンアルカロイド・アトロピン注射液
Opium Alkaloids and Atropine Injection is an aqueous injection. It contains not less than 0.
Opium Alkaloids and Atropine Injection is a colorless or light brown, clear liquid. It is affected by light. pH: 2.5 – 3.5
(1) To 1 mL of Opium Alkaloids and Atro-pine Injection add 1 mL of ethanol (99.5), mix, and use this solution as the sample solution. Proceed with the sample so-lution as directed in the Identification (1) under Opium Alkaloids Hydrochlorides. (2) To 2 mL of Opium Alkaloids and Atropine Injection add 2 mL of ammonia TS, extract with 10 mL of diethyl ether, and filter the diethyl ether layer. Evaporate the filtrate on a water bath to dryness, add 1 mL of ethanol (99.5) to the residue, and heat to dissolve. Allow to stand this solution in an ice water for 30 minutes with occasional shaking. After crystals are formed, use the supernatant liquid as the sample solution. Separately, dissolve 0.03 g of Atropine Sulfate RS in 100 mL of water, proceed with 2 mL of this solution in the same manner as for the sample solution, and use a solution so obtained as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chro-matography. Develop the plate with a mixture of methanol and ammonia water (28) (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: a spot of about 0.2 Rf value among the several spots obtained from the sample solution and an orange colored spot from the standard solution show the same color tone, and have the same Rf value (atropine). Extractable volume <6.05> It meets the requirements.
(1) Morphine—Pipet 2 mL of Opium Alkaloids and Atropine Injection, add exactly 10 mL of the internal standard solution, then add water to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of morphine hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard so-lution, then add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine (C17H19NO3) ＝ MS × QT/QS × 0.887 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios ‌of the peak area of morphine to that of the internal standard is not more than 2.0z. (2) Atropine sulfate hydrate—Pipet 2 mL of Opium Alkaloids and Atropine Injection, add exactly 2 mL of the internal standard solution, and add 10 mL of diluted dilute hydrochloric acid (1 in 10). Shake this solution with two 10-mL portions of dichloromethane. Remove the dichloro-methane layer, to the water layer add 2 mL of ammonia TS, immediately add 20 mL of dichloromethane, shake vigor-ously, filter the dichloromethane extract through filter paper on which 5 g of anhydrous sodium sulfate is placed, and evaporate the filtrate to dryness under reduced pressure. To the residue add 0.5 mL of 1,2-dichloroethane and 0.5 mL of bis-trimethyl silyl acetamide, stopper tightly, warm in a water bath at 609C for 15 minutes, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Atropine Sulfate RS (determine separately the loss on drying <2.41> under the same conditions as Atropine Sulfate Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, and add exactly 2 mL of the internal standard solution. Proceed with this solution in the same manner as directed for the sample solution, and use this solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of atropine to that of the internal standard. Amount (mg) of atropine sulfate hydrate [(C17H23NO3)2.H2SO4.H2O] ＝ MS × QT/QS × 1/50 × 1.027 MS: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis Internal standard solution—A solution of homatropine hydrobromide (1 in 4000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1.5 m in length, packed with 180 to 250 mm siliceous earth for gas chromatography coated in 1 to 3z with 50z phenyl-methyl silicone polymer for gas chromatography. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen or helium. Flow rate: Adjust so that the retention time of atropine is about 5 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the internal standard and atropine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 5 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of atropine to that of the internal standard is not more than 2.0z.
Opium Alkaloids and Scopolamine Injection
アヘンアルカロイド・スコポラミン注射液
Opium Alkaloids and Scopolamine Injection is an aqueous injection. It contains not less than 1.
Opium Alkaloids and Scopolamine Injection is a clear, colorless to light brown liquid. It is affected by light. pH: 2.5 – 3.5
(1) To 1 mL of Opium Alkaloids and Scopolamine Injection add 1 mL of water and 2 mL of ethanol (99.5), mix, and use this solution as the sample solu-tion. Proceed with the sample solution as directed in the Identification (1) under Opium Alkaloids Hydrochlorides. (2) To 1 mL of Opium Alkaloids and Scopolamine Injec-tion add 1 mL of water and 2 mL of ammonia TS, extract with 10 mL of diethyl ether, and filter the diethyl ether layer. Evaporate the filtrate on a water bath to dryness, add 1 mL of ethanol (99.5) to the residue, and heat to dissolve. Allow to stand this solution in an ice water for 30 minutes with oc-casional shaking. After crystals are formed, use the superna-tant liquid as the sample solution. Separately, dissolve 0.03 g of Scopolamine Hydrobromide RS in 100 mL of water. To 2 mL of this solution add 2 mL of ammonia TS, proceed with this solution in the same manner as for the sample solution, and use a solution so obtained as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and ammonia water (28) (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendor-ff's TS for spraying on the plate: a spot with an Rf value of about 0.7 among the several spots obtained from the sample solution and an orange colored spot from the standard solu-tion show the same color tone, and have the same Rf value (scopolamine). Extractable volume <6.05> It meets the requirements.
(1) Morphine—Pipet 1 mL of Opium Alkaloids and Scopolamine Injection, add 10 mL of the internal stand-ard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard solution, add water to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. ‌Amount (mg) of morphine (C17H19NO3) ＝ MS × QT/QS × 0.887 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrin hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 2.0z. (2) Scopolamine hydrobromide hydrate—Pipet 2 mL of Opium Alkaloids and Scopolamine Injection, and add ex-actly 2 mL of the internal standard solution. To this solution add 10 mL of diluted dilute hydrochloric acid (1 in 10), and MS: Amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of homatropine hydrobromide (1 in 4000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1.5 m in length, packed with 180 to 250 mm siliceous earth for gas chromatography coated in 1 to 3z with 50z phenyl-methyl silicone polymer for gas chromatography. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen or helium. Flow rate: Adjust so that the retention time of scopola-mine is about 8 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the internal standard and scopolamine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 5 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of scopolamine to that of the internal standard is not more than 2.0z.
Opium Alkaloids Hydrochlorides Injection
アヘンアルカロイド塩酸塩注射液
Opium Alkaloids Hydrochlorides Injection is an aqueous injection. It contains not less than 0.
Opium Alkaloids Hydrochlorides Injection is a clear, colorless or light brown liquid. It is affected by light. pH: 2.5 – 3.5
To 1 mL of Opium Alkaloids Hydrochlo-rides Injection add 1 mL of ethanol (99.5), mix, and use this solution as the sample solution, and proceed as directed in the Identification (1) under Opium Alkaloids Hydrochlo-rides. Extractable volume <6.05> It meets the requirement.
Pipet 2 mL of Opium Alkaloids Hydrochlorides In-jection, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of morphine hydrochloride hydrate for assay, and dissolve in exactly 10 mL of the internal standard solution, add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine (C17H19NO3) ＝ MS × QT/QS × 0.887 ‌MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrine hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 1.0z.
Opium Ipecac Powder contains not less than 0.
Opium Ipecac Powder occurs as a light brown powder.
(1) Proceed with 1 g of Opium Ipecac Pow-der as directed in the Identification (1) under Powdered Opium. Proceed with 1 g of Opium Ipecac Powder as directed in the Identification (2) under Powdered Opium. Shake frequently a mixture of 3 g of Opium Ipecac Powder and 5 mL of hydrochloric acid, and allow to stand for 1 hour. Filter the solution into an evaporating dish. Add 5 mg of chlorinated lime to the filtrate: an orange color is produced at the circumference of the chlorinated lime (emetine).
Weigh accurately about 50 g of Opium Ipecac Pow-der in a glass stoppered flask, add 250 mL of dilute ethanol, warm in a water bath at 409C for 1 hour with stirring, and filter through a glass filter (G3). Transfer the residue on the filter to the first glass-stoppered flask, add 50 mL of dilute ethanol, warm in a water bath at 409C for 10 minutes with stirring, and filter through the glass filter. Repeat the ex-traction with three 50-mL portions of dilute ethanol. Com-bine all the filtrates in a mortar, evaporate on a water bath to dryness, add 10 mL of ethanol (99.5) to the residue, and evaporate again. After cooling, triturate the residue with an exactly measured 10 mL of water, add 2 g of calcium hy-droxide and an exactly measured 40 mL of water, stir the mixture for 20 minutes, and filter. To 30 mL of the filtrate add 0.1 g of magnesium sulfate heptahydrate, shake for 1 minute, then add 0.3 g of calcium hydroxide, shake for 1 minute, allow to stand for 1 hour, and filter. To an exactly measured 20 mL of the filtrate add 5 mL of sodium hydrox-ide TS, and adjust the pH to between 9.0 and 9.2 with ammonium chloride. Extract the solution successively with 60 mL, 40 mL and 30 mL of a mixture of chloroform and ethanol (95) (3:1). Combine all the extracts, distil, then evaporate the solvent on a water bath. Dissolve the residue in 20 mL of dilute sodium hydroxide TS and 10 mL of diethyl ether with shaking, add 0.5 g of ammonium chloride, shake vigorously with caution, and proceed as directed in the Assay under Powdered Opium. Each mL of 0.05 mol/L sulfuric acid VS ＝ 28.53 mg of C17H19NO3
Orange Peel Syrup is a brownish yellow to red-dish brown liquid. It has a characteristic odor, a sweet taste and a bitter aftertaste. Specific gravity d 20: about 1.25
To 25 mL of Orange Peel Syrup add 50 mL of ethyl acetate, shake for 5 minutes, allow to stand until clear ethyl acetate layer separate, and take the ethyl acetate layer, and evaporate on a water bath to dryness. Dissolve the residue in 10 mL of ethanol (95), filter if necessary, and use this solution as the sample solution. Separately, dissolve 10 mg of naringin for thin-layer chromatography in 10 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and water (8:2:1) to a dis-tance of about 10 cm, and air-dry the plate. Spray evenly dilute 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine TS on the plate, and allow to stand in ammonia gas: one of the several spots obtained from the sample solution has the same color tone and the same Rf value with the spot from the standard solution.
Oxytocin Injection is an aqueous injection. It contains not less than 90.
Oxytocin Injection is a colorless, clear liquid. pH <2.54> 2.5 – 4.5 Bacterial endotoxins <4.01> Less than 10 EU/oxytocin Unit. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to the Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a portion of Oxytocin Injection ac-cording to the labeled Units, dilute with the diluent so that each mL contains about 1 Unit, and use this solution as the sample solution. Separately, dissolve 1 bottle of Oxytocin RS in the mobile phase A to make exactly 20 mL. Pipet a suita-ble volume of this solution, dilute with the diluent to make a known concentration solution so that each mL contains about 1 Unit, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of oxytocin in each so-lution. Units per mL of Oxytocin Injection ＝ MS × AT/AS × b/a MS: Units per mL of the standard solution a: Volume (mL) of Oxytocin Injection taken b: Total volume of the sample solution prepared by dilut-ing with the diluent Diluent: Dissolve 5 g of chlorobutanol, 1.1 g of sodium acetate trihydrate, 5 g of acetic acid (100) and 6 mL of ethanol (99.5) in water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. ‌Mobile phase A: Dissolve 15.6 g of sodium dihydrogen phosphate dihydrate in 1000 mL of water. Mobile phase B: A mixture of water and acetonitrile (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 30 70 → 40 30 → 60 30 – 30.1 40 → 70 60 → 30 30.1 – 45 70 30 Flow rate: 1.0 mL per minute. System suitability— System performance: Dissolve an adequate amount of oxytocin and vasopressin in the mobile phase A, so that each mL contains about 0.02 mg each of them. When the proce-dure is run with 100 mL of this solution under the above op-erating conditions, vasopressin and oxytocin are eluted in this order with the resolution between these peaks being not less than 14, and the symmetry factor of the peak of oxyto-cin is not more than 1.5. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of oxytocin is not more than 2.0z.
Containers—Hermetic containers. Storage—In a cold place, and avoid freezing.
Ozagrel Sodium for Injection is a preparation for injection, which is dissolved before use. It contains not less than 95.
Ozagrel Sodium for Injection occurs as white, masses or powder.
Dissolve an amount of Ozagrel Sodium for Injection, equivalent to 40 mg of Ozagrel Sodium, in water to make 40 mL. To 1 mL of this solution add water to make 200 mL, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 269 nm and 273 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—Dissolve an amount of Ozagrel Sodium for Injection, equivalent to 0.20 g of Ozagrel So-dium, in the mobile phase to make 100 mL. To 5 mL of this solution add the mobile phase to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Purity (4) under Ozagrel Sodium. Bacterial endotoxins <4.01> Less than 3.7 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take a number of Ozagrel Sodium for Injection, equivalent to about 0.4 g of ozagrel sodium (C13H11N2NaO2), and dissolve all the contents in water to make exactly 200 mL. Pipet 5 mL of this solution, add ex-actly 10 mL of the internal standard solution and 5 mL of water, mix, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Ozagrel Sodium RS, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solu-tion, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Then, proceed as directed in the Assay under Ozagrel Sodium. Pancreatin
Ozagrel Sodium Injection is an aqueous injection. It contains not less than 95.
Panipenem and Betamipron for Injection is a prepa-ration for injection which is dissolved before use. It contains not less than 90.
Panipenem and Betamipron for Injection oc-curs as two layers of upper and lower. The former occurs pale yellowish white to light yellow, masses or masses con-taining powder, and the latter white, masses or masses con-taining powder. It is deliquescent.
(1) Powder Panipenem and Betamipron for Injection, weigh a portion of the powder, equivalent to 40 mg (potency) of Panipenem, dissolve in 4 mL of water, add 1 mL of hydroxylammonium chloride-ethanol TS, allow Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 22 22 – 25 100 → 90 0 → 10 25 – 30 30 – 35 90 → 85 10 → 15 35 – 40 85 → 77 15 → 23 40 – 50 77 → 0 23 → 100 50 – 55 to stand for 3 minutes, add 1 mL of acidic ammonium iron Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of panipenem. System suitability— Test for required detectability: Use the diluted sample so-lution (1 in 100) as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add water to make exactly 10 mL. Confirm that the peak area of panipenem obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of panipenem are not less than 4000 and 0.8 to 1.2, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of panipenem is not more than 0.95z. Bacterial endotoxins <4.01> Less than 0.15 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of Content uniformity test. After preparation of the sample solution and standard so-lution, keep them at not exceeding 59C. Dissolve the content of 1 container of Panipenem and Betamipron for Injection in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 500 mL. Take exactly V mL of this solution, equivalent to 5 mg (potency) of Panipenem, add exactly 5 mL of the internal standard solu-tion, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make 20 mL, and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. buffer solution (pH 7.0) to make 20 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg (potency) of Panipenem RS and about 50 mg of betamipron for assay (separately determine the water <2.48> in the same manner as Betamipron), dissolve in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solu-tion (pH 7.0) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT1 and QT2, of the peak areas of panipenem and betamipron to that of the internal standard obtained from the sample solution, and the ratios, QS1, and QS2, of the peak areas of panipenem and be-tamipron to that of the internal standard obtained from the standard solution. Amount [mg (potency)] of panipenem (C15H21N3O4S) ＝ MS1 × QT1/QS1 × 25/V Amount (mg) of betamipron (C10H11NO3) ＝ MS2 × QT2/QS2 × 25/V MS1: Amount [mg (potency)] of Panipenem RS taken MS2: Amount (mg) of betamipron for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium p-styrenesulfonate in 0.02 mol/L 3-(N-morpholino) propanesulfonic acid buffer solution (pH 7.0) (1 in 10,000). Operating conditions— Column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay under Panipenem. Detector: An ultraviolet absorption photometer (wave- Amount [mg (potency)] of panipenem (C ＝ MS1 × QT1/QS1 × 25/V 15H21N3O4S) length: 260 nm). Flow rate: Adjust so that the retention time of panipenem is about 9 minutes. ‌Amount (mg) of betamipron (C10H11NO3) ＝ MS2 × QT2/QS2 × 25/V MS1: Amount [mg (potency)] of Panipenem RS taken MS2: Amount (mg) of betamipron for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium p-styrenesulfonate in 0.02 mol/L 3-(N-morpholino) propanesulfonic acid buffer solution (pH 7.0) (1 in 10,000). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Clarity and color of solution—A solution of an amount of Panipenem and Betamipron for Injection, equivalent to 0.5 g (potency) of Panipenem, in 10 mL of water is clear, and has no more color than Matching Fluid J. (2) Related substances—After preparation of the sample solution, keep it at not exceeding 59C and use within 60 minutes. Take 1 container of Panipenem and Betamipron for Injection, dissolve in water so that each mL contains 1 mg (potency) of panipenem, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method, and calcu-late the amount of them by the area percentage method: the amount of each peak other than panipenem and betamipron is not more than 8.0z, and the total amount of peaks other than panipenem and betamipron is not more than 13.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter, 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of 0.02 mol/L phosphate buffer (pH 8.0) and acetonitrile (100:1). Mobile phase B: A mixture of 0.02 mol/L phosphate buffer (pH 8.0) and acetonitrile (3:1). Flowing of mobile phase: Control the gradient by mixing the mobile phase A and B as directed in the following table. Description Panipenem and Betamipron for Injection oc-curs as two layers of upper and lower. The former occurs pale yellowish white to light yellow, masses or masses con-taining powder, and the latter white, masses or masses con-taining powder. It is deliquescent. Identification (1) Powder Panipenem and Betamipron for Injection, weigh a portion of the powder, equivalent to 40 mg (potency) of Panipenem, dissolve in 4 mL of water, add 1 mL of hydroxylammonium chloride-ethanol TS, allow Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 22 22 – 25 100 → 90 0 → 10 25 – 30 30 – 35 90 → 85 10 → 15 35 – 40 85 → 77 15 → 23 40 – 50 77 → 0 23 → 100 50 – 55 to stand for 3 minutes, add 1 mL of acidic ammonium iron Flow rate: 1.0 mL per minute. Time span of measurement: About 3 times as long as the retention time of panipenem. System suitability— Test for required detectability: Use the diluted sample so-lution (1 in 100) as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, add water to make exactly 10 mL. Confirm that the peak area of panipenem obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of panipenem are not less than 4000 and 0.8 to 1.2, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of panipenem is not more than 0.95z. Bacterial endotoxins <4.01> Less than 0.15 EU/mg (po-tency). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of Content uniformity test. After preparation of the sample solution and standard so-lution, keep them at not exceeding 59C. Dissolve the content of 1 container of Panipenem and Betamipron for Injection in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 500 mL. Take exactly V mL of this solution, equivalent to 5 mg (potency) of Panipenem, add exactly 5 mL of the internal standard solu-tion, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make 20 mL, and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. buffer solution (pH 7.0) to make 20 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg (potency) of Panipenem RS and about 50 mg of betamipron for assay (separately determine the water <2.48> in the same manner as Betamipron), dissolve in 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid buffer solu-tion (pH 7.0) to make 20 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT1 and QT2, of the peak areas of panipenem and betamipron to that of the internal standard obtained from the sample solution, and the ratios, QS1, and QS2, of the peak areas of panipenem and be-tamipron to that of the internal standard obtained from the standard solution. Amount [mg (potency)] of panipenem (C15H21N3O4S) ＝ MS1 × QT1/QS1 × 25/V Amount (mg) of betamipron (C10H11NO3) ＝ MS2 × QT2/QS2 × 25/V MS1: Amount [mg (potency)] of Panipenem RS taken MS2: Amount (mg) of betamipron for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium p-styrenesulfonate in 0.02 mol/L 3-(N-morpholino) propanesulfonic acid buffer solution (pH 7.0) (1 in 10,000). Operating conditions— Column, column temperature, and mobile phase: Proceed as directed in the operating conditions in the Assay under Panipenem. Detector: An ultraviolet absorption photometer (wave- Amount [mg (potency)] of panipenem (C ＝ MS1 × QT1/QS1 × 25/V 15H21N3O4S) length: 260 nm). Flow rate: Adjust so that the retention time of panipenem is about 9 minutes. ‌Amount (mg) of betamipron (C10H11NO3) ＝ MS2 × QT2/QS2 × 25/V MS1: Amount [mg (potency)] of Panipenem RS taken MS2: Amount (mg) of betamipron for assay taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium p-styrenesulfonate in 0.02 mol/L 3-(N-morpholino) propanesulfonic acid buffer solution (pH 7.0) (1 in 10,000). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
After preparation of the sample solution and stand-ard solution, keep them at not exceeding 59C. Dissolve the total amount of the contents of 10 containers of Panipenem and Betamipron for Injection in 0.02 mol/L 3-(N-morpho-lino)propanesulfonic acid buffer solution (pH 7.0) to make exactly 500 mL. Take exactly V mL of this solution, equiva-lent to about 50 mg (potency) of Panipenem, add 0.02 mol/L 3-(N-morpholino)-propanesulfonic acid buffer solu-tion (pH 7.0) to make exactly 50 mL. Pipet 5 mL of this so-lution, add exactly 5 mL of the internal standard solution, add 0.02 mol/L 3-(N-morpholino)propanesulfonic acid System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, betamipron, panipenem and the internal standard are eluted in this order, and the resolutions between the peaks of betamipron and panipenem, and panipenem and the internal standard are not less than 3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of betamipron and panipenem to that of the internal standard are not more than 1.0z, respectively.
Papaverine Hydrochloride Injection is an aqueous injection. It contains not less than 95.
Papaverine Hydrochloride Injection is a clear, colorless liquid. pH: 3.0 – 5.0
(1) To 1 mL of Papaverine Hydrochloride Injection add 3 drops of sodium acetate TS: a white precipi-tate is produced. (2) Dilute a volume of Papaverine Hydrochloride Injec-tion, equivalent to 0.1 g of Papaverine Hydrochloride, with
Paroxetine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of paroxetine (C19H20FNO3: 329.37). Method of preparation Prepare as directed under Tablets, with Paroxetine Hydrochloride Hydrate. Identification Powder Paroxetine Hydrochloride Tablets. To a portion of the powder, equivalent to 10 mg of paroxe-tine (C19H20FNO3), add 140 mL of ethanol (99.5), sonicate for 5 minutes, add ethanol (99.5) to make 200 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 233 nm and 237 nm, between 263 nm and 267 nm, between 269 nm and 273 nm, and between 293 nm and 297 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Paroxetine Hydrochloride Tablets add V/5 mL of 0.1 mol/L hydrochloric acid TS, disintegrate by sonicating for 10 minutes, add 3V/5 mL of a mixture of water and 2-propanol (1:1), and sonicate for 20 minutes. To this solution add a mixture of water and 2-propanol (1:1) to make exactly V mL so that each mL contains about 0.
Powder Paroxetine Hydrochloride Tablets. To a portion of the powder, equivalent to 10 mg of paroxe-tine (C19H20FNO3), add 140 mL of ethanol (99.5), sonicate for 5 minutes, add ethanol (99.5) to make 200 mL, and filter. Determine the absorption spectrum of the filtrate as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 233 nm and 237 nm, between 263 nm and 267 nm, between 269 nm and 273 nm, and between 293 nm and 297 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Paroxetine Hydrochloride Tablets add V/5 mL of 0.1 mol/L hydrochloric acid TS, disintegrate by sonicating for 10 minutes, add 3V/5 mL of a mixture of water and 2-propanol (1:1), and sonicate for 20 minutes. To this solution add a mixture of water and 2-propanol (1:1) to make exactly V mL so that each mL contains about 0.2 mg of paroxetine (C19H20FNO3), filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the fil-trate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of paroxetine (C19H20FNO3) ＝ MS × AT/AS × V/100 × 0.900 MS: Amount (mg) of Paroxetine Hydrochloride RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu- System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of paroxetine are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 2.0z.
Weigh accurately the mass of not less than 20 Parox-etine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of paroxetine (C19H20FNO3), add 20 mL of 0.1 mol/L hydro-chloric acid TS, sonicate for 10 minutes. To this solution add 60 mL of a mixture of water and 2-propanol (1:1), and soni-cate for 20 minutes. Then add a mixture of water and 2-propanol (1:1) to make exactly 100 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 23 mg of Paroxetine Hydrochloride RS (separately determine the water <2.48> in the same manner as Paroxetine Hydrochloride Hydrate), and dissolve in 20 mL of 0.1 mol/L hydrochloric acid TS, add a mixture of water and 2-propanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 25 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of paroxetine in each solution. tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of 5-mg and 10-mg Amount (mg) of paroxetine (C ＝ MS × AT/AS × 0.900 19H20FNO3) tablet is not less than 80z, and of 20-mg tablet is not less than 75z. Start the test with 1 tablet of Paroxetine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of paroxetine (C19H20FNO3), and use this solution as the sample solution. Separately, weigh accurately about 11 mg of Paroxetine Hydrochloride RS (separately determine the water <2.48> in the same manner as Paroxetine Hydrochlo-ride Hydrate), and dissolve in the dissolution medium to make exactly 100 mL. Pipet 3 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of paroxetine in each solution. Dissolution rate (z) with respect to the labeled amount of paroxetine (C19H20FNO3) ＝ MS × AT/AS × V?/V × 1/C × 54 × 0.900 MS: Amount (mg) of Paroxetine Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of paroxetine (C19H20FNO3) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. MS: Amount (mg) of Paroxetine Hydrochloride RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with trimethylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 1000 mL of water, and adjust to pH 4.5 with acetic acid (100). To 600 mL of this solution, add 400 mL of acetonitrile and 10 mL of triethylamine, then adjust to pH 5.5 with acetic acid (100). Flow rate: Adjust so that the retention time of paroxetine is about 9 minutes. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of paroxetine are not less than 5000 and not more than 3.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of paroxetine is not more than 1.0z.
パラアミノサリチル酸カルシウム顆粒
Powder Calcium Paraaminosalicylate Gran-ules, weigh a portion of the powder, equivalent to 50 mg of Calcium Paraaminosalicylate Hydrate, add 100 mL of water, shake, and filter. To 10 mL of the filtrate add 1 mL of 1 mol/L hydrochloric acid TS, shake, and add 1 drop of iron (III) chloride TS: a red-purple color develops. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Calcium Paraaminosalicylate Granules is not less than 75z. Start the test with an accurately weighed amount of Cal-cium Paraaminosalicylate Granules, equivalent to about 0.25 g of calcium paraaminosalicylate hydrate (C7H5CaNO3. 31/2H2O), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of calcium paraaminosalicy-late hydrate for assay (separately determine the water <2.48> in the same manner as Calcium Paraaminosalicylate Hy-drate), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, at 300 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of calcium paraaminosalicylate hydrate (C7H5CaNO3.31/2H2O) ＝ MS/MT × AT/AS × 1/C × 900 × 1.330 MS: Amount (mg) of calcium paraaminosalicylate hydrate for assay taken, calculated on the anhydrous basis MT: Amount (g) of Calcium Paraaminosalicylate Gran-ules taken C: Labeled amount (mg) of calcium paraaminosalicylate hydrate (C7H5CaNO3.31/2H2O) in 1 g
Powder Calcium Paraaminosalicylate Granules, weigh accurately a portion of the powder, equivalent to about 0.2 g of calcium paraaminosalicylate hydrate (C7H5CaNO3.31/2H2O), add 60 mL of water and 0.75 mL of dilute hydrochloric acid, and dissolve by heating on a water bath. After cooling, add water to make exactly 100 mL, and filter. Pipet 30 mL of the filtrate, transfer to an iodine flask, and proceed as directed in the Assay under Calcium Para-aminosalicylate Hydrate. Each mL of 0.05 mol/L bromine VS ＝ 4.238 mg of C7H5CaNO3.31/2H2O
パズフロキサシンメシル酸塩注射液
Pazufloxacin Mesilate Injection is an aqueous injec-tion. It contains not less than 95.
Pazufloxacin Mesilate Injection is a clear, col-orless liquid.
To a volume of Pazufloxacin Mesilate Injec-tion, equivalent to 20 mg of Pazufloxacin Mesilate, add a mixture of methanol and 1 mol/L hydrochloric acid TS (49:1) to make 100 mL. To 5 mL of this solution add a mix-ture of methanol and 1 mol/L hydrochloric acid TS (49:1) to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 237 nm and 241 nm, between 314 nm and 324 nm, between 328 nm and 332 nm, and between 343 nm and 347 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Pazufloxacin Mesilate Injection, equivalent to about 12 mg of pazufloxacin mesilate (C16H15FN2O4.CH4O3S), and add water to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 23 mg of Pazufloxacin Mesilate RS, previously dried at 1059C for 3 hours, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solu-tion, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pazufloxacin to that of the internal standard. Amount (mg) of pazufloxacin mesilate (C16H15FN2O4.CH4O3S) ＝ MS × QT/QS × 1/2 MS: Amount (mg) of Pazufloxacin Mesilate RS taken Internal standard solution—A solution of acetanilide in the mobile phase (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Pazufloxacin Mesilate. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pazufloxacin and acetanilide are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pazufloxacin to that of the internal standard is not more than 1.0z.
シロップ用ペミロラストカリウム
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 255 nm and 259 nm and between 355 nm and 359 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Pemirolast Potassium for Syrup in single-dose packages meet the requirement of the Content uniformity test. Dissolve the total amount of the content of 1 package of Pemirolast Potassium for Syrup in water to make exactly V mL so that each mL contains about 50 mg of pemirolast potassium (C10H7KN6O). Pipet 10 mL of this solution, add water to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Pemirolast Potassium Tablets
Pemirolast Potassium Ophthalmic Solution
Pemirolast Potassium Ophthalmic Solution is an aqueous ophthalmic preparation. It contains not less than 95.
Pemirolast Potassium Ophthalmic Solution is a clear, colorless liquid.
To a volume of Pemirolast Potassium Oph-thalmic Solution, equivalent to 1 mg of Pemirolast Potassi-um, add diluted 0.1 mol/L phosphate buffer solution for an-tibiotics (pH 8.0) (1 in 10) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 255 nm and 259 nm, and between 355 nm and 359 nm. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—To a volume of Pemirolast Potassium Ophthalmic Solution, equivalent to 2 mg of Pemirolast Potassium, add 1 mL of methanol and diluted 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) (1 in 10) to make 5 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add 20 mL of methanol and diluted 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) (1 in 10) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than pemirolast obtained from the sample solution is not larger than 3/10 times the peak area of pemirolast from the standard solution, and the total area of the peaks other than pemirolast from the sample solution is not larger than the peak area of pemirolast from the stand-ard solution. Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of trifluoroacetic acid TS and methanol (4:1). Mobile phase B: A mixture of methanol and trifluoroacet-ic acid TS (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. ing conditions, the relative standard deviation of the peak area of pemirolast is not more than 2.0z. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet a volume of Pemirolast Potassium Ophthalmic Solution, equivalent to 2 mg of pemirolast potassium (C10H7KN6O), add exactly 2 mL of the internal standard so-lution, then add a mixture of diluted 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) (1 in 10) and metha-nol (3:2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Pemirolast Potassium RS (separately determine the water <2.48> in the same manner as Pemirolast Potassium), and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, then add a mixture of diluted 0.1 mol/L phosphate buffer solution for antibiotics (pH 8.0) (1 in 10) and methanol (3:2) to make 20 mL, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Calcu-late the ratios, QT and QS of the peak area of pemirolast to that of the internal standard. Amount (mg) of pemirolast potassium (C10H7KN6O) ＝ MS × QT/QS × 1/25 MS: Amount (mg) of Pemirolast Potassium RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 1000). Operating conditions— Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Detector: An ultraviolet spectrophotometer (wavelength: 260 nm). Column: A stainless steel column 3.9 mm in inside diame- 0 – 60 100 → 0 0 → 100 Flow rate: Adjust so that the retention time of pemirolast is about 19 minutes. Time span of measurement: About 3 times as long as the retention time of pemirolast, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add diluted 0.1 mol/L phosphate buffer solu-tion for antibiotics (pH 8.0) (1 in 10) to make exactly 20 mL. Confirm that the peak area of pemirolast obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 10 mg of pemirolast potas-sium in 10 mL of diluted 0.1 mol/L phosphate buffer solu-tion for antibiotics (pH 8.0) (1 in 10), transfer this solution to a colorless test tube, and illuminate with a D65 fluorescent lamp (3000 lx) for 72 hours. To 2 mL of this solution add 1 mL of methanol and diluted 0.1 mol/L phosphate buffer so-lution for antibiotics (pH 8.0) (1 in 10) to make 5 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak, having the relative retention time about 0.9 to pemirolast, and the peak of pemirolast is not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, methanol and acetic acid (100) (30:20:1). Flow rate: Adjust so that the retention time of pemirolast is about 4 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pemirolast and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pemirolast to that of the internal standard is not more than 1.0z.
Pemirolast Potassium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pemirolast potassium (C10H7KN6O: 266.30). Method of preparation Prepare as directed under Tablets, with Pemirolast Potassium. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 255 nm and 259 nm, and between 355 nm and 359 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pemirolast Potassium Tablets add 50 mL of water for 5 mg of pemirolast potassium (C10H7KN6O), and shake to disintegrate the tablet completely. Then, add water to make exactly V mL so that each mL contains about 50 mg of pemirolast potassium (C10H7KN6O), and filter.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 255 nm and 259 nm, and between 355 nm and 359 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pemirolast Potassium Tablets add 50 mL of water for 5 mg of pemirolast potassium (C10H7KN6O), and shake to disintegrate the tablet completely. Then, add water to make exactly V mL so that each mL contains about 50 mg of pemirolast potassium (C10H7KN6O), and filter. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add 1 mL of diluted potassium hydroxide TS (1 in 100), add water to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pemirolast potassium (C ＝ MS × AT/AS × V/400 10H7KN6O) Amount (mg) of pemirolast potassium (C10H7KN6O) ＝ MS × AT/AS × V/400 MS: Amount (mg) of Pemirolast Potassium RS taken, cal-culated on the anhydrous basis
(1) Heavy metals <1.07>—Proceed with 2.0 g of Penbutolol Sulfate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Penbutolol Sulfate according to Method 4, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.8 g of Penbutolol Sulfate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot Amount (mg) of pemirolast potassium (C ＝ MS × AT/AS × 1/4 10H7KN6O) 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 2- MS: Amount (mg) of Pemirolast Potassium RS taken, cal-culated on the anhydrous basis
Powder Pemirolast Potassium for Syrup. Weigh ac-curately a portion of the powder, equivalent to about 5 mg of pemirolast potassium (C10H7KN6O), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Pemirolast Potassium RS (separately determine the water <2.48> in the same manner as Pemirolast Potassium), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, at 357 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Amount (mg) of pemirolast potassium (C10H7KN6O) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of Pemirolast Potassium RS taken, cal-culated on the anhydrous basis
注射用ベンジルペニシリンカリウム
Benzylpenicillin Potassium for Injection oc-curs as white, crystals or crystalline powder.
Proceed as directed in the Identification (2) under Benzylpenicillin Potassium. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> The pH of a solution prepared by dissolving an amount of Benzylpenicillin Potassium for Injection, equiva-lent to 1.0 × 105 Units of Benzylpenicillin Potassium, in 10 mL of water is 5.0 to 7.5.
Clarity and color of solution—A solution prepared by dissolving an amount of Benzylpenicillin Potassium for Injection, equivalent to 1.0 × 106 Units of Benzylpenicillin Potassium, in 10 mL of water is clear. Perform the test with this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: the absorbance at 400 nm is not more than 0.10. Loss on drying <2.41> Not more than 1.2z (3 g, in vacu-um, below 0.67 kPa, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 1.25 × 10－4 EU/ Unit. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 containers of Benzylpenicillin Potassium for Injec-tion. Weigh accurately an amount of a portion of the con-tents, equivalent to about 6 × 104 Units of Benzylpenicillin Potassium, dissolve in water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Benzylpenicillin Potassium RS, equiva-lent to about 6 × 104 Units, dissolve in water to make ex-actly 20 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine the peak areas, AT and AS, of benzylpenicillin in each solution. Amount (unit) of Benzylpenicillin Potassium (C16H17KN2O4S) ＝ MS × AT/AS MS: Amount (unit) of Benzylpenicillin Potassium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To a mixture of diammonium hydrogen phosphate solution (33 in 5000) and acetonitril (19:6), add phosphoric acid to adjust the pH of this solution to 8.0. Flow rate: Adjust so that the retention time of benzyl-penicillin is about 7.5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of benzylpenicillin are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of benzylpenicillin is not more than 1.0z.
ペントバルビタールカルシウム錠
Pentobarbital Calcium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pentobarbital calcium (C H CaN O : with 20 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak 490.61). 22 34 4 6 areas of pentobarbital is not more than 5z. Loss on drying <2.41> Not more than 7.0z (1 g, 1059C, 5 hours). Assay Weigh accurately about 20 mg of Pentobarbital Cal-cium, dissolve in 5 mL of water, add exactly 5 mL of the in-ternal standard solution and water to make 50 mL. To 5 mL of this solution add water to make 20 mL. To 2 mL of this solution add water to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 18 mg of Pentobarbital RS, previously dried at 1059C for 2 hours, dissolve in 10 mL of acetonitrile for liquid chroma-tography, add exactly 5 mL of the internal standard solu-tion, and add water to make 50 mL. To 5 mL of this solution add water to make 20 mL. To 2 mL of this solution add water to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample so- Method of preparation Prepare as directed under Tablets, with Pentobarbital Calcium. Identification To a quantity of powdered Pentobarbital Calcium Tablets, equivalent to 5.6 mg of Pentobarbital Cal-cium, add 60 mL of water, shake thoroughly, then add water to make 100 mL, and filter. To 6 mL of the filtrate add dilute sodium hydroxide TS to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Pentobarbital Calcium Tablets add exactly V/10 mL of the internal standard solution, add 60 mL of water, shake vigorously until the tablet is completely disin- ‌tegrated, then add water to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, to 2 mL of the subse- uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pentobarbital to that of the internal standard. quent filtrate add water to make V mL so that each mL con-tains about 10 mg of pentobarbital calcium (C22H34CaN4O6), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pentobarbital calcium (C in 1 tablet ＝ MS × QT/QS × V/25 × 1.084 22H34CaN4O6) Amount (mg) of pentobarbital calcium (C22H34CaN4O6) ＝ MS × QT/QS × V/50 × 1.084 MS: Amount (mg) of Pentobarbital Calcium RS taken Internal standard solution—Dissolve 0.5 g of isopropyl par-ahydroxybenzoate in 20 mL of acetonitrile for liquid chro-matography, and add water to make 200 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Pentobarbital Calcium Tablets is not less than 80z. Start the test with 1 tablet of Pentobarbital Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of pen-tobarbital calcium (C22H34CaN4O6).
To a quantity of powdered Pentobarbital Calcium Tablets, equivalent to 5.6 mg of Pentobarbital Cal-cium, add 60 mL of water, shake thoroughly, then add water to make 100 mL, and filter. To 6 mL of the filtrate add dilute sodium hydroxide TS to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Pentobarbital Calcium Tablets add exactly V/10 mL of the internal standard solution, add 60 mL of water, shake vigorously until the tablet is completely disin- ‌tegrated, then add water to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, to 2 mL of the subse- uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pentobarbital to that of the internal standard. quent filtrate add water to make V mL so that each mL con-tains about 10 mg of pentobarbital calcium (C22H34CaN4O6), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pentobarbital calcium (C in 1 tablet ＝ MS × QT/QS × V/25 × 1.084 22H34CaN4O6) Amount (mg) of pentobarbital calcium (C22H34CaN4O6) ＝ MS × QT/QS × V/50 × 1.084 MS: Amount (mg) of Pentobarbital Calcium RS taken Internal standard solution—Dissolve 0.5 g of isopropyl par-ahydroxybenzoate in 20 mL of acetonitrile for liquid chro-matography, and add water to make 200 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Pentobarbital Calcium Tablets is not less than 80z. Start the test with 1 tablet of Pentobarbital Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of pen-tobarbital calcium (C22H34CaN4O6). Pipet 3 mL of this solu-tion, add dilute sodium hydroxide TS to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 26 mg of Pentobarbital RS, previ-ously dried at 1059C for 2 hours, dissolve in 2 mL of ethanol (99.5), and add water to make exactly 100 mL. Pipet 4 mL of this solution, and add the dissolution medium to make ex-actly 20 mL. Pipet 3 mL of this solution, add dilute sodium hydroxide TS to make exactly 10 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 241 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by adding dilute sodium hydroxide TS to 3 mL of the dissolution medium to make 10 mL, as the blank. Dissolution rate (z) with respect to the labeled amount of pentobarbital calcium (C22H34CaN4O6) ＝ MS × AT/AS × V?/V × 1/C × 180 × 1.084 MS: Amount (mg) of Pentobarbital RS taken C: Labeled amount (mg) of pentobarbital calcium (C22H34CaN4O6) in 1 tablet
Weigh accurately about 20 mg of Pentobarbital Cal-cium, dissolve in 5 mL of water, add exactly 5 mL of the in-ternal standard solution and water to make 50 mL. To 5 mL of this solution add water to make 20 mL. To 2 mL of this solution add water to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 18 mg of Pentobarbital RS, previously dried at 1059C for 2 hours, dissolve in 10 mL of acetonitrile for liquid chroma-tography, add exactly 5 mL of the internal standard solu-tion, and add water to make 50 mL. To 5 mL of this solution add water to make 20 mL. To 2 mL of this solution add water to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample so- Method of preparation Prepare as directed under Tablets, with Pentobarbital Calcium. Identification To a quantity of powdered Pentobarbital Calcium Tablets, equivalent to 5.6 mg of Pentobarbital Cal-cium, add 60 mL of water, shake thoroughly, then add water to make 100 mL, and filter. To 6 mL of the filtrate add dilute sodium hydroxide TS to make 20 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Pentobarbital Calcium Tablets add exactly V/10 mL of the internal standard solution, add 60 mL of water, shake vigorously until the tablet is completely disin- ‌tegrated, then add water to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, to 2 mL of the subse- uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pentobarbital to that of the internal standard. quent filtrate add water to make V mL so that each mL con-tains about 10 mg of pentobarbital calcium (C22H34CaN4O6), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pentobarbital calcium (C in 1 tablet ＝ MS × QT/QS × V/25 × 1.084 22H34CaN4O6) Amount (mg) of pentobarbital calcium (C22H34CaN4O6) ＝ MS × QT/QS × V/50 × 1.084 MS: Amount (mg) of Pentobarbital Calcium RS taken Internal standard solution—Dissolve 0.5 g of isopropyl par-ahydroxybenzoate in 20 mL of acetonitrile for liquid chro-matography, and add water to make 200 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Pentobarbital Calcium Tablets is not less than 80z. Start the test with 1 tablet of Pentobarbital Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of pen-tobarbital calcium (C22H34CaN4O6). Pipet 3 mL of this solu-tion, add dilute sodium hydroxide TS to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 26 mg of Pentobarbital RS, previ-ously dried at 1059C for 2 hours, dissolve in 2 mL of ethanol (99.5), and add water to make exactly 100 mL. Pipet 4 mL of this solution, and add the dissolution medium to make ex-actly 20 mL. Pipet 3 mL of this solution, add dilute sodium hydroxide TS to make exactly 10 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 241 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by adding dilute sodium hydroxide TS to 3 mL of the dissolution medium to make 10 mL, as the blank. Dissolution rate (z) with respect to the labeled amount of pentobarbital calcium (C22H34CaN4O6) ＝ MS × AT/AS × V?/V × 1/C × 180 × 1.084 MS: Amount (mg) of Pentobarbital RS taken C: Labeled amount (mg) of pentobarbital calcium (C22H34CaN4O6) in 1 tablet Assay To 20 Pentobarbital Calcium Tablets add 120 mL of water, shake vigorously for 10 minutes, then add water to make exactly 200 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly V/10 mL of the internal standard solution, and add water to make V mL so that each mL contains about 0.5 mg of pentobarbital calcium (C H CaN O ). To 2 mL of this MS: Amount (mg) of Pentobarbital RS taken Internal standard solution—Dissolve 0.5 g of isopropyl par-ahydroxybenzoate in 20 mL of acetonitrile for liquid chro-matography, and add water to make 200 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 4.0 with diluted phosphoric acid (1 in 10). To 650 mL of this solution add 350 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of pentobar-bital is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, pentobarbital and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pentobarbital to that of the internal stand-ard is not more than 1.0z.
Peplomycin Sulfate for Injection is a preparation for injection which is dissolved before use. It contains not less than 90.
Peplomycin Sulfate for Injection occurs as white light masses or powder.
Take an amount of Peplomycin Sulfate for Injection, equivalent to 10 mg (potency) of Peplomycin Sul-fate, and dissolve in 15 mL of Copper (II) sulfate TS and water to make 2 mL. Apply this solution to the column (pre-pared by filling a 15 mm inside diameter and 15 cm long chromatography tube with 15 mL of strongly basic ion ex-change resin (Cl type) for column chromatography (75 – 150 mm in particle diameter) and run off. Then wash the column using water at 2.5 mL per minute, collect about 30 mL of the effluent. Add water to the effluent to make 250 mL, and de-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it ex-hibits maxima between 242 nm and 246 nm, and between 291 nm and 295 nm. Further determine the absorbances A1 and A2, at 243 nm and 293 nm, respectively: the ratio A1/A2 is 1.20 to 1.30. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> The pH of a solution prepared by dissolving an amount of Peplomycin Sulfate for Injection, equivalent to 50 mg (potency) of Peplomycin Sulfate, in 10 mL of water is 4.5 to 6.0.
Clarity and color of solution—A solution prepared by dissolving an amount of Peplomycin Sulfate for Injec-tion, equivalent to 10 mg (potency) of Peplomycin Sulfate, in 10 mL of water is clear and colorless. Loss on drying <2.41> Not more than 4.0z (60 mg, in vacuum, phosphorus (V) oxide, 609C, 3 hours). Perform the sampling preventing from moisture absorption. Bacterial endotoxins <4.01> Less than 1.5 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Mycobacterium smegmatis ATCC 607 Agar media for base layer, seed and transferring test organisms— Glycerin 10.0 g Peptone 10.0 g Meat extract 10.0 g Sodium chloride 3.0 g Agar 15.0 g Water 1000 mL Mix all the ingredients, and sterilize. Adjust to pH 6.9 to 7.1 with sodium hydroxide TS after sterilization. Liquid media for suspending the test organism Glycerin 10.0 g Peptone 10.0 g Meat extract 10.0 g Sodium chloride 3.0 g Water 1000 mL Mix all the components, and sterilize. Adjust to pH 6.9 to 7.1 with sodium hydroxide TS after sterilization. Preparation of seeded agar layer—Cultivate the test organism on the slant of the agar medium for transferring the test organism at 279C for 40 to 48 hours, then inoculate the test organism thus obtained in 100 mL of the liquid me-dia for suspending the test organism, cultivate with shaking at between 259C and 279C for 5 days, and use this as the sus-pension of test organism. Store the suspension of test organ-ism at a temperature not exceeding 59C, and use within 14 days. Add 0.5 mL of the suspension of test organism in 100 mL of the agar medium for seed previously kept at 489C, mix thoroughly, and use as the seeded agar layer. Preparation of cylinder-agar plate—Proceed as di-rected in 1.7. Preparation of cylinder-agar plates under the Microbial Assay for Antibiotics, dispensing 5.0 mL of agar medium for base layer and 8.0 mL of the agar medium for seed into the Petri dish. Standard solutions—Weigh accurately an amount of Peplomycin Sulfate RS, equivalent to about 20 mg (po-tency), dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below, and use within 15 days. Measure exactly a suitable amount of the standard stock solution before use, add 0.1 mol/L phosphate buffer solution (pH 6.8) to make solutions so that each mL contains 4 mg (potency) and 2 mg (potency), and use these solutions as the high concentration standard solution and low concentration standard solution, respectively. Sample solutions—Weigh accurately the mass of the contents of not less than 10 containers of Peplomycin Sul-fate for Injection. Weigh accurately an amount of the con-tents, equivalent to about 10 mg (potency) of Peplomycin Sulfate, dissolve in 0.1 mol/L phosphate buffer solution (pH 6.8) to make exactly 100 mL. Measure exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer so- ‌lution (pH 6.8) to make solutions so that each mL contains 4 mg (potency) and 2 mg (potency), and use these solutions as the high concentration sample solution and low concentra-tion sample solution, respectively.‌
Perphenazine Maleate Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of perphenazine maleate (C21H26ClN3OS. 2C4H4O4: 636.11). Method of preparation Prepare as directed under Tablets, with Perphenazine Maleate. Identification (1) Shake a quantity of powdered Per-phenazine Maleate Tablets, equivalent to 0.04 g of Per-phenazine Maleate, with 3 mL of dilute hydrochloric acid and 30 mL of water, centrifuge. Filter the supernatant liq-uid, add 3 mL of ammonia solution (28) to the filtrate, and extract with three 10-mL portions of chloroform. [Reserve the aqueous layer, and use for test (4).] Wash the combined chloroform extracts with two 5-mL portions of water, and separate the chloroform layer. Evaporate 6 mL of the chlo-roform solution on a water bath to dryness. Proceed with the residue as directed in the Identification (1) under Perphena-zine Maleate. Evaporate 20 mL of the chloroform solution ob-tained in (1) on a water bath to dryness, dissolve the residue in 20 mL of methanol, and filter, if necessary. Warm the fil-trate, add 5 mL of a warm solution of 2,4,6-trinitrophenol in methanol (1 in 25), allow to stand for 4 hours, and proceed as directed in the Identification (2) under Perphenazine sample solution and standard solution at 255 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Amount (mg) of perphenazine maleate (C21H26ClN3OS.2C4H4O4) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of perphenazine maleate for assay taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Perphenazine Maleate Tablets is not less than 70z. Conduct this procedure without exposure to light. Start the test with 1 tablet of Perphenazine Maleate Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 3.
(1) Shake a quantity of powdered Per-phenazine Maleate Tablets, equivalent to 0.04 g of Per-phenazine Maleate, with 3 mL of dilute hydrochloric acid and 30 mL of water, centrifuge. Filter the supernatant liq-uid, add 3 mL of ammonia solution (28) to the filtrate, and extract with three 10-mL portions of chloroform. [Reserve the aqueous layer, and use for test (4).] Wash the combined chloroform extracts with two 5-mL portions of water, and separate the chloroform layer. Evaporate 6 mL of the chlo-roform solution on a water bath to dryness. Proceed with the residue as directed in the Identification (1) under Perphena-zine Maleate. Evaporate 20 mL of the chloroform solution ob-tained in (1) on a water bath to dryness, dissolve the residue in 20 mL of methanol, and filter, if necessary. Warm the fil-trate, add 5 mL of a warm solution of 2,4,6-trinitrophenol in methanol (1 in 25), allow to stand for 4 hours, and proceed as directed in the Identification (2) under Perphenazine sample solution and standard solution at 255 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Amount (mg) of perphenazine maleate (C21H26ClN3OS.2C4H4O4) ＝ MS × AT/AS × V?/V × 1/50 MS: Amount (mg) of perphenazine maleate for assay taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Perphenazine Maleate Tablets is not less than 70z. Conduct this procedure without exposure to light. Start the test with 1 tablet of Perphenazine Maleate Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 3.5 mg of perphenazine male-ate (C21H26ClN3OS.2C4H4O4), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of perphenazine maleate for assay, previously dried at 1059C for 3 hours, dissolve in 10 mL of 0.1 mol/L hydrochloric acid TS, and add the dissolution medium to make exactly 200 mL. Pipet 5 mL of this solution, add the dissolution me-dium to make exactly 200 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 255 nm of the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of perphenazine maleate (C H ClN OS.2C H O ) Maleate. 21 26 3 4 4 To 2 mL of the filtrate obtained in the Assay add water to make 50 mL. Determine the absorption spectrum of the solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits maxima between 253 nm and 257 nm and between 303 nm and 313 nm. Filter, if necessary, the aqueous layer reserved in (1), evaporate the filtrate to make about 5 mL, add 2 mL of di-lute sulfuric acid, and extract with two 10-mL portions of diethyl ether. Combine the diethyl ether extracts, evaporate ＝ MS × AT/AS × V?/V × 1/C × 45/4 MS: Amount (mg) of perphenazine maleate for assay taken C: Labeled amount (mg) of perphenazine maleate (C21H26ClN3OS.2C4H4O4) in 1 tablet
Weigh accurately and powder not less than 20 Per-phenazine Maleate Tablets. Weigh accurately a portion of the powder, equivalent to about 40 mg of perphenazine maleate (C H ClN OS.2C H O ), shake well with 15 mL of on a water bath to dryness, dissolve the residue in 5 mL of 21 26 3 4 4 4 sulfuric acid TS, and add 1 to 2 drops of potassium perman-ganate TS: the red color of potassium permanganate TS 1 mol/L hydrochloric acid TS and 50 mL of methanol, add water to make exactly 100 mL, and filter. Discard the first 20 ‌mL of the filtrate, measure exactly 5 mL of the subsequent filtrate, add water to make exactly 250 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 40 mg of perphenazine maleate for assay, previously dried at 1059C for 3 hours, dissolve in 15 mL of 1 mol/L hy-drochloric acid TS and 50 mL of methanol, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 250 mL, and use this solution as the stand-ard solution. Determine the absorbances, AT and AS, of the sample solution and the standard solution at 255 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank.‌ Amount (mg) of perphenazine maleate (C21H26ClN3OS.2C4H4O4) Identification (1) Determine the absorption spectrum of a solution of Pethidine Hydrochloride (1 in 2000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pethidine Hydrochloride, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Pethidine Hydrochloride (1 in 50) re-sponds to Qualitative Tests <1.09> (2) for chloride. ＝ MS × AT/AS MS: Amount (mg) of perphenazine maleate for assay taken
(1) Shake well a quantity of powdered Per-phenazine Tablets, equivalent to 25 mg of Perphenazine, with 10 mL of methanol, and filter. Evaporate 2 mL of the filtrate on a water bath to dryness. With the residue, proceed as directed in the Identification (1) under Perphenazine. Add 5 mL of the filtrate obtained in (1) to 10 mL of a warm solution of 2,4,6-trinitrophenol in methanol (1 in 25), and proceed as directed in the Identification (2) under Per-phenazine. Determine the absorption spectrum of the filtrate obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 309 nm and 313 nm. Add 30 mL of methanol to another 10 mL of the filtrate, and determine the absorption spectrum: it exhibits a maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Disintegrate 1 tablet of Perphenazine Tablets by shaking with 5 mL of water, shake well with 70 mL of methanol, and ‌add methanol to make exactly 100 mL. Centrifuge this solu-tion, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL of a solution containing about 4 mg of perphenazine (C21H26ClN3OS) in each ml, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Perphenazine RS, previously dried in vacu-um over phosphorus (V) oxide at 659C for 4 hours, dissolve in methanol to make exactly 250 mL. Pipet 5 mL of this so-lution, add methanol to make exactly 50 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 258 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>. Amount (mg) of perphenazine (C21H26ClN3OS) ＝ MS × AT/AS × V?/V × 1/25 MS: Amount (mg) of Perphenazine RS taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 90 minutes of Perphenazine Tablets is not less than 70z. Start the test with 1 tablet of Perphenazine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 10 mg of Perphenazine RS, previously dried in vacu-um with phosphorus (V) oxide at 659C for 4 hours, dissolve in 5 mL of 0.1 mol/L hydrochloric acid TS, and add the dis-solution medium to make exactly 250 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 255 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of perphenazine (C21H26ClN3OS) ＝ MS × AT/AS × 1/C × 18 MS: Amount (mg) of Perphenazine RS taken C: Labeled amount (mg) of perphenazine (C21H26ClN3OS) in 1 tablet
Weigh accurately and powder not less than 20 Perphenazine Tablets. Weigh accurately a portion of the powder, equivalent to about 4 mg of perphenazine (C21H26ClN3OS), add 70 mL of methanol, shake well, and add methanol to make exactly 100 mL. Filter the solution, and discard the first 20 mL of the filtrate. Pipet 5 mL of the subsequent filtrate, add methanol to make exactly 50 mL, and use this solution as the sample solution. Weigh accu-rately about 10 mg of Perphenazine RS, previously dried in vacuum over phosphorus (V) oxide at 659C for 4 hours, and dissolve in methanol to make exactly 250 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and the standard solution at 258 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of perphenazine (C21H26ClN3OS) ＝ MS × AT/AS × 2/5 MS: Amount (mg) of Perphenazine RS taken
Pethidine Hydrochloride Injection is an aqueous in-jection. It contains not less than 95.
Pethidine Hydrochloride Injection is a clear, colorless liquid. It is affected by light. pH 4.0 – 6.0
Take a volume of Pethidine Hydrochloride Injection equivalent to 0.1 g of Pethidine Hydrochloride, and add water to make 200 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 250 nm and 254 nm, between 255 nm and 259 nm, and between 261 nm and 265 nm. Bacterial endotoxins <4.01> Less than 6.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Pethidine Hydrochlo-ride Injection, equivalent to about 0.1 g of pethidine hydro-chloride (C15H21NO2.HCl), add exactly 10 mL of the internal standard solution, and add the mobile phase to make 50 mL. To 5 mL of this solution add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of pethidine hydrochloride for assay, previously dried at 1059C for 3 hours, add exactly 10 mL of the internal standard solution, and add the mobile phase to make 50 mL. To 5 mL of this solution add the mo-bile phase to make 20 mL, and use this solution as the stand-ard solution. Perform the test with 20 mL of the sample solu-tion and standard solution as directed under Liquid Chroma- order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of pethidine to that of the internal standard is not more than 1.0z.
Phellodendron, Albumin Tannate and Bis-muth Subnitrate Powder is brownish yellow in color, and has a bitter taste.
(1) Shake thoroughly 0.1 g of Phelloden- ‌JP XVIII Crude Drugs and Related Drugs / Powdered Picrasma Wood 2099‌ dron, Albumin Tannate and Bismuth Subnitrate Powder with 5 mL of methanol, filter, and use the filtrate as the sam-ple solution. Separately, dissolve 1 mg of Berberine Chloride RS or berberine chloride hydrate for thin-layer chromatogra-phy in 1 mL of methanol, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (7:2:1) to a distance of about 10 cm, air-dry the plate. Exa-mine under ultraviolet light (main wavelength: 365 nm): one spot of the several spots obtained from the sample solution and a spot with yellow to yellow-green fluorescence from the standard solution show the same color tone and the same Rf value (phellodendron bark). To 0.3 g of Phellodendron, Albumin Tannate and Bismuth Subnitrate Powder add 20 mL of ethanol (95), heat in a water bath for 3 minutes with shaking, cool, and filter. To 10 mL of the filtrate add 1 drop of iron (III) chloride TS: a blue-green color is produced. Allow to stand: a bluish black precipitate is produced (albumin tannate). To 0.3 g of Phellodendron, Albumin Tannate and Bismuth Subnitrate Powder add 10 mL of diluted pyridine (1 in 5), warm in a water bath for 3 minutes with shaking, cool, and filter. Add 1 mL of ninhydrin-ascorbic acid TS to the filtrate, and heat in a water bath: a blue color is produced (albumin tannate). To 0.5 g of Phellodendron, Albumin Tannate and Bismuth Subnitrate Powder add 5 mL of dilute hydrochloric acid and 10 mL of water, warm, shake thoroughly, and filter. The filtrate responds to the Qualitative Tests <1.09> for bismuth salt.
Weigh accurately about 0.7 g of Phellodendron, Al-bumin Tannate and Bismuth Subnitrate Powder, shake well with 10 mL of water and 20 mL of diluted nitric acid (1 in 3), add water to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet the subsequent 10 mL of the filtrate, and add water to make exactly 100 mL. Pipet 25 mL of this solution, add diluted nitric acid (1 in 100) to make ex-actly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.23 g of bismuth nitrate pentahydrate, add 20 mL of diluted nitric acid (1 in 3) and water to make exactly 100 mL. Pipet 10 mL of this solution, and add water to make exactly 100 mL. Pipet 25 mL of this solution, add diluted nitric acid (1 in 100) to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions. On the other hand, determine the absorbance A0 of the solution prepared in the same manner using 20 mL of diluted nitric acid (1 in 3) instead of the standard solution. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A bismuth hollow-cathode lamp. Wavelength: 223.1 nm. Amount (mg) of bismuth (Bi) ＝ M × (AT － A0)/(AS － A0) × 0.431 M: Amount (mg) of bismuth nitrate pentahydrate taken
Phenobarbital Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of phenobarbital (C12H12N2O3: 232.24). Method of preparation Prepare as directed under Tablets, with Phenobarbital. Identification To a quantity of powdered Phenobarbital Tablets, equivalent to 20 mg of Phenobarbital, add 20 mL of boric acid-potassium chloride-sodium hydroxide buffer solu-tion (pH 9.6), shake, and centrifuge. To 1 mL of the super-natant liquid add boric acid-potassium chloride-sodium hy-droxide buffer solution (pH 9.6) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed Dissolution rate (z) with respect to the labeled amount of phenobarbital (C12H12N2O3) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of phenobarbital for assay taken C: Labeled amount (mg) of phenobarbital (C12H12N2O3) in 1 tablet Assay Weigh accurately the mass of not less than 20 tablets of Phenobarbital Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 30 mg of phenobarbital (C12H12N2O3), add exactly 30 mL a mixture of water and acetonitrile (1:1), shake for 10 minutes, and cen-trifuge. Pipet 1 mL of the supernatant liquid, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of phenobarbital for assay, previ-ously dried at 1059C for 2 hours, and dissolve in exactly 30 mL of a mixture of water and acetonitrile (1:1). Pipet 1 mL of this solution, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con- under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- its a maximum between 238 nm and 242 nm. Uniformity of dosage units <6.02> Perform the Mass varia- ditions, and determine the peak areas, AT phenobarbital in each solution. Amount (mg) of phenobarbital (C H and AS, of N O ) tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ＝ MS × AT/AS 12 12 2 3 To 1 tablet of Phenobarbital Tablets add exactly V mL of a mixture of water and acetonitrile (1:1) so that each mL contains about 1 mg of phenobarbital (C12H12N2O3), sonicate to disintegrate, shake for 10 minutes, and centri-fuge.
To a quantity of powdered Phenobarbital Tablets, equivalent to 20 mg of Phenobarbital, add 20 mL of boric acid-potassium chloride-sodium hydroxide buffer solu-tion (pH 9.6), shake, and centrifuge. To 1 mL of the super-natant liquid add boric acid-potassium chloride-sodium hy-droxide buffer solution (pH 9.6) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed Dissolution rate (z) with respect to the labeled amount of phenobarbital (C12H12N2O3) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of phenobarbital for assay taken C: Labeled amount (mg) of phenobarbital (C12H12N2O3) in 1 tablet
Weigh accurately the mass of not less than 20 tablets of Phenobarbital Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 30 mg of phenobarbital (C12H12N2O3), add exactly 30 mL a mixture of water and acetonitrile (1:1), shake for 10 minutes, and cen-trifuge. Pipet 1 mL of the supernatant liquid, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of phenobarbital for assay, previ-ously dried at 1059C for 2 hours, and dissolve in exactly 30 mL of a mixture of water and acetonitrile (1:1). Pipet 1 mL of this solution, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq- uid Chromatography <2.01> according to the following con- under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- its a maximum between 238 nm and 242 nm. Uniformity of dosage units <6.02> Perform the Mass varia- ditions, and determine the peak areas, AT phenobarbital in each solution. Amount (mg) of phenobarbital (C H and AS, of N O ) tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ＝ MS × AT/AS 12 12 2 3 To 1 tablet of Phenobarbital Tablets add exactly V mL of a mixture of water and acetonitrile (1:1) so that each mL contains about 1 mg of phenobarbital (C12H12N2O3), sonicate to disintegrate, shake for 10 minutes, and centri-fuge. Pipet 1 mL of the supernatant liquid, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of phenobarbital (C12H12N2O3) ＝ MS × AT/AS × V/30 MS: Amount (mg) of phenobarbital for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Phenobarbital Tablets is not less than 75z. Start the test with 1 tablet of Phenobarbital Tablets, with-draw not less than 20 mL of the medium at the specified MS: Amount (mg) of phenobarbital for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of water and acetonitrile for liq-uid chromatography (11:9). Flow rate: Adjust so that the retention time of phenobar-bital is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of phenobarbital are not less than 3000 ‌and not more than 1.5, respectively.‌‌ System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of phenobarbital is not more than 1.0z.
フェノールスルホンフタレイン注射液
Phenolsulfonphthalein Injection is an aqueous in-jection. It contains not less than 0.
Phenolsulfonphthalein Injection is a clear, orange-yellow to red liquid.
To 1 mL of Phenolsulfonphthalein Injection add 2 to 3 drops of sodium hydroxide TS, and proceed as di-rected in the Identification (1) under Phenolsulfonphthalein. pH <2.54> 6.0 – 7.6 Bacterial endotoxins <4.01> Less than 7.5 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> Perform the test ac-cording to Method 2: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Sensitivity To 1.0 mL of Phenolsulfonphthalein Injection add 5 mL of water. To 0.20 mL of this solution add 50 mL of freshly boiled and cooled water and 0.40 mL of 0.01 mol/L sodium hydroxide VS: a deep red-purple color devel-ops, and it changes to light yellow on the addition of 0.40 mL of 0.005 mol/L sulfuric acid VS.
Pipet 5 mL of Phenolsulfonphthalein Injection, and Amount (mg) of phenolsulfonphthalein (C19H14O5S) ＝ MS × AT/AS MS: Amount (mg) of phenolsulfonphthalein for assay taken
Phenytoin Powder contains not less than 95.
Weigh a portion of Phenytoin Powder, equivalent to 0.3 g of Phenytoin, stir well with two 100-mL portions of diethyl ether, and extract. Combine the diethyl ether extracts, and filter. Evaporate the filtrate on a water bath to dryness, and proceed with the residue as directed in the Identification under Phenytoin. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately an amount of Phenytoin Powder, equivalent to about 50 mg of phenytoin (C15H12N2O2), add 30 mL of methanol, sonicate for 15 minutes with occasional shaking, shake for another 10 minutes, and add methanol to make exactly 50 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal ing conditions, the relative standard deviation of the ratio of the peak area of phenytoin to that of the internal standard is not more than 1.0z.
Phenytoin Sodium for Injection is a preparation for injection which is dissolved before use. When dried, it contains not less than 98.
Phenytoin Sodium for Injection occurs as white, crystals or crystalline powder. It is odorless. It is soluble in water and in ethanol (95), and practically insoluble in chloroform and in diethyl ether. The pH of a solution of 1.0 g of Phenytoin Sodium for In-jection in 20 mL of water is about 12. It is hygroscopic. A solution of Phenytoin Sodium for Injection absorbs car-bon dioxide gradually when exposed to air, and a crystalline precipitate of phenytoin is produced.
(1) With the residue obtained in the Assay, proceed as directed in the Identification under Phenytoin. (2) Ignite 0.5 g of Phenytoin Sodium for Injection, cool, and dissolve the residue in 10 mL of water: the solution changes red litmus paper to blue, and responds to Qualita-tive Tests <1.09> (1) for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Phenytoin Sodium for Injection in 20 mL of freshly boiled and cooled water in a glass-stoppered test tube: the solution is clear and colorless. If any turbidity is produced, add 4.0 mL of 0.1 mol/L sodium hydroxide VS: the solution becomes clear and colorless. (2) Heavy metals <1.07>—Proceed with 1.0 g of Phenytoin Sodium for Injection according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 2.5z (1 g, 1059C, 4 hours).
Weigh accurately the content of not less than 10 containers of Phenytoin Sodium for Injection, transfer about 0.3 g of the content, previously dried and accurately weighed, to a separator, dissolve in 50 mL of water, add 10 mL of dilute hydrochloric acid, and extract with 100 mL of diethyl ether, then with four 25-mL portions of diethyl ether. Combine the diethyl ether extracts, and evaporate on a water bath. Dry the residue at 1059C for 2 hours, and weigh it as ‌the mass of phenytoin (C15H12N2O2: 252.27). pare the control solution with 2.0 mL of Standard Lead So- Amount (mg) of phenytoin sodium (C15H11N2NaO2) ＝ amount (mg) of phenytoin (C15H12N2O2) × 1.087
Phenytoin Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of phenytoin (C15H12N2O2: 252.27). Method of preparation Prepare as directed under Tablets, with Phenytoin. Identification Weigh a portion of powdered Phenytoin Tablets, equivalent to about 0.3 g of Phenytoin, transfer to a separator, and add 1 mL of dilute hydrochloric acid and 10 mL of water. Extract with 100 mL of diethyl ether, then with four 25-mL potions of diethyl ether. Combine the extracts, evaporate the diethyl ether on a water bath, and dry the residue at 1059C for 2 hours. Proceed with the residue as di-rected in the Identification under Phenytoin. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ‌To 1 tablet of Phenytoin Tablets add 3V/5 mL of a mix-ture of water and acetonitrile (1:1), sonicate for 15 minutes with occasional shaking, shake for another 10 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly V mL so that each mL contains about 1 mg of phenytoin (C15H12N2O2).
Weigh a portion of powdered Phenytoin Tablets, equivalent to about 0.3 g of Phenytoin, transfer to a separator, and add 1 mL of dilute hydrochloric acid and 10 mL of water. Extract with 100 mL of diethyl ether, then with four 25-mL potions of diethyl ether. Combine the extracts, evaporate the diethyl ether on a water bath, and dry the residue at 1059C for 2 hours. Proceed with the residue as di-rected in the Identification under Phenytoin. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ‌To 1 tablet of Phenytoin Tablets add 3V/5 mL of a mix-ture of water and acetonitrile (1:1), sonicate for 15 minutes with occasional shaking, shake for another 10 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly V mL so that each mL contains about 1 mg of phenytoin (C15H12N2O2). Centrifuge this solution, pipet 5 mL of the su-pernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Pro-ceed as directed in the Assay. Amount (mg) of phenytoin (C15H12N2O2) ＝ MS × QT/QS × V/25 MS: Amount (mg) of phenytoin for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 25,000). Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Phenytoin Tablets, and powder in an agate mortar. Weigh accurately a portion of the powder, equivalent to about 50 mg of phenytoin (C15H12N2O2), add 30 mL of a mixture of water and acetonitrile (1:1), sonicate for 15 minutes with oc-casional shaking, shake for another 10 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly 50 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of phenytoin for assay, previ-ously dried at 1059C for 2 hours, and dissolve in a mixture of water and acetonitrile (1:1) to make exactly 25 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of phenytoin to that of the internal standard. Amount (mg) of phenytoin (C15H12N2O2) ＝ MS × QT/QS × 2 MS: Amount (mg) of phenytoin for assay taken Internal standard solution—A solution of propyl parahy-droxybenzoate in the mobile phase (1 in 25,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 258 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and 0.02 mol/L phosphate buffer solution (pH 3.5) (11:9). Flow rate: Adjust so that the retention time of phenytoin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, phenytoin and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of phenytoin to that of the internal standard is not more than 1.0z.
Pilocarpine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled ride in 10 mL of water is between 3.5 and 4.5. It is hygroscopic. It is affected by light. amount of pilocarpine hydrochloride (C HCl: 244.72). 11H16N2O2. Identification (1) Dissolve 0.1 g of Pilocarpine Hydro-chloride in 5 mL of water, add 1 drop of dilute nitric acid, 1 mL of hydrogen peroxide TS, 1 mL of chloroform and 1 drop of a potassium dichromate solution (1 in 300), and shake the mixture vigorously: a violet color develops in the chloroform layer while no color or a light yellow color is produced in the aqueous layer. (2) To 1 mL of a solution of Pilocarpine Hydrochloride (1 in 20) add 1 mL of dilute nitric acid and 2 to 3 drops of sil-ver nitrate TS: a white precipitate or opalescence is pro-duced. Melting point <2.60> 200 – 2039C Purity (1) Sulfate—Dissolve 0.5 g of Pilocarpine Hydro-chloride in 20 mL of water, and use this solution as the sam-ple solution. To 5.0 mL of the sample solution add 1 mL of dilute hydrochloric acid and 0.5 mL of barium chloride TS: no turbidity is produced. Nitrate—To 2.0 mL of the sample solution obtained in (1) add 2 mL of iron (II) sulfate TS, and superimpose the mixture upon 4 mL of sulfuric acid: no dark brown color de-velops at the zone of contact. Related substances—Dissolve 0.3 g of Pilocarpine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu- Method of preparation Prepare as directed under Tablets, with Pilocarpine Hydrochloride. Identification Perform the test with 10 mL each of the sam-ple solution and the standard solution, both obtained in the assay, as directed under Liquid Chromatography <2.01> ac-cording to the following conditions: the principal peaks in the chromatograms obtained from the sample solution and standard solution show the same retention time, and both spectra of these peaks in the chromatograms exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: Photodiode array detector (wavelength: 215 nm; spectrum range of measurement: 200 – 370 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add phosphate buffer solution (pH 4.0) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the two peaks, having the relative retention time of about 0.78 and about 0.92 to pilocarpine, obtained from the sample solution is not larger than the peak area of pilocarpine from the standard solu-tion, the area of the peak other than pilocarpine and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of pilocarpine from the standard solution, and the total area of the peaks other than pilocarpine from the sample solution is not larger than 2 times the peak area of pilocarpine from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.3 times as long as the retention time of pilocarpine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add phosphate buffer solution (pH 4.0) to make exactly 20 mL. Confirm that the peak area of pilocar-pine obtained with 10 mL of this solution is equivalent to 7 – 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilocarpine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilocarpine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pilocarpine Hydrochloride Tablets add a suitable amount of phosphate buffer solution (pH 4.0), shake until the tablet is completely disintegrated, then add phosphate buffer solution (pH 4.0) to make exactly V mL so that each mL contains about 0.
(1) Dissolve 0.1 g of Pilocarpine Hydro-chloride in 5 mL of water, add 1 drop of dilute nitric acid, 1 mL of hydrogen peroxide TS, 1 mL of chloroform and 1 drop of a potassium dichromate solution (1 in 300), and shake the mixture vigorously: a violet color develops in the chloroform layer while no color or a light yellow color is produced in the aqueous layer. (2) To 1 mL of a solution of Pilocarpine Hydrochloride (1 in 20) add 1 mL of dilute nitric acid and 2 to 3 drops of sil-ver nitrate TS: a white precipitate or opalescence is pro-duced.
(1) Sulfate—Dissolve 0.5 g of Pilocarpine Hydro-chloride in 20 mL of water, and use this solution as the sam-ple solution. To 5.0 mL of the sample solution add 1 mL of dilute hydrochloric acid and 0.5 mL of barium chloride TS: no turbidity is produced. Nitrate—To 2.0 mL of the sample solution obtained in (1) add 2 mL of iron (II) sulfate TS, and superimpose the mixture upon 4 mL of sulfuric acid: no dark brown color de-velops at the zone of contact. Related substances—Dissolve 0.3 g of Pilocarpine Hydrochloride in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solu- Method of preparation Prepare as directed under Tablets, with Pilocarpine Hydrochloride. Identification Perform the test with 10 mL each of the sam-ple solution and the standard solution, both obtained in the assay, as directed under Liquid Chromatography <2.01> ac-cording to the following conditions: the principal peaks in the chromatograms obtained from the sample solution and standard solution show the same retention time, and both spectra of these peaks in the chromatograms exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: Photodiode array detector (wavelength: 215 nm; spectrum range of measurement: 200 – 370 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. Purity Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Pipet 1 mL of the sample solution, add phosphate buffer solution (pH 4.0) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the two peaks, having the relative retention time of about 0.78 and about 0.92 to pilocarpine, obtained from the sample solution is not larger than the peak area of pilocarpine from the standard solu-tion, the area of the peak other than pilocarpine and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of pilocarpine from the standard solution, and the total area of the peaks other than pilocarpine from the sample solution is not larger than 2 times the peak area of pilocarpine from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.3 times as long as the retention time of pilocarpine, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add phosphate buffer solution (pH 4.0) to make exactly 20 mL. Confirm that the peak area of pilocar-pine obtained with 10 mL of this solution is equivalent to 7 – 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilocarpine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilocarpine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pilocarpine Hydrochloride Tablets add a suitable amount of phosphate buffer solution (pH 4.0), shake until the tablet is completely disintegrated, then add phosphate buffer solution (pH 4.0) to make exactly V mL so that each mL contains about 0.2 mg of pilocarpine hydro-chloride (C11H16N2O2.HCl), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of pilocarpine hydrochloride for assay, previously dried at 1059C for 2 hours, and dissolve in phosphate buffer solu-tion (pH 4.0) to make exactly 100 mL. Pipet 5 mL of this solution, add phosphate buffer solution (pH 4.0) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of pilocarpine in each solution. Amount (mg) of pilocarpine hydrochloride (C11H16N2O2.HCl) ＝ MS × AT/AS × V/200 MS: Amount (mg) of pilocarpine hydrochloride for assay taken Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilocarpine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilocarpine is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Pilocarpine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Pilocarpine Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 3 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of pilocarpine hydrochloride (C11H16N2O2.HCl), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of pilocarpine hydrochloride for assay, previ-ously dried at 1059C for 2 hours, and dissolve in the dissolu-tion medium to make exactly 100 mL. Pipet 2 mL of this so-lution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of pilocarpine in each solution. Dissolution rate (z) with respect to the labeled amount of pilocarpine hydrochloride (C11H16N2O2.HCl) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of pilocarpine hydrochloride for assay taken C: Labeled amount (mg) of pilocarpine hydrochloride (C11H16N2O2.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilocarpine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilocarpine is not more than 1.0z.
To 20 Pilocarpine Hydrochloride Tablets add a suit-able amount of phosphate buffer solution (pH 4.0), shake until the tablets are completely disintegrated, then add phos-phate buffer solution (pH 4.0) to make exactly V mL so that each mL contains about 0.4 mg of pilocarpine hydrochloride (C11H16N2O2.HCl), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 40 mg of pilo-carpine hydrochloride for assay, previously dried at 1059C for 2 hours, dissolve in phosphate buffer solution (pH 4.0) ‌to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of pilocar-pine in each solution. Amount (mg) of pilocarpine hydrochloride (C11H16N2O2.HCl) in 1 tablet ＝ MS × AT/AS × V/2000 MS: Amount (mg) of pilocarpine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 215 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with phenylated silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of 0.05 mol/L potassium dihy-drogen phosphate TS add phosphoric acid to adjust to pH 2.5. To this solution add 5.0 mL of triethylamine, and adjust to pH 2.5 with phosphoric acid. Flow rate: Adjust so that the retention time of pilocarpine is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilocarpine are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilocarpine is not more than 1.0z.
Pilsicainide Hydrochloride Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of pilsicainide hydrochloride hydrate (C17H24N2O.HCl.1/2 H2O: 317.85). Method of preparation Prepare as directed under Cap-sules, with Pilsicainide Hydrochloride Hydrate. Identification Take out the contents of Pilsicainide Hydro-chloride Capsules, to a quantity of the content, equivalent to 50 mg of Pilsicainide Hydrochloride Hydrate, add 10 mL of water, and shake well. Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 1 mL of the filtrate, add 1 mL of 1 mol/L hydrochloric acid TS and 8 mL of water. De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 261 nm and 265 nm, and between 268 nm and 272 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Pilsicainide Hydrochloride Capsules, add water, and shake to disperse the content of the capsule uni-formly while warming in a water bath. After cooling, add ex-actly V mL of the internal standard solution so that 0.2 mL of the internal standard solution is added for each mg of pil- sicainide hydrochloride hydrate (C H N O.HCl.1/ H O), the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of pilsicainide hydro-chloride hydrate (C17H24N2O.
Take out the contents of Pilsicainide Hydro-chloride Capsules, to a quantity of the content, equivalent to 50 mg of Pilsicainide Hydrochloride Hydrate, add 10 mL of water, and shake well. Centrifuge this solution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. To 1 mL of the filtrate, add 1 mL of 1 mol/L hydrochloric acid TS and 8 mL of water. De-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 261 nm and 265 nm, and between 268 nm and 272 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Pilsicainide Hydrochloride Capsules, add water, and shake to disperse the content of the capsule uni-formly while warming in a water bath. After cooling, add ex-actly V mL of the internal standard solution so that 0.2 mL of the internal standard solution is added for each mg of pil- sicainide hydrochloride hydrate (C H N O.HCl.1/ H O), the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 28 mg of pilsicainide hydro-chloride hydrate (C17H24N2O.HCl.1/2H2O), and use this solu-tion as the sample solution. Separately, weigh accurately about 28 mg of pilsicainide hydrochloride hydrate for assay, dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this so-lution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of pilsicainide in each solution. Dissolution rate (z) with respect to the labeled amount of pilsicainide hydrochloride hydrate (C17H24N2O.HCl.1/2 H2O) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of pilsicainide hydrochloride hydrate for assay taken C: Labeled amount (mg) of pilsicainide hydrochloride hy-drate (C17H24N2O.HCl.1/2 H2O) in 1 capsule Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pilsicainide are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pilsicainide is not more than 1.0z.
Take out the contents of not less than 20 Pilsicainide Hydrochloride Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of pilsicainide hydro-chloride hydrate (C17H24N2O.HCl.1/2 H2O), add 50 mL of water and shake well. After adding exactly 10 mL of the in- 17 24 2 2 2 ternal standard solution, add water to make 100 mL. To 5 then, add water so that each mL contains about 0.5 mg of pilsicainide hydrochloride hydrate (C17H24N2O.HCl.1/2H2O). To 5 mL of this solution, add water to make 50 mL, and filter. Discard the first 10 mL of the filtrate, and use the sub-sequent filtrate as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of pilsicainide hydrochloride hydrate (C17H24N2O.HCl.1/2H2O) ＝ MS × QT/QS × V/10 MS: Amount (mg) of pilsicainide hydrochloride hydrate for assay taken Internal Standard Solution—Dissolve 2.5 g of lidocaine for assay in 20 mL of 0.5 mol/L hydrochloric acid TS, and add water to make 1000 mL. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Pilsicainide Hydrochloride Capsules is not less than 85z. Start the test with 1 capsule of Pilsicainide Hydrochloride Capsules, withdraw not less than 20 mL of the medium at mL of this solution add water to make 50 mL, and filter the solution. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of pilsicainide hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard solution, and add water to make 100 mL. To 5 mL of this solution add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pilsicainide to that of the internal standard. Amount (mg) of pilsicainide hydrochloride hydrate (C17H24N2O.HCl.1/2H2O) ＝ MS × QT/QS MS: Amount (mg) of pilsicainide hydrochloride hydrate for assay taken Internal Standard Solution—Dissolve 2.5 g of lidocaine for assay in 20 mL of 0.5 mol/L hydrochloric acid TS, and add water to make 1000 mL. ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of around 409C. Mobile phase: To 750 mL of water add 5 mL of triethyla-mine, adjust the pH to 4.0 with phosphoric acid, and add water to make 1000 mL. To this solution, add 200 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of pilsicainide is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and pilsicainide are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pilsicainide to that of the internal standard is not more than 1.0z.
Pioglitazone Hydrochloride and Glimepiride Tablets
ピオグリタゾン塩酸塩･グリメピリド錠
(1) Powder Pioglitazone Hydrochloride and Glimepiride Tablets, weigh a portion of the powder, equivalent to 33 mg of Pioglitazone Hydrochloride, add 20 mL of 0.1 mol/L hydrochloric acid TS, and disintegrate completely by vigorous shaking for several minutes. Filter 2 mL of this solution through a membrane filter with a pore size not exceeding 0.45 mm. To 1 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 267 nm and 271 nm. (2) Wash the membrane ˆlter obtained in (1) with 100 mL of 0.1 mol/L hydrochloric acid TS, and extract with methanol so that each mL contains about 10 mg of glimepi-ride (C24H34N4O5S). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 227 nm and 231 nm.
Related substances—Powder Pioglitazone Hydro-chloride and Glimepiride Tablets, weigh a portion of the powder, equivalent to 10 mg of Glimepiride, add 30 mL of a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1), shake vigorously for 20 minutes, and add the mobile phase A to make 50 mL. Filter this solution through a mem-brane filter with a pore size not exceeding 0.2 mm, discard the first 4 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 40 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peak, having a relative retention time of about 0.23 to glimepiride obtained from the sample solution is not larger than 2.5 times the peak area of glimepiride from the stand-ard solution. The area of the peak other than glimepiride and other than the peak mentioned above from the sample solution is not larger than 1/2 times the peak area of glimepiride from the standard solution, and the total area of these peaks is not larger than the peak area of glimepiride from the standard solution. The total area of the peaks other than glimepiride from the sample solution is not larger than 3 times the peak area of glimepiride from the standard solu-tion. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: Dissolve 1.1 g of sodium dihydrogen phosphate dihydrate in water to make 1000 mL, and adjust to pH 1.6 with diluted phosphoric acid (1 in 10). To 650 mL of this solution add 600 mL of acetonitrile. Mobile phase B: Dissolve 1.1 g of sodium dihydrogen phosphate dihydrate in water to make 1000 mL, and adjust to pH 1.6 with diluted phosphoric acid (1 in 10). To 300 mL of this solution add 700 mL of acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 15 – 60 100 → 0 0 → 100 Flow rate: 1.0 mL per minute. Time span of measurement: For 60 minutes after injec-tion, beginning after the peak having a relative retention time of about 0.23 to glimepiride. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase A to make exactly 20 mL. Confirm that the peak area of glimepiride obtained with 40 mL of this solution is equivalent to 7 to 13z of that with 40 mL of the standard solution. System performance: When the procedure is run with 40 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 20,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 40 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Pioglitazone hydrochloride—To 1 tablet of Pioglita-zone Hydrochloride and Glimepiride Tablets add 30 mL of a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1), shake vigorously for 20 minutes, and add the mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL. Filter this solution through a mem-brane filter with a pore size not exceeding 0.2 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/10 mL of the internal standard solu-tion, add the mobile phase to make V? mL so that each mL contains about 66 mg of pioglitazone hydrochloride (C19H20N2O3S.HCl), and use this solution as the sample solu-tion. Then, proceed as directed in the Assay (1). Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × QT/QS × V?/V × 1/10 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl benzoate in the mobile phase (1 in 10,000). Glimepiride—To 1 tablet of Pioglitazone Hydrochlo-ride and Glimepiride Tablets add 30 mL of a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1), shake vigorously for 20 minutes, and add the mixture of aceto-nitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make ex-actly 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add ex-actly V?/10 mL of the internal standard solution, add the mobile phase to make V? mL so that each mL contains about 6 mg of glimepiride (C24H34N4O5S), and use this solution as the sample solution. Then, proceed as directed in the Assay (2). Amount (mg) of glimepiride (C24H34N4O5S) ＝ MS × QT/QS × V?/V × 1/100 MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl benzoate in the mobile phase (1 in 10,000). Dissolution <6.10> (1) Pioglitazone hydrochloride—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of a solution, which is prepared by mixing 50 mL of 0.2 mol/L hydrochlo-ric acid TS and 150 mL of potassium chloride solution (3 in 20), adding water to make 1000 mL and adjusting to pH 2.0 with 5 mol/L hydrochloric acid TS, as the dissolution medi-um, the dissolution rate in 45 minutes of Pioglitazone Hy-drochloride and Glimepiride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride and Glimepiride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the disso-lution medium to make exactly V? mL so that each mL con-tains about 18 mg of pioglitazone hydrochloride (C19H20N2O3S.HCl), and use this solution as the sample solu-tion. Separately, weigh accurately about 37 mg of Pioglita-zone Hydrochloride RS (separately determine the water <2.48> in the same manner as Pioglitazone Hydrochloride), dissolve in 20 mL of methanol, and add the dissolution me-dium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of pioglitazone in each solution. Dissolution rate (z) with respect to the labeled amount of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1) (Pioglitazone Hydrochloride). System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pioglitazone are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviation of the peak area of pioglitazone is not more than 1.0 z. (2) Glimepiride—When the test is performed at 50 revo-lutions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citrate buffer solu-tion (pH 7.5) as the dissolution medium, the dissolution rate in 30 minutes of Pioglitazone Hydrochloride and Glimepi-ride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride and Glimepiride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the disso-lution medium to make exactly V? mL so that each mL con-tains about 1.1 mg of glimepiride (C24H34N4O5S), and use this solution as the sample solution. Separately, weigh accurately about 55 mg of Glimepiride RS (separately determine the water <2.48> in the same manner as Glimepiride), dissolve in acetonitrile to make exactly 250 mL. Pipet 10 mL of this so-lution, and add acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of glimepiride in each solution. Dissolution rate (z) with respect to the labeled amount of glimepiride (C24H34N4O5S) ＝ MS × AT/AS × V?/V × 1/C × 9/5 MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of glimepiride (C24H34N4O5S) in 1 tablet Operating conditions— Detector, column, column temperature and mobile phase: Proceed as directed in the operating conditions in the Assay (Pioglitazone Hydrochloride). Flow rate: Adjust so that the retention time of glimepiride is about 5.4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of glimepiride are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviation of the peak area of glimepiride is not more than 2.0z.
(1) Pioglitazone hydrochloride—Weigh accurately the mass of not less than 20 Pioglitazone Hydrochloride and Glimepiride Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 33 mg of pioglita-zone hydrochloride (C19H20N2O3S.HCl), add 30 mL of a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1), shake vigorously for 20 minutes, and add a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL. Filter this solution through a mem-brane filter with a pore size not exceeding 0.2 mm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Pioglitazone Hydrochloride RS (separately deter-mine the water <2.48> in the same manner as Pioglitazone Hydrochloride), dissolve in a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pioglitazone to that of the internal standard. ‌Amount (mg) of pioglitazone hydrochloride area of glimepiride to that of the internal standard. (C19H20N2O3S.HCl) ＝ MS × QT/QS Amount (mg) of glimepiride (C ＝ MS × QT/QS × 1/10 24H34N4O5S) MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl benzoate in the mobile phase (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 228 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.80 g of sodium dihydrogen phos-phate dihydrate in water to make 1000 mL, and adjust to pH 4.0 with diluted phosphoric acid (1 in 10). To 500 mL of this solution add 500 mL of acetonitrile. Flow rate: Adjust so that the retention time of pioglita-zone is about 2.3 minutes. System suitability— System performance: To 33 mg of Pioglitazone Hydro-chloride RS add 5 mL of the glimepiride standard stock solu-tion obtained in (2), and add a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make 50 mL. To 5 mL of this solution add 5 mL of the internal standard solu-tion, and add the mobile phase to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, pioglitazone, the internal standard, and glimepiride are eluted in this order, and the resolutions between the peaks of pioglitazone and the internal standard and between the peaks of the internal standard and glimepi-ride are not less than 4 and not less than 3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of pioglitazone to that of the internal standard is not more than 1.0z. Glimepiride—Weigh accurately the mass of not less than 20 Pioglitazone Hydrochloride and Glimepiride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3 mg of glimepiride (C24H34N4O5S), add 30 mL of a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1), shake vigorously for 20 minutes, and add a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of the in-ternal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Glimepiride RS (separately determine the water <2.48> in the same manner as Glimepi-ride), dissolve in the mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL, and use this solution as the glimepiride standard stock solution. Pipet 10 mL of the glimepiride standard stock solution, and add the mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 100 mL. Pipet 5 mL of this so-lution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak MS: Amount (mg) of Glimepiride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl benzoate in the mobile phase (1 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: To 33 mg of Pioglitazone Hydro-chloride RS add 5 mL of the glimepiride standard stock solution, and add a mixture of acetonitrile and 0.1 mol/L hydrochloric acid TS (9:1) to make 50 mL. To 5 mL of this solution add 5 mL of the internal standard solution, and add the mobile phase to make 50 mL. When the procedure is run with 20 mL of this solution under the above operating condi-tions, pioglitazone, the internal standard and glimepiride are eluted in this order, and the resolutions between the peaks of pioglitazone and the internal standard and between the peaks of the internal standard and glimepiride are not less than 4 and not less than 3, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviation of the ratio of the peak area of glimepiride to that of the internal standard is not more than 1.0z.
Pioglitazone Hydrochloride and Metformin Hydrochloride Tablets
ピオグリタゾン塩酸塩･メトホルミン塩酸塩錠
(1) Shake vigorously a quantity of pow-dered Pioglitazone Hydrochloride and Metformin Hydro-chloride Tablets, equivalent to 0.33 mg of Pioglitazone Hydrochloride, with 10 mL of water, and filter through a membrane filter with a pore size not exceeding 0.45 mm. After washing the membrane filter with 10 mL of water, dissolve the retained substance on the filter by running through 10 mL of 0.1 mol/L hydrochloric acid TS, and determine the absorption spectrum of the filtrate so obtained as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 267 nm and 271 nm. (2) Shake vigorously a quantity of powdered Pioglita-zone Hydrochloride and Metformin Hydrochloride Tablets, equivalent to 20 mg of Metformin Hydrochloride, with 50 mL of water, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 1 mL of the filtrate add water to make 50 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 230 nm and 234 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Pioglitazone hydrochloride—To 1 tablet of Pioglita-zone Hydrochloride and Metformin Hydrochloride Tablets add 40 mL of 0.1 mol/L hydrochloric acid TS, shake vigor-ously for 10 minutes, add 40 mL of methanol, and shake. To this solution add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/20 mL of the internal standard solution, then add a mixture of 0.1 mol/L hydro-chloric acid TS and methanol (1:1) to make V? mL so that each mL contains about 16.5 mg of pioglitazone hydrochlo-ride (C19H20N2O3S.HCl), and use this solution as the sample solution. Then, proceed as directed in the Assay (1). Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × QT/QS × V?/V × 1/20 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) (1 in 2500). Metformin hydrochloride—To 1 tablet of Pioglita-zone Hydrochloride and Metformin Hydrochloride Tablets add 40 mL of 0.1 mol/L hydrochloric acid TS, shake vigor-ously for 10 minutes, add 40 mL of methanol, and shake. To this solution add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/20 mL of the internal standard solution, then add a mixture of 0.1 mol/L hydro-chloric acid TS and methanol (1:1) to make V? mL so that each mL contains about 0.25 mg of metformin hydrochlo-ride (C4H11N5.HCl), and use this solution as the sample solu-tion. Then, proceed as directed in the Assay (2). Amount (mg) of metformin hydrochloride (C4H11N5.HCl) ＝ MS × QT/QS × V?/V × 1/2 MS: Amount (mg) of metformin hydrochloride for assay taken Internal standard solution—A solution of 4?-methoxya-cetophenone in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) (1 in 2000). Dissolution <6.10> (1) Pioglitazone hydrochloride—When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of a solution, which is prepared by mixing 50 mL of 0.2 mol/L hydrochlo-ric acid TS and 150 mL of potassium chloride solution (3 in 20), adding water to make 1000 mL and adjusting to pH 2.0 with 5 mol/L hydrochloric acid TS, as the dissolution me-dium, the dissolution rate in 30 minutes of Pioglitazone Hy-drochloride and Metformin Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride and Metformin Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 18.4 mg of pioglitazone hydrochlo-ride (C19H20N2O3S.HCl), and use this solution as the sample solution. Separately, weigh accurately about 37 mg of Pioglitazone Hydrochloride RS (separately, determine the water <2.48> in the same manner as Pioglitazone Hydrochlo-ride), and dissolve in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of pioglitazone in each solution. Dissolution rate (z) with respect to the labeled amount of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of pioglitazone are not less than 8000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of pioglitazone is not more than 1.0z. (2) Metformin hydrochloride—When the test is per-formed at 50 revolutions per minute according to the Paddle method, using 900 mL of the dissolution medium used in (1), the dissolution rate in 30 minutes of Pioglitazone Hydro-chloride and Metformin Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride and Metformin Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 0.56 mg of metformin hydrochlo-ride (C4H11N5.HCl), and use this solution as the sample solu-tion. Separately, weigh accurately about 28 mg of metformin hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in the dissolution medium to make ex-actly 50 mL, use this solution as the standard solution. Per-form the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of metformin in each so-lution. Dissolution rate (z) with respect to the labeled amount of metformin hydrochloride (C4H11N5.HCl) ＝ MS × AT/AS × V?/V × 1/C × 1800 MS: Amount (mg) of metformin hydrochloride for assay taken C: Labeled amount (mg) of metformin hydrochloride (C4H11N5.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (2). System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of metformin are not less than 6000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of metformin is not more than 1.0z.
(1) Pioglitazone hydrochloride—Weigh accurately the mass of not less than 20 Pioglitazone Hydrochloride and Metformin Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 33 mg of pioglitazone hydrochloride (C19H20N2O3S.HCl), add 40 mL of 0.1 mol/L hydrochloric acid TS, shake vigorously for 10 minutes, add 40 mL of methanol, and shake. Add a mix-ture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent fil-trate, add exactly 5 mL of the internal standard solution, then add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Pioglitazone Hydrochloride RS (separately determine the water <2.48> in the same manner as Pioglitazone Hydrochlo-ride), and dissolve in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, then add a mixture of 0.1 mol/L hydrochloric System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pioglitazone and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pioglitazone to that of the internal standard is not more than 1.0z. (2) Metformin hydrochloride—Weigh accurately the mass of not less than 20 Pioglitazone Hydrochloride and Metformin Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 0.5 g of metformin hydrochloride (C4H11N5.HCl), add 40 mL of 0.1 mol/L hydrochloric acid TS, shake vigorously for 10 minutes, add 40 mL of methanol, and shake. Add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add exactly 5 mL of the internal standard solution, then add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of metfor-min hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make exactly 10 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, then add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) to make 100 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of metformin to that of the internal standard. acid TS and methanol (1:1) to make 100 mL, and use this so- Amount (mg) of metformin hydrochloride (C H N .HCl) lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed ＝ MS × QT/QS × 10 4 11 5 under Liquid Chromatography <2.01> according to the fol- lowing conditions, and calculate the ratios, QT and QS, of the peak area of pioglitazone to that of the internal stand-ard. Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × QT/QS MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.2 g of sodium lauryl sulfate in 1000 mL of a mixture of a solution of ammonium dihydro-gen phosphate (23 in 4000) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of pioglita-zone is about 9 minutes. MS: Amount (mg) of metformin hydrochloride for assay taken Internal standard solution—A solution of 4?-methoxya-cetophenone in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (1:1) (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.2 g of sodium lauryl sulfate in 1000 mL of a mixture of a solution of ammonium dihydro-gen phosphate (23 in 4000) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of metformin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, metformin and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the ratio of the peak area of metformin to that of the internal standard is not more than 1.0z.‌
Pioglitazone Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pioglitazone hydrochloride (C19H20N2O3S. HCl: 392.90). Method of preparation Prepare as directed under Tablets, with Pioglitazone Hydrochloride. Identification To an amount of powdered Pioglitazone Hy-drochloride Tablets, equivalent to 2.8 mg of Pioglitazone Hydrochloride, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 267 nm and 271 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol- ‌lowing method: it meets the requirement. Disintegrate 1 tablet of Pioglitazone Hydrochloride Tablets with 10 mL of 0.1 mol/L hydrochloric acid TS, add 70 mL of methanol, shake vigorously for 10 minutes, then add methanol to make exactly 100 mL, and centrifuge. Take exactly V mL of the supernatant liquid, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly V? mL so that each mL contains about 26 mg of pioglitazone hydrochloride (C19H20N2O3S.
To an amount of powdered Pioglitazone Hy-drochloride Tablets, equivalent to 2.8 mg of Pioglitazone Hydrochloride, add 100 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 267 nm and 271 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol- ‌lowing method: it meets the requirement. Disintegrate 1 tablet of Pioglitazone Hydrochloride Tablets with 10 mL of 0.1 mol/L hydrochloric acid TS, add 70 mL of methanol, shake vigorously for 10 minutes, then add methanol to make exactly 100 mL, and centrifuge. Take exactly V mL of the supernatant liquid, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly V? mL so that each mL contains about 26 mg of pioglitazone hydrochloride (C19H20N2O3S.HCl), and use this solution as the sample solution. Separately, weigh accurately about 33 mg of Pioglitazone Hydrochloride RS (separately, determine the water <2.48> in the same manner as Pioglita-zone Hydrochloride), dissolve in 10 mL of 0.1 mol/L hydro-chloric acid TS, and add methanol to make exactly 100 mL. Pipet 4 mL of this solution, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (9:1) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand-ard solution at 269 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a mixture of methanol and 0.1 mol/L hydrochloric acid TS (9:1) as the blank. Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × AT/AS × V?/V × 2/25 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution, which is prepared by mixing 50 mL of 0.2 mol/L hydrochloric acid TS and 150 mL of potassium chlo-ride solution (3 in 20), adding water to make 1000 mL and adjusting to pH 2.0 with 5 mol/L hydrochloric acid TS, as the dissolution medium, the dissolution rate in 45 minutes of Pioglitazone Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Pioglitazone Hydrochloride Tablets, withdraw 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 18 mg of pioglitazone hydro-chloride (C19H20N2O3S.HCl), and use this solution as the sample solution. Separately, weigh accurately about 23 mg of Pioglitazone Hydrochloride RS (separately determine the water <2.48> in the same manner as Pioglitazone Hydrochlo-ride), dissolve in 10 mL of methanol, and add the dissolution medium to make exactly 50 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 269 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24> using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × AT/AS × V?/V × 1/C × 72 MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis C Labeled amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) in 1 tablet
Accurately weigh the mass of not less than 20 Pioglitazone Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 25 mg of pioglitazone hydrochloride (C19H20N2O3S.HCl), add 45 mL of methanol and exactly 5 mL of the internal standard solution, sonicate, and centrifuge. To 2 mL of the superna-tant liquid add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Pioglitazone Hydrochloride RS (separately, determine the water <2.48> in the same manner as Pioglita-zone Hydrochloride), dissolve in 45 mL of methanol, and add exactly 5 mL of the internal standard solution. Pipet 2 mL of this solution, add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of pioglitazone to that of the in-ternal standard. Amount (mg) of pioglitazone hydrochloride (C19H20N2O3S.HCl) ＝ MS × QT/QS MS: Amount (mg) of Pioglitazone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of benzophenone in methanol (1 in 750). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 269 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of ammonium acetate solution (77 in 10,000), acetonitrile and acetic acid (100) (25:25:1). Flow rate: Adjust so that the retention time of pioglita-zone is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, pioglitazone and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pioglitazone to that of the internal standard is not more than 1.0z.
Piperacillin Sodium for Injection is a white, powder or masses.
Proceed as directed in the Identification under Piperacillin Sodium. pH <2.54> The pH of a solution prepared by dissolving an amount of Piperacillin Sodium for Injection, equivalent to 1.0 g (potency) of Piperacillin Sodium, in 4 mL of water is 5.0 – 7.0.
(1) Clarity and color of solution—Dissolve an amount of Piperacillin Sodium for Injection, equivalent to 4.0 g (potency) of Piperacillin Sodium, in 17 mL of water: the solution is clear and colorless. (2) Related substances—Proceed as directed in the Purity (4) under Piperacillin Sodium. Water <2.48> Not more than 1.0z (3 g, volumetric titra-tion, direct titration). Bacterial endotoxins <4.01> Less than 0.04 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Piperacillin Sodium for Injection. Weigh accurately an amount of the contents, equivalent to about 20 mg (po-tency) of Piperacillin Sodium, dissolve in water to make ex-actly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 20 mg (potency) of Piperacillin RS, and dissolve in the mobile phase to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Proceed as directed in the Assay under Piperacillin Sodium. Amount [mg (potency)] of piperacillin (C23H27N5O7S) ＝ MS × QT/QS MS: Amount [mg (potency)] of Piperacillin RS taken Internal standard solution—A solution of acetanilide in the mobile phase (1 in 5000). Piperazine Adipate
Take a quantity of Piperazine Phosphate Tablets equivalent to 0.1 g of Piperazine Phosphate Hy-drate, previously powdered, add 10 mL of water, shake while warming for 10 minutes, allow to cool, and filter. To 3 mL of the filtrate add 3 drops of Reinecke salt TS: a light red precipitate is formed. Disintegration <6.09> It meets the requirement. The time limit of the test is 10 minutes.
Weigh accurately not less than 20 Piperazine Phos-phate Tablets, and powder. Weigh accurately a quantity of the powder, equivalent to about 0.15 g of piperazine phos-phate hydrate (C4H10N2.H3PO4.H2O). Add 5 mL of formic acid, shake for 5 minutes, centrifuge, and collect the super-natant liquid. To the residue add 5 mL of formic acid, shake for 5 minutes, centrifuge, and collect the supernatant liquid. Repeat twice the same procedure with 5 mL each of acetic acid (100), combine all the supernatant liquids, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determi-nation, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 10.11 mg of C4H10N2.H3PO4.H2O
Pitavastatin Calcium Orally Disintegrating Tablets
ピタバスタチンカルシウム口腔内崩壊錠
Pitavastatin Calcium Orally Disintegrating Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pitavastatin calcium (C50H46CaF2N2O8: 880.98). Method of preparation Prepare as directed under Tablets, with Pitavastatin Calcium Hydrate. Identification To a quantity of Pitavastatin Calcium Orally Disintegrating Tablets, equivalent to 4 mg of pitavastatin calcium (C50H46CaF2N2O8), add 10 mL of methanol, shake thoroughly, and centrifuge. To 1 mL of the supernatant liq-uid add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 243 nm and 247 nm. Purity Related substances—Conduct this procedure using light-resistant vessels. To a quantity of Pitavastatin Calcium Orally Disintegrating Tablets, equivalent to 20 mg of mL of the standard solution under the above operating con- ditions, the internal standard and pitavastatin are eluted in pitavastatin calcium (C50H46 CaF2N2O8), add 60 mL of a mix- this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pitavastatin to that of the internal standard is not more than 1.0z. Containers and storage Containers—Tight containers. Storage—Light-resistant. ture of acetonitrile and water (3:2), sonicate to disintegrate, add a mixture of acetonitrile and water (3:2) to make 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the amount of related substance A having the relative retention times of about 1.1 to pitavastatin is not more than 0.5z, the amount of related substance B having the relative retention times of about 1.5 is not more than 0.2z, the amount of related sub-stance TA having the relative retention times of about 1.7 is not more than 0.5z, and the amount of the peak other than pitavastatin and the peaks mentioned above is not more than 0.1z. Furthermore, the total amount of the peaks other than pitavastatin is not more than 2.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: To 10 mL of dilute acetic acid add water to make 1000 mL. To 800 mL of this solution add diluted so-dium acetate TS (1 in 100) to adjust to pH 3.8. Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 20 20 – 40 60 → 30 40 → 70 40 – 65 Flow rate: Adjust so that the retention time of pitavastatin is about 23 minutes. Time span of measurement: About 2.7 times as long as the retention time of pitavastatin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add a mixture of acetonitrile and water (3:2) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Confirm that the peak area of pitavastatin obtained with 10 mL of this solution is equiva-lent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pitavastatin are not less than 7500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of pitavastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Pitavastatin Calcium Orally Disintegrating Tablets add exactly V mL of the internal standard solution so that each mL contains about 0.
To a quantity of Pitavastatin Calcium Orally Disintegrating Tablets, equivalent to 4 mg of pitavastatin calcium (C50H46CaF2N2O8), add 10 mL of methanol, shake thoroughly, and centrifuge. To 1 mL of the supernatant liq-uid add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 243 nm and 247 nm.
Related substances—Conduct this procedure using light-resistant vessels. To a quantity of Pitavastatin Calcium Orally Disintegrating Tablets, equivalent to 20 mg of mL of the standard solution under the above operating con- ditions, the internal standard and pitavastatin are eluted in pitavastatin calcium (C50H46 CaF2N2O8), add 60 mL of a mix- this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pitavastatin to that of the internal standard is not more than 1.0z.
Conduct this procedure using light-resistant vessels. To not less than 20 tablets of Pitavastatin Calcium Orally Disintegrating Tablets add exactly V mL of a mixture of acetonitrile and water (3:2) so that each mL contains about 0.2 mg of pitavastatin calcium (C50H46CaF2N2O8), and sonicate to disintegrate the tablets. Pipet 5 mL of this solu-tion, add exactly 5 mL of the internal standard solution, shake, then filter through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately weigh accurately about 24 mg of Pitavastatin Methylbenzylamine RS (separately determine the water <2.48> by coulometric titration using 0.1 g), dis-solve in a mixture of acetonitrile and water (3:2) to make ex-actly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the ‌sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pitavastatin to that of the internal standard. Purity Related substances—Conduct this procedure using light-resistant vessels. Take a quantity of Pitavastatin Cal-cium Tablets, equivalent to 20 mg of pitavastatin calcium (C50H46CaF2N2O8), add 60 mL of a mixture of acetonitrile Amount (mg) of pitavastatin calcium (C H CaF N O ) and water (3:2), sonicate to disintegrate, and add a mixture in 1 tablet 50 46 2 2 8 of acetonitrile and water (3:2) to make 100 mL. Filter this solution through a membrane filter with a pore size not ＝ MS × QT/QS × V/N × 1/100 × 0.812 MS: Amount (mg) of Pitavastatin Methylbenzylamine RS taken, calculated on the anhydrous basis N: Number of tablets taken Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of acetonitrile and water (3:2) (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Pitavastatin Calcium Hydrate. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pitavastatin are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pitavastatin to that of the internal standard is not more than 1.0z.
Pitavastatin Calcium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pitavastatin calcium (C50H46CaF2N2O8: 880.98). Method of preparation Prepare as directed under Tablets, with Pitavastatin Calcium Hydrate. Identification Powder Pitavastatin Calcium Tablets. Weigh a portion of the powder, equivalent to 4 mg of pitavastatin calcium (C50H46CaF2N2O8), add 10 mL of meth-anol and shake well, and centrifuge. To 1 mL of the super-natant liquid, add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 242 nm and 246 nm. Flow rate: Adjust so that the retention time of pitavastatin is about 23 minutes. Time span of measurement: About 2.7 times as long as the retention time of pitavastatin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of acetonitrile and water (3:2) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Confirm that the peak area of pitavastatin obtained with 50 mL of this solution is equiva-lent to 7 to 13z of that with 50 mL of the solution for system suitability test. System performance: When the procedure is run with 50 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pitavastatin are not less than 7500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of pitavastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Pitavastatin Calcium Tablets add exactly V mL of the internal standard solution so that each mL contains about 0.
Powder Pitavastatin Calcium Tablets. Weigh a portion of the powder, equivalent to 4 mg of pitavastatin calcium (C50H46CaF2N2O8), add 10 mL of meth-anol and shake well, and centrifuge. To 1 mL of the super-natant liquid, add methanol to make 50 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 242 nm and 246 nm. Flow rate: Adjust so that the retention time of pitavastatin is about 23 minutes. Time span of measurement: About 2.7 times as long as the retention time of pitavastatin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add a mixture of acetonitrile and water (3:2) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 5 mL of the solution for system suitability test, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Confirm that the peak area of pitavastatin obtained with 50 mL of this solution is equiva-lent to 7 to 13z of that with 50 mL of the solution for system suitability test. System performance: When the procedure is run with 50 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pitavastatin are not less than 7500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of pitavastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Pitavastatin Calcium Tablets add exactly V mL of the internal standard solution so that each mL contains about 0.2 mg of pitavastatin calcium (C50H46CaF2N2O8), and add V mL of a mixture of acetonitrile and water (3:2), shake well until the tablet is disintegrated completely. Filter this so-lution through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of pitavastatin calcium (C50H46CaF2N2O8) ＝ MS × QT/QS × V/100 × 0.812 MS: Amount (mg) of Pitavastatin Methylbenzylamine RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of acetonitrile and water (3:2) (3 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of Pitavastatin Calcium Tablets is not less than 85z. Conduct this procedure using light-resistant vessels. Start the test with 1 tablet of Pitavastatin Calcium Tablets, with-draw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.1 mg of pitavastatin calcium (C50H46CaF2N2O8), and use this solution as the sample solu-tion. Separately, weigh accurately about 24 mg of Pitavasta-tin Methylbenzylamine RS (separately determine the water <2.48> by coulometric titration using 0.1 g), and dissolve in a mixture of acetonitrile and water (3:2) to make exactly 200 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of pitavastatin in each solution. Dissolution rate (z) with respect to the labeled amount of pitavastatin calcium (C50H46CaF2N2O8) ＝ MS × AT/AS × V?/V × 1/C × 9/2 × 0.812 MS: Amount (mg) of Pitavastatin Methylbenzylamine RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of pitavastatin calcium (C50H46CaF2N2O8) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pitavastatin are not less than 4500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pitavastatin is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 Pitavastatin Calcium Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of pitavastatin cal-cium (C50H46CaF2N2O8), add 30 mL of a mixture of aceto-nitrile and water (3:2), and sonicate for 10 minutes. To this solution, add a mixture of acetonitrile and water (3:2) to make exactly 50 mL. Filter this solution through a mem-brane filter with a pore size not exceeding 0.45 mm. Pipet 5 mL of this filtrate, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Sepa-rately, weigh accurately about 24 mg of Pitavastatin Methyl-benzylamine RS (separately determine the water <2.48> by coulometric titration using 0.1 g), and dissolve in a mixture of acetonitrile and water (3:2) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pitavastatin to that of the internal standard. Amount (mg) of pitavastatin calcium (C50H46CaF2N2O8) ＝ MS × QT/QS × 1/2 × 0.812 MS: Amount (mg) of Pitavastatin Methylbenzylamine RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of acetonitrile and water (3:2) (3 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 10 mL of dilute acetic acid add water to make 1000 mL. To 350 mL of this solution add 650 mL of methanol, and dissolve 0.29 g of sodium chloride in this solution. Flow rate: Adjust so that the retention time of pitavastatin is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pitavastatin are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pitavastatin to that of the internal standard is not more than 1.0z.
Powder Pivmecillinam Hydrochloride Disintegration <6.09> Perform the test using the disk: it meets the requirement.
Weigh accurately the mass of not less than 20 Piv-mecillinam Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 0.1 g (potency) of Pivmecillinam Hydrochloride, add 50 mL of the mobile phase, shake vigorously for 10 minutes, and add the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution and the mobile phase to make 50 mL, filter through a mem-brane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Pivmecillinam Hydrochloride RS, equivalent to about 20 mg (potency), dissolve in the mobile phase, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Pivmecillinam Hydrochloride. Tablets, dissolve a portion of the powder, equivalent to 35 mg (potency) of Pivmecillinam Hydrochloride, in 4 mL of a mixture of acetonitrile and acetic acid (100) (97:3), and filter Amount [mg (potency)] of mecillinam (C ＝ MS × QT/QS × 5 15H23N3O3S) through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately dis-solve 25 mg of Pivmecillinam Hydrochloride RS in 2 mL of a mixture of acetonitrile and acetic acid (100) (97:3), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 2 mL each of the sample solution and stand-ard solution on a plate of silica gel for thin-layer chromatog-raphy, and immediately develop the plate with a mixture of acetone, water and acetic acid (100) (10:1:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for 10 minutes: the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. Water <2.48> Not more than 3.0z (1 g of powdered Piv-mecillinam Hydrochloride Tablets, volumetric titration, direct titration). Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pivmecillinam Hydrochloride Tablets add 40 mL of the mobile phase, shake vigorously for 10 minutes, and add the mobile phase to make exactly 50 mL. Pipet V mL, equivalent to about 10 mg (potency) of Pivmecillinam Hydrochloride, add exactly 5 mL of the internal standard so-lution and the mobile phase to make 50 mL, filter through a membrane filter with a pore size not exceeding 0.45 mm, dis-card the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately an amount of Pivmecillinam Hydrochloride RS, equivalent to about 20 mg (potency), dissolve in the mobile phase, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Pivmecillinam Hydrochloride. MS: Amount [mg (potency)] of Pivmecillinam Hydrochlo-ride RS taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 12,500).
Weigh accurately an amount of Polaprezinc Granules, equivalent to about 0.1 g of polaprezinc [(C9H12N4O3Zn)n], add exactly 20 mL of 0.2 mol/L hydro-chloric acid TS, shake vigorously for 10 minutes, and centri-fuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of L-Carnosine RS, previously dried at 1059C for 3 hours, dissolve in 5 mL of 0.2 mol/L hydrochloric acid TS, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of L-carnosine to that of the internal standard. mity test. To the total content of 1 package of Polaprezinc Granules add exactly V mL of 0.2 mol/L hydrochloric acid TS so that Amount (mg) of polaprezinc [(C9H ＝ MS × QT/QS × 4 × 1.292 12N4O3Zn)n] each mL contains about 5 mg of polaprezinc [(C9H12N4O3Zn)n], shake vigorously for 10 minutes, and cen-trifuge. Pipet 5 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of polaprezinc [(C9H12N4O3Zn)n] ＝ MS × QT/QS × V/5 × 1.292 MS: Amount (mg) of L-Carnosine RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buŠer solution (pH 4.0) as the dissolution medium, the dissolution rate in 15 minutes of Polaprezinc Granules is not less than 80z. Start the test with an accurately weighed amount of Polaprezinc Granules, equivalent to about 75 mg of polaprezinc [(C9H12N4O3Zn)n], withdraw not less than 20 mL of the medium at the speciˆed minute after starting the test, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the ˆrst ˆltrate, pipet 1 mL of the subsequent ˆltrate, add diluted nitric acid (77 in 10,000) to make exactly 25 mL, and use this solution as the sample solution. Separately, pipet suitable volumes of Standard Zinc Stock Solution, to each solution add diluted nitric acid (77 in 10,000) so that each mL con-tains 0.4 to 0.8 mg of zinc (Zn: 65.38), and use these solu-tions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and calculate the amount of zinc in the sample solution using a calibration curve obtained from the absorbances of the standard solutions. Dissolution rate (z) with respect to the labeled amount of polaprezinc [(C9H12N4O3Zn)n] ＝ Content (mg/mL) of zinc in the sample solution/MT × 1/C × 2250 × 4.429 MS: Amount (mg) of L-Carnosine RS taken Internal standard solution—Dissolve 0.25 g of 4-aminoa-cetophenone in 5 mL of acetonitrile, and add the mobile phase to make 100 mL. Operating conditions— Proceed as directed in the operating conditions in the Assay (1) under Polaprezinc. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, 4-aminoacetophenone and L-carnosine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of L-carnosine to that of the internal standard is not more than 1.0z.
Macrogol Ointment is white in color. It has a faint, characteristic odor.
Dissolve 50 mg of Macrogol Ointment in 5 mL of dilute hydrochloric acid, add 1 mL of barium chloride TS, shake, filter if necessary, and add 1 mL of a solution of phosphomolybdic acid n-hydrate (1 in 10) to the filtrate: a yellow-green precipitate is formed.
Powdered Cellulose occurs as a white pow-der. It is practically insoluble in water, in ethanol (95) and in diethyl ether.◆
(1) Dissolve 20 g of zinc chloride and 6.5 g of potassium iodide in 10.5 mL of water, add 0.5 g of iodine, ether through the column. Evaporate the eluate to dryness in a previously dried and tared evaporation dish. Dry the residue at 1059C for 30 minutes, and weigh after allowing to cool in a desiccator: the difference between the mass of the residue and the mass obtained from a blank determination does not exceed 15.0 mg (0.15z). Loss on drying <2.41> Not more than 6.5z (1 g, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g calcu-lated on the dried basis). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 103 CFU/g and 102 CFU/g, respectively. Es-cherichia coli, Salmonella, Pseudomonas aeruginosa and Staphylococcus aureus are not observed.◆ ◆Containers and storage Containers—Tight containers.◆ Celmoleukin (Genetical Recombination)
Pravastatin Sodium Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of pravastatin sodium (C23H35NaO7: 446.51). Method of preparation Prepare as directed under Gran-ules, with Pravastatin Sodium. Identification To an amount of Pravastatin Sodium Fine Granules, equivalent to 10 mg of Pravastatin Sodium, add 20 mL of water, sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the fil-trate, and add water to 1 mL of the subsequent filtrate to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 237 nm and 241 nm. Purity Related substances—The sample solution and the standard solution are stored at not exceeding 59C after preparation. To an amount of Pravastatin Sodium Fine Granules, equivalent to 25 mg of Pravastatin Sodium, add 25 mL of a mixture of water and methanol (1:1), sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, dis-card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of water and methanol (1:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the area of the peaks, having the relative retention time of about 0.36 and about 1.9 to pravastatin, obtained from the sample solution is not larger than 1/2 times and 3 times the peak area of pravastatin from the standard solution, respec-tively, the area of the peak other than pravastatin and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks other than pravastatin from the sample solution is not larger than 4.5 times the peak area of pravastatin from the standard solu-tion. For the area of the peaks, having the relative retention time of about 0.28, about 0.36 and about 0.88, multiply their correction factors, 1.16, 1.72 and 1.22, respectively. Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, methanol, acetic acid (100) and triethylamine (750:250:1:1). Mobile phase B: A mixture of methanol, water, acetic acid (100) and triethylamine (650:350:1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: 1.3 mL per minute. Time span of measurement: For 75 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of pravastatin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pravastatin is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Pravastatin Sodium Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Pravastatin Sodium Fine Granules add exactly V mL of the internal standard solution so that each mL contains 0.
To an amount of Pravastatin Sodium Fine Granules, equivalent to 10 mg of Pravastatin Sodium, add 20 mL of water, sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the fil-trate, and add water to 1 mL of the subsequent filtrate to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 237 nm and 241 nm.
Related substances—The sample solution and the standard solution are stored at not exceeding 59C after preparation. To an amount of Pravastatin Sodium Fine Granules, equivalent to 25 mg of Pravastatin Sodium, add 25 mL of a mixture of water and methanol (1:1), sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, dis-card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of water and methanol (1:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the area of the peaks, having the relative retention time of about 0.36 and about 1.9 to pravastatin, obtained from the sample solution is not larger than 1/2 times and 3 times the peak area of pravastatin from the standard solution, respec-tively, the area of the peak other than pravastatin and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks other than pravastatin from the sample solution is not larger than 4.5 times the peak area of pravastatin from the standard solu-tion. For the area of the peaks, having the relative retention time of about 0.28, about 0.36 and about 0.88, multiply their correction factors, 1.16, 1.72 and 1.22, respectively. Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, methanol, acetic acid (100) and triethylamine (750:250:1:1). Mobile phase B: A mixture of methanol, water, acetic acid (100) and triethylamine (650:350:1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Flow rate: 1.3 mL per minute. Time span of measurement: For 75 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of pravastatin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pravastatin is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the Pravastatin Sodium Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Pravastatin Sodium Fine Granules add exactly V mL of the internal standard solution so that each mL contains 0.25 mg of pravastatin sodium (C23H35NaO7), sonicate for 15 min-utes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the filtrate, pipet 2 mL of the subsequent filtrate add a mixture of water and methanol (1 in 1) to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ MS × QT/QS × V/100 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (3 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Pravastatin Sodium Fine Granules is not less than 80z. Start the test with an accurately weighed amount of Pravastatin Sodium Fine Granules, equivalent to about 5 mg of pravastatin sodium (C23H35NaO7), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample so-lution. Separately, weigh accurately about 23 mg of Pravastatin 1,1,3,3-Tetramethylbutylammonium RS (sepa-rately determine the water <2.48> in the same manner as Pravastatin Sodium), and dissolve in water to make exactly 100 mL. Pipet 3 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. ‌Determine the absorbances, AT1 and AS1, at 238 nm and AT2 and AS2 at 265 nm of the sample solution and standard solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of pravastatin sodium (C23H35NaO7) ＝ MS/MT × (AT1 － AT2)/(AS1 － AS2) × 1/C × 27 × 0.806 MS: Amount (mg) of Pravastatin 1,1,3,3-Tetramethyl-butylammonium RS taken, calculated on the anhy-drous basis MT: Amount (g) of Pravastatin Sodium Fine Granules taken C: Labeled amount (mg) of pravastatin sodium (C23H35NaO7) in 1 g
Weigh accurately an amount of Pravastatin Sodium Fine Granules, equivalent to about 5 mg of pravastatin so-dium (C23H35NaO7), add exactly 20 mL of the internal stand-ard solution, sonicate for 15 minute, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the filtrate, to 2 mL of the subsequent filtrate add a mixture of water and methanol (1:1) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 32 mg of Pravastatin 1,1,3,3-Tetramethylbutylammonium RS (separately determine the water <2.48> in the same manner as Pravastatin Sodium), and dissolve in a mixture of water and methanol (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, then add a mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pravastatin to that of the internal standard. Pravastatin Sodium Solution
Pravastatin Sodium Solution contains not less than 95.
Pass a volume of Pravastatin Sodium Solu-tion, equivalent to 1 mg of Pravastatin Sodium, through a column [5.5 mm in inside diameter, packed with 30 mg of divinylbenzene-N-vinyl pyrrolidone copolymer for column chromatography (30 mm in particle size), and washed with 1 mL of methanol and 1 mL of water]. Then wash with 1 mL of water, and elute with 1 mL of methanol. To 0.1 mL of the eluate add water to make 10 mL, and determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 237 nm and 241 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—The sample solution and the standard solution are stored at not exceeding 159C after preparation. To a volume of Pravastatin Sodium Solution, equivalent to 2 mg of Pravastatin Sodium, add a mixture of methanol and water (5:3) to make 10 mL, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of water and methanol (1:1) to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma- Amount (mg) of pravastatin sodium (C ＝ MS × QT/QS × 1/5 × 1.052 23H 35NaO7) tography <2.01> according to the following conditions. De- termine each peak area of both solutions by the automatic integration method: the area of the peaks, having the relative MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Pravastatin Sodium. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pravastatin are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pravastatin to that of the internal standard is not more than 1.0z.
To an exact volume of Pravastatin Sodium Solution, equivalent to 2 mg of pravastatin sodium (C23H35NaO7), add exactly 5 mL of the internal standard solution, add water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Pravastatin 1,1,3,3-Tetramethylbutylammonium RS (separately deter-mine the water <2.48> in the same manner as Pravastatin Sodium), and dissolve in a solution of disodium hydrogen phosphate dodecahydrate (1 in 200) to make exactly 50 mL. Pipet 6 mL of this solution, add exactly 5 mL of the internal standard solution, add water to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QT and QS, of the peak area of pravastatin to that of the internal standard. Amount (mg) of pravastatin sodium ＝ MS × QT/QS × 3/25 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in methanol (3 in 10,000). Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 238 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of water, methanol, acetic acid (100) and triethylamine (500:500:1:1). Flow rate: Adjust so that the retention time of pravastatin is about 20 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pravastatin are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pravastatin to that of the internal standard is not more than 1.0z.
Pravastatin Sodium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of pravastatin sodium (C23H35NaO7: 446.51). Method of preparation Prepare as directed under Tablets, with Pravastatin Sodium. Identification To a quantity of powdered Pravastatin So-dium Tablets, equivalent to 10 mg of Pravastatin Sodium, add 20 mL of water, sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the fil-trate, and add water to 1 mL of the subsequent filtrate to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 237 nm and 241 nm. Purity Related substances—The sample solution and the standard solution are stored at not exceeding 159C after preparation. To an amount of powdered Pravastatin Sodium Tablets, equivalent to 50 mg of Pravastatin Sodium, add 40 mL of a mixture of water and methanol (1:1), sonicate, then add a mixture of water and methanol (1:1) to make 50 mL, centrifuge, and use the supernatant liquid as the sample solu-tion. Pipet 1 mL of the sample solution, add a mixture of water and methanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peaks, having the relative retention time about 0.36 and about 1.9 to pravastatin obtained from the sample solution is not larger than 3/10 times and 2 times the peak area of pravastatin from the standard solution, respectively, the area of the peak other than pravastatin and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks other than pravastatin from the sample solution is not larger than 3 times the peak area of pravastatin from the standard solution. For the area of the peaks, having the relative retention time about 0.28, about 0.36 and about 0.88, multiply their correction factors, 1.16, 1.72 and 1.22, respectively. Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, methanol, acetic acid (100) and triethylamine (750:250:1:1). Mobile phase B: A mixture of methanol, water, acetic acid (100) and triethylamine (650:350:1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 50 50 – 75 50 → 0 50 → 100 Flow rate: 1.3 mL per minute. Time span of measurement: For 75 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of pravastatin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pravastatin is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pravastatin Sodium Tablets add exactly V mL of the internal standard solution so that each mL con-tains 0.25 mg of pravastatin sodium (C23H35NaO7), sonicate for 15 minutes, and centrifuge. To 2 mL of the supernatant liquid add a mixture of water and methanol (1:1) to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ MS × QT/QS × V/100 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (3 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Pravastatin Sodium Tablets is not less than 85z. Start the test with 1 tablet of Pravastatin Sodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.
To a quantity of powdered Pravastatin So-dium Tablets, equivalent to 10 mg of Pravastatin Sodium, add 20 mL of water, sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the fil-trate, and add water to 1 mL of the subsequent filtrate to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits a maximum between 237 nm and 241 nm.
Related substances—The sample solution and the standard solution are stored at not exceeding 159C after preparation. To an amount of powdered Pravastatin Sodium Tablets, equivalent to 50 mg of Pravastatin Sodium, add 40 mL of a mixture of water and methanol (1:1), sonicate, then add a mixture of water and methanol (1:1) to make 50 mL, centrifuge, and use the supernatant liquid as the sample solu-tion. Pipet 1 mL of the sample solution, add a mixture of water and methanol (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area by the automatic integration method: the area of the peaks, having the relative retention time about 0.36 and about 1.9 to pravastatin obtained from the sample solution is not larger than 3/10 times and 2 times the peak area of pravastatin from the standard solution, respectively, the area of the peak other than pravastatin and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks other than pravastatin from the sample solution is not larger than 3 times the peak area of pravastatin from the standard solution. For the area of the peaks, having the relative retention time about 0.28, about 0.36 and about 0.88, multiply their correction factors, 1.16, 1.72 and 1.22, respectively. Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, methanol, acetic acid (100) and triethylamine (750:250:1:1). Mobile phase B: A mixture of methanol, water, acetic acid (100) and triethylamine (650:350:1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 50 50 – 75 50 → 0 50 → 100 Flow rate: 1.3 mL per minute. Time span of measurement: For 75 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and methanol (1:1) to make exactly 10 mL. Confirm that the peak area of pravastatin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pravastatin is not more than 1.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Pravastatin Sodium Tablets add exactly V mL of the internal standard solution so that each mL con-tains 0.25 mg of pravastatin sodium (C23H35NaO7), sonicate for 15 minutes, and centrifuge. To 2 mL of the supernatant liquid add a mixture of water and methanol (1:1) to make 20 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ MS × QT/QS × V/100 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (3 in 10,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Pravastatin Sodium Tablets is not less than 85z. Start the test with 1 tablet of Pravastatin Sodium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.5 mg of pravastatin (C23H36O7), and use this solution as the sample solution. Separately, weigh accurately about 23 mg of Pravastatin 1,1,3,3-Tetramethylbutylammo-nium RS (separately determine the water <2.48> in the same manner as Pravastatin Sodium), and dissolve in water to make exactly 100 mL. Pipet 3 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Determine the absorbances, AT1 and AS1, at 238 nm and AT2 and AS2 at 256 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of pravastatin sodium (C23H35NaO7) ＝ MS × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 27 × 0.806 MS: Amount (mg) of Pravastatin 1,1,3,3-Tetramethyl-butylammonium RS taken, calculated on the anhy-drous basis C: Labeled amount (mg) of pravastatin sodium (C23H35NaO7) in 1 tablet
Weigh accurately and powder not less than 20 Pravastatin Sodium Tablets. Weigh accurately a portion of the powder, equivalent to about 10 mg of pravastatin so-dium (C23H35NaO7), add exactly 40 mL of the internal stand-ard solution, sonicate for 15 minutes, and centrifuge. Filter the supernatant liquid, discard the first 5 mL of the filtrate, to 2 mL of the subsequent filtrate add a mixture of water and methanol (1:1) to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 32 mg of Pravastatin 1,1,3,3-Tetramethylbutylammonium RS (separately determine the water <2.48> in the same manner as Pravastatin Sodium), and dissolve in a mixture of water and methanol (1:1) to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, then add a mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QT and QS, of the peak area of pravastatin to that of the internal standard. Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ MS × QT/QS × 2/5 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Pravastatin Sodium. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pravastatin are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pravastatin to that of the internal standard is not more than 1.0z.
Prazepam Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of prazepam (C19H17ClN2O: 324.80). Method of preparation Prepare as directed under Tablets, with Prazepam. Identification (1) To a quantity of powdered Prazepam Tablets, equivalent to 0.05 g of Prazepam, add 25 mL of acetone, shake well, and filter. Take 5 mL of the filtrate, evaporate on a water bath to dryness, and dissolve the residue in 3 mL of sulfuric acid. With this solution, proceed as directed in the Identification (1) under Prazepam. To a quantity of powdered Prazepam Tablets, equiv-alent to 0.02 g of Prazepam, add 200 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000), shake well, and filter. To 5 mL of the filtrate add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 50 mL, and determine the absorption spectrum as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 241 nm and 245 nm, between 283 nm and 287 nm and between 363 nm and 367 nm, and minima between 263 nm and 267 nm and between 334 nm and 338 nm. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 900 mL of 0.1 mol/L hydrochloric acid TS as the dissolution medium, the dissolution rate in 30 minutes of Prazepam Tablets is not less than 80z. Start the test with 1 tablet of Prazepam Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, measure exactly the subsequent V mL of the filtrate, add the dissolution medium to make ex- ‌actly V? mL so that each mL contains about 5 mg of prazep-am (C19H17ClN2O), and use this solution as the sample solu-tion.
(1) To a quantity of powdered Prazepam Tablets, equivalent to 0.05 g of Prazepam, add 25 mL of acetone, shake well, and filter. Take 5 mL of the filtrate, evaporate on a water bath to dryness, and dissolve the residue in 3 mL of sulfuric acid. With this solution, proceed as directed in the Identification (1) under Prazepam. To a quantity of powdered Prazepam Tablets, equiv-alent to 0.02 g of Prazepam, add 200 mL of a solution of sulfuric acid in ethanol (99.5) (3 in 1000), shake well, and filter. To 5 mL of the filtrate add a solution of sulfuric acid in ethanol (99.5) (3 in 1000) to make 50 mL, and determine the absorption spectrum as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 241 nm and 245 nm, between 283 nm and 287 nm and between 363 nm and 367 nm, and minima between 263 nm and 267 nm and between 334 nm and 338 nm. Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 900 mL of 0.1 mol/L hydrochloric acid TS as the dissolution medium, the dissolution rate in 30 minutes of Prazepam Tablets is not less than 80z. Start the test with 1 tablet of Prazepam Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, measure exactly the subsequent V mL of the filtrate, add the dissolution medium to make ex- ‌actly V? mL so that each mL contains about 5 mg of prazep-am (C19H17ClN2O), and use this solution as the sample solu-tion. Separately, weigh accurately about 5 mg of prazepam for assay, previously dried at 1059C for 2 hours, add 200 mL of the dissolution medium and dissolve with shaking, or by sonicating if necessary, add the dissolution medium to make exactly 1000 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 240 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of a solution of Prazosin Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Prazosin Hydrochloride as directed in the potassium chlo-ride disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Prazosin Hydrochloride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. To 0.1 g of Prazosin Hydrochloride add 5 mL of of prazepam (C 19H 17ClN2O) water and 1 mL of ammonia TS, shake, allow to stand for 5 minutes, and filter. Render the filtrate acid with acetic acid ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of prazepam for assay taken C: Labeled amount (mg) of prazepam (C19H17ClN2O) in 1 tablet
Weigh accurately not less than 20 Prazepam Tablets, and powder. Weigh accurately a quantity of the powder, equivalent to about 50 mg of prazepam (C19H17ClN2O), add 30 mL of acetone, shake well, centrifuge, and separate the supernatant liquid. Repeat the same procedure twice with 30 mL each of acetone, combine all the supernatants liquid, and evaporate on a water bath to dryness. Dissolve the residue in 50 mL of a mixture of acetic anhydride and acetic acid (100) (7:3), and titrate <2.50> with 0.02 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.02 mol/L perchloric acid VS ＝ 6.496 mg of C19H17ClN2O
Precipitated Calcium Carbonate Fine Granules
(1) To a quantity of powdered Precipitated Calcium Carbonate Fine Granules, equivalent to 0.5 g of Precipitated Calcium Carbonate, add 10 mL of dilute hydro-chloric acid, shake thoroughly, and filter. Boil the filtrate, then cool, and neutralize with ammonia TS: the solution re-sponds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. (2) Powdered Precipitated Calcium Carbonate Fine Granules responds to Qualitative Tests <1.09> (1) for carbon-ate. Uniformity of dosage units <6.02> The granules in single-dose packages meet the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 10 minutes of Precipitated Cal-cium Carbonate Fine Granules is not less than 80z. Start the test with an accurately weighed amount of ‌JP XVIII Official Monographs / Precipitated Calcium Carbonate Tablets 595 Precipitated Calcium Carbonate Fine Granules, equivalent to about 0.5 g of calcium carbonate (CaCO3), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 2 mL of the subsequent filtrate, add the mobile phase to make exactly 20 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 28 mg of calcium carbonate for assay, previously dried at 1809C for 4 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 10 mL of this solu-tion, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of calcium in each solution. Dissolution rate (z) with respect to the labeled amount of calcium carbonate (CaCO3) ＝ MS/MT × AT/AS × 1/C × 1800 MS: Amount (mg) of calcium carbonate for assay taken MT: Amount (mg) of Precipitated Calcium Carbonate Fine Granules taken C: Labeled amount (mg) of calcium carbonate (CaCO3) in 1 g Operating conditions— Detector: An electric conductivity detector. Column: A polyether ether ketone tube 4.6 mm in inside diameter and 10 cm in length, packed with slightly acidic ion-exchange silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of tartaric acid (3 in 2000) and a solution of dipicolinic acid (1 in 3000) (1:1). Flow rate: Adjust so that the retention time of calcium is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, sodium and calcium are eluted in this order with the resolution between these peaks being not less than 4.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of calcium is not more than 2.0z.
Weigh accurately a quantity of powdered Precipitat-ed Calcium Carbonate Fine Granules, equivalent to about 0.12 g of calcium carbonate (CaCO3), add 20 mL of water and 3 mL of dilute hydrochloric acid, and agitate for 15 minutes with the aid of ultrasonic waves. Then, add 80 mL of water, 15 mL of a solution of potassium hydroxide (1 in 10) and 50 mg of NN indicator, and titrate <2.50> immedi-ately with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 5.005 mg of CaCO3
Precipitated Calcium Carbonate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of calcium carbonate (CaCO3: 100.09). Method of preparation Prepare as directed under Tablets, with Precipitated Calcium Carbonate. Identification (1) To a quantity of powdered Precipitated Calcium Carbonate Tablets, equivalent to 0.5 g of Precipi-tated Calcium Carbonate, add 10 mL of dilute hydrochloric acid, shake throughly, and filter, if necessary. Boil, then cool, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. (2) Powdered Precipitated Calcium Carbonate Tablets responds to Qualitative Tests <1.09> (1) for carbonate. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Disintegration <6.09> Apply to the preparation intended to be used as antacid. Perform the test using the disk: it meets the requirement. Dissolution <6.10> Apply to the preparation intended to be used as hyperphosphatemia. When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolu-tion rate in 10 minutes of Precipitated Calcium Carbonate Tablets is not less than 80z. Start the test with 1 tablet of Precipitated Calcium Car-bonate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 56 mg of calcium carbonate (CaCo3), and use this solution as the sample solution.
(1) To a quantity of powdered Precipitated Calcium Carbonate Tablets, equivalent to 0.5 g of Precipi-tated Calcium Carbonate, add 10 mL of dilute hydrochloric acid, shake throughly, and filter, if necessary. Boil, then cool, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. (2) Powdered Precipitated Calcium Carbonate Tablets responds to Qualitative Tests <1.09> (1) for carbonate. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Disintegration <6.09> Apply to the preparation intended to be used as antacid. Perform the test using the disk: it meets the requirement. Dissolution <6.10> Apply to the preparation intended to be used as hyperphosphatemia. When the test is performed at 50 revolutions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution medium, the dissolu-tion rate in 10 minutes of Precipitated Calcium Carbonate Tablets is not less than 80z. Start the test with 1 tablet of Precipitated Calcium Car-bonate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 56 mg of calcium carbonate (CaCo3), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of calcium carbonate for assay, previously dried at 1809C for 4 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of calcium in each solution. Dissolution rate (z) with respect to the labeled amount of calcium carbonate (CaCO3) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of calcium carbonate for assay taken C: Labeled amount (mg) of calcium carbonate (CaCO3) in 1 tablet Operating conditions— Detector: An electric conductivity detector. Column: A polyether ether ketone tube 4.6 mm in inside diameter and 10 cm in length, packed with slightly acidic ion-exchange silica gel for liquid chromatography (7 mm in particle diameter). ‌Column temperature: A constant temperature of about 409C.‌ Mobile phase: A mixture of a solution of tartaric acid (3 in 2000) and a solution of dipicolinic acid (1 in 3000) (1:1). Flow rate: Adjust so that the retention time of calcium is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, sodium and calcium are eluted in this order with the resolution between these peaks being not less than 4.5. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of calcium is not more than 2.0z. Acid-neutralizing capacity <6.04> Apply to the preparation intended to be used as antacid. Weigh accurately and powder not less than 40 Precipitated Calcium Carbonate Tablets. Perform the test with an accu-rately weighed amount of the powder, equivalent to about 0.25 g of Calcium Carbonate: the amount of 0.1 mol/L hy-drochloric acid VS consumed per 1 g of Precipitated Cal-cium Carbonate is not less than 190 mL.
Weigh accurately and powder not less than 20 Precipitated Calcium Carbonate Tablets. To an accurately weighed portion of the powder, equivalent to about 0.12 g of calcium carbonate (CaCO3), add 20 mL of water, 3 mL of dilute hydrochloric acid, and agitate, if necessary, for 15 minutes with the aid of ultrasonic waves. Then, add 80 mL of water, 15 mL of a solution of potassium hydroxide (1 in 10) and 50 mg of NN indicator, and titrate <2.50> immedi-ately with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 5.005 mg of CaCO3
Prednisolone Sodium Succinate for Injection
注射用プレドニゾロンコハク酸エステルナトリウム
Prednisolone Sodium Succinate for Injection is a preparation for injection which is dissolved before use. It contains not less than 72.
Prednisolone Sodium Succinate for Injection occurs as a white, powder or porous, friable mass. It is freely soluble in water. It is hygroscopic. in a desiccator for 6 hours (in vacuum, phosphorus (V) oxide, 609C), dissolve in methanol to make exactly 25 mL. Pipet 5 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, mix, and use this so-lution as the standard solution. Perform the test with 10 mL of the sample solution and standard solution as directed under Liquid Chromatography according <2.01> to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of prednisolone succinate to that of the inter-nal standard. Amount (mg) of prednisolone sodium succinate (C25H31NaO8)
(1) To 2 mg of Prednisolone Sodium Suc-cinate for Injection add 2 mL of sulfuric acid, and allow to stand for 2 to 3 minutes: a deep red color, without fluores- ＝ MS × QT/QS × 5 × 1.048 Amount (mg) of prednisolone (C21 ＝ MS × QT/QS × 5 × 0.783 H28O5) cence, develops. To this solution add 10 mL of water cau-tiously: the color disappears and a gray, flocculent precipi-tate is formed. Dissolve 0.01 g of Prednisolone Sodium Succinate for Injection in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: an orange to red precipitate is formed. Dissolve 0.1 g of Prednisolone Sodium Succinate for Injection in 2 mL of sodium hydroxide TS, allow to stand for 10 minutes, and filter. Add 1 mL of dilute hydrochloric acid to the filtrate, shake, and filter if necessary. Adjust the solution with diluted ammonia TS (1 in 10) to a pH of about 6, and add 2 to 3 drops of iron (III) chloride TS: a brown precipitate is formed. Prednisolone Sodium Succinate for Injection re-sponds to Qualitative Tests <1.09> (1) for sodium salt. pH <2.54> Dissolve 1.0 g of Prednisolone Sodium Suc-cinate for Injection in 40 mL of water: the pH of the solu-tion is between 6.5 and 7.2.
Clarity and color of solution—Dissolve 0.25 g of Prednisolone Sodium Succinate for Injection in 10 mL of water: the solution is clear and colorless. Loss on drying <2.41> Not more than 2.0z (0.15 g, in vacuum, phosphorus (V) oxide, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 2.4 EU/mg of pred-nisolone (C21H28O5). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take a quantity of sealed containers of Prednisolone Sodium Succinate for Injection, equivalent to about 0.1 g of prednisolone (C21H28O5), and dissolve the contents in a suita-ble amount of diluted methanol (1 in 2), and transfer to a 100-mL volumetric flask. Wash each container with diluted methanol (1 in 2), collect the washings in the volumetric flask, and add diluted methanol (1 in 2) to make volume. Pipet 4 mL of this solution, add diluted methanol (1 in 2) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, mix, and use this so-lution as the sample solution. Separately, weigh accurately about 25 mg of Prednisolone Succinate RS, previously dried MS: Amount (mg) of Prednisolone Succinate RS taken Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted methanol (1 in 2) (1 in 25,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.32 g of tetra n-butylammonium bromide, 3.22 g of disodium hydrogen phosphate dodacahy-drate and 6.94 g of potassium dihydrogen phosphate in 1000 mL of water. To 840 mL of this solution add 1160 mL of methanol. Flow rate: Adjust so that the retention time of predniso-lone succinate is about 15 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, prednisolone succinate and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of prednisolone succinate to that of the in-ternal standard is not more than 1.0z.
(1) Weigh a quantity of powdered Pred-nisolone Tablets, equivalent to 0.05 g of Prednisolone, add 10 mL of chloroform, shake for 15 minutes, and filter. Evaporate the filtrate on a water bath to dryness. Dry the residue at 1059C for 1 hour, and proceed as directed in the Identification (1) under Prednisolone. rately about 10 mg of Prednisolone RS, previously dried at 1059C for 3 hours, dissolve in 10 mL of water and 50 mL of methanol, and add methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand-ard solution at 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of prednisolone (C21H28O5) ＝ MS × AT/AS × V?/V × 1/10 MS: Amount (mg) of Prednisolone RS taken Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Prednisolone Tablets is not less than 70z. Start the test with 1 tablet of Prednisolone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 10 mg of Prednisolone RS, previously dried at 1059C for 3 hours, and dissolve in ethanol (95) to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at the maximum wavelength at about 242 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of prednisolone (C21H28O5) ＝ MS × AT/AS × 1/C × 45 MS: Amount (mg) of Prednisolone RS taken C: Labeled amount (mg) of prednisolone (C21H28O5) in 1 tablet
Weigh accurately and powder not less than 20 Pred-nisolone Tablets using an agate mortar. Weigh accurately a portion of the powder, equivalent to about 5 mg of predniso-lone (C21H28O5), add 1 mL of water, and shake gently. Add exactly 5 mL of the internal standard solution and 15 mL of methanol, and shake vigorously for 20 minutes. To 1 mL of this solution add the mobile phase to make 10 mL, and filter through a membrane filter with pore size of 0.45 mm. Dis- ‌card the first 3 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 25 mg of Prednisolone RS, previously dried at 1059C for 3 hours, dissolve in 50 mL of methanol, add exactly 25 mL of the internal standard solution, and add methanol to make 100 mL. To 1 mL of this solution add the mobile phase to make 10 mL, and use this solution as the standard solu-tion. Proceed as directed in the Assay under Prednisolone with these solutions. tion. Separately, dissolve 20 mg each of prednisolone, corti-sone acetate and hydrocortisone acetate in exactly 10 mL of a mixture of chloroform and methanol (9:1). Pipet 1 mL of this solution, add a mixture of chloroform and methanol (9:1) to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer Amount (mg) of prednisolone (C ＝ MS × QT/QS × 1/5 21H28O5) chromatography. Develop the plate with a mixture of dichloromethane, diethyl ether, methanol and water (385:75:40:6) to a distance of about 15 cm, and air-dry the MS: Amount (mg) of Prednisolone RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 2000).
Probenecid Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of Tablets is not less than 80z. Start the test with 1 tablet of Probenecid Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 14 mg of probenecid (C13H19 NO4S), and use this solution as the sample solution.
(1) Weigh a quantity of powdered Probenecid Tablets, equivalent to 0.5 g of Probenecid, add 50 mL of ethanol (95) and 1 mL of 1 mol/L hydrochloric acid TS, shake, and filter. Evaporate the filtrate on a water bath to about 20 mL. After cooling, collect produced crys-tals, recrystallize with 50 mL of dilute ethanol, and dry at 1059C for 4 hours: it melts <2.60> between 1969C and 2009C. With the crystals so obtained, proceed as directed in the Identification (1) under Probenecid. (2) Determine the absorption spectrum of a solution of the dried crystals obtained in (1) in ethanol (99.5) (1 in 50,000) as directed under Ultraviolet-visible Spectropho- solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 244 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of probenecid (C13H19NO4S) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Probenecid RS taken C: Labeled amount (mg) of probenecid (C13H19NO4S) in 1 tablet
Weigh accurately, and powder not less than 20 Probenecid Tablets. Weigh accurately a portion of the pow- ‌der, equivalent to about 0.25 g of probenecid (C13H19NO4S), add 30 mL of water and 2 mL of 1 mol/L hydrochloric acid TS, shake, add 30 mL of ethanol (99.5), disperse the parti-cles by sonicating, and add ethanol (99.5) to make exactly 100 mL. Centrifuge the solution, pipet 3 mL of the superna-tant liquid, add 1 mL of 1 mol/L hydrochloric acid TS, and add ethanol (99.5) to make exactly 50 mL. Pipet 5 mL of the solution, add ethanol (99.5) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 0.125 g of Probenecid RS, previously dried at 1059C for 4 hours, add 15 mL of water and 1 mL of 1 mol/L hydrochloric acid TS, then add ethanol (99.5) to make ex-actly 50 mL. Pipet 3 mL of this solution, add 1 mL of 1 mol/L hydrochloric acid TS, and add ethanol (99.5) to make exactly 50 mL. Pipet 5 mL of the solution, add ethanol (99.5) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 248 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by mixing 1 mL of 0.1 mol/L hy-drochloric acid TS and sufficient ethanol (99.5) to make ex-actly 50 mL, as the blank. Amount (mg) of probenecid (C13H19NO4S) ＝ MS × AT/AS × 2 MS: Amount (mg) of Probenecid RS taken
To an amount of powdered Probucol Fine Granules, equivalent to 50 mg of Probucol, add 100 mL of methanol, shake, and filter. To 2 mL of the filtrate add methanol to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: the granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Probucol Fine Granules add 70 mL of methanol, shake thor-oughly, and add methanol to make exactly 100 mL. Centri- fuge, pipet V mL of the supernatant liquid, equivalent to about 5 mg of probucol (C31H48O2S2), add exactly 5 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS × 10/V MS: Amount (mg) of Probucol RS taken Internal standard solution—A solution of bis(cis-3,3,5-trimethylcyclohexyl) phthalate in methanol (1 in 250).
Weigh accurately an amount of powdered Probucol Fine Granules, equivalent to about 0.25 g of probucol (C31H48O2S2), add 70 mL of methanol, shake thoroughly, and add methanol to make exactly 100 mL. Centrifuge, pipet 2 mL of the supernatant liquid, add exactly 5 mL of the in-ternal standard solution, add methanol to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Probucol RS, previously dried under reduced pressure at 809C for 1 hour, and dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solu-tion, add exactly 5 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of probucol to that of the internal standard. Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS × 5 MS: Amount (mg) of Probucol RS taken Internal standard solution—A solution of bis(cis-3,3,5-trimethylcyclohexyl) phthalate in methanol (1 in 250). Operating conditions— Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Probucol. Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and probucol are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of probucol to that of the internal standard is not more than 1.0z.
To an amount of powdered Probucol Tablets, equivalent to 50 mg of Probucol, add 100 mL of methanol, shake, and filter. To 2 mL of the filtrate add methanol to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 240 nm and 244 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Shake 1 tablet of Probucol Tablets with a suitable amount of methanol until the tablet is disintegrated, and add metha-nol to make exactly V mL so that each mL of the solution contains about 2.5 mg of probucol (C31H48O2S2). Centrifuge the solution, pipet 2 mL of the supernatant liquid, add ex-actly 5 mL of the internal standard solution, then add meth-anol to make 100 mL, and use this solution as the sample so-lution. Then, proceed as directed in the Assay. Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS × V/20 MS: Amount (mg) of Probucol RS taken Internal standard solution—A solution of bis(cis-3,3,5-trimethylcyclohexyl) phthalate in methanol (1 in 250). Disintegration <6.09> It meets the requirement.
Weigh accurately the mass of 20 Probucol Tablets, and powder the tablets. Weigh accurately a portion of the powder, equivalent to about 0.25 g of probucol (C31H48O2S2), add 70 mL of methanol, shake thoroughly, and add methanol to make exactly 100 mL. Centrifuge, pipet 2 mL of the supernatant liquid, add exactly 5 mL of the in-ternal standard solution, add methanol to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Probucol RS, previously dried under reduced pressure at 809C for 1 hour, and dissolve in methanol to make exactly 20 mL. Pipet 2 mL of this solu-tion, add exactly 5 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of probucol to that of the internal standard. Amount (mg) of probucol (C31H48O2S2) ＝ MS × QT/QS × 5 MS: Amount (mg) of Probucol RS taken Internal standard solution—A solution of bis(cis-3,3,5-trimethylcyclohexyl) phthalate in methanol (1 in 250). Operating conditions— Detector, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Probucol. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of probucol to that of the internal standard is not more than 1.0z.
Procainamide Hydrochloride Injection is a clear, colorless or light yellow liquid. pH: 4.0 – 6.0
(1) To a volume of Procainamide Hydro-chloride Injection, equivalent to 10 mg of Procainamide Hy-drochloride, add 1 mL of dilute hydrochloric acid and water to make 5 mL: the solution responds to Qualitative Tests <1.09> (1) for primary aromatic amines. To a volume of Procainamide Hydrochloride Injec-tion, equivalent to 0.1 g of Procainamide Hydrochloride, add water to make 100 mL. To 1 mL of this solution add water to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 277 nm and 281 nm. Procainamide Hydrochloride Injection responds to Qualitative Tests <1.09> (2) for chloride. Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Dilute an accurately measured volume of Procain-amide Hydrochloride Injection, equivalent to about 0.5 g of procainamide hydrochloride (C13H21N3O.HCl), with 5 mL of hydrochloric acid and water to 50 mL, add 10 mL of potas-sium bromide solution (3 → 10), cool to 159C or lower, and titrate <2.50> with 0.1 mol/L sodium nitrite VS (potentiome-tric titration method or amperometric titration). Each mL of 0.1 mol/L sodium nitrite VS ＝ 27.18 mg of C13H21N3O.HCl
To a quantity of powdered Procainamide Hydrochloride Tablets, equivalent to 1.5 g of Procainamide Hydrochloride, add 30 mL of water, shake well, filter, and use the filtrate as the sample solution. To 0.2 mL of the sam-ple solution add 1 mL of dilute hydrochloric acid and 4 mL of water: the solution responds to Qualitative Tests <1.09> for primary aromatic amines. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Procainamide Hydrochloride Tablets add 3V/5 mL of 0.02 mol/L phosphate buffer solution (pH 3.0), sonicate to disintegrate the tablet completely, add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly V mL so that each mL contains about 2.5 mg of procainamide hydrochloride (C13H21N3O.HCl), and shake for 5 minutes. Centrifuge this solution, pipet 1 mL of the supernatant liq-uid, add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 250 mL, and use this solution as the sample so-lution. Proceed as directed in the Assay. Amount (mg) of procainamide hydrochloride (C13H21N3O.HCl) ＝ MS × AT/AS × V/20 MS: Amount (mg) of procainamide hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Procainamide Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Procainamide Hydrochloride Tablets, withdraw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 2nd fluid for dissolution test to make exactly V? mL so that each mL contains about 7 mg of procainamide hydrochloride (C13H21N3O.HCl), and use this solution as the sample solution. Separately, weigh accurately about 0.125 g of procainamide hydrochloride for assay, pre-viously dried at 1059C for 4 hours, and dissolve in water to make exactly 1000 mL. Pipet 5 mL of this solution, add 2nd fluid for dissolution test to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 278 nm. Dissolution rate (z) with respect to the labeled amount of procainamide hydrochloride (C13H21N3O.HCl) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of procainamide hydrochloride for assay taken C: Labeled amount (mg) of procainamide hydrochloride (C13H21N3O.HCl) in 1 tablet
To 10 Procainamide Hydrochloride Tablets add about 300 mL of 0.02 mol/L phosphate buffer solution (pH 3.0) and sonicate to disintegrate the tablets completely. To this solution add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 500 mL, and stir for 5 minutes. Centri-fuge this solution, pipet V mL of the supernatant liquid, and add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly V? mL so that each mL contains about 10 mg of procainamide hydrochloride (C13H21N3O.HCl). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 50 mg of procainamide hydro-chloride for assay, previously dried at 1059C for 4 hours, dissolve in 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 100 mL. Pipet 2 mL of this solution, add 0.02 mol/L phosphate buffer solution (pH 3.0) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of procainamide in each solution. Amount (mg) of procainamide hydrochloride (C13H21N3O.HCl) ＝ MS × AT/AS × V?/V × 1/10 MS: Amount (mg) of procainamide hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of 0.02 mol/L phosphate buffer solution (pH 3.0) and methanol (9:1). Flow rate: Adjust so that the retention time of procainamide is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of procainamide are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of procainamide is not more than 1.0z.
Procaine Hydrochloride Injection is an aqueous in-jection. It contains not less than 95.
Procaine Hydrochloride Injection is a clear, colorless liquid.
(1) To a volume of Procaine Hydrochlo-ride Injection, equivalent to 0.01 g of Procaine Hydrochlo-ride, add water to make 1000 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 219 nm and 223 nm, and between 289 nm and 293 nm. (2) Procaine Hydrochloride Injection responds to Quali-tative Tests <1.09> (2) for chloride. pH <2.54> 3.3 – 6.0 Bacterial endotoxins <4.01> Less than 0.02 EU/unit. Apply to the preparations intended for intraspinal administration. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
To an exactly measured volume of Procaine Hydro-chloride Injection, equivalent to about 20 mg of procaine hydrochloride (C13H20N2O2.HCl), add the mobile phase to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution and the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of procaine hydro-chloride for assay, previously dried in a desiccator (silica gel) for 4 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution and the mobile phase to make 20 mL, and use this solution as the standard solution. Perform ‌the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of procaine hydrochlo-ride to that of the internal standard. Amount (mg) of procaine hydrochloride (C13H20N2O2.HCl) ＝ MS × QT/QS × 2/5 MS: Amount (mg) of procaine hydrochloride for assay taken Internal standard solution—A solution of caffeine in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column about 6 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust the pH of 0.05 mol/L potassium dihydrogen phosphate TS to 3.0 with phosphoric acid, and add an amount of sodium 1-pentane sulfonate to make a so-lution so that containing 0.1z. To 800 mL of this solution add 200 mL of methanol. Flow rate: Adjust so that the retention time of procaine is about 10 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, procaine and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of procaine to that of the internal standard is not more than 1.0z.
プロクロルペラジンマレイン酸塩錠
(1) Weigh a quantity of powdered Pro-chlorperazine Maleate Tablets, equivalent to 5 mg of Pro-chlorperazine Maleate, add 15 mL of acetic acid (100), shake, and filter. To 5 mL of the filtrate add 3 mL of sulfu-ric acid, and shake: a light red color develops. To this solu-tion add 1 drop of potassium dichromate TS: a green-brown color is produced and changes to brown on standing. Weigh a quantity of powdered Prochlorperazine Maleate Tablets, equivalent to 0.08 g of Prochlorperazine Maleate, add 15 mL of methanol and 1 mL of dimethyla-mine, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 0.08 g of Prochlor-perazine Maleate RS in 15 mL of methanol and 1 mL of dimethylamine, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol and ammonia TS (15:2) to a distance of about 10 cm, and air-dry the plate. Spray evenly palladium (II) chloride TS on the plate: the spots obtained from the sample solution and standard solution show a red-purple color, and has the same Rf value. To a quantity of powdered Prochlorperazine Maleate Tablets, equivalent to 0.04 g of Prochlorperazine Maleate, add 10 mL of 1 mol/L hydrochloric acid TS and 20 mL of diethyl ether, shake, and centrifuge. Transfer the diethyl ether layer to a separator, wash with 5 mL of 0.05 mol/L sulfuric acid TS, and evaporate on a water bath to dryness. Dissolve the residue in 5 mL of sulfuric acid TS, filter, if necessary, and add 1 to 2 drops of potassium permanganate TS: the red color of the test solution is discharged immedi-ately. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Prochlorperazine Maleate Tablets add 3V/5 mL of a mixture of dilute phosphoric acid (1 in 500) and ethanol (99.5) (1:1), sonicate until the tablet is disintegrated, and shake vigorously for 10 minutes. Add exactly V/20 mL of ‌the internal standard solution, and a mixture of dilute phos-phoric acid (1 in 500) and ethanol (99.5) (1:1) to make V mL so that each mL contains about 80 mg of prochlorperazine maleate (C20H24ClN3S.2C4H4O4). Centrifuge this solution, ditions, and calculate the ratios, QT and QS, of the peak area of prochlorperazine to that of the internal standard. Amount (mg) of prochlorperazine maleate (C H ClN S.2C H O ) and use the supernatant liquid as the sample solution. Pro- 20 24 3 4 4 4 ceed as directed in the Assay. Amount (mg) of prochlorperazine maleate (C20H24ClN3S.2C4H4O4) ＝ MS × QT/QS × V/250 MS: Amount (mg) of Prochlorperazine Maleate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 45 minutes of Prochlorperazine Maleate Tablets is not less than 75z. Start the test with 1 tablet of Prochlorperazine Maleate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 9 mg of prochlorperazine maleate (C20H24ClN3S.2C4H4O4), and use this solution as the sample solution. Separately, weigh accu-rately about 18 mg of Prochlorperazine Maleate RS, previ-ously dried at 1059C for 3 hours, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using the dissolu-tion medium as the blank, and determine the absorbances, A T and AS, at 255 nm. Dissolution rate (z) with respect to the labeled amount of prochlorperazine maleate (C20H24ClN3S.2C4H4O4) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Prochlorperazine Maleate RS taken C: Labeled amount (mg) of prochlorperazine maleate (C20H24ClN3S.2C4H4O4) in 1 tablet
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 Prochlorpera-zine Maleate Tablets, and powder in an agate mortar. Weigh accurately a portion of the powder, equivalent to about 8 mg of prochlorperazine maleate (C20H24ClN3S.2C4H4O4), add 60 mL of a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1), and shake vigorously for 10 minutes. Add exactly 5 mL of the internal standard solution, and add a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 100 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 20 mg of Prochlorperazine Maleate RS, previously dried at 1059C for 3 hours, and dissolve in a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make exactly 25 mL. Pipet 10 mL of this solu-tion, add exactly 5 mL of the internal standard solution and a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con- ＝ MS × QT/QS × 2/5 MS: Amount (mg) of Prochlorperazine Maleate RS taken Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of diluted phosphoric acid (1 in 500) and ethanol (99.5) (1:1) (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 257 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted 0.05 mol/L sodium di-hydrogen phosphate TS (1 in 2) and acetonitrile (11:9). Flow rate: Adjust so that the retention time of prochlor-perazine is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, prochlorperazine and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of prochlorperazine to that of the internal stand-ard is not more than 1.0z.
Progesterone Injection is an oily solution for injec-tion. It contains not less than 95.
Progesterone Injection is a clear, colorless to pale yellow, oily liquid. MS: Amount (mg) of Progesterone RS taken Internal standard solution—A solution of testosterone propionate in ethanol (99.5) (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 241 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of acetonitrile and water (7:3). Flow rate: Adjust so that the retention time of progester- one is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, progesterone and the internal standard are eluted in this order with the resolution between these peaks being not less than 9. ‌System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of progesterone to that of the internal standard is not more than 1.0z.‌
To a quantity of Propafenone Hydrochlo-ride Tablets, equivalent to 0.3 g of Propafenone Hydrochlo-ride, add 60 mL of water, and disintegrate by warming. After cooling, centrifuge, and to 3 mL of the supernatant liquid add water to make 500 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 247 nm and 251 nm, and between 302 nm and 306 nm. Separately, determine the both maximal absorbances, A1 and A2, of the solution, the ratio of A1/A2 is between 2.30 and 2.55. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Propafenone Hydrochloride Tablets add 30 mL of a mixture of water and acetonitrile (1:1), shake well to disintegrate, add a mixture of water and acetonitrile (1:1) to make exactly 50 mL, and centrifuge. Pipet V mL of the su-pernatant liquid, equivalent to about 6 mg of propafenone hydrochloride (C21H27NO3.HCl), add exactly 5 mL of the in-ternal standard solution, add methanol to make 50 mL, and use this solution as the sample solution. Proceed as directed in the Assay. ‌Amount (mg) of propafenone hydrochloride (C21H27NO3.HCl) ＝ MS × QT/QS × 10/V MS: Amount (mg) of propafenone hydrochloride for assay taken Internal standard solution—A solution of isopropyl benzo-ate in methanol (1 in 200). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Propafenone Hydrochloride Tables is not less than 75z. Start the test with 1 tablet of Propafenone Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 67 mg of propafenone hydro-chloride (C21H27NO3.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 13 mg of propafenone hydrochloride for assay, previously dried at 1059C for 2 hours, dissolve in water to make exactly 200 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 305 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of propafenone hydrochloride (C21H27NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 450 MS: Amount (mg) of propafenone hydrochloride for assay taken C: Labeled amount (mg) of propafenone hydrochloride (C21H27NO3.HCl) in 1 tablet
To a quantity of Propafenone Hydrochloride Tablets, equivalent to 1.5 g of propafenone hydrochloride (C21H27NO3.HCl), add 70 mL of a mixture of water and acetonitrile (1:1), shake well to disintegrate, shake well for another 5 minutes, add a mixture of water and acetonitrile (1:1) to make exactly 100 mL, and centrifuge. Pipet 4 mL of the supernatant liquid, and add methanol to make exactly 50 mL. Pipet 5 mL of the solution, add exactly 5 mL of the in-ternal standard solution, add methanol to make 50 mL, and Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.6 g of sodium 1-nonanesulfonate and 2.3 g of phosphoric acid in water to make 1000 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 900 mL of the filtrate add 600 mL of acetonitrile. Flow rate: Adjust so that the retention time of propafe-none is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, propafenone and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of propafenone to that of the internal standard is not more than 1.0z.
Propiverine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of propiverine hydrochloride (C23H29NO3.HCl: 403.94). Method of preparation Prepare as directed under Tablets, with Propiverine Hydrochloride. Identification Shake vigorously a quantity of powdered Propiverine Hydrochloride Tablets, equivalent to 50 mg of Propiverine Hydrochloride, with 20 mL of water. Add acetonitrile to make 100 mL, centrifuge, and filter the super-natant liquid, if necessary. Determine the absorption spec-trum of the supernatant liquid or the filtrate under Ultravio-let-visible Spectrophotometry <2.24>: it exhibits a maximum between 257 nm and 261 nm. Purity Related substances—Shake vigorously a quantity of powdered Propiverine Hydrochloride Tablets, equivalent to 50 mg of Propiverine Hydrochloride, with the mobile phase, add the mobile phase to make 100 mL, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the area of the peak, having the relative retention time about 0.28 to propiverine, obtained from the sample solution is not larger than 3/10 times the peak area of propiverine from the standard solution, the area of the peak other than propiver-ine and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of propiverine from the standard solution, and the total area of the peaks other than propiverine from the sample solution is not larger than 7/10 times the peak area of propiverine from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Propiverine Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of propiverine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of propiverine obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propiverine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propiverine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Propiverine Hydrochloride Tablets add the mobile phase, shake vigorously, add the mobile phase to make exactly V mL so that each mL contains about 0.
Shake vigorously a quantity of powdered Propiverine Hydrochloride Tablets, equivalent to 50 mg of Propiverine Hydrochloride, with 20 mL of water. Add acetonitrile to make 100 mL, centrifuge, and filter the super-natant liquid, if necessary. Determine the absorption spec-trum of the supernatant liquid or the filtrate under Ultravio-let-visible Spectrophotometry <2.24>: it exhibits a maximum between 257 nm and 261 nm.
Related substances—Shake vigorously a quantity of powdered Propiverine Hydrochloride Tablets, equivalent to 50 mg of Propiverine Hydrochloride, with the mobile phase, add the mobile phase to make 100 mL, centrifuge, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 15 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the area of the peak, having the relative retention time about 0.28 to propiverine, obtained from the sample solution is not larger than 3/10 times the peak area of propiverine from the standard solution, the area of the peak other than propiver-ine and the peak mentioned above from the sample solution is not larger than 1/5 times the peak area of propiverine from the standard solution, and the total area of the peaks other than propiverine from the sample solution is not larger than 7/10 times the peak area of propiverine from the stand-ard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Propiverine Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of propiverine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of propiverine obtained with 15 mL of this solution is equivalent to 3.5 to 6.5z of that with 15 mL of the standard solution. System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propiverine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of propiverine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Propiverine Hydrochloride Tablets add the mobile phase, shake vigorously, add the mobile phase to make exactly V mL so that each mL contains about 0.1 mg of propiverine hydrochloride (C23H29NO3.HCl), centrifuge, and use the supernatant liquid as the sample solution. Sepa-rately, weigh accurately about 50 mg of Propirevine Hydro-chloride RS, previously dried at 1059C for 1 hour, and dis-solve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Propiverine Hydro-chloride. Amount (mg) of propiverine hydrochloride (C23H29NO3.HCl) ＝ MS × AT/AS × V/500 MS: Amount (mg) of Propiverine Hydrochloride RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, ‌the dissolution rate in 20 minutes of Propiverine Hydrochlo-ride Tablets is not less than 85z. Start the test with 1 tablet of Propiverine Hydrochloride Tablets, withdraw not less than 25 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of propiverine hydrochloride (C23H29NO3.HCl). Pipet 15 mL of this solution, add exactly 2 mL of 0.1 mol/L hydrochloric acid TS, and use this solution as the sample solution. Sepa-rately, weigh accurately about 28 mg of Propiverine Hydro-chloride RS, previously dried at 1059C for 1 hour, and dis-solve in the dissolution medium to make exactly 100 mL. Pipet 4 mL of this solution, and add the dissolution medium to make exactly 100 mL. Further, pipet 15 mL of this solu-tion, add exactly 2 mL of 0.1 mol/L hydrochloric acid TS, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of propiverine in each solution. Dissolution rate (z) with respect to the labeled amount of propiverine hydrochloride (C23H29NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of Propiverine Hydrochloride RS taken C: Labeled amount (mg) of propiverine hydrochloride (C23H29NO3.HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To diluted 0.02 mol/L potassium dihydro-gen phosphate TS (1 → 2) add phosphoric acid, and adjust to pH 2.0. To 560 mL of this solution add 440 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of propirevine is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of propiverine are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above opera-tions conditions, the relative standard deviation of the peak area of propiverine is not more than 2.0z.
Weigh accurately and powder not less than 20 Propiverine Hydrochloride Tablets. Weigh accurately a por-tion of the powder, equivalent to about 50 mg of propiverine hydrochloride (C23H29NO3.HCl), add the mobile phase, shake vigorously, and add the mobile phase to make exactly 100 mL. Centrifuge this solution, pipet 10 mL of the super-natant liquid, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Propiverine Hydrochloride RS, previously dried at 1059C for 1 hour, and dissolve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Propiverine Hydrochloride. Amount (mg) of propiverine hydrochloride (C23H29NO3.HCl) ＝ MS × AT/AS MS: Amount (mg) of Propiverine Hydrochloride RS taken
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 288 nm and 292 nm, and between 317 nm and 321 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Propranolol Hydrochloride Tablets add 20 mL of water, and shake until the tablet is completely disinte-grated. Add 50 mL of methanol, shake vigorously for 10 minutes, then add methanol to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet V mL of the subsequent filtrate, add methanol to make exactly V? mL so that each mL contains about 20 mg of propranolol hydro-chloride (C16H21NO2.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 50 mg of propranolol hydrochloride for assay, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 290 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of propranolol hydrochloride (C16H21NO2.HCl) ＝ MS × AT/AS × V?/V × 1/25 MS: Amount (mg) of propranolol hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Propranolol Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Propranolol Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 10 mg of propranolol hydro-chloride (C16H21NO2.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 50 mg of propranolol hydrochloride for assay, previously dried at 1059C for 4 hours, and dissolve in water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, of the sample solution and standard solution at 290 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>. ‌Dissolution rate (z) with respect to the labeled amount of propranolol hydrochloride (C16H21NO2.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of propranolol hydrochloride for assay taken C: Labeled amount (mg) of propranolol hydrochloride (C16H21NO2.HCl) in 1 tablet
Weigh accurately the mass of not less than 20 Propranolol Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 20 mg of propranolol hydrochloride (C16H21NO2.HCl), add 60 mL of methanol, shake for 10 minutes, and add methanol to make exactly 100 mL. Filter, discard the first 20 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add methanol to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of propranolol hydrochloride for assay, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 50 mL. Pipet 2 mL of this solution, add methanol to make exactly 100 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, of the sample solution and standard solution at 290 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of propranolol hydrochloride (C16H21NO2.HCl) ＝ MS × AT/AS × 2/5 MS: Amount (mg) of propranolol hydrochloride for assay taken
Propylthiouracil Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of propylthiouracil (C7H10N2OS: 170.23). Method of preparation Prepare as directed under Tablets, with Propylthiouracil. Identification To a quantity of powdered Propylthiouracil Tablets, equivalent to 0.3 g of Propylthiouracil, add 5 mL of ammonia TS, allow to stand for 5 minutes with occasional shaking, add 10 mL of water, and centrifuge. To the super-natant liquid add acetic acid (31), collect the precipitate pro-duced, recrystallize from water, and dry at 1059C for 1 hour: it melts <2.60> between 2189C and 2219C. Proceed with the residue as directed in the Identification under Propylthiouracil. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Propylthiouracil Tablets add 3V/4 mL of 2nd fluid for dissolution test, sonicate until the tablet is dis-integrated, and add 2nd fluid for dissolution test to make ex-actly V mL so that each mL contains about 0.
To a quantity of powdered Propylthiouracil Tablets, equivalent to 0.3 g of Propylthiouracil, add 5 mL of ammonia TS, allow to stand for 5 minutes with occasional shaking, add 10 mL of water, and centrifuge. To the super-natant liquid add acetic acid (31), collect the precipitate pro-duced, recrystallize from water, and dry at 1059C for 1 hour: it melts <2.60> between 2189C and 2219C. Proceed with the residue as directed in the Identification under Propylthiouracil. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Propylthiouracil Tablets add 3V/4 mL of 2nd fluid for dissolution test, sonicate until the tablet is dis-integrated, and add 2nd fluid for dissolution test to make ex-actly V mL so that each mL contains about 0.25 mg of propylthiouracil (C7H10N2OS). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, dis-card the first 5 mL of the filtrate, pipet 2 mL of the subse-quent filtrate, add 2nd fluid for dissolution test to make ex-actly 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of propylthiouracil (C7H10N2OS) ＝ MS × AT/AS × V/200 MS: Amount (mg) of propylthiouracil for assay taken Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Propylthiouracil Tablets is not less than 80z. Start the test with 1 tablet of Propylthiouracil Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 5.6 mg of propylthiouracil (C7H10N2OS), and use this solution as the sample solution. Separately, weigh about 50 mg of propylthiouracil for assay, previously dried at 1059C for 3 hours, and dissolve in the dissolution medium to make exactly 1000 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbance at 274 mm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of propylthiouracil (C7H10N2OS) ＝ MS × AT/AS × V?/V × 1/C × 9 MS: Amount (mg) of propylthiouracil for assay taken C: Labeled amount (mg) of propylthiouracil (C7H10N2OS) in 1 tablet
Weigh accurately the mass of not less than 20 Propylthiouracil Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of propylthiouracil (C7H10N2OS), add 150 mL of 2nd fluid for dissolution test, disperse finely the particles by sonicating, and add 2nd fluid for dissolution test to make exactly 200 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add 2nd fluid for dissolution test to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 50 mg of propylthiouracil for assay, previously dried at 1059C for 2 hours, and dissolve in 2nd fluid for dissolu-tion test to make exactly 200 mL. Pipet 2 mL of this solu-tion, add 2nd fluid for dissolution test to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbance at 274 nm, AT and AS, of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of propylthiouracil (C7H10N2OS) ＝ MS × AT/AS MS: Amount (mg) of propylthiouracil for assay taken
Protamine Sulfate Injection is an aqueous injection. It contains not less than 92.
Protamine Sulfate Injection is a colorless liq-uid. It is odorless or has the odor of preservatives.
(1) Dilute a volume of Protamine Sulfate Injection, equivalent to 1 mg of Protamine Sulfate, with water to make 2 mL, and proceed as directed in the Identifi-cation (1) under Protamine Sulfate. (2) Dilute a volume of Protamine Sulfate Injection, equivalent to 5 mg of Protamine Sulfate, with water to make 1 mL, and proceed as directed in the Identification (2) under Protamine Sulfate. pH <2.54> 5.0 – 7.0 Bacterial endotoxins <4.01> Less than 6.0 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Protein—Pipet a volume of Protamine Sulfate Injection, equivalent to about 10 mg of Protamine Sulfate, transfer to a Kjeldahl flask, evaporate on a water bath to dryness with the aid of a current of air, determine the nitro-gen as directed under Nitrogen Determination <1.08>, and calculate the amount of protein by converting 0.24 mg of nitrogen (N: 14.01) to 1 mg of protein. Heparin-binding activity—Proceed the test as di-rected in the Heparin-binding capacity under Protamine Sulfate, changing the sample solution (a) as below, and de-termine the amount of heparin bound to 1 mg of protein by dividing by the amount of protein. Sample solution (a)—Pipet a volume of Protamine Sulfate Injection, equivalent to 15.0 mg of Protamine Sul-fate, and add water to make exactly 100 mL. Repeat this procedure two more times, and designate the solutions so obtained as the sample solutions (a1), (a2) and (a3).
精製ヒアルロン酸ナトリウム注射液
Purified Sodium Hyaluronate Injection is an aque-ous injection. It contains not less than 90.
Purified Sodium Hyaluronate Injection occurs as a clear, colorless, and viscous liquid.
(1) To 1 mL of a solution of Purified So-dium Hyaluronate Injection (1 in 10) add 6 mL of sulfuric acid, and heat in a water bath for 10 minutes. After cooling, add 0.2 mL of carbazole TS, and allow to stand at room temperature: a red to red-purple color develops. To 1 mL of a solution of Purified Sodium Hyaluronate Injection (1 in 10) add 0.2 mL of 1 mol/L acetic acid-sodium acetate buffer solution (pH 6.0) and 5 units of hyaluronidase, and allow to stand at 509C for 1 hour. To this solution add 1 mL of a solution of dipotassium tetraborate tetrahydrate (1 in 20), heat in a water bath for Average molecular mass ＝ 0.78 7 minutes. After cooling, add 6 mL of acetic acid (100) and In the case of the labeled average molecular mass of between 1,500,000 and 3,900,000. Calculate the average molecular mass of Purified Sodium Hyaluronate according to the following equation: it is be-tween 1,500,000 and 3,900,000. For [h], use the maximum viscosity under Viscosity. Ø[h] × 105 » 1 2.4 mL of 4-dimethylaminobenzaldehyde-hydrochloric acid-acetic acid TS, and allow to stand at room temperature: a yellowish red to red color develops. To 1 mL of a solution of Purified Sodium Hyaluronate Injection (1 in 10) add 2 to 3 drops of a solution of cetylpyridinium chloride monohydrate (1 in 20): a white precipitate is formed. Average molecular mass ＝ 22.8 0.816 Viscosity <2.53> Apply to the preparation which labeled average mo- ‌lecular mass of sodium hyaluronate is 600,000 to 1,200,000. Weigh accurately an amount of Purified Sodium Hyaluronate Injection, equivalent to about 10 mg of Puri-fied Sodium Hyaluronate, add 0.2 mol/L sodium chloride TS to make exactly 20 mL, and use this solution as the sam-ple solution. Perform the test with the sample solution at 30 ± 0.19C according to Method 1, using an Ubbelohde-type
Weigh accurately an amount of Purified Sodium Hyaluronate Injection, equivalent to about 10 mg of Puri-fied Sodium Hyaluronate, and add 0.2 mol/L sodium chlo-ride TS to make exactly 20 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay under Purified Sodium Hyaluronate. viscometer showing the downflowing time of 0.2 mol/L so- dium chloride TS is between 200 and 300 seconds. Calculate Content (mg) of sodium hyaluronate [(C14H20 NNaO11)n] the intrinsic viscosity [h] according to the following equa-tion, where c is the content obtained in the Assay expressed as the concentration (g/dL): 11.8 – 19.5 dL/g. per mL of Purified Sodium Hyaluronate Injection ＝ MS/MT × AT/AS × 1/5 × r × 2.279 MS: Amount (mg) of D-Glucuronolactone RS taken [h] ＝ 2(hsp － ln h rel)/c × 0.87 ＋ 1.33 MT: Amount (g) of Purified Sodium Hyaluronate Injec-tion taken hsp (specific viscosity) ＝ hrel － 1 hrel (relative viscosity) ＝ t/t0 Apply to the preparation which labeled average molecular mass of sodium hyaluronate is 1,500,000 to 2,000,000. Weigh accurately an amount of Purified Sodium Hyaluronate Injection, equivalent to about 4 mg of Purified Sodium Hyaluronate, add 0.2 mol/L sodium chloride TS to make exactly 20 mL, and use this solution as the sample solution. Perform the test with the sample solution at 30 ± 0.19C according to Method 1, using an Ubbelohde-type vis-cometer showing the downflowing time of 0.2 mol/L sodium chloride TS is between 200 and 300 seconds. Calculate the intrinsic viscosity [h] according to the following equation: 24.5 – 31.5 dL/g. r: Density (g/mL) of Purified Sodium Hyaluronate Injec-tion measured as directed under Determination of Specific Gravity and Density <2.56>
Purified Sodium Hyaluronate Ophthalmic Solution
精製ヒアルロン酸ナトリウム点眼液
Purified Sodium Hyaluronate Ophthalmic Solution 1 － 0.432・ln hrel [h] ＝ (1 － hrel (relative viscosity) ＝ t/t0 )/(0.0108 × M ) is an aqueous ophthalmic preparation. It contains not less than 90.
Purified Sodium Hyaluronate Ophthalmic So-lution occurs as a clear, colorless, and viscous liquid.
(1) To 1 mL of Purified Sodium Hyaluronate Ophthalmic Solution add 0.2 mL of 1 mol/L acetic acid-sodium acetate buffer solution (pH 6.0) and 5 units of hyaluronidase, and allow to stand at 509C for 1 hour. Add 1 mL of a solution of dipotassium tetraborate tetrahydrate (1 in 20), and heat in a water bath for 7 minutes. After cooling, add 6 mL of acetic acid (100) and 2.4 mL of 4-dimethylaminobenzaldehyde-hydrochloric acid-acetic acid TS, and allow to stand at room temperature: a yellowish red to red color develops. (2) To 1 volume of Purified Sodium Hyaluronate Oph-thalmic Solution, equivalent to 7.5 mg of purified sodium hyaluronate [(C14H20NNaO11)n], add 2 volumes of acetone, shake well, and centrifuge at 3000 rpm for 10 minutes. Re-move the acetone, wash the precipitate with a mixture of ace-tone and water (5:1), dry the precipitate under reduced pres-sure (not exceeding 0.67 kPa) at 609C for 5 hours using phosphorus (V) oxide as a desiccant, and determine the in- Average molecular mass ＝ 0.78 frared absorption spectrum as directed in ATR method Apply to the preparation which labeled average mo-lecular mass of sodium hyaluronate is 1,500,000 to 2,000,000. Calculate the average molecular mass by the following equa-tion, where [h] is the intrinsic viscosity obtained in the Vis-cosity: it is 1,500,000 to 2,000,000. Ø[h] × 105 » 1 under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 1605 cm－1, 1404 cm－1, 1375 cm－1, 1150 cm－1, 1025 cm－1 and 945 cm－1. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law. Average molecular mass ＝ 22.8 0.816 ‌Viscosity <2.53> Perform the test according to Method 1 at 30 ± 0.19C: the kinematic viscosity is 3.0 to 4.0 mm2/s or 17 to 30 mm2/s. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. Average molecular mass When determined by the follow-ing method it is between 600,000 and 1,200,000. Determination of viscosity <2.53> Weigh accurately an amount of Purified Sodium Hyaluronate Ophthalmic Solution, equivalent to about 15 mg of purified sodium hyaluronate [(C14H20NNaO11)n], add 0.2 mol/L sodium chloride TS to make exactly 30 mL, and use this solution as the sample solution. Perform the test with the sample solution according to Method 1 at 30 ± 0.19C, using an Ubbelohde-type viscometer with the down-flowing time of 0.2 mol/L sodium chloride TS is between 200 and 300 seconds. Calculate the intrinsic viscosity [h] according to the following equation, where c is the content obtained in the Assay expressed as the concentration (g/dL): 11.8 – 19.5 dL/g. [h] ＝ 2(hsp － ln hrel)/c × 0.87 ＋ 1.33 hsp (specific viscosity) ＝ hrel － 1 hrel (relative e viscosity) ＝ t/t0 Calculation of average molecular mass Calculate by the following equation, using the intrinsic viscosity obtained in (i) for [h]. Ø[h] × 105 » 1 Flow rate: Adjust so that the retention time of hyaluronic acid is about 5 minutes. System suitability— System performance: Dissolve 50 mg of purified sodium hyaluronate in 50 mL of sodium chloride solution (9 in 1000). To 1 mL of this solution and 2 mL of a solution of e-aminocaproic acid (1 in 500) add the mobile phase to make 20 mL, and use this solution as the solution for system suitability test. When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, hyaluronic acid and e-aminocaproic acid are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hyaluronic acid is not more than 2.0z.
Powder Quetiapine Fumarate Fine Granules. To a portion of the powder, equivalent to 12.5 mg of quetia-pine (C21H25N3O2S), add 60 mL of a mixture of water and acetonitrile (1:1), shake, then add the same mixture to make 100 mL, and filter. To 3 mL of the filtrate add the same mix-ture to make 25 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 290 nm and 296 nm. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Quetiapine Fumarate Fine Granules is not less than 80z. Start the test with an accurately weighed amount of Quetiapine Fumarate Fine Granules, equivalent to about 0.1 g of quetiapine (C21H25N3O2S), withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 1.0 mm. Discard not less than 5 mL of the first filtrate, pipet 4 mL of the subsequent filtrate, add water to make exactly 20 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 32 mg of Quetia-pine Fumarate RS (separately determine the water <2.48> in the same manner as Quetiapine Fumarate), and dissolve in water to make exactly 50 mL. Pipet 4 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and A S, at 289 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of quetiapine (C21H25N3O2S) ＝ MS/MT × AT/AS × 1/C × 360 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis MT: Amount (g) of Quetiapine Fumarate Fine Granules taken C: Labeled amount (mg) of quetiapine (C21H25N3O2S) in 1 g
To an accurately weighed amount of Quetiapine Fumarate Fine Granules, equivalent to about 0.25 g of quetiapine (C21H25N3O2S), add 10 mL of water, and allow to stand for 15 minutes. Add 100 mL of the mobile, shake for 15 minutes, then add the mobile phase to make exactly 200 mL, and stir the solution thoroughly. After standing for 15 minutes, pipet 6 mL of the supernatant liquid, add the mo-bile phase to make exactly 50 mL, and filter through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 17 mg of Quetiapine Fumarate RS (separately determine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase, dissolve by sonicating, then add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 50 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of quetiapine in each solution. Amount (mg) of quetiapine (C21H25N3O2S) ＝ MS × AT/AS × 50/3 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, diammonium hydrogen phosphate solution (33 in 12,500) and acetonitrile (54:39:7). Flow rate: Adjust so that the retention time of quetiapine is about 15 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 1.0z.
Quetiapine Fumarate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of quetiapine (C21H25N3O2S: 383.51). Method of preparation Prepare as directed under Tablets, with Quetiapine Fumarate. Identification Powder Quetiapine Fumarate Tablets. To a portion of the powder, equivalent to about 12.5 mg of quetiapine (C H N O S), add 5 mL of water, shake, add 60 21 25 3 2 mL of a mixture of water and acetonitrile (1:1), shake, then add the same mixture to make 100 mL, and filter. To 3 mL of the filtrate add the same mixture to make 25 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 290 nm and 296 nm. Purity Related substances—To 10 Quetiapine Fumarate Tablets add 10 mL of water, allow to stand for 15 minutes, then shake for 25 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly 200 mL. Stir this solution for 4 hours, and allow to stand for 15 minutes. Pipet 3 mL of this solution, add the mobile phase so that each mL con- tains about 0.15 mg of quetiapine (C H N O S), and filter card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 18 mg of Quetiapine Fumarate RS (separately deter-mine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase, dissolve by sonicating, then add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of quetiapine (C21H25N3O2S) ＝ MS × AT/AS × V/16 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis 21 25 3 2 through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak with the relative retention time of about 0.6 to quetiapine obtained from the sample solution is not larger than 1/5 times the peak area of quetiapine from the standard solution, the area of the peak other than quetiapine and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of quetiapine from the stand-ard solution, and the total area of the peaks other than quetiapine and the peak with the relative retention time of about 0.6 is not larger than 1/5 times the peak area of quetiapine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.3 times as long as the retention time of quetiapine, beginning after the peak of fu-maric acid. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of quetiapine obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Quetiapine Fumarate Tablets add 5 mL of water, allow to stand for 15 minutes, then shake for 25 minutes, add 30 mL of a mixture of water and acetonitrile (1:1), shake, and add the same mixture to make exactly 50 mL. Stir this solution for 4 hours, and allow to stand for 15 minutes. To exactly 8 mL of this solution, add the mobile phase to make exactly V mL so that each mL contains about 0.
Powder Quetiapine Fumarate Tablets. To a portion of the powder, equivalent to about 12.5 mg of quetiapine (C H N O S), add 5 mL of water, shake, add 60 21 25 3 2 mL of a mixture of water and acetonitrile (1:1), shake, then add the same mixture to make 100 mL, and filter. To 3 mL of the filtrate add the same mixture to make 25 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 290 nm and 296 nm.
Related substances—To 10 Quetiapine Fumarate Tablets add 10 mL of water, allow to stand for 15 minutes, then shake for 25 minutes, and add a mixture of water and acetonitrile (1:1) to make exactly 200 mL. Stir this solution for 4 hours, and allow to stand for 15 minutes. Pipet 3 mL of this solution, add the mobile phase so that each mL con- tains about 0.15 mg of quetiapine (C H N O S), and filter card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 18 mg of Quetiapine Fumarate RS (separately deter-mine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase, dissolve by sonicating, then add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. Amount (mg) of quetiapine (C21H25N3O2S) ＝ MS × AT/AS × V/16 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis 21 25 3 2 through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak with the relative retention time of about 0.6 to quetiapine obtained from the sample solution is not larger than 1/5 times the peak area of quetiapine from the standard solution, the area of the peak other than quetiapine and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of quetiapine from the stand-ard solution, and the total area of the peaks other than quetiapine and the peak with the relative retention time of about 0.6 is not larger than 1/5 times the peak area of quetiapine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.3 times as long as the retention time of quetiapine, beginning after the peak of fu-maric acid. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of quetiapine obtained with 50 mL of this solution is equivalent to 7 to 13z of that with 50 mL of the standard solution. System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 2.0z. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Quetiapine Fumarate Tablets add 5 mL of water, allow to stand for 15 minutes, then shake for 25 minutes, add 30 mL of a mixture of water and acetonitrile (1:1), shake, and add the same mixture to make exactly 50 mL. Stir this solution for 4 hours, and allow to stand for 15 minutes. To exactly 8 mL of this solution, add the mobile phase to make exactly V mL so that each mL contains about 0.16 mg of quetiapine (C21H25N3O2S), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis- Dissolution <6.10> When the test is performed at 50 revolu- tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Quetiapine Fumarate Tablets is not less than 75z. Start the test with 1 tablet of Quetiapine Fumarate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the mobile phase to make exactly V? mL so that each mL contains about 14 mg of quetiapine (C21H25N3O2S), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Quetiapine Fumarate RS (separately determine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase, sonicate to dissolve, and add the mobile phase to make exactly 100 mL. Pipet 8 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of quetiapine in each solution. Dissolution rate (z) with respect to the labeled amount of quetiapine (C21H25N3O2S) ＝ MS × AT/AS × V?/V × 1/C × 72 × 0.869 MS: Amount (mg) of Quetiapine Fumarate RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of quetiapine (C21H25N3O2S) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4 mm in inside diameter and 8 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol, a solution of diam-monium hydrogen phosphate (33 in 12,500) and acetonitrile (54:39:7). Flow rate: Adjust so that the retention time of quetiapine is about 4 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quetiapine are not less than 1400 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the peak area of quetiapine is not more than 2.0z.
To 20 Quetiapine Fumarate Tablets add 20 mL of water, allow to stand for 15 minutes, shake for 25 minutes, and add a mixture of water and acetonitrile (1:1) to make ex-actly 500 mL. Stir the solution for 4 hours. After standing for 15 minutes, pipet 4 mL of this solution, and add the mo-bile phase to make exactly V mL so that each mL contains about 0.16 mg of quetiapine (C21H25N3O2S). Filter this solu-tion through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 18 mg of Quetiapine Fumarate RS (separately determine the water <2.48> in the same manner as Quetiapine Fumarate), add 60 mL of the mobile phase, dissolve by sonicating, then add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of quetiapine in each solution. Quinapril Hydrochloride
Quinapril Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of quinapril hydrochloride (C25H30N2O5.HCl: 474.98). Method of preparation Prepare as directed under Tablets, with Quinapril Hydrochloride. Identification To a quantity of powdered Quinapril Hydro-chloride Tablets, equivalent to 20 mg of Quinapril Hydro-chloride, add 10 mL of methanol, shake for 5 minutes, and centrifuge. To 5 mL of the supernatant liquid add 0.5 mL of dilute hydrochloric acid, and add methanol to make 20 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 256 nm and 260 nm, between 262 nm and 266 nm, and between 269 nm and 273 nm. Purity To an amount of the supernatant liquid obtained in the Assay add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) so that each mL contains 0.
To a quantity of powdered Quinapril Hydro-chloride Tablets, equivalent to 20 mg of Quinapril Hydro-chloride, add 10 mL of methanol, shake for 5 minutes, and centrifuge. To 5 mL of the supernatant liquid add 0.5 mL of dilute hydrochloric acid, and add methanol to make 20 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 256 nm and 260 nm, between 262 nm and 266 nm, and between 269 nm and 273 nm.
To an amount of the supernatant liquid obtained in the Assay add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) so that each mL contains 0.2 mg of Quinapril Hydrochloride, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method: the area of the peak having the relative retention time of about 0.5 to quinapril obtained from the sample solution is not larger than 2 times the peak area of quinapril from the standard solution, and the area of the peak, having the relative retention time of about 2.0 from the sample solution is not larger than the peak area of quinapril from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (2) under Quinapril Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quinapril are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quinapril is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Quinapril Hydrochloride Tablets add 3V/ 5 mL of a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1), shake vigorously to disintegrate the tablet, shake again for 10 minutes, add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly V mL so that each mL contains about 0.22 mg of quinapril hydrochloride (C25H30N2O5.HCl), and centri-fuge. Pipet 15 mL of the supernatant liquid, add exactly 2 mL of the internal standard solution, add a mixture of phos-phate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of quinapril hydrochloride (C25H30N2O5.HCl) ＝ MS × QT/QS × V/120 MS: Amount (mg) of quinapril hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) (1 in 800). Dissolution <6.10> When the test is performed at 75 revolu- tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Quinapril Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Quinapril Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly V? mL so that each mL contains about 1.2 mg of quinapril hydrochloride (C25H30N2O5.HCl), and use this solution as the sample solution. Separately, weigh accurately about 24 mg of quinapril hydrochloride for assay (separately, determine the water <2.48> in the same manner as Quinapril Hydrochloride), and dissolve in a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liq-uid chromatography (1:1) to make exactly 200 mL. Pipet 2 tant liquid, equivalent to about 6.5 mg of quinapril hydro-chloride (C25H30N2O5.HCl), add exactly 4 mL of the internal standard solution, add a mixture of phosphate buffer solu-tion (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of quinapril hydrochloride for assay (separately, determine the water <2.48> in the same manner as Quinapril Hydrochlo-ride), and dissolve in a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly 100 mL. Pipet 25 mL of this solution, add ex-actly 4 mL of the internal standard solution, add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions. Calculate the ratios, QT and QS, of the peak area of quinapril to that of the internal standard. mL of this solution, add a mixture of phosphate buffer solu-tion (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each Amount (mg) of quinapril hydrochloride (C in 1 tablet ＝ MS × QT/QS × 1/V × 25/4 25H30N2O5.HCl) of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of quinapril in each solution. Dissolution rate (z) with respect to the labeled amount of quinapril hydrochloride (C25H30N2O5.HCl) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of quinapril hydrochloride for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of quinapril hydrochloride (C25H30N2O5.HCl) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: While keeping the temperature not below 259C, adjust to pH 2.0 of 0.1 mol/L potassium dihydrogen phosphate TS with perchloric acid. To 1000 mL of this solu-tion add 1500 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of quinapril is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of quinapril are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of quinapril is not more than 2.0z.
To 20 Quinapril Hydrochloride Tablets add 300 mL of a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1), shake vigor-ously to disintegrate the tablets, shake again for 10 minutes, and add a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) to make exactly 500 mL. Centrifuge this solution, pipet V mL of the superna- MS: Amount (mg) of quinapril hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of phosphate buffer solution (pH 7.0) and acetonitrile for liquid chromatography (1:1) (1 in 800). Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 214 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: While keeping the temperature not below 259C, adjust to pH 2.0 of 0.2 mol/L potassium dihydrogen phosphate TS with perchloric acid. To 1000 mL of this solu-tion add 1000 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of quinapril is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, quinapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of quinapril to that of the internal standard is not more than 1.0z.
Rebamipide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of rebamipide (C19H15ClN2O4: 370.79). Method of preparation Prepare as directed under Tablets, with Rebamipide. Identification To a quantity of powdered Rebamipide Tablets, equivalent to 30 mg of Rebamipide, add 5 mL of a mixture of methanol and ammonia solution (28) (9:1), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 30 mg of rebami-pide for assay in 5 mL of a mixture of methanol and ammo-nia solution (28) (9:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol and formic acid (75:25:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Rebamipide Tablets add 10 mL of water, shake well for 10 minutes, add exactly 10 mL of the internal standard solution, add 10 mL of N,N-dimethylformamide, shake well for 5 minutes, and add N,N-dimethylformamide to make 50 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, equivalent to 3 mg of rebamipide (C19H15ClN2O4), and add 20 mL of N,N-dimethylformamide and water to make 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, dis-card the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 0.1 g of rebamipide for assay, previously dried at 1059C for 2 hours, dissolve in N,N-dimethylformamide, and add exactly 10 mL of the internal standard solution and N,N-dimethylformamide to make 50 mL. Pipet 1.5 mL of this solution, add 20 mL of N,N-dimethylformamide, add water to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay. ‌Amount (mg) of rebamipide (C19H15ClN2O4) ＝ MS × QT/QS × 3/2V MS: Amount (mg) of rebamipide for assay taken Internal standard solution—A solution of acetanilide in N,N-dimethylformamide (1 in 150). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of diluted disodium hydrogen phosphate-citric acid buffer solution (pH 6.0) (1 in 4), as the dissolution medium, the dissolution rate in 60 minutes of Rebamipide Tablets is not less than 75z. Start the test with 1 tablet of Rebamipide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg of rebamipide (C19H15ClN2O4), and use this solution as the sample solution.
To a quantity of powdered Rebamipide Tablets, equivalent to 30 mg of Rebamipide, add 5 mL of a mixture of methanol and ammonia solution (28) (9:1), shake for 10 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 30 mg of rebami-pide for assay in 5 mL of a mixture of methanol and ammo-nia solution (28) (9:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatogra-phy. Develop the plate with a mixture of ethyl acetate, meth-anol and formic acid (75:25:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution has the same Rf value as the spot from the standard solution. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Rebamipide Tablets add 10 mL of water, shake well for 10 minutes, add exactly 10 mL of the internal standard solution, add 10 mL of N,N-dimethylformamide, shake well for 5 minutes, and add N,N-dimethylformamide to make 50 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, equivalent to 3 mg of rebamipide (C19H15ClN2O4), and add 20 mL of N,N-dimethylformamide and water to make 50 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.5 mm, dis-card the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 0.1 g of rebamipide for assay, previously dried at 1059C for 2 hours, dissolve in N,N-dimethylformamide, and add exactly 10 mL of the internal standard solution and N,N-dimethylformamide to make 50 mL. Pipet 1.5 mL of this solution, add 20 mL of N,N-dimethylformamide, add water to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay. ‌Amount (mg) of rebamipide (C19H15ClN2O4) ＝ MS × QT/QS × 3/2V MS: Amount (mg) of rebamipide for assay taken Internal standard solution—A solution of acetanilide in N,N-dimethylformamide (1 in 150). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of diluted disodium hydrogen phosphate-citric acid buffer solution (pH 6.0) (1 in 4), as the dissolution medium, the dissolution rate in 60 minutes of Rebamipide Tablets is not less than 75z. Start the test with 1 tablet of Rebamipide Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg of rebamipide (C19H15ClN2O4), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of rebamipide for assay, previously dried at 1059C for 2 hours, and dissolve in N,N-dimethylformamide to make exactly 25 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank, and deter- mine the absorbances, AT and AS, at 326 nm. Dissolution rate (z) with respect to the labeled amount of rebamipide (C19H15ClN2O4) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of rebamipide for assay taken C: Labeled amount (mg) of rebamipide (C19H15ClN2O4) in 1 tablet
To 10 Rebamipide Tablets add exactly V/5 mL of the internal standard solution and 50 mL of N,N-dimethyl-formamide, and disintegrate the tablets by sonicating. Shake this solution for 5 minutes, add N,N-dimethylformamide to make V mL so that each mL contains about 10 mg of re-bamipide (C19H15ClN2O4). Centrifuge this solution, and to 5 mL of the supernatant liquid add N,N-dimethylformamide to make 50 mL. To 2 mL of this solution add 20 mL of N,N-dimethylformamide and water to make 50 mL. Filter, if nec-essary, through a membrane filter with a pore size not exceeding 0.5 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 0.1 g of rebamipide for assay, previously dried at 1059C for 2 hours, dissolve in N,N-dimethylformamide, and add exactly 2 mL of the inter-nal standard solution and N,N-dimethylformamide to make 100 mL. To 2 mL of this solution, add 20 mL of N,N-dimethylformamide and water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and Q S, of the peak area of rebamipide to that of the internal standard. Amount (mg) of rebamipide (C19H15ClN2O4) ＝ MS × QT/QS × V/100 MS: Amount (mg) of rebamipide for assay taken Internal standard solution—A solution of acetanilide in N,N-dimethylformamide (1 in 20). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 300 mL of phosphate buffer solution (pH 6.2) add 750 mL of water. To 830 mL of this solution add 170 mL of acetonitrile. Flow rate: Adjust so that the retention time of rebamipide is about 20 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and rebamipide are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of rebamipide to that of the internal standard is not more than 1.0z.
Reserpine Injection is an aqueous injection. It contains not less than 90.
Reserpine Injection is a clear, colorless or pale yellow liquid. pH: 2.5 – 4.0
Measure a volume of Reserpine Injection, equivalent to 1.5 mg of Reserpine, add 10 mL of diethyl ether, shake for 10 minutes, and take the aqueous layer. If necessary, add 10 mL of diethyl ether to the aqueous layer, and shake for 10 minutes to repeat the process. To the aqueous layer add water to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 265 nm and 269 nm. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay Measure exactly a volume of Reserpine Injection, equivalent to about 4 mg of reserpine (C33H40N2O9). Sepa-rately, weigh accurately about 4 mg of Reserpine RS, previ-ously dried in vacuum at 609C for 3 hours. Transfer them to separate separator, add 10 mL each of water and 5 mL each of ammonia TS, and extract with one 20-mL portion of chlo-roform, then with three 10-mL portions of chloroform with shaking vigorously. Combine the chloroform extracts, wash with two 50-mL portions of diluted hydrochloric acid (1 in 1000), and combine the washings. Then wash the chloroform extract with two 50-mL portions of a solution of sodium hydrogen carbonate (1 in 100), and combine the all wash-ings. Extract the combined washing with two 10-mL portions of chloroform, and combine the washings with the former chloroform extract. Transfer the chloroform solution to a 100-mL volumetric flask through a pledget of absorbent cot-ton previously wetted with chloroform, wash with a small amount of chloroform, dilute with chloroform to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Determine the absorbances, AT and AS, of the sample solution and the standard solution, respectively, at 295 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>.‌ Amount (mg) of reserpine (C33H40N2O9) ＝ MS × AT/AS MS: Amount (mg) of Reserpine RS taken
Take a portion of powdered Reserpine Tablets, equivalent to 0.4 mg of Reserpine, add 20 mL of acetonitrile, shake for 30 minutes, and centrifuge. Determine the absorption spectrum of the supernatant liquid as directed under Ultraviolet-visible Spectrophotometry <2.24>: it ex-hibits maxima between 265 nm and 269 nm, and between 294 nm and 298 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure without exposure to light, using light-resistant vessels. To 1 tablet of Reserpine Tablets add 2 mL of water, disintegrate by warming at 509C for 15 minutes while shaking. After cooling, add exactly 2 mL of the internal standard solution per 0.1 mg of reserpine (C33H40N2O9), add 2 mL of acetonitrile, warm at 509C for 15 minutes while shaking, and after cooling, add water to make 10 mL. Centrifuge the solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 10 mg of Reserpine RS, previously dried at 609C in vacuum for 3 hours, dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution add exactly 10 mL of the internal standard solution, 5 mL of acetonitrile and water to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Reserpine. Amount (mg) of reserpine (C33H40N2O9) ＝ MS × QT/QS × C/10 MS: Amount (mg) of Reserpine RS taken C: Labeled amount (mg) of reserpine (C33H40N2O9) in 1 tablet Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 50,000). Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Paddle method, using 500 mL of a solution prepared by dissolving 1 g of polysor- ‌bate 80 in diluted dilute acetic acid (1 in 200) to make 20 L as the dissolution medium, the dissolution rate in 30 minutes of Reserpine Tablets is not less than 70z. Start the test with 1 tablet of Reserpine Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, filter through a filter laminated with polyester fibers, discard not less than 10 mL of the first fil-trate, and use the subsequent filtrate as the sample solution. Separately, dry Reserpine RS at 609C in vacuum for 3 hours, weigh accurately an amount 100 times the labeled amount of Reserpine Tablets, dissolve in 1 mL of chloroform and 80 mL of ethanol (95), and add the dissolution medium to make exactly 200 mL. Pipet 1 mL of this solution, add the dissolu-tion medium to make exactly 250 mL, and use this solution as the standard solution. Pipet 5 mL each of the sample solution and standard solution, transfer to glass-stoppered brown test tubes T and S, respectively, add exactly 5 mL each of ethanol (99.5), shake well, add exactly 1 mL each of diluted vanadium (V) oxide TS (1 in 2), shake vigorously, and allow to stand for 30 minutes. Perform the test with these solutions as directed under Fluorometry <2.22>, and determine the intensity of fluorescence, FT and FS, at the wavelength of excitation at 400 nm and at the wavelength of fluorescence at 500 nm. Dissolution rate (z) with respect to the labeled amount of reserpine (C33H40N2O9) ＝ MS × FT/FS × 1/C MS: Amount (mg) of Reserpine RS taken C: Labeled amount (mg) of reserpine (C33H40N2O9) in 1 tablet
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately and powder not less than 20 Reserpine Tablets. Weigh accurately a quan-tity of the powder, equivalent to about 0.5 mg of reserpine (C33H40N2O9), add 3 mL of water, and warm at 509C for 15 minutes while shaking. After cooling, add exactly 10 mL of the internal standard solution, 10 mL of acetonitrile and warm at 509C for 15 minutes while shaking. After cooling, add water to make 50 mL, centrifuge, and use the superna-tant liquid as the sample solution. Separately, weigh accu-rately about 10 mg of Reserpine RS, previously dried at 609C in vacuum for 3 hours, and dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add ex-actly 10 mL of the internal standard solution, 5 mL of aceto-nitrile and water to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Reserpine. Amount (mg) of reserpine (C33H40N2O9) ＝ MS × QT/QS × 1/20 MS: Amount (mg) of Reserpine RS taken Internal standard solution—A solution of butyl parahydrox-ybenzoate in acetonitrile (1 in 50,000).
Ribavirin Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of ribavirin (C8H12N4O5: 244.20). Method of preparation Prepare as directed under Cap-sules, with Ribavirin. Identification Take out the content of Ribavirin Capsules. Shake thoroughly an amount of the content, equivalent to 0.1 g of Ribavirin, with 10 mL of water, allow to stand for 1 minute, filter, and use the filtrate as the sample solution. Separately, dissolve 50 mg of ribavirin in 5 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of acetonitrile and diluted ammonium chlo-ride TS (1 in 20) (9:2) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution have the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Ribavirin Capsules add 250 mL of water previously warmed to 379C, shake in a water bath of 379C for 15 minutes, then allow standing to cool to room tempera-ture, add water to make exactly 500 mL, and filter. Discard the first 3 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg of ribavirin (C8H12N4O5), and use this solution as the sample solution.
Take out the content of Ribavirin Capsules. Shake thoroughly an amount of the content, equivalent to 0.1 g of Ribavirin, with 10 mL of water, allow to stand for 1 minute, filter, and use the filtrate as the sample solution. Separately, dissolve 50 mg of ribavirin in 5 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of acetonitrile and diluted ammonium chlo-ride TS (1 in 20) (9:2) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wave-length: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution have the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Ribavirin Capsules add 250 mL of water previously warmed to 379C, shake in a water bath of 379C for 15 minutes, then allow standing to cool to room tempera-ture, add water to make exactly 500 mL, and filter. Discard the first 3 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 20 mg of ribavirin (C8H12N4O5), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Ribavirin RS, previously dried at 1059C for 5 hours, dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ribavirin in each solution. Amount (mg) of ribavirin (C8H12N4O5) ＝ MS × AT/AS × V?/V × 1/2 MS: Amount (mg) of Ribavirin RS taken Operating conditions— Proceed as directed in the operating conditions in the Dis-solution. System suitability— Proceed as directed in the system suitability in the Dissolu-tion. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Ribavirin Capsules is not less than 85z. Start the test with 1 capsule of Ribavirin Capsules, with-draw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 3 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of ribavirin (C8H12N4O5), and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Ribavirin RS, previously dried at 1059C for sulfuric acid TS. Flow rate: Adjust so that the retention time of ribavirin is about 4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ribavirin are not less than 500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ribavirin is not more than 2.0z.
Cut and open the capsules of not less than 20 Ribavirin Capsules, take out the contents and weigh the mass accurately, and mix uniformly. Weigh accurately an amount of the content, equivalent to about 0.1 g of ribavirin (C8H12N4O5), add 100 mL of water, shake for 30 minutes, then add water to make exactly 200 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 25 mg of Ribavirin RS, previously dried at 1059C for 5 hours, dissolve in water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of ribavirin in each solution. Amount (mg) of ribavirin (C8H12N4O5) ＝ MS × AT/AS × 4 MS: Amount (mg) of Ribavirin RS taken Operating conditions— Detector, column, column temperature, mobile phase A, and flow rate: Proceed as directed in the operating condi-tions in the Assay under Ribavirin. Mobile phase B: A mixture of mobile phase A and aceto-nitrile for liquid chromatography (9:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 5 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ribavirin in each solution. Dissolution rate (z) with respect to the labeled amount of ribavirin (C8H12N4O5) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Ribavirin RS taken C: Labeled amount (mg) of ribavirin (C8H12N4O5) in 1 capsule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 207 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 10 cm in length, packed with strongly acidic ion-exchange resin for liquid chromatography composed with sulfonic acid group bound styrene-divinylbenzene copolymer (9 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Adjust to pH 2.5 of water with 0.5 mol/L 0 – 15 100 0 15 – 20 100 → 0 0 → 100 System suitability— System performance: To 5 mL of the standard solution add 1 mL of sodium hydroxide TS, allow to stand for 30 minutes, and add 1 mL of 1 mol/L hydrochloric acid TS. When the procedure is run with 5 mL of this solution under the above operating conditions, the resolution between the peak having the relative retention time of about 0.85 to ribavirin and the peak of ribavirin is not less than 4.0. Fur-thermore, when the procedure is run with 5 mL of the stand-ard solution under the above operating conditions, the sym-metry factor of the peak of ribavirin is not more than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of ribavirin is not more than 1.0z.
Rifampicin Capsules contain not less than 93.0z and not more than 105.0z of the labeled potency of rifampicin (C43H58N4O12: 822.94). Method of preparation Prepare as directed under Cap-sules, with Rifampicin. Identification Take out the content of Rifampicin Cap-sules, mix well, and powder, if necessary. Dissolve an amount of the content, equivalent to 20 mg (potency) of Rifampicin, in methanol to make 100 mL, and filter. To 5 mL of the filtrate add 0.05 mol/L phosphate buffer solution (pH 7.0) to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 234 nm and 238 nm, between 252 nm and 256 nm, between 331 nm and 335 nm, and between 472 nm and 476 nm. Purity Related substances—Perform the test quickly after the sample solution and standard solution are prepared. Open the capsules of not less than 20 Rifampicin Capsules, carefully take out the content, weigh accurately, and pow-der. Weigh accurately a portion of the powder, equivalent to about 20 mg (potency) of Rifampicin, and dissolve in aceto-nitrile to make exactly 10 mL. Pipet 2 mL of this solution, add a mixture of acetonitrile and methanol (1:1) to make ex-actly 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Rifampicin RS, equivalent to about 20 mg (potency), and dissolve in aceto- Amount [ mg (potency)] of rifampicin (C ＝ MS × AT/AS × 1000 43H58N4O12) nitrile to make exactly 10 mL. Pipet 2 mL of this solution, and add the mixture of acetonitrile and methanol (1:1) to make exactly 20 mL. Pipet 1 mL of this solution, add the MS: Amount [mg (potency)] of Rifampicin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.2 g of citric acid monohydrate and 1.4 g of sodium perchlorate in 1000 mL of a mixture of water, acetonitrile and phosphate buffer solution (pH 3.1) (11:7:2). mixture of acetonitrile and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the amount of the peaks of quinone substance and N-oxide substance, having the relative retention time of about 0.5 and about 1.2 to rifampicin, obtained from the sample solu-tion are not more than 4.0z and not more than 1.5z, re-spectively. The amount of the peak other than the peaks mentioned above is not more than 1.0z, and the total amount of these related substances is not more than 2.0z. For the areas of the peaks of the quinone substance and N-oxide substance, multiply their correction factors, 1.24 and 1.16, respectively. Amount (mg) of quinone substance ＝ MS/MT × ATa/AS × 2.48 Amount (mg) of N-oxide substance ＝ MS/MT × ATb/AS × 2.32 Each amount (mg) of related substances other than quinone and N-oxide substances ＝ MS/MT × ATi/AS × 2 MS: Amount [mg (potency)] of Rifampicin RS taken MT: Amount [mg (potency)] of sample taken AS: Peak area of the standard solution ATa: Peak area of quinone substance ATb: Peak area of N-oxide substance ATi: Each peak area of related substances other than qui-none and N-oxide substances Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.1 g of sodium perchlorate, 6.5 g of citric acid monohydrate and 2.3 g of potassium dihy-drogen phosphate in 1100 mL of water, and add 900 mL of acetonitrile. Flow rate: Adjust so that the retention time of rifampicin is about 12 minutes. Time span of measurement: About 2.5 times as long as the retention time of rifampicin. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and methanol (1:1) to make exactly 20 mL. Confirm that the peak area of rifampicin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 dissolve in 5 mL of methanol, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make ex-actly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 334 nm of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of rifampicin (C43H58N4O12) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Rifampicin RS taken C: Labeled amount [mg (potency)] of rifampicin (C43H58N4O12) in 1 capsule Assay Open the capsules of not less than 20 Rifampicin Capsules, take out the content, weigh accurately the mass of the content, and powder. Weigh accurately a portion of the powder, equivalent to about 75 mg (potency) of Rifampicin, dissolve in a mixture of acetonitrile and methanol (1:1) to make exactly 50 mL. Pipet 10 mL of this solution, and add acetonitrile to make exactly 50 mL. Pipet 5 mL of this solu-tion, add a solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodium hydrogen phosphate dodecahydrate and 3.1 g of potassium dihydrogen phosphate in 1000 mL of a mixture of water and acetonitrile (3:1) to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Rifampi-cin RS, equivalent to about 30 mg (potency), dissolve in 20 mL of a mixture of acetonitrile and methanol (1:1), and add acetonitrile to make exactly 100 mL. Pipet 5 mL of this solu-tion, add the solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodium hydrogen phosphate dodecahydrate and 3.1 g of potassium dihydrogen phosphate in 1000 mL of the mixture of water and acetonitrile (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of rifam-picin in each solution. Amount [mg (potency)] of rifampicin (C H N O ) mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry ＝ MS × AT/AS × 5/2 43 58 4 12 factor of the peak of rifampicin is not less than 2500 and not more than 4.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rifampicin is not more than 2.0z. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method using a sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Rifampicin Capsules is not less than 80z. Start the test with 1 capsule of Rifampicin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 17 mg (potency) of rifampicin (C43H58N4O12), and use this solution as the sample solution.
Take out the content of Rifampicin Cap-sules, mix well, and powder, if necessary. Dissolve an amount of the content, equivalent to 20 mg (potency) of Rifampicin, in methanol to make 100 mL, and filter. To 5 mL of the filtrate add 0.05 mol/L phosphate buffer solution (pH 7.0) to make 100 mL. Determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 234 nm and 238 nm, between 252 nm and 256 nm, between 331 nm and 335 nm, and between 472 nm and 476 nm.
Related substances—Perform the test quickly after the sample solution and standard solution are prepared. Open the capsules of not less than 20 Rifampicin Capsules, carefully take out the content, weigh accurately, and pow-der. Weigh accurately a portion of the powder, equivalent to about 20 mg (potency) of Rifampicin, and dissolve in aceto-nitrile to make exactly 10 mL. Pipet 2 mL of this solution, add a mixture of acetonitrile and methanol (1:1) to make ex-actly 20 mL, and use this solution as the sample solution. Separately, weigh accurately an amount of Rifampicin RS, equivalent to about 20 mg (potency), and dissolve in aceto- Amount [ mg (potency)] of rifampicin (C ＝ MS × AT/AS × 1000 43H58N4O12) nitrile to make exactly 10 mL. Pipet 2 mL of this solution, and add the mixture of acetonitrile and methanol (1:1) to make exactly 20 mL. Pipet 1 mL of this solution, add the MS: Amount [mg (potency)] of Rifampicin RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.2 g of citric acid monohydrate and 1.4 g of sodium perchlorate in 1000 mL of a mixture of water, acetonitrile and phosphate buffer solution (pH 3.1) (11:7:2). mixture of acetonitrile and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the amount of the peaks of quinone substance and N-oxide substance, having the relative retention time of about 0.5 and about 1.2 to rifampicin, obtained from the sample solu-tion are not more than 4.0z and not more than 1.5z, re-spectively. The amount of the peak other than the peaks mentioned above is not more than 1.0z, and the total amount of these related substances is not more than 2.0z. For the areas of the peaks of the quinone substance and N-oxide substance, multiply their correction factors, 1.24 and 1.16, respectively. Amount (mg) of quinone substance ＝ MS/MT × ATa/AS × 2.48 Amount (mg) of N-oxide substance ＝ MS/MT × ATb/AS × 2.32 Each amount (mg) of related substances other than quinone and N-oxide substances ＝ MS/MT × ATi/AS × 2 MS: Amount [mg (potency)] of Rifampicin RS taken MT: Amount [mg (potency)] of sample taken AS: Peak area of the standard solution ATa: Peak area of quinone substance ATb: Peak area of N-oxide substance ATi: Each peak area of related substances other than qui-none and N-oxide substances Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.1 g of sodium perchlorate, 6.5 g of citric acid monohydrate and 2.3 g of potassium dihy-drogen phosphate in 1100 mL of water, and add 900 mL of acetonitrile. Flow rate: Adjust so that the retention time of rifampicin is about 12 minutes. Time span of measurement: About 2.5 times as long as the retention time of rifampicin. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add a mixture of acetonitrile and methanol (1:1) to make exactly 20 mL. Confirm that the peak area of rifampicin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 dissolve in 5 mL of methanol, and add water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make ex-actly 20 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 334 nm of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using water as the blank. Dissolution rate (z) with respect to the labeled amount of rifampicin (C43H58N4O12) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount [mg (potency)] of Rifampicin RS taken C: Labeled amount [mg (potency)] of rifampicin (C43H58N4O12) in 1 capsule
Open the capsules of not less than 20 Rifampicin Capsules, take out the content, weigh accurately the mass of the content, and powder. Weigh accurately a portion of the powder, equivalent to about 75 mg (potency) of Rifampicin, dissolve in a mixture of acetonitrile and methanol (1:1) to make exactly 50 mL. Pipet 10 mL of this solution, and add acetonitrile to make exactly 50 mL. Pipet 5 mL of this solu-tion, add a solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodium hydrogen phosphate dodecahydrate and 3.1 g of potassium dihydrogen phosphate in 1000 mL of a mixture of water and acetonitrile (3:1) to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately an amount of Rifampi-cin RS, equivalent to about 30 mg (potency), dissolve in 20 mL of a mixture of acetonitrile and methanol (1:1), and add acetonitrile to make exactly 100 mL. Pipet 5 mL of this solu-tion, add the solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodium hydrogen phosphate dodecahydrate and 3.1 g of potassium dihydrogen phosphate in 1000 mL of the mixture of water and acetonitrile (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of rifam-picin in each solution. Amount [mg (potency)] of rifampicin (C H N O ) mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry ＝ MS × AT/AS × 5/2 43 58 4 12 factor of the peak of rifampicin is not less than 2500 and not more than 4.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rifampicin is not more than 2.0z. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method using a sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Rifampicin Capsules is not less than 80z. Start the test with 1 capsule of Rifampicin Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 17 mg (potency) of rifampicin (C43H58N4O12), and use this solution as the sample solution. Separately, weigh accurately about 17 mg (potency) of Rifampicin RS, MS: Amount [mg (potency)] of Rifampicin RS taken Operating conditions— Proceed as directed in the operating conditions in the Assay under Rifampicin. System suitability— System performance: Dissolve 30 mg (potency) of Rifam-picin RS in 20 mL of the mixture of acetonitrile and metha-nol (1:1), and add acetonitrile to make 100 mL. To 5 mL of this solution add 2 mL of a solution of butyl parahydrox-ybenzoate in the mixture of acetonitrile and methanol (1:1) (1 in 5000), then add the solution prepared by dissolving 2.1 g of citric acid monohydrate, 27.6 g of disodium hydrogen phosphate dodecahydrate and 3.1 g of potassium dihydrogen phosphate in 1000 mL of a mixture of water and acetonitrile (3:1) to make exactly 50 mL. When the procedure is run with 50 mL of this solution under the above operating conditions, butyl parahydroxybenzoate and rifampicin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 5 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rifampicin is not more than 1.0z. ‌Containers and storage Containers—Tight containers. Rilmazafone Hydrochloride Hydrate
To a quantity of powdered Rilmazafone Hy-drochloride Tablets, equivalent to 10 mg of Rilmazafone Hydrochloride Hydrate, add 5 mL of methanol, shake for 10 minutes, and centrifuge. Filter the supernatant liquid with a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, dissolve 2 mg of rilmazafone hydrochloride hydrate in 1 mL of metha-nol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solu-tion and standard solution on a plate of silica gel with fluo-rescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, acetonitrile, water and acetic acid (100) (8:4:3:3) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Rilmazafone Hydrochloride Tablets add V mL of water so that each mL contains about 0.2 mg of rilmazafone hydrochloride hydrate (C21H20Cl2N6O3.HCl. 2H2O). Add exactly 2V mL of the internal standard solution, shake vigorously for 10 minutes, and filter with a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of Rilmazafone Hydrochloride RS (separately determine the water <2.48> in the same manner as Rilmazafone Hydro-chloride Hydrate), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 20 mL of the internal standard solution, and use this solution as the stand-ard solution. Then, proceed as directed in the Assay under Rilmazafone Hydrochloride Hydrate. Amount (mg) of rilmazafone hydrochloride hydrate (C21H20Cl2N6O3.HCl.2H2O) ＝ MS × QT/QS × V/100 × 1.070 MS: Amount (mg) of Rilmazafone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and acetonitrile (1:1) (3 in 100,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Rilmazafone Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Rilmazafone Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.1 mg of rilmazafone hydro-chloride hydrate (C21H20Cl2N6O3.HCl.2H2O), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Rilmazafone Hydrochloride RS (separately determine the water <2.48> in the same manner as Rilmaza-fone Hydrochloride Hydrate), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL. Then, pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of rilmazafone in each solution. Dissolution rate (z) with respect to the labeled amount of rilmazafone hydrochloride hydrate (C21H20Cl2N6O3.HCl.2H2O) ＝ MS × AT/AS × V?/V × 1/C × 9/2 × 1.070 MS: Amount (mg) of Rilmazafone Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of rilmazafone hydrochloride hy-drate (C21H20Cl2N6O3.HCl.2H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the ‌Assay under Rilmazafone Hydrochloride Hydrate. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rilmazafone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rilmazafone is not more than 2.0z.
(1) Heavy metals <1.07>—Evaporate 100 mL of Ringer's Solution to about 40 mL on a water bath. Add 2 mL of dilute acetic acid and water to make 50 mL, and per- of rilmazafone hydrochloride hydrate (C21H20 Cl2N6O3.HCl. form the test using this solution as the test solution. Control solution: To 3.0 mL of Standard Lead Solution 2H2O), add 10 mL of water and exactly 20 mL of the inter-nal standard solution, shake vigorously for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of Rilmazafone Hydro-chloride RS (separately determine the water <2.48> in the same manner as Rilmazafone Hydrochloride Hydrate), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 20 mL of the internal standard, and use this solution as the standard solution. Then, proceed as directed in the Assay under Rilmazafone Hydrochloride Hydrate. Amount (mg) of rilmazafone hydrochloride hydrate (C21H20Cl2N6O3.HCl.2H2O) add 2 mL of dilute acetic acid and water to make 50 mL (not more than 0.3 ppm). (2) Arsenic <1.11>—Perform the test with 20 mL of Rin- ger's Solution as the test solution (not more than 0.1 ppm). Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ＝ MS × QT/QS × 1/10 × 1.070 MS: Amount (mg) of Rilmazafone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and acetonitrile (1:1) (3 in 100,000).
Weigh accurately the mass of not less than 20 tablets of Rilmazafone Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 2 mg Identification (1) Evaporate 10 mL of Ringer's Solution to 5 mL: the solution responds to Qualitative Tests <1.09> for potassium salt. Evaporate 10 mL of Ringer's Solution to 5 mL: the solution responds to Qualitative Test <1.09> for calcium salt. Ringer's Solution responds to Qualitative Tests <1.09> for sodium salt. Ringer's Solution responds to Qualitative Tests <1.09> for chloride. pH <2.54> 5.0 – 7.5 Purity (1) Heavy metals <1.07>—Evaporate 100 mL of Ringer's Solution to about 40 mL on a water bath. Add 2 mL of dilute acetic acid and water to make 50 mL, and per- of rilmazafone hydrochloride hydrate (C21H20 Cl2N6O3.HCl. form the test using this solution as the test solution. Control solution: To 3.0 mL of Standard Lead Solution 2H2O), add 10 mL of water and exactly 20 mL of the inter-nal standard solution, shake vigorously for 10 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of Rilmazafone Hydro-chloride RS (separately determine the water <2.48> in the same manner as Rilmazafone Hydrochloride Hydrate), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 20 mL of the internal standard, and use this solution as the standard solution. Then, proceed as directed in the Assay under Rilmazafone Hydrochloride Hydrate. Amount (mg) of rilmazafone hydrochloride hydrate (C21H20Cl2N6O3.HCl.2H2O) add 2 mL of dilute acetic acid and water to make 50 mL (not more than 0.3 ppm). (2) Arsenic <1.11>—Perform the test with 20 mL of Rin- ger's Solution as the test solution (not more than 0.1 ppm). Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ＝ MS × QT/QS × 1/10 × 1.070 MS: Amount (mg) of Rilmazafone Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in a mixture of water and acetonitrile (1:1) (3 in 100,000).
Ringer's Solution is an aqueous injection. It contains not less than 0.
Ringer's Solution is a clear and colorless liq-uid. It has a slightly saline taste.
(1) Chlorine—To 20 mL of Ringer's Solution, accurately measured, add 30 mL of water. Titrate <2.50> with 0.1 mol/L silver nitrate VS while shaking vigorously (indicator: 3 drops of fluorescein sodium TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 3.545 mg of Cl (2) Calcium chloride Hydrate—To 50 mL of Ringer's Solution, exactly measured, add 2 mL of 8 mol/L potassium hydroxide TS and 50 mg of NN indicator, and titrate <2.50> immediately with 0.01 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS, until the color of the solution changes from red-purple to blue. Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.470 mg of CaCl2.2H2O
Containers—Hermetic containers. Plastic containers for aqueous infusions may be used.
To an amount of Risperidone Fine Granules, equivalent to 2 mg of Risperidone, add 100 mL of 2-propanol, shake thoroughly, and filter. Determine the ab-sorption spectrum of the filtrate as directed under Ultravio-let-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 277 nm and 281 nm and between 283 nm and 287 nm.
Related substances—To an amount of Risperidone Fine Granules, equivalent to 2 mg of Risperidone, add 20 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Pipet 1 mL of the sample solution, add a mix-ture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than risperi-done obtained from the sample solution is not larger than 1/2 times the peak area of risperidone from the standard so-lution, and the total area of the peaks other than risperidone from the sample solution is not larger than the peak area of risperidone from the standard solution. For the area of the peaks, having the relative retention time of about 0.4 and about 1.6 to risperidone, multiply their correction factors, 1.9 and 1.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of risperidone, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 50 mL. Confirm that the peak area of risperidone obtained with 10 mL of this solution is equivalent to 7.5 to 12.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.5z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Risperidone Fine Granules is not less than 75z. Start the test with an accurately weighed amount of Risperidone Fine Granules, equivalent to about 3 mg of risperidone (C23H27FN4O2), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add diluted hydrochloric acid (1 in 137) to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of risperidone for assay (separately deter-mine the loss on drying <2.41> under the same conditions as Risperidone), and dissolve in methanol to make exactly 50 mL. Pipet 15 mL of this solution, add methanol to make ex-actly 25 mL. Pipet 2 mL of this solution, and add water to make exactly 200 mL. Pipet 3 mL of this solution, add ex-actly 3 mL of diluted hydrochloric acid (1 in 137), and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac- cording to the following conditions, and determine the peak areas, AT and AS, of risperidone in each solution. Dissolution rate (z) with respect to the labeled amount of risperidone (C23H27FN4O2) ＝ MS/MT × AT/AS × 1/C × 54/5 MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis MT: Amount (g) of Risperidone Fine Granules taken C: Labeled amount (mg) of risperidone (C23H27FN4O2) in 1 g Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (13:7) add 1 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 3 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 3500 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.0z.
If necessary powder Risperidone Fine Granules, and weigh accurately an amount, equivalent to about 2 mg of risperidone (C23H27FN4O2), add 8 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and add a mixture of 0.1 mol/L hydrochloric acid TS and metha-nol (3:2) to make exactly 20 mL. Filter the solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of risperidone for assay (separately determine the loss on drying <2.41> under the same conditions as Risperidone), and dissolve in a mixture of 0.1 mol/L hydro-chloric acid TS and methanol (3:2) to make exactly 50 mL. Pipet 10 mL of this solution, add a mixture of 0.1 mol/L hy-drochloric acid TS and methanol (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of risperidone in each solution. Amount (mg) of risperidone (C23H27FN4O2) ＝ MS × AT/AS × 1/25 MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). ‌Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (4:1) add 1.5 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 1.0z.
Risperidone Oral Solution contains not less than 95.
Risperidone Oral Solution occurs as a clear and colorless liquid.
To a volume of Risperidone Oral Solution, equivalent to 2 mg of Risperidone, add 50 mg of sodium hydrogen carbonate and 10 mL of diethyl ether, shake, cen-trifuge, and evaporate the supernatant liquid to dryness in lukewarm water. Determine the absorption spectrum of a so-lution of the residue in 100 mL of 2-propanol as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of risperidone for assay (separately determine the loss on drying <2.41> under the same conditions as Risperidone), and dissolve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, add 10 mL of water, then add methanol to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of risperi-done in each solution. its maxima between 277 nm and 281 nm and between 283 nm Amount (mg) of risperidone (C H FN O ) and 287 nm. ＝ MS × AT/AS × 1/25 23 27 4 2 pH Being specified separately when the drug is granted ap-proval based on the Law.
Related substances—To a volume of Risperidone Oral Solution, equivalent to 2 mg of Risperidone, add meth-anol to make 20 mL, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than risperidone obtained from the sample solution is not larger than 1/2 times the peak area of risperi-done from the standard solution, and the total area of the peaks other than risperidone from the sample solution is not larger than the peak area of risperidone from the standard solution. For the area of the peaks, having the relative reten-tion time of about 0.4 and about 1.6 to risperidone, multiply their correction factors, 1.9 and 1.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (4:1) add 1.5 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the peak area of risperidone is not more than 1.0z.
To an exact volume of Risperidone Oral Solution, equivalent to about 2 mg of risperidone (C H FN O ), add 23 27 4 2 Method of preparation Prepare as directed under Liquids and Solutions for Oral Administration, with Risperidone. Description Risperidone Oral Solution occurs as a clear and colorless liquid. Identification To a volume of Risperidone Oral Solution, equivalent to 2 mg of Risperidone, add 50 mg of sodium hydrogen carbonate and 10 mL of diethyl ether, shake, cen-trifuge, and evaporate the supernatant liquid to dryness in lukewarm water. Determine the absorption spectrum of a so-lution of the residue in 100 mL of 2-propanol as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib- methanol to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of risperidone for assay (separately determine the loss on drying <2.41> under the same conditions as Risperidone), and dissolve in methanol to make exactly 50 mL. Pipet 10 mL of this solution, add 10 mL of water, then add methanol to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of risperi-done in each solution. its maxima between 277 nm and 281 nm and between 283 nm Amount (mg) of risperidone (C H FN O ) and 287 nm. ＝ MS × AT/AS × 1/25 23 27 4 2 pH Being specified separately when the drug is granted ap-proval based on the Law. Purity Related substances—To a volume of Risperidone Oral Solution, equivalent to 2 mg of Risperidone, add meth-anol to make 20 mL, and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add a mixture of methanol and water (9:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than risperidone obtained from the sample solution is not larger than 1/2 times the peak area of risperi-done from the standard solution, and the total area of the peaks other than risperidone from the sample solution is not larger than the peak area of risperidone from the standard solution. For the area of the peaks, having the relative reten-tion time of about 0.4 and about 1.6 to risperidone, multiply their correction factors, 1.9 and 1.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 275 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (4:1) add 1.5 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 13 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the peak area of risperidone is not more than 1.0z.
Risperidone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of risperidone (C23H27FN4O2: 410.48). Method of preparation Prepare as directed under Tablets, with Risperidone. Identification Powder Risperidone Tablets. To a portion of the powder, equivalent to 2 mg of Risperidone, add 100 mL of 2-propanol, shake thoroughly, and filter. Determine the absorption spectrum of the filtrate as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 277 nm and 281 nm and between 283 nm and 287 nm. Purity Related substances—Powder Risperidone Tablets. To a portion of the powder, equivalent to 2 mg of Risperi-done, add 20 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 1 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than risperidone obtained from the sample solution is not larger than 1/2 times the peak area of risperidone from the stand-ard solution, and the total area of the peaks other than risperidone from the sample solution is not larger than the peak area of risperidone from the standard solution. For the area of the peaks, having the relative retention time of about 0.4 and about 1.6 to risperidone, multiply their correction factors, 1.9 and 1.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of risperidone, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 50 mL. Confirm that the peak area of risperidone obtained with 10 mL of this solution is equivalent to 7.5 to 12.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Risperidone Tablets add 3V/5 mL of a mix-ture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly V mL so that each mL contains 0.
Powder Risperidone Tablets. To a portion of the powder, equivalent to 2 mg of Risperidone, add 100 mL of 2-propanol, shake thoroughly, and filter. Determine the absorption spectrum of the filtrate as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 277 nm and 281 nm and between 283 nm and 287 nm.
Related substances—Powder Risperidone Tablets. To a portion of the powder, equivalent to 2 mg of Risperi-done, add 20 mL of a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 1 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than risperidone obtained from the sample solution is not larger than 1/2 times the peak area of risperidone from the stand-ard solution, and the total area of the peaks other than risperidone from the sample solution is not larger than the peak area of risperidone from the standard solution. For the area of the peaks, having the relative retention time of about 0.4 and about 1.6 to risperidone, multiply their correction factors, 1.9 and 1.5, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of risperidone, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 50 mL. Confirm that the peak area of risperidone obtained with 10 mL of this solution is equivalent to 7.5 to 12.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 4000 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Risperidone Tablets add 3V/5 mL of a mix-ture of 0.1 mol/L hydrochloric acid TS and methanol (3:2), shake, and add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly V mL so that each mL contains 0.1 mg of risperidone (C23H27FN4O2). Filter this so-lution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of risperidone (C23H27FN4O2) ＝ MS × AT/AS × V/500 MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Risperidone Tablets is not less than 75z. Start the test with 1 tablet of Risperidone Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add diluted hydrochloric acid (1 in 137) to make exactly V? mL so that each mL contains about 0.56 mg of risperidone (C23H27FN4O2), and use this solution as the sam-ple solution. Separately, weigh accurately about 28 mg of risperidone for assay (separately determine the loss on drying <2.41> under the same conditions as Risperidone), and dis- solve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add methanol to make exactly 25 mL. Pipet 2 mL of this solution, and add water to make exactly 200 mL. Pipet 3 mL of this solution, add exactly 3 mL of diluted hy-drochloric acid (1 in 137), and use this solution as the stand-ard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of risperidone in each solution. Dissolution rate (z) with respect to the labeled amount of risperidone (C23H27FN4O2) ＝ MS × AT/AS × V?/V × 1/C × 9/5 MS: Amount (mg) of risperidone for assay taken, calcu-lated on the dried basis C: Labeled amount (mg) of risperidone (C23H27FN4O2) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 1000 mL of a mixture of water and acetonitrile (13:7) add 1 mL of trifluoroacetic acid, and adjust to pH 3.0 with ammonia solution (28). Flow rate: Adjust so that the retention time of risperidone is about 3 minutes. ‌System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risperidone are not less than 3500 and not more than 2.5, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risperidone is not more than 2.0z.
Weigh accurately the mass of not less than 20 Risperidone Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 2 mg of risperidone (C23H27FN4O2), add 8 mL of a mixture of 0.1 mol/L hydro-chloric acid TS and methanol (3:2), shake, and add a mix-ture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 20 mL. Filter the solution through a mem-brane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of risperidone for assay (separately determine the loss on drying <2.41> under the same conditions as Risperidone), and dissolve in a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 50 mL. Pipet 10 mL of this solution, add a mixture of 0.1 mol/L hydrochloric acid TS and methanol (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of risperidone in each solution. Ritodrine Hydrochloride
Ritodrine Hydrochloride Injection is an aqueous in-jection. It contains not less than 95.
Ritodrine Hydrochloride Injection is a clear and colorless liquid. Identiˆcation To a volume of Ritodrine Hydrochloride In-jection, equivalent to 50 mg of Ritodrine Hydrochloride, add 0.01 mol/L hydrochloric acid TS to make 100 mL. To 10 mL of this solution add 0.01 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 272 nm and 276 nm. pH Being speciˆed separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 25 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Pipet a volume of Ritodrine Hydrochloride Injec-tion, equivalent to about 20 mg of ritodrine hydrochloride ‌(C17H21NO3.HCl), and add a mixture of 0.02 mol/L sodium dihydrogen phosphate dihydrate solution and methanol (7:3) to make exactly 250 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Rito-drine Hydrochloride RS, previously dried at 1059C for 2 hours, dissolve in a mixture of 0.02 mol/L sodium dihydro-gen phosphate dihydrate solution and methanol (7:3) to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions. Determine the peak areas, AT and AS, of ritodrine in each solution. Amount (mg) of ritodrine hydrochloride (C17H21NO3.HCl) ＝ MS × AT/AS MS: Amount (mg) of Ritodrine Hydrochloride RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 272 nm and 276 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Ritodrine Hydrochloride Tablets add 9 mL of 0.01 mol/L hydrochloric acid TS, shake until the tablet is completely disintegrated, add 0.01 mol/L hydrochloric acid TS to make exactly 10 mL, and filter through a membrane filter having pore size of 0.45 mm. Pipet 3 mL of the filtrate, add exactly 1 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Ritodrine Hydrochloride RS, previ-ously dried at 1059C for 2 hours, and dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 3 mL of this solution, add exactly 1 mL of the internal standard solu-tion, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ritodrine to that of the internal standard. Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.6 g of diammonium hydrogen Amount (mg) of ritodrine hydrochloride (C ＝ MS × QT/QS × 1/5 17H 21NO3.HCl) phosphate and 1.1 g of sodium 1-heptansulfonate in 840 mL of water, add 160 mL of acetonitrile for liquid chromatogra-phy, and adjust to pH 3.0 with phosphoric acid. Flow rate: Adjust so that the retention time of ritodrine is about 19 minutes. System Suitability— System performance: Dissolve 10 mg of ritodrine hydro-chloride in 50 mL of dilute sulfuric acid. Heat a portion of this solution in a water bath for about 30 minutes, and allow to cool. Measure a portion of this solution, and add the same volume of 2 mol/L sodium hydroxide TS. Dissolve 2 mg of ritodrine hydrochloride in 10 mL of this solution, and add a mixture of 0.02 mol/L sodium dihydrogen phosphate dihydrate solution and methanol (7:3) to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ritodrine and ritodrine threo-isomer are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ritodrine is not more than 1.0z.
Ritodrine Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of ritodrine hydrochloride (C17H21NO3.HCl: 323.81). Method of preparation Prepare as directed under Tablets, with Ritodrine Hydrochloride. Identification To 10 mL of the filtrate obtained in the Assay add 0.01 mol/L hydrochloric acid TS to make 100 MS: Amount (mg) of Ritodrine Hydrochloride RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ritodrine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ritodrine to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Ritodrine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Ritodrine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.
To 10 mL of the filtrate obtained in the Assay add 0.01 mol/L hydrochloric acid TS to make 100 MS: Amount (mg) of Ritodrine Hydrochloride RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (3 in 10,000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ritodrine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ritodrine to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Ritodrine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Ritodrine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of ritodrine hydro-chloride (C17H21NO3.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 28 mg of Ritodrine Hydrochloride RS, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 80 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ritodrine in each solution. ‌Dissolution rate (z) with respect to the labeled amount of ritodrine hydrochloride (C17H21NO3.HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of Ritodrine Hydrochloride RS taken C: Labeled amount (mg) of ritodrine hydrochloride (C17H21NO3.HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 80 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ritodrine are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 80 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ritodrine is not more than 1.5z.
To 20 Ritodrine Hydrochloride Tablets add 150 mL of 0.01 mol/L hydrochloric acid TS, shake for 20 minutes, and add 0.01 mol/L hydrochloric acid TS to make exactly 200 mL. Filter through a glass filter (G4), and discard the first 20 mL of the filtrate. Pipet 30 mL of the subsequent fil-trate, add exactly 5 mL of the internal standard solution and 0.01 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Ritodrine Hydrochloride RS, previ-ously dried at 1059C for 2 hours, and dissolve in 0.01 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 30 mL of this solution, add exactly 5 mL of the internal standard solu-tion and 0.01 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of ritodrine to that of the inter-nal standard. with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ritodrine to that of the internal standard is not more than 1.0z.
Rosuvastatin Calcium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of rosuvastatin (C22H28FN3O6S: 481.54). Method of preparation Prepare as directed under Tablets, with Rosuvastatin Calcium. Identification Perform the test with 10 mL each of the sam-ple solution and standard solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions: the principal peaks in the chro-matograms obtained from the sample solution and the stand-ard solution show the same retention time, and both spectra of these peaks in the chromatograms exhibit similar intensi-ties of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: A photodiode array detector (wavelength: 242 nm; spectrum range of measurement: 220 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. Purity Related substances—To an amount of Rosuvastatin Calcium Tablets, equivalent to 0.1 g of rosuvastatin (C22H28FN3O6S), add 50 mL of water, shake for 30 minutes, then add 25 mL of acetonitrile, and shake for 30 minutes. To this solution add water to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of water and acetonitrile (3:1) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the peak area of related substance C, hav-ing the relative retention time of about 1.6 to rosuvastatin, obtained from the sample solution is not larger than 1.4 times the peak area of rosuvastatin from the standard solu-tion, the peak area of related substance D, having the rela-tive retention time of about 2.3, from the sample solution is not larger than 7/10 times the peak area of rosuvastatin from the standard solution, and the area of the peak other than the peaks of rosuvastatin and related substance B (di-astereomer) having a relative retention time of about 1.1 and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of rosuvastatin from the standard solution. Furthermore, the total area of the peaks other than rosuvastatin from the sample solution is not larger than 2.1 times the peak area of rosuvastatin from the standard solution. For the area of the peak of related sub-stance C, multiply the correction factor 1.4. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of rosuvastatin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, add a mixture of water and acetonitrile (3:1) to make exactly 100 mL. Confirm that the peak area of rosuvastatin obtained with 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 10 mL of the standard solu-tion. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Rosuvastatin Calcium Tablets add 3V/4 mL of 0.1 mol/L phosphate buffer solution (pH 7), and shake for 45 minutes. To this solution add 0.1 mol/L phosphate buffer solution (pH 7) to make exactly V mL so that each mL contains about 25 mg of rosuvastatin (C22H28FN3O6S), and filter through a membrane filter with a pore size not exceeding 0.
Perform the test with 10 mL each of the sam-ple solution and standard solution obtained in the Assay, as directed under Liquid Chromatography <2.01> according to the following conditions: the principal peaks in the chro-matograms obtained from the sample solution and the stand-ard solution show the same retention time, and both spectra of these peaks in the chromatograms exhibit similar intensi-ties of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: A photodiode array detector (wavelength: 242 nm; spectrum range of measurement: 220 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay.
Related substances—To an amount of Rosuvastatin Calcium Tablets, equivalent to 0.1 g of rosuvastatin (C22H28FN3O6S), add 50 mL of water, shake for 30 minutes, then add 25 mL of acetonitrile, and shake for 30 minutes. To this solution add water to make 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card the first 5 mL of the filtrate, and use the subsequent fil-trate as the sample solution. Pipet 1 mL of the sample solu-tion, add a mixture of water and acetonitrile (3:1) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the peak area of related substance C, hav-ing the relative retention time of about 1.6 to rosuvastatin, obtained from the sample solution is not larger than 1.4 times the peak area of rosuvastatin from the standard solu-tion, the peak area of related substance D, having the rela-tive retention time of about 2.3, from the sample solution is not larger than 7/10 times the peak area of rosuvastatin from the standard solution, and the area of the peak other than the peaks of rosuvastatin and related substance B (di-astereomer) having a relative retention time of about 1.1 and the peaks mentioned above from the sample solution is not larger than 1/5 times the peak area of rosuvastatin from the standard solution. Furthermore, the total area of the peaks other than rosuvastatin from the sample solution is not larger than 2.1 times the peak area of rosuvastatin from the standard solution. For the area of the peak of related sub-stance C, multiply the correction factor 1.4. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of rosuvastatin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, add a mixture of water and acetonitrile (3:1) to make exactly 100 mL. Confirm that the peak area of rosuvastatin obtained with 10 mL of this solution is equiva-lent to 3.5 to 6.5z of that with 10 mL of the standard solu-tion. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Rosuvastatin Calcium Tablets add 3V/4 mL of 0.1 mol/L phosphate buffer solution (pH 7), and shake for 45 minutes. To this solution add 0.1 mol/L phosphate buffer solution (pH 7) to make exactly V mL so that each mL contains about 25 mg of rosuvastatin (C22H28FN3O6S), and filter through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 0.1 g of Rosuvastatin Calcium RS (separately determine the water <2.48> in the same man-ner as Rosuvastatin Calcium), add 0.1 mol/L phosphate buffer solution (pH 7) to make exactly 250 mL. Pipet 15 mL of this solution, add 0.1 mol/L phosphate buffer solution (pH 7) to make exactly 250 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 241 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of rosuvastatin (C22H28FN3O6S) ＝ MS × AT/AS × 3V/12,500 × 0.962 MS: Amount (mg) of Rosuvastatin Calcium RS taken, cal-culated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L citrate buffer solution (pH 6.6) as the dis-solution medium, the dissolution rate in 30 minutes of Rosuvastatin Calcium Tablets is not less than 80z. Start the test with 1 tablet of Rosuvastatin Calcium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 2.8 mg of rosuvastatin (C22H28FN3O6S), and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Rosuvastatin Calcium RS (separately determine the water <2.48> in the same manner as Rosuvastatin Calcium), add 50 mL of water, sonicate, add 25 mL of acetonitrile to dissolve, and add water to make exactly 100 mL. Pipet 10 mL of this solution, add the dissolution medium to make exactly 200 mL. Pipet 10 mL of this solution, and add the dissolution medium to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of rosuvastatin in each solution. Dissolution rate (z) with respect to the labeled amount of rosuvastatin (C22H28FN3O6S) ＝ MS × AT/AS × V?/V × 1/C × 9/4 × 0.962 MS: Amount (mg) of Rosuvastatin Calcium RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of rosuvastatin (C22H28FN3O6S) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ‌259C. Mobile phase: A mixture of water, acetonitrile and phos-phoric acid (600:400:1). Flow rate: Adjust so that the retention time of rosuvasta-tin is about 2 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rosuvastatin are not less than 1900 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 1.5z.
To 10 tablets of Rosuvastatin Calcium Tablets add exactly 300 mL of water, and shake for 30 minutes. To this solution add 125 mL of acetonitrile, shake for 15 minutes, and add water to make exactly 500 mL. Pipet 5 mL of this solution, add a mixture of water and acetonitrile (3:1) to make exactly V mL so that each mL contains about 25 mg of rosuvastatin (C22H28FN3O6S), and filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 0.1 g of Rosuvastatin Calcium RS (separately deter-mine the water <2.48> in the same manner as Rosuvastatin Calcium), add 50 mL of water, sonicate, add 25 mL of acetonitrile, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of water and acetonitrile (3:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of rosuvastatin in each solution. Amount (mg) of rosuvastatin (C22H28FN3O6S) in 1 tablet of Rosuvastatin Calcium Tablets ＝ MS × AT/AS × V/400 × 0.962 MS: Amount (mg) of Rosuvastatin Calcium RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 3.2 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and diluted trifluoroacetic acid (1 in 100) (62:37:1). Flow rate: Adjust so that the retention time of rosuvasta-tin is about 13 minutes. System suitability— System performance: To 10 mg of rosuvastatin calcium add 100 mL of water and 20 mL of 1 mol/L hydrochloric acid TS, heat on a water bath of 609C for 2 hours, and neu-tralize with sodium hydroxide TS. After cooling, add 50 mL of acetonitrile and water to make 200 mL. To 10 mL of this solution add 10 mL of a mixture of water and acetonitrile (3:1). When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution be-tween rosuvastatin and related substance B (diastereomer) is not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 1.5z.
Roxatidine Acetate Hydrochloride Extended-release Capsules
ロキサチジン酢酸エステル塩酸塩徐放カプセル
Roxatidine Acetate Hydrochloride Extended-release Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of roxatidine ace-tate hydrochloride (C19H28N2O4.HCl: 384.90). Method of preparation Prepare as directed under Cap-sules, with Roxatidine Acetate Hydrochloride. Identification To 1 mL of the filtrate obtained in the Assay add ethanol (99.5) to make 20 mL, and determine the ab-sorption spectrum as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 275 nm and 278 nm, and between 282 nm and 285 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the contents of 1 capsule of Roxatidine Acetate Hydrochloride Extended-release Capsules, add exactly V mL of ethanol (99.5) so that each mL contains about 2.
To 1 mL of the filtrate obtained in the Assay add ethanol (99.5) to make 20 mL, and determine the ab-sorption spectrum as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 275 nm and 278 nm, and between 282 nm and 285 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take out the contents of 1 capsule of Roxatidine Acetate Hydrochloride Extended-release Capsules, add exactly V mL of ethanol (99.5) so that each mL contains about 2.5 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl), disperse the particles by sonicating, and filter through a membrane filter with a pore size of not more than 1.0 mm. To exactly 8 mL of the filtrate add exactly 2 mL of the internal standard solution, mix, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of roxatidine acetate hydrochloride (C19H28N2O4.HCl) ＝ MS × QT/QS × V/20 MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken Internal standard solution—A solution of benzoic acid in ethanol (99.5) (1 in 500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rates of a 37.5-mg capsule in 45 minutes, in 90 minutes and in 8 hours are 10 – 40z, 35 – 65z, and not less than 70z, respectively, and of a 75-mg capsule in 60 minutes, in 90 minutes and in 8 hours are 20 – 50z, 35 – 65z, and not less than 70z, respectively. Start the test with 1 capsule of Roxatidine Acetate Hydrochloride Extended-release Capsules, withdraw exactly 20 mL of the medium at the specified minute after starting the test, and supply exactly 20 mL of warmed water to 37 ± 0.59C immediately after withdrawing of the medium every time, and filter the media withdrawn through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 42 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl), and use this solution as the sample solu-tion. Separately, weigh accurately about 21 mg of Roxati-dine Acetate Hydrochloride RS, previously dried in a desic-cator (in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this so-lution as the standard solution. Perform the test with exactly 100 mL each of the sample solutions and standard solution as directed under Liquid Chromatography <2.01> according to ‌the following conditions, and determine the peak areas, AT(n) and AS, of roxatidine acetate in each solution. Dissolution rate (z) with respect to the labeled amount of length: 274 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with cyanopropylsilanized roxatidine acetate hydrochloride (C19H nth medium withdrawing (n ＝ 1,2,3) 28N2O4.HCl) on the silica gel for liquid chromatography (5 mm in particle diame- ter). Column temperature: A constant temperature of about «AT(n) n－1 ØAT(i) 1 »$ V? 1 359C. A S ＝ MS × ＋∑ S i＝1 A × 45 × V × C × 180 Mobile phase: A mixture of hexane, ethanol (99.5), triethylamine and acetic acid (100) (384:16:2:1). MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken C: Labeled amount (mg) of roxatidine acetate hydrochlo-ride (C19H28N2O4.HCl) in 1 capsule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile, triethyla-mine and acetic acid (100) (340:60:2:1). Flow rate: Adjust so that the retention time of roxatidine acetate is about 5 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of roxatidine acetate are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of roxatidine acetate is not more than 1.0z.
Take out the contents of not less than 20 Roxatidine Acetate Hydrochloride Extended-release Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 75 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl), add exactly 30 mL of ethanol (99.5), shake, and filter through a membrane filter with a pore size of not more than 1.0 mm. To exactly 8 mL of the filtrate add exactly 2 mL of the internal standard solution, mix, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Roxatidine Acetate Hydrochloride RS, previously dried in a desiccator (in vacuum, phosphorus (V) oxide) for 4 hours, and dissolve in ethanol (99.5) to make exactly 20 mL. To exactly 8 mL of this solution add exactly 2 mL of the in-ternal standard solution, mix, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of roxatidine acetate to that of the internal standard. Amount (mg) of roxatidine acetate hydrochloride (C19H28N2O4.HCl) ＝ MS × QT/QS × 3/2 MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken Internal standard solution—A solution of benzoic acid in ethanol (99.5) (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave- Flow rate: Adjust so that the retention time of roxatidine acetate is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and roxatidine acetate are elute in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of roxatidine acetate to that of the internal standard is not more than 1.0z.
Roxatidine Acetate Hydrochloride Extended-release Tablets
ロキサチジン酢酸エステル塩酸塩徐放錠
Powder Roxatidine Acetate Hydrochloride Extended-release Tablets. To a portion of the powder, equivalent to 37.5 mg of Roxatidine Acetate Hydrochloride, add 40 mL of ethanol (99.5), and disperse the particles by sonicating for 10 minutes with occasional shaking. After shaking thoroughly, add ethanol (99.5) to make 50 mL. Filter the solution, and to 4 mL of the filtrate add ethanol (99.5) to make 25 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 274 nm and 278 nm and between 281 nm and 285 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Roxatidine Acetate Hydrochloride Exten-ded-release Tablets add 5 mL of a mixture of water, triethylamine and acetic acid (100) (340:2:1), sonicate for 5 minutes with occasional shaking, then add 7.5 mL of aceto-nitrile, then sonicate again for 5 minutes. Add 5 mL of a mixture of water, triethylamine and acetic acid (100) (340:2:1), sonicate for 5 minutes, shake thoroughly, add a mixture of water, triethylamine and acetic acid (100) (340:2:1) to make exactly 50 mL, centrifuge, and filter the supernatant liquid. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, equivalent to 6 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl), add ex-actly 3 mL of the internal standard solution, then add the mobile phase to make 20 mL, and use this solution as the ‌sample solution. Then, proceed as directed in the Assay. Amount (mg) of roxatidine acetate hydrochloride (C19H28N2O4.HCl) ＝ MS × QT/QS × 8/V MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken Internal standard solution—A solution of sodium benzoate in the mobile phase (3 in 2000). Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Rox-atidine Acetate Hydrochloride Extended-release Tablets, and powder. Weigh accurately a portion of the powder, equiva-lent to about 37.5 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl), add 40 mL of the mobile phase, and sonicate for 10 minutes with occasional shaking. Further shake thoroughly, add the mobile phase to make exactly 50 mL, centrifuge, and filter the supernatant liquid. Discard the first 10 mL of the filtrate, pipet 8 mL of the subsequent fil-trate, add exactly 3 mL of the internal standard solution, add the mobile phase to make 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 38 mg of Roxatidine Acetate Hydrochloride RS, previously dried in a desiccator (in vacuum, phosphorous (V) oxide) for 4 hours, dissolve in the mobile phase to make exactly 50 mL. Pipet 8 mL of this solution, add exactly 3 mL of the internal standard solution, then add the mobile phase to make 20 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> under the following conditions, and calculate the ratios, QT and QS, of the peak area of roxatidine acetate to that of the internal standard. Amount (mg) of roxatidine acetate hydrochloride (C19H28N2O4.HCl) ＝ MS × QT/QS MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken Internal standard solution—A solution of sodium benzoate in the mobile phase (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile, triethyla-mine and acetic acid (100) (340:60:2:1). Flow rate: Adjust so that the retention time of roxatidine acetate is about 8 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and roxatidine acetate are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of roxatidine acetate to that of the internal standard is not more than 1.0z.
Roxatidine Acetate Hydrochloride for Injection
注射用ロキサチジン酢酸エステル塩酸塩
Roxatidine Acetate Hydrochloride for Injection is a preparation for injection which is dissolved before use. It contains not less than 95.
To an amount of Roxatidine Acetate Hydro-chloride for Injection, equivalent to 75 mg of Roxatidine Acetate Hydrochloride, add 30 mL of ethanol (99.5), shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. To 1 mL of the filtrate add ethanol (99.5) to make 20 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 275 nm and 279 nm and between 282 nm and 286 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
Clarity and color of solution Dissolve an amount of Roxatidine Acetate Hydrochloride for Injection, equiva-lent to 75 mg of Roxatidine Acetate Hydrochloride, in 20 mL of isotonic sodium chloride solution: the solution is clear and colorless. Bacterial endotoxins <4.01> Less than 4.0 EU/mg. Uniformity of dosage units <6.02> It meets the require-ments of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Dissolve with water each content of 10 Roxatidine Acetate Hydrochloride for Injection, wash the containers with water, combine the solution of the content and wash-ings, and add water to make exactly V mL so that each mL contains about 3.75 mg of roxatidine acetate hydrochloride (C19H28N2O4.HCl). Pipet 5 mL of this solution, add water to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Roxatidine Acetate Hydrochloride RS, previously dried in a desiccator (in vacuum, phosphorous (V) oxide) for 4 hours, dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the standard solution. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of roxatidine acetate ‌to that of the internal standard. Amount (mg) of roxatidine acetate hydrochloride (C19H28N2O4.HCl) in 1 Roxatidine Acetate Hydrochloride for Injection ＝ MS × QT/QS × V/50 MS: Amount (mg) of Roxatidine Acetate Hydrochloride RS taken Internal standard solution—Dissolve 20 mg of guanine in 10 mL of 2 mol/L hydrochloric acid TS, add 50 mL of water, then add 20 mL of a solution of sodium hydroxide (1 in 25) and water to make 100 mL. To 10 mL of this solution add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile, triethyla-mine and acetic acid (100) (340:60:2:1). Flow rate: Adjust so that the retention time of roxatidine acetate is about 14 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and roxatidine acetate are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of roxatidine acetate to that of the internal standard is not more than 1.0z.
Roxithromycin Tablets contain not less than 95.0z and not more than 110.0z of the labeled potency of roxithromycin (C41H76N2O15: 837.05). Method of preparation Prepare as directed under Tablets, with Roxithromycin. Identification To a quantity of powdered Roxithromycin Tablets, equivalent to 0.3 g (potency) of Roxithromycin, add 10 mL of acetonitrile, shake, and centrifuge. Evaporate the supernatant liquid on a water bath under reduced pressure, dry the residue at 609C under reduced pressure for 1 hour, and determine the infrared absorption spectrum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 3460 cm－1, 2940 cm－1, 1728 cm－1, 1633 cm－1 and 1464 cm－1. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Roxithromycin Tablets add 7V/10 mL of the mobile phase, sonicate to disintegrate the tablet, shake, add exactly V/25 mL of the internal standard solution, and add the mobile phase to make V mL so that each mL contains about 1.
To a quantity of powdered Roxithromycin Tablets, equivalent to 0.3 g (potency) of Roxithromycin, add 10 mL of acetonitrile, shake, and centrifuge. Evaporate the supernatant liquid on a water bath under reduced pressure, dry the residue at 609C under reduced pressure for 1 hour, and determine the infrared absorption spectrum of the residue as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 3460 cm－1, 2940 cm－1, 1728 cm－1, 1633 cm－1 and 1464 cm－1. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Roxithromycin Tablets add 7V/10 mL of the mobile phase, sonicate to disintegrate the tablet, shake, add exactly V/25 mL of the internal standard solution, and add the mobile phase to make V mL so that each mL contains about 1.5 mg (potency) of roxithromycin (C41H76N2O15). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Then, proceed as directed in the Assay. ‌Amount [mg (potency)] of roxithromycin (C41H76N2O15)‌‌ ＝ MS × QT/QS × V/25 MS: Amount [mg (potency)] of Roxithromycin RS taken Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 800). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Roxithromycin Tablets is not less than 80z. Start the test with 1 tablet of Roxithromycin Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 0.17 mg (potency) of rox-ithromycin (C41H76N2O15), and use this solution as the sam-ple solution. Separately, weigh accurately about 33 mg (po-tency) of Roxithromycin RS, dissolve in the dissolution me-dium to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of roxithromycin in each solution. Dissolution rate (z) with respect to the labeled amount [mg (potency)] of roxithromycin (C41H76N2O15) ＝ MS × AT/AS × V?/V × 1/C × 450 MS: Amount [mg (potency)] of Roxithromycin RS taken C: Labeled amount [mg (potency)] of roxithromycin (C41H76N2O15) in 1 tablet Operating conditions— Detector, column temperature and mobile phase: Proceed as directed in the operating conditions in the Assay. Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Flow rate: Adjust so that the retention time of rox-ithromycin is about 5 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of roxithromycin are not less than 2300 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of roxithromycin is not more than 1.0z.
Weigh accurately the mass of not less than 20 Rox-ithromycin Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 38 mg (potency) of rox-ithromycin (C41H76N2O15), add 20 mL of the mobile phase, shake vigorously, add exactly 1 mL of the internal standard solution, and then add the mobile phase to make 25 mL. Filter the solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 38 mg (potency) of Rox-ithromycin RS, dissolve in the mobile phase, add exactly 1 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of rox-ithromycin to that of the internal standard. Amount [mg (potency)] of roxithromycin (C41H76N2O15) ＝ MS × QT/QS MS: Amount [mg (potency)] of Roxithromycin RS taken Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 49.1 g of ammonium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.3 with 2 mol/L sodium hydroxide TS. To 690 mL of this solu-tion add 310 mL of acetonitrile. Flow rate: Adjust so that the retention time of rox-ithromycin is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, roxithromycin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of roxithromycin to that of the internal stand-ard is not more than 1.0z.
Salicylated Alum Powder contains not less than 2.
Salicylated Alum Powder occurs as a white powder.
(1) The colored solution obtained in the Assay has a red-purple color. Determine the absorption spec-trum of the solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 520 nm and 535 nm (salicylic acid). (2) Shake 0.3 g of Salicylated Alum Powder with 5 mL of methanol, filter, and use the filtrate as the sample solu-tion. Separately, dissolve 0.01 g of salicylic acid in 5 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on the plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of chloroform, acetone and ace-tic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample so-lution and that from the standard solution show the same Rf value. Spray evenly iron (III) chloride TS upon the plate: the spot from the standard solution and the corresponding spot from the sample solution reveal a purple color.
Weigh accurately about 0.33 g of Salicylated Alum Powder, add 80 mL of ethanol (95), and shake vigorously. Dilute with ethanol (95) to make exactly 100 mL, filter, and discard the first 10 mL of the filtrate. Use the subsequent fil-trate as the sample solution. Dissolve about 0.1 g of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours and accurately weighed, in sufficient ethanol (95) to make exactly 100 mL. Pipet 10 mL of this solution, dilute with ethanol (95) to make exactly 100 mL, and use the solu-tion as the standard solution. Pipet 10 mL each of the sam-ple solution and standard solution, to each add exactly 5 mL of a solution of iron (III) nitrate enneahydrate (1 in 200), and dilute with hydrochloric acid-potassium chloride buffer solution (pH 2.0) to make exactly 25 mL. Determine the ab-sorbances, AT and AS, of both solutions at 530 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared in the same manner with 10 mL of ethanol (95) as the blank. Amount (mg) of salicylic acid (C7H6O3) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of salicylic acid for assay taken
Saponated Cresol Solution contains not less than 42 volz and not more than 52 volz of cresol.
Saponated Cresol Solution is a yellow-brown to red-brown, viscous liquid. It has the odor of cresol. It is miscible with water, with ethanol (95) and with glyce-rin. It is alkaline.
Proceed as directed in the Identification un-der Cresol, using the distillate in the Purity (3).
(1) Alkalinity—Mix well 0.50 mL of Saponated Cresol Solution with 10 mL of neutralized ethanol, add 2 to 3 drops of phenolphthalein TS and 0.10 mL of 1 mol/L hy-drochloric acid VS: no red color develops. Unsaponified matter—To 1.0 mL of Saponated Cresol Solution add 5 mL of water, and shake: the solution is clear. ‌JP XVIII Official Monographs / Croconazole Hydrochloride 817 Cresol fraction—Transfer 180 mL of Saponated Cresol Solution to a 2000-mL distilling flask, add 300 mL of water and 100 mL of dilute sulfuric acid, and distil with steam until the distillate becomes clear. Draw off the water from the condenser, and continue the distillation until water vapor begins to come out of the tip of the condenser. Cool the condenser again, and continue distillation for 5 minutes. Dissolve 20 g of sodium chloride per 100 mL of the distillate, allow to stand, and collect the separated clear oil layer. After adding about 15 g of powdered calcium chloride for drying in small portions with frequent shaking, allow to stand for 4 hours. Filter, and distil exactly 50 mL of the filtrate: the dis-tillate is not less than 43 mL between 1969C and 2069C.
Transfer 5 mL of Saponated Cresol Solution, ex-actly measured, to a 500-mL distilling flask, holding the pipet vertically for 15 minutes to draw off the solution into the flask. Add 200 mL of water, 40 g of sodium chloride and 3 mL of dilute sulfuric acid, connect the distilling apparatus with the distilling flask, and distil into a cassia flask which contains 30 g of powdered sodium chloride and exactly 3 mL of kerosene, until the distillate reaches 90 mL. Draw off the water from the condenser, and continue the distillation until water vapor begins to come out of the tip of the condenser. Allow the cassia flask to stand in warm water for 15 minutes to dissolve the sodium chloride with frequent shaking. Cool to 159C, add a saturated solution of sodium chloride, and allow to stand for more than 3 hours with occasional shak-ing. Allow to stand for 1 to 2 minutes with gentle shaking, and combine the separated oil drops with the oil layer. The volume (mL) subtracted 3 (mL) from the oil layer measured represents the amount (mL) of cresol.
Sarpogrelate Hydrochloride Fine Granules
To an amount of Sarpogrelate Hydrochlo-ride Fine Granules, equivalent to 50 mg of Sarpogrelate Hy-drochloride, add 10 mL of 0.01 mol/L hydrochloric acid TS, allow to stand at room temperature for 10 minutes, then add 0.01 mol/L hydrochloric acid TS to make 100 mL, and dis-perse the particles by sonicating. Centrifuge this solution, and to 5 mL of the supernatant liquid add 0.01 mol/L hy- drochloric acid TS to make 50 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 269 nm and 273 nm and between 274 nm and 278 nm.
Related substances—Conduct this procedure within 3 hours after preparation of the sample solution. Powder Sarpogrelate Hydrochloride Fine Granules. To a portion of the powder, equivalent to 0.10 g of Sarpogrelate Hydrochlo-ride, add 50 mL of the mobile phase, and disperse the parti-cles by sonicating. Filter thorough a membrane filter with a pore size not exceeding 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solu-tion. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of decomposed substance A, having the relative retention time about 0.82 to sarpogrelate, obtained from the sample solution is not larger than 2.5 times that of sarpogrelate from the standard solu-tion, and the area of the peak other than sarpogrelate and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of sarpogrelate from the standard solution. For the peak area of the decomposed sub-stance A, multiply the correction factor, 0.78. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Sarpogrelate Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of sarpogrelate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of sarpogrelate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of sarpogrelate hy-drochloride in 20 mL of water, and use this solution as the sarpogrelate hydrochloride stock solution. To 1 mL of the sarpogrelate hydrochloride stock solution add 2 mL of so-dium hydroxide TS, shake thoroughly, allow to stand for 10 minutes, and add 3 mL of 1 mol/L hydrochloric acid TS. To this solution add 1 mL of the sarpogrelate hydrochloride stock solution, and add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the decomposed substance A and sarpogrelate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sarpogrelate is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Sarpogrelate Hydrochloride Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Sarpogrelate Hydrochloride Fine Granules add exactly V/10 mL of the internal standard solution, and add 4V/5 mL of the mobile phase, disperse the particles by sonicating, then add the mobile phase to make V mL so that each mL con-tains about 1 mg of sarpogrelate hydrochloride (C24H31NO6. HCl), and centrifuge. To 5 mL of the supernatant liquid add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × QT/QS × V/50 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Sarpogrelate Hydrochloride Fine Granules is not less than 85z. Start the test with an accuretely weighed amount of Sar-pogrelate Hydrochloride Fine Granules, equivalent to about 50 mg of sarpogrelate hydrochloride (C24H31NO6.HCl), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 25 mg of Sarpogrelate Hydrochloride RS (separately determine the water <2.48> in the same manner as Sarpogre-late Hydrochloride), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make ex-actly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 270 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS/MT × AT/AS × 1/C × 180 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (g) of Sarpogrelate Hydrochloride Fine Granules taken C: Labeled amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) in 1 g
Weigh accurately about 50 mg each of Sarpogrelate Method of preparation Prepare as directed under Gran-ules, with Sarpogrelate Hydrochloride. Identification To an amount of Sarpogrelate Hydrochlo-ride Fine Granules, equivalent to 50 mg of Sarpogrelate Hy-drochloride, add 10 mL of 0.01 mol/L hydrochloric acid TS, allow to stand at room temperature for 10 minutes, then add 0.01 mol/L hydrochloric acid TS to make 100 mL, and dis-perse the particles by sonicating. Centrifuge this solution, and to 5 mL of the supernatant liquid add 0.01 mol/L hy- drochloric acid TS to make 50 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 269 nm and 273 nm and between 274 nm and 278 nm. Purity Related substances—Conduct this procedure within 3 hours after preparation of the sample solution. Powder Sarpogrelate Hydrochloride Fine Granules. To a portion of the powder, equivalent to 0.10 g of Sarpogrelate Hydrochlo-ride, add 50 mL of the mobile phase, and disperse the parti-cles by sonicating. Filter thorough a membrane filter with a pore size not exceeding 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solu-tion. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of decomposed substance A, having the relative retention time about 0.82 to sarpogrelate, obtained from the sample solution is not larger than 2.5 times that of sarpogrelate from the standard solu-tion, and the area of the peak other than sarpogrelate and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of sarpogrelate from the standard solution. For the peak area of the decomposed sub-stance A, multiply the correction factor, 0.78. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Sarpogrelate Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of sarpogrelate, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of sarpogrelate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of sarpogrelate hy-drochloride in 20 mL of water, and use this solution as the sarpogrelate hydrochloride stock solution. To 1 mL of the sarpogrelate hydrochloride stock solution add 2 mL of so-dium hydroxide TS, shake thoroughly, allow to stand for 10 minutes, and add 3 mL of 1 mol/L hydrochloric acid TS. To this solution add 1 mL of the sarpogrelate hydrochloride stock solution, and add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the decomposed substance A and sarpogrelate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sarpogrelate is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Sarpogrelate Hydrochloride Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Sarpogrelate Hydrochloride Fine Granules add exactly V/10 mL of the internal standard solution, and add 4V/5 mL of the mobile phase, disperse the particles by sonicating, then add the mobile phase to make V mL so that each mL con-tains about 1 mg of sarpogrelate hydrochloride (C24H31NO6. HCl), and centrifuge. To 5 mL of the supernatant liquid add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × QT/QS × V/50 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Sarpogrelate Hydrochloride Fine Granules is not less than 85z. Start the test with an accuretely weighed amount of Sar-pogrelate Hydrochloride Fine Granules, equivalent to about 50 mg of sarpogrelate hydrochloride (C24H31NO6.HCl), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent fil-trate as the sample solution. Separately, weigh accurately about 25 mg of Sarpogrelate Hydrochloride RS (separately determine the water <2.48> in the same manner as Sarpogre-late Hydrochloride), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make ex-actly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solu-tion and standard solution at 270 nm as directed under Ul-traviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS/MT × AT/AS × 1/C × 180 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis MT: Amount (g) of Sarpogrelate Hydrochloride Fine Granules taken C: Labeled amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) in 1 g Assay Powder Sarpogrelate Hydrochloride Fine Granules. Weigh accurately a portion of the powder, equivalent to about 0.25 g of sarpogrelate hydrochloride (C24H31NO6. HCl), add exactly 25 mL of the internal standard solution, add 200 mL of the mobile phase, and disperse the particles by sonicating. To this solution add the mobile phase to make 250 mL, and centrifuge. To 5 mL of the supernatant liquid add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Sarpogrelate Hydrochloride RS (separately deter-mine the water <2.48> in the same manner as Sarpogrelate Hydroxide), add exactly 5 mL of the internal standard solu-tion, and add the mobile phase to make 50 mL. To 5 mL of this solution add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of sarpogrelate to that of the internal standard. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × QT/QS × 5 ‌MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Sarpogrelate Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sarpogrelate and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of sarpogrelate to that of the internal standard is not more than 1.0z.
Sarpogrelate Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled standard solution. For the peak area of the decomposed sub-stance A, multiply the correction factor, 0.78. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Sarpogrelate Hydrochloride. Time span of measurement: About 2.5 times as long as the retention time of sarpogrelate, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mobile phase to make exactly 50 mL. Confirm that the peak area of sarpogrelate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of sarpogrelate hy-drochloride in 20 mL of water, and use this solution as the sarpogrelate hydrochloride stock solution. To 1 mL of the sarpogrelate hydrochloride stock solution add 2 mL of so-dium hydroxide TS, shake thoroughly, allow to stand for 10 minutes, and add 3 mL of 1 mol/L hydrochloric acid TS. To this solution add 1 mL of the sarpogrelate hydrochloride stock solution, and add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the decomposed substance A and sarpogrelate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times amount of sarpogrelate hydrochloride (C HCl: 465.97). 24H 31NO6. with 10 mL of the standard solution under the above operat- ing conditions, the relative standard deviation of the peak area of sarpogrelate is not more than 2.0z. Method of preparation Prepare as directed under Tablets, with Sarpogrelate Hydrochloride. Identification Powder Sarpogrelate Hydrochloride Tablets. To a portion of the powder, equivalent to 50 mg of Sarpogrelate Hydrochloride, add 10 mL of 0.01 mol/L hy-drochloric acid TS, allow to stand at room temperature for 10 minutes, then add 0.01 mol/L hydrochloric acid TS to make 100 mL, and disperse the particles by sonicating. Cen-trifuge this solution, and to 5 mL of the supernatant liquid add 0.01 mol/L hydrochloric acid TS to make 50 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 269 nm and 273 nm, and between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sarpogrelate Hydrochloride Tablets add ex-actly V/10 mL of the internal standard solution, and disinte-grate the tablet. Add 4V/5 mL of the mobile phase, disperse the particles by sonicating, then add the mobile phase to make V mL so that each mL contains about 1 mg of sar-pogrelate hydrochloride (C24H31NO6.
Powder Sarpogrelate Hydrochloride Tablets. To a portion of the powder, equivalent to 50 mg of Sarpogrelate Hydrochloride, add 10 mL of 0.01 mol/L hy-drochloric acid TS, allow to stand at room temperature for 10 minutes, then add 0.01 mol/L hydrochloric acid TS to make 100 mL, and disperse the particles by sonicating. Cen-trifuge this solution, and to 5 mL of the supernatant liquid add 0.01 mol/L hydrochloric acid TS to make 50 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its maxima between 269 nm and 273 nm, and between 274 nm and 278 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sarpogrelate Hydrochloride Tablets add ex-actly V/10 mL of the internal standard solution, and disinte-grate the tablet. Add 4V/5 mL of the mobile phase, disperse the particles by sonicating, then add the mobile phase to make V mL so that each mL contains about 1 mg of sar-pogrelate hydrochloride (C24H31NO6.HCl), and centrifuge. To 5 mL of the supernatant liquid add the mobile phase to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl)
Related substances—Conduct this procedure within 12 hours after preparation of the sample solution. Powder Sarpogrelate Hydrochloride Tablets. To a portion of the powder, equivalent to 0.10 g of Sarpogrelate Hydrochloride, add 50 mL of the mobile phase, and disperse the particles by sonicating. Filter the solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Pipet 2 mL of the sample solution, add the mo-bile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of decomposed substance A, having the relative retention time about 0.82 to sarpogrelate, obtained from the sample solution is not larger than 1.5 times that of sarpogrelate from the standard solu-tion, and the area of the peak other than sarpogrelate and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of sarpogrelate from the ＝ MS × QT/QS × V/50 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Sarpogrelate Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Sarpogrelate Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 55.6 mg of sarpogrelate hydro-chloride (C24H31NO6.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of ‌Sarpogrelate Hydrochloride RS (separately determine the water <2.48> in the same manner as Sarpogrelate Hydrochlo-ride), and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, of the sample solution and standard solution at 270 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) in 1 tablet
Weigh accurately the mass of not less than 20 Sar-pogrelate Hydrochloride Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.25 g of sarpogrelate hydrochloride (C24H31NO6.HCl), add exactly 25 mL of the internal standard solution, add about 200 mL of the mobile phase, and disperse the particles by sonicating. To this solution add the mobile phase to make 250 mL, and centrifuge. To 5 mL of the supernatant liquid add the mobile phase to make 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of Sar-pogrelate Hydrochloride RS (separately determine the water <2.48> in the same manner as Sarpogrelate Hydroxide), add exactly 5 mL of the internal standard solution, and add the mobile phase to make 50 mL. To 5 mL of this solution add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> under the following conditions, and calculate the ratios, QT and QS, of the peak area of sar-pogrelate to that of the internal standard. Amount (mg) of sarpogrelate hydrochloride (C24H31NO6.HCl) ＝ MS × QT/QS × 5 MS: Amount (mg) of Sarpogrelate Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (1 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Sarpogrelate Hydrochloride. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sarpogrelate and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of sarpogrelate to that of the internal standard is not more than 1.0z.
Scopolia Extract and Carbon Powder is easily dustable and black in color. It is tasteless.
Scopolia Extract and Ethyl Aminobenzoate Powder contains not less than 22.
Scopolia Extract and Ethyl Aminobenzoate Powder is slightly brownish white in color. It has a slightly bitter taste, leaving a sensation of numbness on the tongue. ‌Identification (1) To 2 g of Scopolia Extract and Ethyl Aminobenzoate Powder add 20 mL of diethyl ether, shake, and filter through a glass filter (G4). Wash the residue with three 10-mL portions of diethyl ether, combine the filtrate and the washings, evaporate to dryness, and perform the fol-lowing test with the residue (ethyl aminobenzoate). Dissolve 0.01 g of the residue in 1 mL of dilute hydro-chloric acid and 4 mL of water: the solution responds to Qualitative Tests <1.09> for primary aromatic amines. Dissolve 0.1 g of the residue in 5 mL of water with the aid of dilute hydrochloric acid added dropwise, and add iodine TS dropwise: a brown precipitate is produced. Warm 0.05 g of the residue with 2 drops of acetic acid (31) and 5 drops of sulfuric acid: the odor of ethyl ace-tate is perceptible. To the diethyl ether-insoluble residue obtained in (1) add 30 mL of water, shake gently, and filter: the filtrate responds to Qualitative Tests <1.09> for sodium salt and for bicarbonate. To the water-insoluble residue obtained in (2) add 10 mL of dilute hydrochloric acid, shake, and filter: the filtrate responds to Qualitative Tests <1.09> for magnesium salt. Place 30 g of Scopolia Extract and Ethyl Aminoben-zoate Powder in a glass-stoppered conical flask, add 100 mL of water, shake for 30 minutes, and filter immediately by suction through a glass filter (G3). Transfer the residue in the flask to the same glass filter with the filtrate, and filter the residue by suction while pressing vigorously the residue on the same glass filter. Place 75 mL of the filtrate in a 300-mL beaker, and add cautiously 10 mL of diluted sulfuric acid (1 in 3). Add 0.2 mL of bromocresol green TS to this so-lution, and add dilute sulfuric acid dropwise while shaking thoroughly, until the color of the solution changes from green to yellow-green. After cooling, place this solution in a separator, wash with two 25-mL portions of a mixture of hexane and diethyl ether (1:1) by shaking well, and place the aqueous layer in another separator. Make slightly alkaline with ammonia TS, add immediately 30 mL of diethyl ether, and shake well. Wash the diethyl ether layer with two 10-mL portions of a saturated solution of sodium chloride, separate the diethyl ether layer, add 3 g of anhydrous sodium sulfate, shake, and filter through a pledget of cotton. Evaporate the filtrate to dryness, dissolve the residue in 0.2 mL of ethanol (95), and use this solution as the sample solution. Separately, dissolve 2 mg of Atropine Sulfate RS or atropine sulfate hy-drate for thin-layer chromatography and 1 mg of Scopola-mine Hydrobromide RS or scopolamine hydrobromide hy-drate for thin-layer chromatography in 1 mL each of ethanol (95), and use these solutions as standard solution (1) and standard solution (2), respectively. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution, standard solution (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ace-tone, water and ammonia solution (28) (90:7:3) to a distance of about 10 cm, and dry the plate at 809C for 10 minutes. After cooling, spray evenly Dragendorff's TS for spraying on the plate: two principal spots obtained from the sample solution show the same color tone and the same Rf value with each spot from the standard solutions, respectively.
Weigh accurately about 0.3 g of Scopolia Extract and Ethyl Aminobenzoate Powder, transfer to a Soxhlet ex-tractor, extract with 100 mL of diethyl ether for 1 hour, and evaporate the solvent on a water bath. Dissolve the residue in 25 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 250 mL, and use this solution as the sample solution. Weigh accurately about 75 mg of Ethyl Aminoben-zoate RS, previously dried in a desiccator (silica gel) for 3 hours, dissolve in 25 mL of 1 mol/L hydrochloric acid TS, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 250 mL, and use this so-lution as the standard solution. Pipet 5 mL each of the sample solution and standard solution, to each add 10 mL of 1 mol/L hydrochloric acid TS, then add 1 mL of a solution of sodium nitrite (1 in 200), prepared before use, and allow to stand for 5 minutes with occasional shaking. Add 5 mL of ammonium amidosulfate TS, shake well, and allow to stand for 10 minutes. Add 2 mL of N-N-diethyl-N?-1-naphthylethylenediamine oxalate-acetone TS, mix immedi-ately, and add water to make exactly 50 mL. Allow to stand for 2 hours, determine the absorbances, AT and AS, of these solutions at 550 nm, as directed under Ultraviolet-visible Spectrophotometry <2.24> using a blank prepared in the same manner with 5 mL of water in place of the sample solu-tion. Amount (mg) of ethyl aminobenzoate (C9H11NO2) ＝ MS × AT/AS MS: Amount (mg) of Ethyl Aminobenzoate RS taken
Scopolia Extract Powder contains not less than 0.
Scopolia Extract Powder is a brownish yellow to grayish yellow-brown powder. It has a faint, characteristic odor and a slightly bitter taste.
(1) To 20 g of Scopolia Extract Powder add 15 mL of water and 8 mL of ammonia TS, mix homoge-neously, add 100 mL of diethyl ether and 7 g of sodium chlo-ride, stopper tightly, shake for 1 hour, add 5 g of powdered tragacanth, and shake vigorously. Allow to stand for 5 minutes, take the clearly separated diethyl ether layer, and filter. Proceed with the filtrate as directed in the Identifica-tion (1) under Scopolia Extract. (2) Place 5.0 g of Scopolia Extract Powder in a glass-stoppered centrifuge tube, add 30 mL of ammonia TS, soni-cate for 5 minutes, and centrifuge. Transfer the supernatant liquid to a separator, add 40 mL of ethyl acetate, and shake. Drain off the ethyl acetate layer, add 3 g of anhydrous so-dium sulfate to the ethyl acetate, shake, and filter after the ethyl acetate becomes clear. Evaporate the solvent of the fil-trate under low pressure (in vacuo), dissolve the residue in 1 mL of ethanol (95), and use this solution as the sample solu-tion. Proceed as directed in the Identification (2) under Scopolia Rhizome.
Weigh accurately about 4 g of Scopolia Extract Powder, place in a glass-stoppered centrifuge tube, add 15 mL of ammonia TS, and shake. Add 25 mL of diethyl ether, stopper tightly, shake for 15 minutes, centrifuge to take the diethyl ether layer. To the aqueous layer add 25 mL of diethyl ether, proceed in the same manner, and repeat this procedure three times. Combine all the extracts, and evapo-rate the solvent on a water bath. Dissolve the residue in 5 mL of the mobile phase, add exactly 3 mL of the internal stand- ‌2136‌ Scopolia Extract and Carbon Powder / Crude Drugs and Related Drugs ard solution, and add the mobile phase to make 25 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.8 mm, discard the first 2 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 25 mg of Atropine Sul-fate RS (separately determine the loss on drying <2.41> under the same manner as Atropine Sulfate Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solu-tion as standard stock solution A. Weigh accurately about 25 mg of Scopolamine Hydrobromide RS (separately determine the loss on drying <2.41> under the same manner as Scopola-mine Hydrobromide Hydrate), dissolve in the mobile phase to make exactly 25 mL, and use this solution as standard stock solution B. Pipet 5 mL of the standard stock solution A and 1 mL of the standard stock solution B, add exactly 3 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions. Calculate the ratios, QTA and QSA, of the peak area of hyoscyamine (atropine), and ratios, QTS and QSS, of the peak area of scopolamine to that of the internal standard in each solution, calculate the amounts of hyoscyamine and scopola-mine by the following equation, and designate the total as the amount of total alkaloids. Amount (mg) of hyoscyamine (C17H23NO3) ＝ MSA × QTA/QSA × 1/5 × 0.855 Amount (mg) of scopolamine (C17H21NO4) ＝ MSS × QTS/QSS × 1/25 × 0.789 MSA: Amount (mg) of Atropine Sulfate RS taken, calcu-lated on the dried basis MSS: Amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of brucine dihydrate in the mobile phase (1 in 2500). Operating conditions— Detector: An ultraviolet absorption spectrometer (wave-length: 210 nm). Column: A stainless steel column about 4 mm in inside di-ameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of a solution obtained by dis-solving 6.8 g of potassium dihydrogenphosphate in 900 mL of water, adding 10 mL of triethylamine, adjusting the pH to 3.5 with phosphoric acid, and adding water to make 1000 mL, and acetonitrile (9:1). Flow rate: Adjust so that the retention time of scopola-mine is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, scopolamine, atropine and the internal standard are eluted in this order with the resolution between the peaks of scopolamine and atropine being not less than 11, and the resolution between the peaks of atropine and the internal standard being not less than 4.
Senega Syrup is a yellow-brown, viscous liq-uid. It has a characteristic odor resembling methyl salicylate and a sweet taste.
Add 5 mL of water to 1 mL of Senega Syrup, and shake: lasting small bubbles are produced.
‌Silodosin Orally Disintegrating Tablets
Conduct this procedure using light-resistant vessels. To 1 tablet of Silodosin Orally Disintegrating Tablets add 15 mL of a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) per 1 mg of Silodosin, and sonicate until the tablet is completely disintegrated while oc-casional shaking. Add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) so that each mL contains about 40 mg of Silodosin, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, dissolve 20 mg of Silodosin RS in a mixture of methanol and a solution of sodium chlo-ride (1 in 200) (7:3) to make 50 mL. To 5 mL of this solution add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions: the retention times of the principal peaks in the chromatograms obtained from the sample solution and standard solution are the same, and both absorption spectra of these peaks exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: A photodiode array detector (wavelength: 270 nm, spectrum range of measurement: 200 – 370 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay.
Related substances—Conduct this procedure using light-resistant vessels. To a number of Silodosin Orally Dis-integrating Tablets, equivalent to 20 mg of Silodosin, add 60 mL of a mixture of methanol and a solution of sodium chlo-ride (1 in 200) (7:3), sonicate until the tablet is completely disintegrated while occasional shaking, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 100 mL. Centrifuge this solution, and filter the su-pernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine each peak area by the automatic integration method: the peak area of related substance A, having the relative retention time of about 1.3 to silodosin, obtained from the sample solution is not lager than the peak area of silodosin from the standard solution, the area of the peak other than silodosin and the peak mentioned above from the sample so-lution is not larger than 1/4 times the peak area of silodosin from the standard solution. Furthermore, the total area of the peaks other than silodosin from the sample solution is not larger than 2 times the peak area of silodosin from the standard solution. For the peak area of related substance A, multiply the correction factor 0.6. Operating conditions— Detector, column, column temperature, and mobile phases A and B: Proceed as directed in the Purity (2) under Silodosin. Flowing of the mobile phase: Control the gradient by mix-ing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 15 – 47 75 → 35 25 → 65 47 – 53 Flow rate: Adjust so that the retention time of silodosin is about 13 minutes. Time span of measurement: About 3.5 times as long as the retention time of silodosin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 20 mL. Con-firm that the peak area of silodosin obtained with 25 mL of this solution is equivalent to 3.5 to 6.5z of that with 25 mL of the standard solution. System performance: Thinly spread out an amount of silodosin in a petri dish, exposure to a 4000 lx light for not less than 24 hours using a D65 fluorescent lamp, and dissolve 4 mg of this sample in a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 20 mL. When the procedure is run with 25 mL of this solution under the above operating conditions, the resolution between the peaks of silodosin and related substance A is not less than 6. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Silodosin Orally Disintegrating Tablets add 3V/5 mL of a mixture of methanol and a solution of sodium chlo-ride (1 in 200) (7:3), sonicate until the tablet is completely disintegrated while occasional shaking. Add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly V mL so that each mL contains about 40 mg of silodosin (C25H32F3N3O4), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of silodosin (C25H32F3N3O4) ＝ MS × AT/AS × V/500 MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis ‌Disintegration Being specified separately when the drug is granted approval based on the Law. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 15 minutes of Silodosin Orally Disintegrating Tablets is not less than 80z. Start the test with 1 tablet of Silodosin Orally Disintegrat-ing Tablets, withdraw not less than 9 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.2 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 1.1 mg of silodosin (C25H32F3N3O4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Silodosin RS (separately determine the water <2.48> in the same manner as Silodosin), and dissolve in 0.1 mol/L hydro-chloric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make ex-actly 50 mL. Pipet 5 mL of this solution, add 0.1 mol/L hy-drochloric acid TS to make exactly 100 mL, and use this so-lution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of silodosin in each solution. Dissolution rate (z) with respect to the labeled amount of silodosin (C25H32F3N3O4) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of silodosin (C25H32F3N3O4) in 1 tablet Operating conditions— Proceed as directed in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of silodosin are not less than 3000 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.0z.
Conduct this procedure using light-resistant vessels. To 20 tablets of Silodosin Orally Disintegrating Tablets add 3V/5 mL of a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3), and sonicate until the tablet is com-pletely disintegrated while occasional shaking. Add a mix-ture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly V mL so that each mL contains about 160 mg of silodosin (C25H32F3N3O4). Pipet 5 mL of this solu-tion, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 20 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of Silodosin RS (separately deter-mine the water <2.48> in the same manner as Silodosin), and dissolve in a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of silodosin in each solution. Amount (mg) of silodosin (C25H32F3N3O4) in 1 tablet ＝ MS × AT/AS × V/2500 MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.9 g of sodium dihydrogen phos-phate dihydrate in 1000 mL of water, and adjust to pH 3.4 with diluted phosphoric acid (1 in 10). To 730 mL of this so-lution add 270 mL of acetonitrile. Flow rate: Adjust so that the retention time of silodosin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of silodosin are not less than 3000 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 1.0z.
Silodosin Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of silodosin (C25H32F3N3O4: 495.53). Method of preparation Prepare as directed under Tablets, with Silodosin. Identification Conduct this procedure using light-resistant vessels. To an amount of powdered Silodosin Tablets, equivalent to 2 mg of Silodosin, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3), sonicate with occasional shaking, then add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, dissolve 20 mg of Silodosin RS in a mixture of metha-nol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL. To 5 mL of this solution add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and use this solution as the standard solu- tion. Perform the test with 25 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions: the retention times of the principal peaks in the chromatograms obtained from these solutions are the same, and the absorp-tion spectra of these peaks exhibit similar intensities of ab-sorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Silodosin. Detector: A photodiode array detector (wavelength: 270 nm, spectrum measuring range: 220 – 370 nm). System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of silodosin are not less than 3000 and not more than 1.6, respectively. Purity Related substances—Conduct this procedure using light-resistant vessels. Powder not less than 10 Silodosin Tablets. To a portion of the powder, equivalent to 20 mg of Silodosin, add a mixture of methanol and a solution of so-dium chloride (1 in 200) (7:3), sonicate with occasional shak-ing, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 100 mL. Centrifuge this so-lution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of related sub-stance A, having the relative retention time of about 1.3 to silodosin, obtained from the sample solution is not larger than the peak area of silodosin from the standard solution, the area of the peak other than silodosin and the peak men-tioned above from the sample solution is not larger than 1/4 times the peak area of silodosin from the standard solution. Furthermore, the total area of the peaks other than silodosin from the sample solution is not larger than 2 times the peak area of silodosin from the standard solution. For the peak System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 20 mL. Con-firm that the peak area of silodosin obtained with 25 mL of this solution is equivalent to 3.5 to 6.5z of that with 25 mL of the standard solution. System performance: Thinly spread out an amount of silodosin in a petri dish, exposure it to a 4000 lx light for not less than 24 hours using a D65 lamp, and dissolve 4 mg of this sample in a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 20 mL. When the procedure is run with 25 mL of this solution under the above operating conditions, the resolution between the peak of silodosin and related substance A is not less than 6. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Silodosin Tablets add exactly 2V/25 mL of the in-ternal standard solution, then add a suitable amount of a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3), sonicate until the tablet is completely disintegrat-ed with occasional stirring, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make V mL so that each mL contains about 40 mg of silodosin (C25H 32F3N3O4), and filter through a membrane filter with a pore size not exceeding 0.
Conduct this procedure using light-resistant vessels. To an amount of powdered Silodosin Tablets, equivalent to 2 mg of Silodosin, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3), sonicate with occasional shaking, then add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, dissolve 20 mg of Silodosin RS in a mixture of metha-nol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL. To 5 mL of this solution add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and use this solution as the standard solu- tion. Perform the test with 25 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions: the retention times of the principal peaks in the chromatograms obtained from these solutions are the same, and the absorp-tion spectra of these peaks exhibit similar intensities of ab-sorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay under Silodosin. Detector: A photodiode array detector (wavelength: 270 nm, spectrum measuring range: 220 – 370 nm). System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of silodosin are not less than 3000 and not more than 1.6, respectively.
Related substances—Conduct this procedure using light-resistant vessels. Powder not less than 10 Silodosin Tablets. To a portion of the powder, equivalent to 20 mg of Silodosin, add a mixture of methanol and a solution of so-dium chloride (1 in 200) (7:3), sonicate with occasional shak-ing, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 100 mL. Centrifuge this so-lution, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Pipet 1 mL of the sample solution, add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the peak area of related sub-stance A, having the relative retention time of about 1.3 to silodosin, obtained from the sample solution is not larger than the peak area of silodosin from the standard solution, the area of the peak other than silodosin and the peak men-tioned above from the sample solution is not larger than 1/4 times the peak area of silodosin from the standard solution. Furthermore, the total area of the peaks other than silodosin from the sample solution is not larger than 2 times the peak area of silodosin from the standard solution. For the peak System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make exactly 20 mL. Con-firm that the peak area of silodosin obtained with 25 mL of this solution is equivalent to 3.5 to 6.5z of that with 25 mL of the standard solution. System performance: Thinly spread out an amount of silodosin in a petri dish, exposure it to a 4000 lx light for not less than 24 hours using a D65 lamp, and dissolve 4 mg of this sample in a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 20 mL. When the procedure is run with 25 mL of this solution under the above operating conditions, the resolution between the peak of silodosin and related substance A is not less than 6. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.5z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. To 1 tablet of Silodosin Tablets add exactly 2V/25 mL of the in-ternal standard solution, then add a suitable amount of a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3), sonicate until the tablet is completely disintegrat-ed with occasional stirring, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make V mL so that each mL contains about 40 mg of silodosin (C25H 32F3N3O4), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of Silodosin RS (separately determine the water <2.48> in the same manner as Silodosin), and dissolve in a mixture of methanol and a solu-tion of sodium chloride (1 in 200) (7:3) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 4 mL of the in-ternal standard solution, then add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of silodosin to that of the internal standard. area of related substance A, multiply the correction factor 0.6. Operating conditions— Amount (mg) of silodosin (C25H ＝ MS × QT/QS × V/500 32F3N3O4) Detector, column, column temperature, mobile phase A and mobile phase B: Proceed as directed in the operating conditions in the Purity (2) under Silodosin. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) (1 in 8000). Operating conditions— Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Proceed as directed in the operating conditions in the Assay under Silodosin. 0 – 15 75 25 15 – 47 75 → 35 25 → 65 47 – 53 35 65 Flow rate: Adjust so that the retention time of silodosin is about 13 minutes. Time span of measurement: About 3.5 times as long as the retention time of silodosin, beginning after the solvent peak. System suitability— Proceed as directed in the system suitability in the Assay. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Silodosin Tablets is not less than 80z. Start the test with 1 tablet of Silodosin Tablets, withdraw not less than 9 mL of the medium at the specified minute ‌after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add 0.2 mol/L hydrochloric acid TS to make exactly V? mL so that each mL contains about 1.1 mg of silodosin (C25H32F3N3O4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Silodosin RS (separately determine the water <2.48> in the same manner as Silodosin), and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add 0.1 mol/L hydrochloric acid TS to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of silodosin in each solution. Dissolution rate (z) with respect to the labeled amount of silodosin (C25H32F3N3O4) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of silodosin (C25H32F3N3O4) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay under Silodosin. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of silodosin are not less than 3000 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of silodosin is not more than 2.0z.
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of not less than 20 Silodosin Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 40 mg of silodosin (C25H32F3N3O4), add exactly 8 mL of the internal standard solution, then add a suitable amount of a mixture of metha-nol and a solution of sodium chloride (1 in 200) (7:3), soni-cate with occasional shaking, and add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 100 mL. To 5 mL of this solution add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 3 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of Silodosin RS (separately determine the water <2.48> in the same manner as Silodosin), add exactly 4 mL of the internal standard solu-tion and a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL. To 5 mL of this so-lution add a mixture of methanol and a solution of sodium chloride (1 in 200) (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of silodosin to that of the internal standard. Amount (mg) of silodosin (C25H32F3N3O4) ＝ MS × QT/QS × 2 MS: Amount (mg) of Silodosin RS taken, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of methanol and a sodium chlo-ride solution (1 in 200) (7:3) (1 in 800). Operating conditions— Proceed as directed in the operating conditions the Assay under Silodosin. System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, silodosin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of silodosin to that of the internal standard is not more than 1.0z.
Silver Nitrate Ophthalmic Solution is a clear, colorless liquid.
Silver Nitrate Ophthalmic Solution responds to Qualitative Tests <1.09> for silver salt and for nitrate.
Measure accurately 20 mL of Silver Nitrate Ophthal-mic Solution, add 30 mL of water and 2 mL of nitric acid, and titrate <2.50> with 0.1 mol/L ammonium thiocyanate VS (indicator: 2 mL of ammonium iron (III) sulfate TS). Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 16.99 mg of AgNO3
Silver Protein Solution contains not less than 0.
Silver Protein Solution is a clear, brown liq-uid, having the odor of mentha oil.
(1) To 1 mL of Silver Protein Solution add 10 mL of ethanol (95), mix, and add 2 mL of sodium hy-droxide TS. Add immediately 1 mL of a solution of copper (II) chloride dihydrate in ethanol (95) (1 in 10), shake, and filter: the filtrate is blue in color (glycerin). To 3 mL of Silver Protein Solution add water to make 10 mL, add 2 mL of dilute hydrochloric acid, shake frequently for 5 minutes, and filter. Add 5 mL of a solution of sodium hydroxide (1 in 10) to the filtrate, and add 2 mL of diluted copper (II) sulfate TS (2 in 25): a purple color de-velops (silver protein). To 5 mL of the sample solution obtained in (2) add iron (III) chloride TS dropwise: a brown precipitate is formed (silver protein). Place 3 mL of Silver Protein Solution in a crucible, heat cautiously, and evaporate almost to dryness. Then in-cinerate gradually by strong heating, dissolve the residue in 1 mL of nitric acid by warming, and add 10 mL of water: the solution responds to Qualitative Tests <1.09> (1) for silver salt.
Pipet 25 mL of Silver Protein Solution into a 250-mL Kjeldahl flask, and heat cautiously until a white gas of glycerin is evolved. After cooling, add 25 mL of sulfuric acid, cover the flask with a funnel, and heat gently for 5 minutes. After cooling, drop gradually 5 mL of nitric acid, heat with occasional shaking in a water bath for 45 minutes, and cool. Add 2 mL of nitric acid, boil gently, and repeat this operation until the solution becomes colorless upon ‌cooling. Transfer cautiously the cooled content in the flask into a 500-mL conical flask with 250 mL of water. Boil gently for 5 minutes, cool, and titrate <2.50> with 0.1 mol/L ammonium thiocyanate VS (indicator: 3 mL of ammonium iron (III) sulfate TS).‌ Each mL of 0.1 mol/L ammonium thiocyanate VS ＝ 10.79 mg of Ag
Simple Ointment is yellow in color. It has a slight, characteristic odor.
Simple Syrup is a clear, colorless to pale yel-low, viscous liquid. It is odorless and has a sweet taste.
(1) Evaporate Simple Syrup on a water bath to dryness. 1 g of the residue so obtained, when ignited, melts to swell, and decomposes, emitting an odor of caramel, to bulky charcoal. (2) To 0.1 g of the residue obtained in (1) add 2 mL of dilute sulfuric acid, boil, add 4 mL of sodium hydroxide TS and 3 mL of Fehling's TS, and heat to boiling: a red to dark red precipitate is produced. Specific gravity <2.56> d 20: 1.310 – 1.325
(1) Artificial sweetening agents—To 100 mL of Simple Syrup add 100 mL of water, shake, acidify a 50-mL portion of the solution with dilute sulfuric acid, and make another 50-mL portion alkaline with sodium hydroxide TS. To each portion add 100 mL of diethyl ether, shake, separate the diethyl ether layer, and evaporate the combined diethyl ether extract on a water bath to dryness: the residue has no sweet taste. (2) Salicylic acid—To the residue obtained in (1) add 2 to 3 drops of dilute iron (III) chloride TS: no purple color de-velops.
Simvastatin Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of sim-vastatin (C25H38O5: 418.57). Method of preparation Prepare as directed under Tablets, with Simvastatin. Identification To an amount of powdered Simvastatin Tablets, equivalent to about 2.5 mg of Simvastatin, add 25 mL of acetonitrile, sonicate for 15 minutes, and centrifuge. To 2 mL of the supernatant liquid add acetonitrile to make 20 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 229 nm and 233 nm, be- tween 236 nm and 240 nm, and between 245 nm and 249 nm. Purity Related substances—Powder not less than 20 Sim-vastatin Tablets. To a portion of the powder, equivalent to about 50 mg of Simvastatin, add 200 mL of a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1), and sonicate for 15 minutes. After cooling, add the same mixture to make 250 mL, and centrifuge. To 5 mL of the supernatant liquid add the same mixture to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.5 to simvastatin obtained from the sample solution is not larger than 1.6 times the peak area of simvastatin from the standard solution, the area of the peak having the rela-tive retention time of about 2.0 from the sample solution is not larger than the peak area of simvastatin from the stand- tin), and dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the following condi-tions, and determine the peak areas, AT and AS, of simvasta-tin in each solution. Dissolution rate (z) with respect to the labeled amount ard solution, and the total area of the peaks other than sim- vastatin is not larger than 4 times the peak area of simvasta- of simvastatin (C 25H 38O5) tin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of simvastatin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of simvastatin obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of simvastatin are not less than 6000 and 0.9 – 1.1, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of simvastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Simvastatin Tablets add V/20 mL of water, and disintegrate the tablet by sonicating. Add a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1) to make 3V/4 mL, and sonicate for 15 minutes. After cooling, add the same mixture to make exactly V mL so that each mL contains about 0.
To an amount of powdered Simvastatin Tablets, equivalent to about 2.5 mg of Simvastatin, add 25 mL of acetonitrile, sonicate for 15 minutes, and centrifuge. To 2 mL of the supernatant liquid add acetonitrile to make 20 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 229 nm and 233 nm, be- tween 236 nm and 240 nm, and between 245 nm and 249 nm.
Related substances—Powder not less than 20 Sim-vastatin Tablets. To a portion of the powder, equivalent to about 50 mg of Simvastatin, add 200 mL of a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1), and sonicate for 15 minutes. After cooling, add the same mixture to make 250 mL, and centrifuge. To 5 mL of the supernatant liquid add the same mixture to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1) to make exactly 200 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.5 to simvastatin obtained from the sample solution is not larger than 1.6 times the peak area of simvastatin from the standard solution, the area of the peak having the rela-tive retention time of about 2.0 from the sample solution is not larger than the peak area of simvastatin from the stand- tin), and dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the following condi-tions, and determine the peak areas, AT and AS, of simvasta-tin in each solution. Dissolution rate (z) with respect to the labeled amount ard solution, and the total area of the peaks other than sim- vastatin is not larger than 4 times the peak area of simvasta- of simvastatin (C 25H 38O5) tin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of simvastatin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of simvastatin obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of simvastatin are not less than 6000 and 0.9 – 1.1, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of simvastatin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Simvastatin Tablets add V/20 mL of water, and disintegrate the tablet by sonicating. Add a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1) to make 3V/4 mL, and sonicate for 15 minutes. After cooling, add the same mixture to make exactly V mL so that each mL contains about 0.1 mg of simvastatin (C25H38O5), centrifuge, and use the supernatant liquid as the sample solu-tion. Then, proceed as directed in the Assay. Amount (mg) of simvastatin (C25H38O5) ＝ MS × AT/AS × V/200 MS: Amount (mg) of Simvastatin RS taken, calculated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution of polysorbate 80, prepared by dissolving 3 g in water to make 1000 mL, as the dissolution medium, the dissolution rate in 45 minutes of Simvastatin Tablets is not less than 70z. ＝ MS × AT/AS × V?/V × 1/C × 45/2 MS: Amount (mg) of Simvastatin RS taken, calculated on the dried basis C: Labeled amount (mg) of simvastatin (C25H38O5) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of methanol and 0.02 mol/L po-tassium dihydrogen phosphate TS (4:1). Flow rate: Adjust so that the retention time of simvastatin is about 4 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of simvastatin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of simvastatin is not more than 1.0z.
Weigh accurately the mass of not less than 20 Sim-vastatin Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of simvastatin (C25H38O5), add 200 mL of a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1), and sonicate for 15 minutes. After cooling, add the same mixture to make exactly 250 mL, and centrifuge. Pipet 5 mL of the superna-tant liquid, add the same mixture to make exactly 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Simvastatin RS (separately determine the loss on drying <2.41> under the same condi-tions as Simvastatin), dissolve in a mixture of acetonitrile and 0.05 mol/L acetate buffer solution (pH 4.0) (4:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of simvastatin in each solution. Start the test with 1 tablet of Simvastatin Tablets, with-draw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane Amount (mg) of simvastatin (C ＝ MS × AT/AS × 5/2 25H38O5) filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of simvastatin (C25H38O5), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Simvastatin RS (separately determine the loss on drying <2.41> under the same conditions as Simvasta- MS: Amount (mg) of Simvastatin RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame- ‌ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 3.90 g of sodium dihydrogen phos-phate dihydrate in 900 mL of water, adjust to pH 4.5 with sodium hydroxide TS or phosphoric acid, and add water to make 1000 mL. To 700 mL of this solution add 1300 mL of acetonitrile. Flow rate: Adjust so that the retention time of simvastatin is about 9 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of simvastatin are not less than 6000 and 0.9 – 1.1, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of simvastatin is not more than 1.0z.
Sitagliptin Phosphate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of sitagliptin (C16H15F6N5O: 407.31). Method of preparation Prepare as directed under Tablets, with Sitagliptin Phosphate Hydrate. ＝ MS × AT/AS MS: Amount (mg) of Sitagliptin Phosphate RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with cyanopropylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.0 with phos-phoric acid, and add water to make 1000 mL. To 850 mL of this solution add 150 mL of acetonitrile for liquid chroma-tography. Flow rate: 1.0 mL per minute. System suitability— System performance: Place 10 mg of Sitagliptin Phos-phate RS and 1 mg of sodium stearyl fumarate in a vial, and add 1 mL of water. Stopper the vial tightly, and heat at 809C for 20 to 48 hours. Take out the contents of the vial, wash the vial 3 times with a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1), combine the washings and the content, and add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make 100 mL. Stir this solution for 1 hour, and centrifuge for 10 minutes or until the solu- Manufacture The management strategy of Sitagliptin Phosphate Tablets is based on systematic development methods, which put emphasis on prior setting targets, understanding of products and processes, and process con-trol, and which is based on quality risk management and proven science. In addition when it can be scientiˆcally pos-sible to explain that a disintegration test ensure quality with distinguishability equal or better than a dissolution test, the following disintegration is alternative for the estimation of dissolution. Disintegration <6.09> Perform the test for 5 minutes: it meets the requirement. Identiˆcation (1) To 1 tablet of Sitagliptin Phosphate Tablets add water so that each mL contains about 0.
Related substances—Use the sample solution ob-tained in the Assay as the sample solution. Separately, pipet 1 mL of the standard solution obtained in the Assay, add a mixture of diluted phosphoric acid (1 in 1000) and aceto- nitrile for liquid chromatography (19:1) to make exactly 500 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak area, AT, of related substance obtained from the sam-ple solution and the peak area, AS, of sitagliptin from the standard solution, and calculate the amount of related sub-stances by the following equation: the total amount of related substances is not more than 0.2z. For this calcula-tion the peak area of the related substance not more than 0.1 z is excluded. Amount (z) of related substance ＝ MS × AT/AS × V?/V × 1/C × 1/50 × 0.806 MS: Amount (mg) of Sitagliptin Phosphate RS taken, cal-culated on the anhydrous basis V?/V: Dilution factor for the sample solution in the Assay C: Labeled amount (mg) of sitagliptin (C16H15F6N5O) in 1 tablet Operating conditions— Detector, column, column temperature, mobile phase and ‰ow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5.5 times as long as the retention time of sitagliptin, beginning after the solvent peak. System suitability— System performance and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: To 5 mL of the standard solution add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make exactly 10 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the SN ratio of the peak of sitagliptin is not less than 10. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sitagliptin Phosphate Tablets add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make exactly 25 mL, and stir thoroughly. Pipet V mL of this solution, and add a mixture of diluted phosphoric acid (1 in 1000) and aceto-nitrile for liquid chromatography (19:1) to make exactly V? mL so that each mL contains about 80 mg of sitagliptin (C16H15F6N5O). Centrifuge this solution, and use the super-natant liquid as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of sitagliptin (C16H15F6N5O) ＝ MS × AT/AS × V?/V × 1/10 × 0.806 MS: Amount (mg) of Sitagliptin Phosphate RS taken, cal-culated on the anhydrous basis Dissolution <6.10> When the test is performed at 100 revo-lutions per minute according to the Basket method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Sitagliptin Phosphate Tablets is not less than 85z. Start the test with 1 tablet of Sitagliptin Phosphate Tablets, withdraw not less than 4 mL of the medium at the speciˆed minute after starting the test, and ˆlter through a membrane ˆlter with a pore size of 0.45 mm. Discard not less than 2 mL of the ˆrst ˆltrate, pipet V mL of the subsequent ˆltrate, add water to make exactly V? mL so that each mL contains about 14 mg of sitagliptin (C16H15F6N5O), and use this solution as the sample solution. Separately, weigh accu-rately about 29 mg of Sitagliptin Phosphate RS (separately determine the water <2.48> in the same manner as Sitagliptin Phosphate Hydrate), and dissolve in a solution of sodium chloride (37 in 25,000) to make exactly 100 mL. Pipet 6 mL of this solution, and add a solution of sodium chloride (37 in 25,000) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of sitagliptin in each solution. Dissolution rate (z) with respect to the labeled amount of sitagliptin (C16H15F6N5O) ＝ MS × AT/AS × V?/V × 1/C × 54 × 0.806 MS: Amount (mg) of Sitagliptin Phosphate RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of sitagliptin (C16H15F6N5O) in 1 tablet Operating conditions— Column, column temperature and ‰ow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 267 nm). Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.0 with phos-phoric acid, and add water to make 1000 mL. To 750 mL of this solution add 250 mL of acetonitrile for liquid chroma-tography. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sitagliptin are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sitagliptin is not more than 1.0z.
To 10 Sitagliptin Phosphate Tablets add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make exactly 250 mL, and stir thoroughly. Pipet V mL of this solution, and add a mixture of diluted phosphoric acid (1 in 1000) and aceto-nitrile for liquid chromatography (19:1) to make exactly V? mL so that each mL contains about 80 mg of sitagliptin (C16H15F6N5O). Centrifuge this solution, and use the super-natant liquid as the sample solution. Separately, weigh accu-rately about 26 mg of Sitagliptin Phosphate RS (separately determine the water <2.48> in the same manner as Sitagliptin Phosphate Hydrate), dissolve in a mixture of diluted phos-phoric acid (1 in 1000) and acetonitrile for liquid chromatog-raphy (19:1) to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of sitagliptin in each solution. Amount (mg) of sitagliptin (C16H15F6N5O) in 1 tablet of Sitagliptin Phosphate Tablets ＝ MS × AT/AS × V?/V × 1/ 10 × 0.806 MS: Amount (mg) of Sitagliptin Phosphate RS taken, cal-culated on the anhydrous basis ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 205 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with cyanopropylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 900 mL of water, adjust to pH 2.0 with phos-phoric acid, and add water to make 1000 mL. To 850 mL of this solution add 150 mL of acetonitrile for liquid chroma-tography. Flow rate: 1.0 mL per minute. System suitability— System performance: Proceed as directed in the system suitability in the Assay under Sitagliptin Phosphate Hydrate. The following method can be applied when sodium stearyl fumarate is contained in the additive of the tablet. Crush 1 tablet of Sitagliptin Phosphate Tablets, transfer to a vial, and add 1 mL of water. Stopper the vial tightly, and heat at 809C for 20 to 48 hours. Take out the contents of the vial, wash the vial 3 times with a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chro-matography (19:1), combine the washings and the content, and add a mixture of diluted phosphoric acid (1 in 1000) and acetonitrile for liquid chromatography (19:1) to make 100 mL. Stir this solution for 1 hour, and centrifuge for 10 minutes or until the solution becomes clear. When the proce-dure is run with 20 mL of the supernatant liquid under the above operating conditions, the resolution between sitaglip-tin and the peak having the relative retention time of about 1.2 to sitagliptin is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sitagliptin is not more than 1.0z.
注射用シベレスタットナトリウム
Sivelestat Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 95.
Related substances—Dissolve an amount of Sivelestat Sodium for Injection, equivalent to 1.0 g of Sivelestat Sodium Hydrate, in water to make 100 mL. To 1 mL of this solution add 9 mL of a mixture of acetonitrile and water (5:4), and use the solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.25 to sivelestat, obtained from the sample solution is not larger than 3 times the peak area of sivelestat from the standard so-lution. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay under Sivelestat Sodium Hydrate. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). System suitability— Proceed as directed in the system suitability in the Purity (2) under Sivelestat Sodium Hydrate. Bacterial endotoxins <4.01> Less than 25 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem- ‌brane filtration method: it meets the requirement.‌‌
Take a number of Sivelestat Sodium for Injection, equivalent to about 1 g of sivelestat sodium hydrate (C20H21N2NaO7S.4H2O), and dissolve all the contents in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL. Pipet 10 mL of this solu-tion, add exactly 5 mL of the internal standard solution and 5 mL of acetonitrile. To 2 mL of this solution add 3 mL of a mixture of water and acetonitrile (1:1), and use the solution as the sample solution. Then, proceed as directed in the Assay under Sivelestat Sodium Hydrate. Amount (mg) of sivelestat sodium hydrate (C20H21N2NaO7S.4H2O) ＝ MS × QT/QS × 20 × 1.216 MS: Amount (mg) of Sivelestat RS taken Internal standard solution—A solution of propyl para-hydroxybenzoate in acetonitrile (1 in 2500).
Sodium Bicarbonate Injection is an aqueous injec-tion. It contains not less than 95.
Sodium Bicarbonate Injection is a clear, color-less liquid.
To a volume of Sodium Bicarbonate Injec-tion, equivalent to 1 g of Sodium Bicarbonate, add water to make 30 mL: the solution responds to Qualitative Tests <1.09> for sodium salt and for bicarbonate. pH <2.54> 7.0 – 8.5 Bacterial endotoxins <4.01> Less than 5.0 EU/mEq. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require- ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Sodium Bicarbonate Injection, equivalent to about 2 g of sodium hydrogen car-bonate (NaHCO3), titrate with 0.5 mol/L sulfuric acid VS, and proceed as directed in the Assay under Sodium Bicar-bonate. Each mL of 0.5 mol/L sulfuric acid VS ＝ 84.01 mg of NaHCO3
Sodium Chromate (51Cr) Injection is a clear, light yellow liquid. It is odorless or has an odor of the preser-vatives. Sodium Citrate Hydrate
Sodium Citrate Injection for Transfusion
輸血用クエン酸ナトリウム注射液
Sodium Citrate Injection for Transfusion is an aqueous injection. It contains not less than 9.
Sodium Citrate Injection for Transfusion is a clear, colorless liquid.
Sodium Citrate Injection for Transfusion re-sponds to Qualitative Tests <1.09> for sodium salt and for citrate. pH <2.54> 7.0 – 8.5 Bacterial endotoxins <4.01> Less than 5.6 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. ‌Assay Pipet 5 mL of Sodium Citrate Injection for Transfu-sion, and add water to make exactly 25 mL. Evaporate 10 mL of this solution, exactly measured, on a water bath to dryness, dry the residue at 1809C for 2 hours, and dissolve in 30 mL of acetic acid (100) by warming. Cool, titrate <2.50> with 0.1 mol/L perchloric acid VS (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction.‌ Each mL of 0.1 mol/L perchloric acid VS ＝ 9.803 mg of C6H5Na3O7.2H2O
Sodium Iodide (131I) Solution contains iodine-131 (131I) in the form of sodium iodide.
Sodium Iodide (131I) Solution is a clear, color-less liquid. It is odorless, or has an odor due to the preserva-tives or stabilizers. Sodium Iodohippurate (131I) Injection
Sodium Iotalamate Injection is an aqueous injec-tion. It contains not less than 95.
Sodium L-Lactate Ringer's Solution is an aqueous injec-tion. It contains not less than 0.
Sodium L-Lactate Ringer's Solution occurs as a clear and colorless liquid.
(1) Sodium L-Lactate Ringer's Solution re-sponds to Qualitative Tests <1.09> (1) for sodium salt. A solution, obtained by concentrating 10 mL of So-dium L-Lactate Ringer's Solution to 5 mL by heating on a water bath, responds to Qualitative Tests <1.09> (1) for po-tassium salt. A solution, obtained by concentrating 10 mL of So-dium L-Lactate Ringer's Solution to 5 mL by heating on a water bath, responds to Qualitative Tests <1.09> (3) for cal-cium salt. Sodium L-Lactate Ringer's Solution responds to Qualitative Tests <1.09> (2) for chloride. Sodium L-Lactate Ringer's Solution responds to Qualitative Tests <1.09> for lactate. pH <2.54> 6.0 – 7.5
Heavy metals <1.07>—Concentrate 100 mL of So- dium L-Lactate Ringer's Solution on a water bath to about 40 mL, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test with this solution as the test solu-tion. Prepare the control solution with 3.0 mL of Standard Lead Solution by adding 2 mL of dilute acetic acid and water to make 50 mL (not more than 0.3 ppm). Bacterial endotoxins <4.01> Less than 0.25 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Sodium, potassium, and calcium—Pipet 10 mL of Sodium L-Lactate Ringer's Solution, add exactly 5 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, pipet 10 mL of standard stock solution, add exactly 5 mL of the in-ternal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QTa, QTb and QTc, of respective peak area of sodium, potassium and calcium to that of the internal standard in the sample solu-tion, and the ratios, QSa, QSb and QSc, of respective peak area of sodium, potassium and calcium to that of the inter-nal standard in the standard solution. Amount (w/vz) of sodium (Na) Mobile phase: To 4 mL of methanesulfonic acid add water to make 3000 mL. Flow rate of mobile phase: Adjust so that the retention time of potassium is about 6 minutes. Suppressor: An anion elimination device with anion-exchange membrane. Refreshing liquid: Diluted 40z tetrabutylammonium hy-droxide TS (1 in 40). Flow rate of refreshing liquid: 2 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, sodium, potassium, the internal standard and cal-cium are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of sodium, potassium and calcium to that of the internal standard is not more than 1.0z. Chlorine—Pipet 1 mL of Sodium L-Lactate Ringer's Solution, add exactly 5 mL of the internal standard solution and water to make 100 mL, and use this solution as the sam-ple solution. Separately, pipet 10 mL of standard stock solu-tion obtained in (1), and add water to make exactly 50 mL. Take exactly 4 mL and 6 mL of this solution, add exactly 5 mL of the internal standard solution to them and water to make 100 mL, and use these solutions as the low concentra-tion standard solution and the high concentration standard solution, respectively. Perform the test with 20 mL each of the sample solution, the low concentration standard solution and the high concentration standard solution as directed under Liquid Chromatography <2.01> according to the fol- ＝ (M Sa1 × f/100 × 0.205 ＋ M Sa2 × 0.393) lowing conditions, and calculate the ratios, QT, QSL and QSH, of the peak area of chlorine to that of the internal standard. × QTa/QSa × 1/10 Amount (w/vz) of potassium (K) Amount (w/vz) of chlorine (Cl) ＝ MSb × QTb /QSb × 1/10 × 0.524 ＝ (MSa × 0.607 ＋ MSb × 0.476 ＋ MSc × 0.482) × (QT － 3QSL ＋ 2QSH)/(QSH － QSL) × 1/25 Amount (w/vz) of calcium (Ca) ＝ MSc × QTc/QSc × 1/10 × 0.273 MSa1: Amount (g) of sodium L-lactate solution for assay taken f: Content (z) of sodium L-lactate solution for assay MSa2: Amount (g) of sodium chloride for assay taken MSb: Amount (g) of potassium chloride for assay taken MSc: Amount (g) of calcium chloride hydrate for assay taken Standard stock solution: Weigh accurately an amount of so-dium L-lactate solution for assay equivalent to about 3.1 g of sodium L-lactate (C3H5NaO3), about 6 g of dried sodium chloride for assay, about 0.3 g of dried potassium chloride for assay and about 0.2 g of calcium chloride hydrate for assay, respectively, and dissolve in water to make exactly 1000 mL. Internal standard solution—A solution of rubidium chloride (1 in 200). Operating conditions— Detector: A conductivity detector. Column: A plastic column 4 mm in inside diameter and 25 cm in length, packed with a weakly acidic ion-exchange resin for liquid chromatography composed with carboxylic acid and phosphonic acid groups combining ethylvinylbenzene-divinylbenzene copolymer (8.5 mm in particle diameter). Column temperature: A constant temperature of about 259C. MSa: Amount (g) of sodium chloride for assay taken MSb: Amount (g) of potassium chloride for assay taken MSc: Amount (g) of calcium chloride hydrate for assay taken Internal standard solution—A solution of sodium bromide (1 in 500). Operating conditions— Detector: A conductivity detector. Column: A plastic column 4 mm in inside diameter and 25 cm in length, packed with a strongly basic ion-exchange resin for liquid chromatography composed with quaternary am-monium group combining ethylvinylbenzene-divinylbenzene copolymer (9 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.25 g of sodium hydrogen car-bonate and 0.64 g of anhydrous sodium carbonate in 2000 mL of water. Flow rate of mobile phase: Adjust so that the retention time of chlorine is about 4 minutes. Suppressor: A cation elimination device with cation-exchange membrane. Refreshing liquid: Diluted sulfuric acid (3 in 4000). Flow rate of refreshing liquid: 2 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the low concentration standard solution under the ‌above operating conditions, lactic acid, chlorine and the in-ternal standard are eluted in this order and the resolution be-tween the peaks of lactic acid and chlorine is not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the low concentration standard solution under the above operating conditions, the relative standard devia-tion of the ratio of the peak area of chlorine to that of the in-ternal standard is not more than 1.0z. L-Lactic acid—Pipet 20 mL of Sodium L-Lactate Ringer's Solution, add exactly 5 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, pipet 20 mL of standard stock solution obtained in (1), add exactly 5 mL of the inter-nal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of lactic acid to that of the internal standard. Amount (w/vz) of L-lactic acid (C3H5O3) ＝ MS × f/100 × QT/QS × 1/10 × 0.795 MS: Amount (g) of sodium L-lactate solution for assay taken f: Content (z) of sodium L-lactate solution for assay Internal standard solution—A solution of sodium acetate trihydrate (1 in 50). Operating conditions— Detector: A conductivity detector. Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with a strongly acidic ion-exchange resin for liquid chromatography composed with sulfonic acid group combining styrene-divinylbenzene co-polymer (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 3000 mL of water add 0.5 mL of hep-tafluorobutylic acid. Flow rate of mobile phase: Adjust so that the retention time of lactic acid is about 9 minutes. Suppressor: A cation elimination device with cation-exchange membrane. Refreshing liquid: Diluted 40z tetrabutylammonium hy-droxide TS (13 in 2000). Flow rate of refreshing liquid: 2 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, lactic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of lactic acid to that of the internal standard is not more than 1.0z.
Sodium L-Lactate Solution is an aqueous solution of sodium salt of L-lactic acid. It contains not less than 95.
Sodium L-Lactate Solution occurs as a clear and colorless viscous liquid. It has no odor or has a slight characteristic odor, and has a slight saline taste. It is miscible with water or with ethanol (99.5).
To an amount of Sodium L-Lactate Solu-tion, equivalent to 1 g of sodium L-lactate (C3H5NaO3), add water to make 50 mL. This solution responds to Qualitative Tests <1.09> for sodium salt and for lactate. D Optical rotation <2.49> [a]20: －38 – －449 To an exact amount of Sodium L-Lactate Solution, equivalent to 2.5 g of sodium L-lactate (C3H5NaO3), add 30 mL of water and 5.0 g of hexaammonium heptamolybdate tetrahydrate, then add water to make exactly 50 mL, and determine using a 100-mm cell. pH <2.54> To an amount of Sodium L-Lactate Solution, equivalent to 5 g of sodium L-lactate (C3H5NaO3), add water to make 50 mL: the pH of this solution is between 6.5 and 7.5.
(1) Chloride <1.03>—Perform the test with an amount of Sodium L-Lactate Solution, equivalent to 1.0 g of sodium L-lactate (C3H5NaO3). Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Sulfate <1.14>—To an amount of Sodium L-Lactate Solution, equivalent to 2.0 g of sodium L-lactate (C3H5NaO3), add 7 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.010z). Heavy metals <1.07>—To an amount of Sodium L-Lactate Solution, equivalent to 2.0 g of sodium L-lactate (C3H5NaO3), add 5 mL of dilute hydrochloric acid, 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Iron <1.10>—Prepare the test solution with an amount of Sodium L-Lactate Solution, equivalent to 2.0 g of sodium L-lactate (C3H5NaO3), according to Method 1, and perform the test according to Method A. Prepare the control solution with 1.0 mL of Standard Iron Solution (not more than 5 ppm). Arsenic <1.11>—To an amount of Sodium L-Lactate Solution, equivalent to 2.5 g of sodium L-lactate (C3H5NaO3), and add water to make 10 mL. Perform the test using 2 mL of this solution as the test solution (not more than 4 ppm). Sugars—To an amount of Sodium L-Lactate Solu-tion, equivalent to 1.0 g of sodium L-lactate (C3H5NaO3), add 10 mL of water and 10 mL of Fehling's TS, and boil for 5 minutes: no red precipitate is produced. ‌Citric, oxalic, phosphoric and L-tartaric acids—To an amount of Sodium L-Lactate Solution, equivalent to 1.0 g of sodium L-lactate (C3H5NaO3), add 1 mL of water and 1 mL of dilute hydrochloric acid, then add 40 mL of calcium hydroxide TS, and boil for 2 minutes: the solution is not changed. Volatile fatty acids—To an amount of Sodium L-Lactate Solution, equivalent to 3.0 g of sodium L-lactate (C3H5NaO3), add 2 mL of dilute sulfuric acid, and heat on a water bath: no acetic acid like nor lactic acid like odor is pro-duced. Cyanide—Transfer an amount of Sodium L-Lactate Solution, equivalent to 1.0 g of sodium L-lactate (C3H5NaO3), to a Nessler tube, add 10 mL of water and 1 drop of phenolphthalein TS, then add dropwise a solution of sodium hydroxide (1 in 10) while shaking until a pale red color appears. Add further 1.5 mL of a solution of sodium hydroxide (1 in 10) and water to make 20 mL, and heat in a water bath for 10 minutes. After cooling, add dropwise dilute hydrochloric acid until a red color of the solution dis-appears, then add 1 drop of acetic acid (31), 10 mL of phos-phate buffer solution (pH 6.8) and 0.25 mL of sodium tolu-enesulfonchloramide TS, stopper immediately, mix gently, and allow to stand for 5 minutes. Add 15 mL of pyridine-pyrazolone TS and water to make 50 mL, and allow to stand at 259C for 30 minutes: the color of the solution is not more intense than that of the following control solution. Control solution: To 1.0 mL of Standard Cyanide Solu-tion add water to make 20 mL. Transfer 1.0 mL of this solu-tion to a Nessler tube, add 10 mL of water and 1 drop of phenolphthalein TS, then proceed in the same manner as descried above. Methanol—Transfer an amount of Sodium L-Lactate Solution, equivalent to 5.0 g of sodium L-lactate (C3H5NaO3), to a distilling flask of the apparatus for alcohol number determination <1.01>, add 10 mL of water, and dis-till. Pipet 5 mL of the distillate, add water to make exactly 10 mL, and use this solution as the sample solution. Sepa-rately, to exactly 1.0 mL of methanol add water to make ex-actly 100 mL. Pipe 5 mL of this solution, add water to make exactly 200 mL. Pipet 5 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following condi-tions: the peak area of methanol obtained from the sample solution is not larger than that from the standard solution (not more than 0.025z). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1.5 m in length, packed with porous ethyl vinylbenzene-divinyl-benzene copolymer for gas chromatography (149 – 177 mm in particle diameter). Column temperature: A constant temperature of about 1209C. Injection port and detector temperature: A constant tem-perature of about 1259C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of methanol is about 2 minutes. System suitability— System performance: To 1 mL of methanol and 1 mL of ethanol (99.5) add water to make 100 mL. To 5 mL of this solution add water to make 200 mL. To 5 mL of this solu-tion add water to make 10 mL. When the procedure is run with 10 mL of this solution under the above operating condi- tions, methanol and ethanol are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of methanol is not more than 5z.
Weigh accurately an amount of Sodium L-Lactate Solution, equivalent to about 0.25 g of sodium L-lactate (C3H5NaO3), dry at 1059C for 4 hours, add 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of solution changes from purple to yellow-green through blue-green (indicator: 2 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 11.21 mg of C3H5NaO3
Sodium Risedronate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of sodium risedronate (C H NNaO P : mL of the standard solution under the above operating con-ditions, the internal standard and risedronic acid are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of risedronic acid to that of the internal stand-ard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 20 minutes of Sodium Risedronate Tablets is not less than 305.09). 7 10 7 2 80z. Start the test with 1 tablet of Sodium Risedronate Tablets, Method of preparation Prepare as directed under Tablets, with Sodium Risedronate Hydrate. Identification Powder Sodium Risedronate Tablets. To a portion of the powder, equivalent to 2.5 mg of sodium risedronate (C7H10NNaO7P2), add 50 mL of diluted dilute sodium hydroxide TS (1 in 20), shake, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 2 mL of the fil-trate, and determine the absorption spectrum of the subse-quent filtrate as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sodium Risedronate Tablets add exactly 10 mL of the mobile phase, shake, and allow to stand for 10 minutes. Disperse the particles by sonicating for 10 minutes with occasional shaking, then centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 1 mL of the fil-trate, pipet exactly V mL of the subsequent filtrate, equiva-lent to about 1.75 mg of sodium risedronate (C7H10NNaO7P2), add exactly 1 mL of the internal standard solution and the mobile phase to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 70 mg of Risedronic Acid RS (separately determine the water <2.48> using 80 mg, in the same manner as Sodium Risedronate Hydrate), dissolve in 3 mL of 0.2 mol/L sodium hydroxide TS, and add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 2 mL of the in-ternal standard solution, then add the mobile phase to make 20 mL, and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of risedronic acid to that of the internal standard. Amount (mg) of sodium risedronate (C7H10NNaO7P2) ＝ MS × QT/QS × 1/V × 1/4 × 1.078 MS: Amount (mg) of Risedronic Acid RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium benzoate in the mobile phase (7 in 2000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Sodium Risedronate Hydrate. System suitability— System performance: When the procedure is run with 20 withdraw 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.
Powder Sodium Risedronate Tablets. To a portion of the powder, equivalent to 2.5 mg of sodium risedronate (C7H10NNaO7P2), add 50 mL of diluted dilute sodium hydroxide TS (1 in 20), shake, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 2 mL of the fil-trate, and determine the absorption spectrum of the subse-quent filtrate as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits a maximum between 260 nm and 264 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sodium Risedronate Tablets add exactly 10 mL of the mobile phase, shake, and allow to stand for 10 minutes. Disperse the particles by sonicating for 10 minutes with occasional shaking, then centrifuge, and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 1 mL of the fil-trate, pipet exactly V mL of the subsequent filtrate, equiva-lent to about 1.75 mg of sodium risedronate (C7H10NNaO7P2), add exactly 1 mL of the internal standard solution and the mobile phase to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 70 mg of Risedronic Acid RS (separately determine the water <2.48> using 80 mg, in the same manner as Sodium Risedronate Hydrate), dissolve in 3 mL of 0.2 mol/L sodium hydroxide TS, and add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 2 mL of the in-ternal standard solution, then add the mobile phase to make 20 mL, and use this solution as the standard solution. Per-form the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of risedronic acid to that of the internal standard. Amount (mg) of sodium risedronate (C7H10NNaO7P2) ＝ MS × QT/QS × 1/V × 1/4 × 1.078 MS: Amount (mg) of Risedronic Acid RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium benzoate in the mobile phase (7 in 2000). Operating conditions— Proceed as directed in the operating conditions in the Assay under Sodium Risedronate Hydrate. System suitability— System performance: When the procedure is run with 20 withdraw 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 2 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 2.8 mg of sodium risedronate (C7H10NNaO7P2), and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Risedronic Acid RS (separately determine the water <2.48> using 80 mg, in the same manner as Sodium Risedronate Hydrate), dissolve in 3 mL of 0.2 mol/L sodium hydroxide TS, and add water to make exactly 50 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of risedronic acid in each solution. Dissolution rate (z) with respect to the labeled amount of sodium risedronate (C7H10NNaO7P2) ＝ MS × AT/AS × V?/V × 1/C × 9/2 × 1.078 MS: Amount (mg) of Risedronic Acid RS taken, calcu-lated on the anhydrous basis C: Labeled amount (mg) of sodium risedronate (C7H10NNaO7P2) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.15 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate, 3.36 g of tetradecyl trimethylammonium bromide, 5.11 g of ammonium dihy-drogen phosphate and 3.11 g of diammonium hydrogen phosphate in 1360 mL of water, and add 640 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of risedronic acid is about 12 minutes. System suitability— System performance: When the procedure is run with 200 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risedronic acid are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 200 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the peak area of risedronic acid is not more than 2.0z.‌
Weigh accurately the mass of not less than 20 So-dium Risedronate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of sodium risedronate (C7H10NNaO7P2), add exactly 10 mL of the in-ternal standard solution, add 190 mL of the mobile phase, shake, and allow to stand for 10 minutes. Disperse the parti-cles by sonicating with occasional shaking, then centrifuge, and filter through a membrane filter with a pore size not exceeding 0.2 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 50 mg of Risedronic Acid RS (separately determine the water <2.48> using 80 mg, in the same manner as Sodium Risedronate Hydrate), dissolve in 3 mL of 0.2 mol/L sodium hydroxide TS, add exactly 10 mL of the internal standard solution, then add the mobile phase to make 200 mL, and use this solution as the standard solu-tion. Then, proceed as directed in the Assay under Sodium Risedronate Hydrate. Amount (mg) of sodium risedronate (C7H10NNaO7P2) ＝ MS × QT/QS × 1.078 MS: Amount (mg) of Risedronic Acid RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium benzoate in the mobile phase (1 in 100).
Sodium Thiosulfate Injection is an aqueous injec-tion. It contains not less than 95.
Sodium Thiosulfate Injection is a clear, color-less liquid.
Sodium Thiosulfate Injection responds to Qualitative Tests <1.09> for sodium salt and for thiosulfate. ‌Bacterial endotoxins <4.01> Less than 0.01 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Sodium Thiosulfate In-jection, equivalent to about 0.5 g of sodium thiosulfate hy-drate (Na2S2O3.5H2O), add water to make 30 mL, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 24.82 mg of Na2S2O3.5H2O
‌Sodium Valproate Extended-release Tablets A
Weigh accurately the mass of not less than 20 So-dium Valproate Extended-release Tablets A, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of sodium valproate (C8H15NaO2), add about 80 mL of the mobile phase, shake thoroughly, add the mobile phase to make exactly 100 mL, and centrifuge. Pipet 20 mL of the supernatant liquid, add exactly 5 mL of the internal standard solution, and use this solution as the sample solu-tion. Separately, weigh accurately about 0.1 g of sodium val-proate for assay, previously dried at 1059C for 3 hours, dis-solve in the mobile phase to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 5 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of valproic acid to that of the internal standard. Amount (mg) of sodium valproate (C8H15NaO2) ＝ MS × QT/QS MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of ethyl para-hydroxybenzoate in the mobile phase (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS (pH 3.0) and acetonitrile for liquid chro-matography (1:1). Flow rate: Adjust so that the retention time of valproic acid is about 6 minutes. ‌System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and valproic acid are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valproic acid to that of the internal standard is not more than 1.0z.
Sodium Valproate Extended-release Tablets B
Sodium Valproate Extended-release Tablets B con- ＝ MS × A ＋ ∑ i＝1 ×  AS 45 × × × 360 V C tain not less than 95.0z and not more than 105.0z of the labeled amount of sodium valproate (C8H15NaO2: 166.19). Method of preparation Prepare as directed under Tablets, with Sodium Valproate. Identiˆcation To a quantity of the powdered Sodium Val-proate Extended-release Tablets B, equivalent to 1.0 g of So-dium Valproate, add 10 mL of water, heat on a water bath for 30 minutes, and ˆlter. To 2.5 mL of the ˆltrate add 2.5 mL of water and 1 mL of a solution of cobalt (II) nitrate hexahydrate (1 in 20), and heat on a water bath: a purple precipitate is formed. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sodium Valproate Extended-release Tablets B add 150 mL of the mobile phase, allow to stand for not less than 16 hours, shake until the ˆlm is disintegrated, and add the mobile phase to make exactly 200 mL. Pipet V mL of this solution, and add the mobile phase to make exactly V? mL so that each mL contains about 1 mg of sodium val-proate (C8H15NaO2).
To 20 Sodium Valproate Extended-release Tablets B add 150 mL of the mobile phase, allow to stand for not less than 16 hours, shake until the ˆlm is disintegrated, and add the mobile phase to make exactly 200 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly V mL so that each mL contains about 1 mg of sodium valproate (C8H15NaO2). Centrifuge this solution, and ˆlter the super-natant liquid through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard the ˆrst 5 mL of the ˆltrate, pipet 20 mL of the subsequent ˆltrate, add exactly 5 mL of the internal standard solution, and use this solution as the Amount (mg) of sodium valproate (C8H ＝ MS × QT/QS × V?/V × 2 15NaO2) sample solution. Separately, weigh accurately about 0.1 g of sodium valproate for assay, previously dried at 1059C for 3 hours, dissolve in the mobile phase to make exactly 100 mL. MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of methyl para-hydroxybenzoate in the mobile phase (1 in 50,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates Pipet 20 mL of this solution, add exactly 5 mL of the inter-nal standard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of val-proic acid to that of the internal standard. of a 200-mg tablet in 8 hours, in 11 hours and in 20 hours are Amount (mg) of sodium valproate (C H NaO ) in 1 tablet 15 to 45z, 35 to 65z, and not less than 70z, respectively, and those of a 400-mg tablet in 9 hours, in 12 hours and in ＝ MS × QT/QS × V/50 8 15 2 21 hours are 15 to 45z, 35 to 65z, and not less than 70z, respectively. Start the test with 1 tablet of Sodium Valproate Extended-release Tablets B, withdraw exactly 20 mL of the medium at MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of methyl para-hydroxybenzoate in the mobile phase (1 in 50,000). ‌Operating conditions—‌ Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS (pH 3.0) and acetonitrile for liquid chro-matography (1:1). Flow rate: Adjust so that the retention time of valproic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and valproic acid are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valproic acid to that of the internal standard is not more than 1.0z.
Sodium Valproate Syrup contains not less than 95.
To a volume of Sodium Valproate Syrup, equivalent to 50 mg of Sodium Valproate, add water to make 10 mL. To 5 mL of this solution add 1 mL of a solu-tion of cobalt (II) nitrate hexahydrate (1 in 20) and warm on a water bath: a purple precipitate is formed. Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/mL and 101 CFU/mL, respectively. Escherichia coli is not observed.
Pipet a volume of Sodium Valproate Syrup, equiva-lent to about 0.1 g of sodium valproate (C8H15NaO2) and add water to make exactly 100 mL. Pipet 20 mL of this solu-tion, add exactly 5 mL of internal standard solution, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of sodium valproate for assay, previously dried at 1059C for 3 hours, dissolve in water to make exactly 50 mL. Pipet 20 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of valproic acid to that of the internal standard. Amount (mg) of sodium valproate (C8H15NaO2) ＝ MS × QT/QS × 2 MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of ethyl parahy- droxybenzoate in the mobile phase (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-genphosphate TS (pH 3.0) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of valproic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and valproic acid are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valproic acid to that of the internal standard is not more than 1.0z.
Sodium Valproate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of sodium valproate (C8H15NaO2: 166.19). Method of preparation Prepare as directed under Tablets, with Sodium Valproate. Identification To a quantity of powdered Sodium Valpro-ate Tablets, equivalent to 0.5 g of Sodium Valproate, add 10 mL of water, shake well, and centrifuge. To 5 mL of the su-pernatant liquid add 1 mL of a solution of cobalt (II) nitrate hexahydrate (1 in 20) and warm on a water bath: a purple precipitate is formed. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sodium Valproate Tablets add 7V/10 mL of the mobile phase, shake vigorously, add the mobile phase to make exactly V mL so that each mL contains about 1 mg of sodium valproate (C8H15NaO2), and centrifuge.
To a quantity of powdered Sodium Valpro-ate Tablets, equivalent to 0.5 g of Sodium Valproate, add 10 mL of water, shake well, and centrifuge. To 5 mL of the su-pernatant liquid add 1 mL of a solution of cobalt (II) nitrate hexahydrate (1 in 20) and warm on a water bath: a purple precipitate is formed. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sodium Valproate Tablets add 7V/10 mL of the mobile phase, shake vigorously, add the mobile phase to make exactly V mL so that each mL contains about 1 mg of sodium valproate (C8H15NaO2), and centrifuge. Filter the su-pernatant liquid, pipet 20 mL of the filtrate, add exactly 5 mL of the internal standard solution, shake vigorously, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of sodium valproate (C8H15NaO2) ＝ MS × QT/QS × V/100 MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mobile phase (1 in 50,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Sodium Valproate Tablets ‌is not less than 85z. Start the test with 1 tablet of Sodium Valproate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.11 mg of sodium valproate (C8H15NaO2), and use this solution as the sample solution. Separately, weigh accurately about 56 mg of sodium valproate for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL of the solution, and use this so-lution as the standard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of valproic acid in each solution. Dissolution rate (z) with respect to the labeled amount of sodium valproate (C8H15NaO2) Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS (pH 3.0) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of valproic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and valproic acid are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valproic acid to that of the internal standard is not more than 1.0z. ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of sodium valproate for assay taken C: Labeled amount (mg) of sodium valproate (C8H15NaO2) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of valproic acid are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valproic acid is not more than 1.5z.
Weigh accurately the mass of not less than 20 So-dium Valproate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.2 g of sodium valproate (C8H15NaO2), add about 160 mL of the mobile phase, shake well, add the mobile phase to make exactly 200 mL, and centrifuge. Filter the supernatant liquid, pipet 20 mL of the filtrate, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.1 g of sodium valproate for assay, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 5 mL of the internal standard solu-tion, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of valproic acid to that of the internal standard. Amount (mg) of sodium valproate (C8H15NaO2) ＝ MS × QT/QS × 2 MS: Amount (mg) of sodium valproate for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mobile phase (1 in 50,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm).
Spectinomycin Hydrochloride for Injection
注射用スペクチノマイシン塩酸塩
Spectinomycin Hydrochloride for Injection is a preparation for injection which is suspended before use. It contains not less than 97.
Spectinomycin Hydrochloride for Injection oc-curs as a white to light yellow-white crystalline powder.
Proceed as directed in the Identification (2) under Spectinomycin Hydrochloride Hydrate. pH <2.54> Dissolve an amount of Spectinomycin Hydro-chloride for Injection, equivalent to 70 mg (potency) of Spectinomycin Hydrochloride Hydrate, in 10 mL of water: the pH of the solution is between 4.0 and 5.6.
Clarity and color of solution—A solution dissolved an amount of Spectinomycin Hydrochloride for Injection, equivalent to 0.70 g (potency) of Spectinomycin Hydrochlo-ride Hydrate, in 10 mL of water is clear, and its absorbance at 425 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.10. Water <2.48> 16.0 – 20.0z (0.3 g, volumetric titration, direct titration). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: 107.5z). Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the mass of the contents of not less than 10 Spectinomycin Hydrochloride for Injection. Weigh accurately a portion of the content, equivalent to about 20 mg (potency) of Spectinomycin Hydrochloride Hydrate, dis-solve in exactly 10 mL of the internal standard solution, add 1 mL of 1,1,1,3,3,3-hexamethyldisilazan, allow to stand at room temperature for 1 hour, and use this solution as the sample solution. Separately, weigh accurately an amount of Spectinomycin Hydrochloride RS, equivalent to about 20 mg (potency), dissolve in exactly 10 mL of the internal standard solution, add 1 mL of 1,1,1,3,3,3-hexamethyldisilazan, allow to stand at room temperature for 1 hour, and use this solution as the standard solution. Then, proceed as directed in the Assay under Spectinomycin Hydrochloride Hydrate. Amount [mg (potency)] of spectinomycin (C14H24N2O7) ＝ MS × QT/QS MS: Amount [mg (potency)] of Spectinomycin Hydrochlo-ride RS taken Internal standard solution—A solution of triphenylantimony in N, N-dimethylformamide (1 in 500).
Spironolactone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of It shows crystal polymorphism. spironolactone (C 24H32 O4S: 416.57). Identification (1) Determine the absorption spectrum of a solution of Spironolactone in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Spironolactone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Spironolactone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Spironolactone RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve Spironolactone and Spironolactone RS in methanol, Method of preparation Prepare as directed under Tablets, with Spironolactone. Identification To an amount of powdered Spironolactone Tablets, equivalent to 10 mg of Spironolactone, add 100 mL of methanol, shake vigorously, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 50 mL, and de-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 236 nm and 240 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Spironolactone Tablets add a mixture of water and acetonitrile (1:1) to make exactly V mL so ‌that each mL contains about 0.
(1) Determine the absorption spectrum of a solution of Spironolactone in methanol (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Spironolactone RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Spironolactone, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Spironolactone RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve Spironolactone and Spironolactone RS in methanol, Method of preparation Prepare as directed under Tablets, with Spironolactone. Identification To an amount of powdered Spironolactone Tablets, equivalent to 10 mg of Spironolactone, add 100 mL of methanol, shake vigorously, and centrifuge. To 5 mL of the supernatant liquid add methanol to make 50 mL, and de-termine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 236 nm and 240 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Spironolactone Tablets add a mixture of water and acetonitrile (1:1) to make exactly V mL so ‌that each mL contains about 0.5 mg of spironolactone (C24H32O4S). After stirring for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. 259C. Mobile phase: A mixture of methanol and water (3:2). Flow rate: Adjust so that the retention time of spironolac- tone is about 11 minutes. Amount (mg) of spironolactone (C ＝ MS × AT/AS × V/50 24H 32O4S) System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con- MS: Amount (mg) of Spironolactone RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of a solution prepared by dissolving 1 g of polysorbate 80 in water to make 500 mL as the dissolution medium, the dissolution rate in 30 minutes of a 25-mg tablet and a 50-mg tablet are not less than 80z and not less than 70z, respec-tively. Start the test with 1 tablet of Spironolactone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 14 mg of spironolactone (C24H32O4S), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Spironolactone RS, previously dried at 1059C for 2 hours, dissolve in 20 mL of ethanol (95), and add the dissolution medium to make ex-actly 100 mL. Pipet 5 mL of this solution, add the dissolu-tion medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 243 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dissolution medium as the blank. Dissolution rate (z) with respect to the labeled amount of spironolactone (C24H32O4S) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of Spironolactone RS taken C: Labeled amount (mg) of spironolactone (C24H32O4S) in 1 tablet
Weigh accurately the mass of not less than 10 Spironolactone Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of spironolactone (C24H32O4S), add a mixture of water and acetonitrile (1:1) to make exactly 100 mL. After stirring this solution for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of Spironolactone RS, previously dried at 1059C for 2 hours, dissolve in a mixture of water and aceto-nitrile (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of spironolactone in each solution. Amount (mg) of spironolactone (C24H32O4S) ＝ MS × AT/AS × 2 MS: Amount (mg) of Spironolactone RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ditions, the number of theoretical plates and the symmetry factor of the peak of spironolactone are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of spironolactone is not more than 1.0z.
Sterile Water for Injection in Containers
Sterile Water for Injection in Containers is a clear and colorless liquid, having no odor.
Potassium permanganate-reducing substances—To 100 mL of Sterile Water for Injection in Containers add 10 mL of dilute sulfuric acid, boil, then add 0.10 mL of 0.02 mol/L potassium permanganate VS, and boil again for 10 minutes: the red color of the solution does not disappear. Conductivity <2.51> When the test is performed according to the following method, the conductivity (259C) is not more than 25 mS・cm－1 for containers with a nominal volume of 10 mL or less, and not more than 5 mS・cm－1 for containers with a nominal volume greater than 10 mL. Transfer a suitable amount of Sterile Water for Injection in Containers to a beaker, and stir the water specimen. Adjust the temperature to 25 ± 19C, and begin agitating the water specimen vigorously, while observing its conductivity periodically. When the change in conductivity becomes not greater than 0.1 mS・cm－1 per 5 minutes, adopt the observed value as the conductivity of the water specimen. Bacterial endotoxins <4.01> Less than 0.25 EU/mL. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> It meets the requirement.
注射用ストレプトマイシン硫酸塩
Streptomycin Sulfate for Injection is a preparation for injection, which is dissolved before use. It contains not less than 90.
Streptomycin Sulfate for Injection occurs as a white or light yellow-white, masses or powder.
Perform the test as directed in the Identifica-tion (2) under Streptomycin Sulfate. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> The pH of a solution prepared by dissolving an amount of Streptomycin Sulfate for Injection, equivalent to 2.0 g (potency) of Streptomycin Sulfate, in 10 mL of water is 4.5 to 7.0.
Clarity and color of solution—Dissolve an amount of Streptomycin Sulfate for Injection, equivalent to 1.0 g (potency) of Streptomycin Sulfate, in 3 mL of water: The so-lution is clear, and the absorbance of this solution at 400 nm, determined as directed under Ultraviolet-visible Spectro-photometry <2.24>, is not more than 0.50. Loss on drying <2.41> Not more than 4.0z (0.5 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours). Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organisms, culture medium and standard solu-tions—Proceed as directed in the Assay under Streptomycin Sulfate. Sample solution—Weigh accurately the contents of not less than 10 Streptomycin Sulfate for Injection. Weigh accurately an amount of the contents, equivalent to 1 g (po-tency) of Streptomycin Sulfate, and dissolve in water to make exactly 200 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make a solutions so that each mL contains 8 mg (potency) and 2 mg (potency), and use these solutions as the high con-centration sample solution and the low concentration sample solution, respectively.
スルホブロモフタレインナトリウム注射液
Sulfobromophthalein Sodium Injection is a clear and colorless or pale yellow liquid. pH: 5.0 – 6.0
(1) Measure a volume of Sulfobromo-phthalein Sodium Injection, equivalent to 0.02 g of Sulfo-bromophthalein Sodium, and proceed as directed in the Identification (1) under Sulfobromophthalein Sodium. (2) Measure a volume of Sulfobromophthalein Sodium Injection, equivalent to 0.1 g of Sulfobromophthalein So-dium, add 0.5 g of anhydrous sodium carbonate, and evapo-rate on a water bath to dryness. Ignite the residue until it is charred. Proceed as directed in the Identification (2) under Sulfobromophthalein Sodium. Extractable volume <6.05> It meets the requirement. Pyrogen <4.04> Add isotonic sodium chloride solution to Sulfobromophthalein Sodium Injection to make a 0.5 w/vz solution of Sulfobromophthalein Sodium. Inject into each of the rabbits 5 mL of this solution per kg of body mass: it meets the requirement.
Measure exactly a volume of Sulfobromophthalein Sodium Injection, equivalent to about 0.1 g of sulfobromo-phthalein sodium (C20H8Br4Na2O10S2), add water to make exactly 500 mL, and proceed as directed in the Assay under Sulfobromophthalein Sodium. Amount (mg) of sulfobromophthalein sodium (C20H8Br4Na2O10S2) ＝ A/881 × 200,000
Sulfur, Salicylic Acid and Thianthol Ointment
イオウ・サリチル酸・チアントール軟膏
Sulfur, Salicylic Acid and Thianthol Ointment is light yellow in color.
(1) Stir well 0.5 g of Sulfur, Salicylic Acid and Thianthol Ointment with 10 mL of water while heating, cool, and filter. To 1 mL of the filtrate add 5 mL of iron (III) nitrate TS: a purple color is produced (salicylic acid). Shake 1 g of Sulfur, Salicylic Acid and Thianthol Ointment with 20 mL of diethyl ether, remove the superna-tant liquid and floating materials. Wash the residue with 10 mL of diethyl ether, and remove the diethyl ether by suction. To the residue add 2 mL of pyridine and 0.2 mL of sodium hydrogen carbonate TS, and boil: a light blue to blue color is produced (sulfur). To 1 g of Sulfur, Salicylic Acid and Thianthol Oint-ment add 15 mL of ethanol (95), stir well while warming on a water bath, cool, and filter. Use the filtrate as the sample so-lution. Dissolve 0.01 g each of salicylic acid and thianthol in mL of ethanol (95), and use these solutions as the standard solution (1) and standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solutions (1) and (2) on a plate of silica gel with fluores-cent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, acetone and acetic acid (100) (45:5:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots of each component obtained from the sample solution and standard solutions (1) and (2) show the same Rf value. Spray iron (III) chloride TS upon the plate evenly: the spot from the standard solution (1) and that from the cor-responding sample solution reveal a purple color.
Sulpiride Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of sulpi-ride (C15H23N3O4S: 341.43). Method of preparation Prepare as directed under Cap-sules, with Sulpiride. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank: it exhibits a maximum between 289 nm and 293 ‌nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Sulpiride Capsules add 30 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, add 0.05 mol/L sulfuric acid TS to make exactly V mL so that each mL contains about 1 mg of sulpiride (C15H23N3O4S), and filter the solution.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank: it exhibits a maximum between 289 nm and 293 ‌nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 capsule of Sulpiride Capsules add 30 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, add 0.05 mol/L sulfuric acid TS to make exactly V mL so that each mL contains about 1 mg of sulpiride (C15H23N3O4S), and filter the solution. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add water to make ex-actly 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. tion. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the sample solution. Proceed as di-rected in the Assay. Amount (mg) of sulpiride (C15H23N3O4S) ＝ MS × AT/AS × V/50 MS: Amount (mg) of sulpiride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medi-um, the dissolution rate of a 50-mg tablet in 30 minutes is Amount (mg) of sulpiride (C ＝ MS × AT/AS × V/50 15H23N3O4S) not less than 80z, that of a 100-mg tablet in 45 minutes is not less than 75z, and that of a 200-mg tablet in 45 minutes is not less than 70z. MS: Amount (mg) of sulpiride for assay taken Dissolution Being specified separately when the drug is granted approval based on the Law.
Cut the capsule of not less than 20 Sulpiride Cap-sules, weigh accurately the mass of the contents, and pow-der. Weigh accurately a portion of the powder, equivalent to about 0.1 g of sulpiride (C15H23N3O4S), add 70 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, and add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet 5 mL of the sub-sequent filtrate, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg of sulpiride for assay, previously dried at 1059C for 3 hours, and dissolve in 0.05 m/L sulfuric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 291 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank. Start the test with 1 tablet of Sulpiride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of sulpiride (C15H23N3 O4S), and use this solution as the sample solution. Sepa-rately, weigh accurately about 28 mg of sulpiride for assay, previously dried at 1059C for 3 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 25 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 291 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of sulpiride (C15H23N3O4S) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of sulpiride for assay taken Amount (mg) of sulpiride (C ＝ MS × AT/AS × 2 15H23N3O4S) C: Labeled amount (mg) of sulpiride (C15H23N3O4S) in 1 tablet MS: Amount (mg) of sulpiride for assay taken
Sulpiride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of sulpi-ride (C15H23N3O4S: 341.43). Method of preparation Prepare as directed under Tablets, with Sulpiride. Identification Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank: it exhibits a maximum between 289 nm and 293 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sulpiride Tablets add 30 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, add 0.05 mol/L sul-furic acid TS to make exactly V mL so that each mL contains about 1 mg of sulpiride (C15H23N3O4S), and filter the solu- Assay Weigh accurately, and powder not less than 20 Sul-piride Tablets.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank: it exhibits a maximum between 289 nm and 293 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Sulpiride Tablets add 30 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, add 0.05 mol/L sul-furic acid TS to make exactly V mL so that each mL contains about 1 mg of sulpiride (C15H23N3O4S), and filter the solu-
Weigh accurately, and powder not less than 20 Sul-piride Tablets. Weigh accurately a portion of the powder, equivalent to about 0.1 g of sulpiride (C15H23N3O4S), add 70 mL of 0.05 mol/L sulfuric acid TS, shake for 30 minutes, and add 0.05 mol/L sulfuric acid TS to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of sulpiride for assay, previ-ously dried at 1059C for 3 hours, and dissolve in 0.05 mol/L sulfuric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this so-lution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 291 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using water as the blank. Amount (mg) of sulpiride (C15H23N3O4S) ＝ MS × AT/AS × 2 MS: Amount (mg) of sulpiride for assay taken
Sulpyrine Injection is an aqueous injection. It contains not less than 95.
Sulpyrine Injection is a clear, colorless or pale yellow liquid. pH: 5.0 – 8.5
(1) To a volume of Sulpyrine Injection, equivalent to 0.2 g of Sulpyrine Hydrate, add water to make 3 mL, then add 2 drops of dilute sulfuric acid and 1 mL of chlorinated lime TS: a deep blue color develops at first, and the color immediately turns red and gradually changes to yel-low. (2) To a volume of Sulpyrine Injection, equivalent to 0.2 g of Sulpyrine Hydrate, add water to make 5 mL, and boil this solution with 3 mL of dilute hydrochloric acid: the odor of sulfur dioxide is perceptible at first, and on further boiling the odor of formaldehyde is perceptible. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet 2 mL of Sulpyrine Injection, dilute with water to exactly 100 mL. Measure exactly a volume (V mL) of this solution, equivalent to about 50 mg of sulpyrine hydrate (C13H16N3NaO4S.H2O), and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to exactly 100 mL, and use this solution as the sample solution. Weigh accurately about 50 mg of sulpyrine for assay (previously de-termine the loss on drying <2.41> under the same conditions as Sulpyrine Hydrate), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to exactly 100 mL, and use this solution as the standard solution. Pipet 2 mL each of the sample solution and standard solution into separate 25-mL volumetric flasks, add 5 mL of ethanol (95), 2 mL of a solution of 4-dimethylaminocinnamaldehyde in ethanol (95) (1 in 250) and 2 mL of acetic acid (100) to each ‌of these solutions, shake well, allow to stand for 15 minutes, and add water to make 25 mL. Perform the test with these solutions as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared with 2 mL of water in the same manner as the blank. Determine the absor-bances, AT and AS, of the subsequent solutions of the sam-ple solution and the standard solution at 510 nm. Amount (mg) of sulpyrine hydrate (C13H16N3NaO4S.H2O) in 1 mL of Sulpyrine Injection ＝ MS × AT/AS × 50/V × 1.054 MS: Amount (mg) of sulpyrine for assay taken, calculated on the dried basis
Sultamicillin Tosilate Tablets contains not less than 90.
Powder Sultamicillin Tosilate Tablets, take a portion of the powder, equivalent to 7 mg (potency) of Sultamicillin Tosilate Hydrate, add 2 mL of methanol and shake well, then centrifuge this solution. To 1 mL of the su-pernatant liquid add 1 mL of hydroxylammonium chloride-ethanol TS, allow to stand for 3 minutes, then add 1 mL of acidic ammonium iron (III) sulfate TS: a red-brown color is produced.
Penicilloic acid—Weigh accurately the mass of not less than 5 Sultamicillin Tosilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 30 mg (potency) of Sultamicillin Tosilate Hydrate, add 0.02 mol/L phosphate buffer (pH 3.0) and sonicate for 5 minutes with occasional shaking, then add 0.02 mol/L phos-phate buffer (pH 3.0) to make exactly 50 mL. Filter this so-lution through a membrane filter with a pore size not exceeding 0.45 mm, and discard the first 5 mL of the filtrate. Pipet 10 mL of the subsequent filtrate into a glass-stoppered flask, add exactly 5 mL of 0.005 mol/L iodine VS, and stop- per tightly. After standing for 5 minutes, titrate <2.50> this solution with 0.005 mol/L sodium thiosulfate VS (indicator: 1.0 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction: the amount of penicilloic acid (C25H34N4O11S2: 630.69) is not more than 5.5z. Each mL of 0.005 mol/L sodium thiosulfate VS ＝ 0.2585 mg of C25H34N4O11S2 Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Perform the procedure within 2 hours after preparation of the sample solution and standard solution. To 1 tablet of Sultamicillin Tosilate Tablets add a suitable amount of the mobile phase, disperse the tablet by sonicating, and add the mobile phase to make exactly 200 mL. If it is necessary, filter or centrifuge. Pipet V mL of this solution, equivalent to about 5.6 mg (potency) of Sultamicillin Tosilate Hydrate, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 47 mg (potency) of Sultamicillin Tosilate RS, dissolve in the mobile phase to make exactly 25 mL. Pipet 3 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 25 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Sultamicillin Tosilate Hydrate. Amount [mg (potency)] of Sultamicillin (C25H30N4O9S2) ＝ MS × QT/QS × 24/V MS: Amount [mg (potency)] of Sultamicillin Tosilate RS taken Internal standard solution: A solution of isopropyl-4-aminobenzoate in the mobile phase (1 in 2500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Sultamicillin Tosilate Tablets is not less than 75z. Start the test with 1 tablet of Sultamicillin Tosilate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.42 mg (potency) of sul-tamicillin (C25H30N4O9S2), and use this solution as the sam-ple solution. Separately, weigh accurately about 27 mg of p-toluenesulfonic acid monohydrate, previously dried in a desiccator using sulfuric acid as desiccant for 18 hours, dis-solve in water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of p-toluenesulfonic acid in each solution. Dissolution rate (z) with respect to the labeled amount of sultamicillin (C25H30N4O9S2) ＝ MS × AT/AS × V?/V × 1/C × 450 × 3.126 MS: Amount (mg) of p-toluenesulfonic acid monohydrate taken C: Labeled amount [mg (potency)] of sultamicillin (C25H30 N4O9S2) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 222 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 13.6 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 5.5 with potassium hydroxide TS. To 950 mL of this solution add 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of p-tol-uenesulfonic acid is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of p-toluenesulfonic acid are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of p-toluenesulfonic acid is not more than 1.5z.
Perform the procedure within 2 hours after the preparation of the sample solution and standard solution. Weigh accurately the mass of not less than 20 tablets of Sultamicillin Tosilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg (po-tency) of Sultamicillin Tosilate Hydrate, add 40 mL of the mobile phase, sonicate, and add the mobile phase to make exactly 50 mL. If it is necessary, filter or centrifuge. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 50 mg (potency) of Sultamicillin Tosilate RS, dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add the mobile phase to make 25 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay under Sultamicillin Tosilate Hydrate. Amount [mg (potency)] of sultamicillin (C25H30N4O9S2) ＝ MS × QT/QS MS: Amount [mg (potency)] of Sultamicillin Tosilate RS taken Internal standard solution: A solution of isopropyl-4-aminobenzoate in the mobile phase (1 in 2500).
Suxamethonium Chloride for Injection occurs as a white, crystalline powder or mass.
Take an amount of Suxamethonium Chlo-ride for Injection, equivalent to 0.05 g of Suxamethonium Chloride Hydrate, dissolve in water to make 10 mL, and use this solution as the sample solution. Separately, dissolve 0.05 g of suxamethonium chloride for thin-layer chromatog-raphy in 10 mL of water, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solution on a plate of cellulose for thin-layer chromatography. Develop the plate with a mixture of a solution of ammonium acetate (1 in 100), acetone, 1-butanol and formic acid (20:20:20:1) to a distance of about 10 cm, and dry the plate at 1059C for 15 minutes. Spray evenly hydrogen hexachloroplatinate (IV)-potassium iodide TS on the plate: the spots obtained from the sample solution and standard solution are blue-purple in color and have similar Rf value. pH <2.54> The pH of a solution of 0.1 g of Suxamethoni-um Chloride for Injection in 10 mL of water is between 4.0 and 5.0.
Related substances—Take an amount of Sux-amethonium Chloride for Injection, equivalent to 0.25 g of Suxamethonium Chloride Hydrate, and proceed as directed in the Purity (2) under Suxamethonium Chloride Hydrate. Bacterial endotoxins <4.01> Less than 1.5 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Weigh accurately the contents of not less than 10 preparations of Suxamethonium Chloride for Injection. Weigh accurately about 0.5 g of the contents, and proceed as directed in the Assay under Suxamethonium Chloride Hy-drate. Each mL of 0.1 mol/L perchloric acid VS Tacalcitol Hydrate
Suxamethonium Chloride Injection is an aqueous injection. It contains not less than 93.
Suxamethonium Chloride Injection is a clear, colorless liquid.
Take a volume of Suxamethonium Chloride Injection, equivalent to 0.05 g of Suxamethonium Chloride Hydrate, add water to make 10 mL, and use this solution as accurately measured 25 mL of 0.1 mol/L sodium hydroxide VS, and boil for 40 minutes under a reflux condenser, and cool. Titrate <2.50> the excess sodium hydroxide with 0.1 mol/L hydrochloric acid VS. Transfer 50 mL of the freshly boiled and cooled water to a flask, add 2 drops of bromo-thymol blue TS, neutralize the solution with 0.1 mol/L sodi-um hydroxide VS, and perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 18.07 mg of C14H30Cl2N2O4
Containers—Hermetic containers. Storage—Not exceeding 59C, and avoid freezing.
Swertia and Sodium Bicarbonate Powder oc-curs as a light grayish yellow powder, having a bitter taste. Identiˆcation (1) To 10 g of Swertia and Sodium Bicar-bonate Powder add 10 mL of ethanol (95), shake for 15 minutes, ˆlter, and use the ˆltrate as the sample solution. Separately, dissolve 2 mg of Swertiamarin RS or swertiama-rin for thin-layer chromatography in 1 mL of ethanol (95), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 10 mL of the sample solution and 5 mL of the standard solution on a plate of silica gel for thin-layer chromatography. Proceed as directed in the Identiˆca- ‌JP XVIII Crude Drugs and Related Drugs / Toad Cake 2159‌ tion under Powdered Swertia Herb. To 0.5 g of Swertia and Sodium Bicarbonate Powder add 10 mL of water. After stirring, centrifuge the mixture with 500 revolutions per minute. Smear, using a small glass rod, the slide glass with a small amount of the precipitate, add 1 drop of a mixture of water and glycerin (1:1), and put a cover glass on it so that the tissue section spreads evenly without overlapping each other, taking precaution against inclusion of bubbles, and use this as the preparation for microscopic examination. If the precipitate separates into two layers, proceed with the upper layer in the same manner, and use as the preparation for microscopic examination. Heat the preparation for microscopic examination in a short time: the preparation reveals the yellow-green to yellow-brown, approximately spherical pollen grains with granular patterns under a microscope <5.01>. The pollen grains are 25 – 34 mm in diameter. The supernatant liquid obtained in (2) by centrifug-ing responds to Qualitative Tests <1.09> (1) for bicarbonate.
Tacalcitol Ointment contains not less than 90.
Perform the test with 30 mL each of the sam-ple solution and standard solution, both are obtained in the Assay, as directed under Liquid Chromatography <2.01> ac-cording to the following conditions: the retention time of the principal peaks in the chromatograms obtained from the sample solution and standard solution is the same, and both adsorption spectra of these peaks exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: A photodiode array detector (wavelength: 265 nm, spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay.
Related substances—This test is only applied to the preparations of 20 mg/g. Conduct this procedure using light-resistant vessels. To an amount of Tacalcitol Ointment, equivalent to about 20 mg of tacalcitol (C27H44O3), add 5 mL of hexane and 5 mL of methanol, shake thoroughly for 15 minutes, and centrifuge. Discard the upper layer, pipet 5 mL of the lower layer, and evaporate the solvents in vacuum. Dissolve the residue in 1 mL of methanol, filter this solution through a membrane filter with a pore size not exceeding 0.2 mm, and use the fil-trate as the sample solution. Perform the test with 30 mL of the sample solution as directed under Liquid Chromatogra-phy <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak other than tacalcitol and pre-tacalcitol, having a relative retention time of about 0.83 to tacalcitol, is not more than 0.8z, and the total amount of the peaks other than tacalcitol and pre-tacalcitol is not more than 2.0z. Operating conditions— Detector, column and column temperature: Proceed as di-rected in the operating conditions in the Assay. Mobile phase A: Water. Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 30 30 – 50 40 → 0 60 → 100 50 – 60 Flow rate: Adjust so that the retention time of tacalcitol is about 24 minutes. Time span of measurement: For 60 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 0.5 mL of the sample solution add methanol to make 50 mL, and use this solution as the solution for system suitability test. Pipet 4 mL of the solution for system suitability test, and add methanol to make exactly 10 mL. Confirm that the peak area of tacalcitol obtained with 30 mL of this solution is equivalent to 28 to 52z of that with 30 mL of the solution for system suitability test. System performance: When the procedure is run with 30 mL of the sample solution under the above operating condi-tions, pre-tacalcitol and tacalcitol are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 30 mL of the solution for system suitability test under the above operating conditions, the relative standard devia- tion of the peak area of tacalcitol is not more than 10z.
Weigh accurately an amount of Tacalcitol Oint-ment, equivalent to about 2 mg of tacalcitol (C27H44O3), add exactly 5 mL of hexane, exactly 4 mL of methanol, and ex-actly 1 mL of the internal standard solution, shake thor-oughly for 15 minutes, and centrifuge. Filter the lower layer through a membrane filter with a pore size not exceeding 0.2 ‌mm, and use the filtrate as the sample solution. Separately, weigh accurately about 1 mg of Tacalcitol RS (separately de-termine the water <2.48> in the same manner as Tacalcitol Hydrate), and dissolve in methanol to make exactly 20 mL. Pipet 1 mL of this solution, and add methanol to make ex-actly 100 mL. Pipet 4 mL of this solution, add exactly 1 mL of the internal standard solution and exactly 5 mL of hexane, shake thoroughly for 15 minutes, and centrifuge. Filter the lower layer through a membrane filter with a pore size not exceeding 0.2 mm, and use the filtrate as the stand-ard solution. Perform the test with 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of tacalcitol to that of the internal standard. Amount (mg) of tacalcitol (C27H44O3) ＝ MS × QT/QS × 2 MS: Amount (mg) of Tacalcitol RS taken, calculated on the anhydrous basis Internal standard solution—A solution of hexyl para-hydroxybenzoate in methanol ( 3 in 2,500,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 265 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of acetonitrile for liquid chroma-tography and diluted 0.25 mol/L acetic acid TS (1 in 10) (13:7). Flow rate: Adjust so that the retention time of tacalcitol is about 18 minutes. System suitability— System performance: When the procedure is run with 30 mL of the standard solution under the above operating con-ditions, the internal standard and tacalcitol are eluted in this order with the resolution between these peaks being not less than 14. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tacalcitol to that of the internal standard is not more than 2.0z.
Take out the contents of Tacrolimus Cap-sules, to a quantity of the contents, equivalent to 5 mg of (separately determine the water <2.48> in the same manner as Tacrolimus Hydrate), and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, add 5 mL of water, allow to stand for 6 hours, and use this solution as the stand-ard solution. Then, proceed as directed in the Assay. Amount (mg) of tacrolimus (C44H69NO12) ＝ MS × QT/QS × V/250 MS: Amount (mg) of Tacrolimus RS taken, calculated on the anhydrous basis Internal standard solution—A solution of heptyl parahy-droxybenzoate in ethanol (99.5) (1 in 20,000). Dissolution Being specified separately when the drug is granted approval based on the Law.
Related substances—Being specified separately when the drug is granted approval based on the Law. Isomer Being specified separately when the drug is granted approval based on the Law. gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, 2-propanol for liquid chromatography and tetrahydrofuran for liquid chromatog-raphy (5:2:2). Flow rate: Adjust so that the retention time of tacrolimus is about 10 minutes. ‌System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, tacrolimus and the internal standard are eluted in this order with the resolution between these peaks being not less than 6, and the number of theoretical plates and the symmetry factor of the peak of tacrolimus are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tacrolimus to that of the internal standard is not more than 1.0z.
Take out the contents of not less than 20 Tacrolimus Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equiva-lent to about 25 mg of tacrolimus (C44H69NO12), add 15 mL of ethanol (99.5) and exactly 10 mL of the internal standard solution, and sonicate for 10 minutes with occasional shak-ing. Centrifuge this solution, to 5 mL of the supernatant liq-uid add 5 mL of water, allow to stand for 6 hours, and use this solution as the sample solution. Separately, weigh accu-rately about 25 mg of Tacrolimus RS (separately determine the water <2.48> in the same manner as Tacrolimus Hydrate), dissolve in 15 mL of ethanol (99.5), add exactly 10 mL of the internal standard solution, add 25 mL of water, allow to stand for 6 hours, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of tacrolimus to that of the internal standard. Amount (mg) of tacrolimus (C44H69NO12) ＝ MS × QT/QS MS: Amount (mg) of Tacrolimus RS taken, calculated on the anhydrous basis Internal standard solution—A solution of heptyl parahy-droxybenzoate in ethanol (99.5) (3 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). tacrolimus (C 44H 69NO12 ), add 2 mL of ethanol (95), shake Column: A stainless steel column 4.6 mm in inside diame- ter and 15 cm in length, packed with octadecylsilanized silica for 10 minutes, and centrifuge. To 1 mL of the supernatant liquid add 0.5 mL of 1,3-dinitrobenzene TS and 0.5 mL of sodium hydroxide TS, shake, and allow to stand for 3 minutes: a light red-purple develops. Purity Related substances—Being specified separately when the drug is granted approval based on the Law. Isomer Being specified separately when the drug is granted approval based on the Law. gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase: A mixture of water, 2-propanol for liquid chromatography and tetrahydrofuran for liquid chromatog-raphy (5:2:2). Flow rate: Adjust so that the retention time of tacrolimus is about 10 minutes. ‌System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, tacrolimus and the internal standard are eluted in this order with the resolution between these peaks being not less than 6, and the number of theoretical plates and the symmetry factor of the peak of tacrolimus are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of tacrolimus to that of the internal standard is not more than 1.0z.
Taltirelin Orally Disintegrating Tablets
Taltirelin Orally Disintegrating Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of taltirelin hydrate (C17H23N7O5. 4H2O: 477.47). Method of preparation Prepare as directed under Tablets, with Taltirelin Hydrate. Identification Powder Taltirelin Orally Disintegrating mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak with the relative retention time of about 0.7 to tal-tirelin is not more than 0.7z and the peaks with the relative retention time of about 0.8 and about 0.9, respectively, are not more than 0.3z, and the peak other than taltirelin and the peaks mentioned above is not more than 0.1z. And the total amount of the peaks other than taltirelin is not more than 1.0z. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of taltirelin, beginning from 1/3 times the retention time of taltirelin. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of taltirelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of taltirelin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Taltirelin Orally Disintegrating Tablets add V/2 mL of the mobile phase and exactly V/10 mL of the internal standard solution, and shake vigorously for 5 minutes. Then, add the mobile phase to make V mL so that each mL contains about 0.
Powder Taltirelin Orally Disintegrating mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak with the relative retention time of about 0.7 to tal-tirelin is not more than 0.7z and the peaks with the relative retention time of about 0.8 and about 0.9, respectively, are not more than 0.3z, and the peak other than taltirelin and the peaks mentioned above is not more than 0.1z. And the total amount of the peaks other than taltirelin is not more than 1.0z. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of taltirelin, beginning from 1/3 times the retention time of taltirelin. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of taltirelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of taltirelin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Taltirelin Orally Disintegrating Tablets add V/2 mL of the mobile phase and exactly V/10 mL of the internal standard solution, and shake vigorously for 5 minutes. Then, add the mobile phase to make V mL so that each mL contains about 0.1 mg of taltirelin hydrate (C17H23N7O5.4H2O), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Then, proceed as directed in the Assay. Tablets. To a portion of the powder, equivalent to 30 mg of Taltirelin Hydrate, add 10 mL of water, shake for 5 minutes, and filter. To 0.5 mL of the filtrate add 2 mL of a solution Amount (mg) of taltirelin hydrate (C17H ＝ MS × QT/QS × V/500 × 1.178 23N7O5.4H2O) of 4-nitrobenzenediazonium fluoroborate (1 in 2000) and 3 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 9.0): a red color is produced.
Related substances—Powder Taltirelin Orally Disin-tegrating Tablets. To a portion of the powder, equivalent to 5 mg of Taltirelin Hydrate, add 20 mL of the mobile phase, shake for 5 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of o-acetanisidide (1 in 2500). Disintegration Being specified separately when the drug is granted approval based on the Law. Dissolution <6.10> When the test is performed at 50 revolu- ‌tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Taltirelin Orally Disintegrating Tablets is not less than 85z. Start the test with 1 tablet of Taltirelin Orally Disintegrat-ing Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 5.6 mg of taltirelin hydrate (C17H23N7O5.4H2O), and use this solution as the sample solu-tion. Separately, weigh accurately about 28 mg of taltirelin hydrate for assay (separately determine the water <2.48> in the same manner as Taltirelin Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the fol-lowing conditions, and determine the peak areas, AT and AS, of taltirelin in each solution. Dissolution rate (z) with respect to the labeled amount of taltirelin hydrate (C17H23N7O5.4H2O) ＝ MS × AT/AS ×V?/V × 1/C × 18 × 1.178 MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of taltirelin hydrate (C17H23N7O5.4H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of taltirelin is not more than 2.0z.
Weigh accurately, and powder not less than 20 Tal-tirelin Orally Disintegrating Tablets. Weigh accurately a portion of the powder, equivalent to about 5 mg of taltirelin hydrate (C17H23N7O5.4H2O), add 25 mL of the mobile phase and exactly 5 mL of the internal standard solution, shake for 5 minutes, add the mobile phase to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of taltirelin hydrate for assay (separately determine the water <2.48> in the same manner as Taltirelin Hydrate), and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solu-tion. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01>, according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of taltirelin to that of the internal standard. MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of o-acetanisidide (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 850 mL of this solution add 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, taltirelin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of taltirelin to that of the internal standard is not more than 1.0z.
Taltirelin Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of tal-tirelin hydrate (C17H23N7O5.4H2O: 477.47). Method of preparation Prepare as directed under Tablets, with Taltirelin Hydrate. Identification Powder Taltirelin Tablets. To a portion of the powder, equivalent to 30 mg of Taltirelin Hydrate, add 10 mL of water, shake for 15 minutes, and filter. To 0.5 mL of the filtrate add 2 mL of a solution of 4-nitrobenzendiazo-nium fluoroborate (1 in 2000) and 3 mL of boric acid-potas-sium chloride-sodium hydroxide buffer solution (pH 9.0): a red color is produced. Purity Related substances—Powder Taltirelin Tablets. To a portion of the powder, equivalent to 5 mg of Taltirelin Hy-drate, add 20 mL of the mobile phase, shake for 20 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate these amounts by the area percentage method: the amount of the peak with the relative retention time of about 0.7 to taltirelin is not more than 0.7z and the peaks with the relative retention time of about Amount (mg) of taltirelin hydrate (C ＝ MS × QT/QS × 1/10 × 1.178 17H23N7O5.4H2O) 0.8 and about 0.9, respectively, are not more than 0.3z, and the peak other than taltirelin and the peaks mentioned above is not more than 0.1z. And the total amount of the peaks other than taltirelin is not more than 1.0z. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of taltirelin, beginning from 1/3 times the retention time of taltirelin. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of taltirelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of taltirelin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Taltirelin Tablets add V/2 mL of the mobile phase and exactly V/10 mL of the internal standard solu-tion, and sonicate for 10 minutes while occasional shaking. Then, add the mobile phase to make V mL so that each mL contains about 0.
Powder Taltirelin Tablets. To a portion of the powder, equivalent to 30 mg of Taltirelin Hydrate, add 10 mL of water, shake for 15 minutes, and filter. To 0.5 mL of the filtrate add 2 mL of a solution of 4-nitrobenzendiazo-nium fluoroborate (1 in 2000) and 3 mL of boric acid-potas-sium chloride-sodium hydroxide buffer solution (pH 9.0): a red color is produced.
Related substances—Powder Taltirelin Tablets. To a portion of the powder, equivalent to 5 mg of Taltirelin Hy-drate, add 20 mL of the mobile phase, shake for 20 minutes, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic in-tegration method, and calculate these amounts by the area percentage method: the amount of the peak with the relative retention time of about 0.7 to taltirelin is not more than 0.7z and the peaks with the relative retention time of about Amount (mg) of taltirelin hydrate (C ＝ MS × QT/QS × 1/10 × 1.178 17H23N7O5.4H2O) 0.8 and about 0.9, respectively, are not more than 0.3z, and the peak other than taltirelin and the peaks mentioned above is not more than 0.1z. And the total amount of the peaks other than taltirelin is not more than 1.0z. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To 900 mL of this solution add 100 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of taltirelin, beginning from 1/3 times the retention time of taltirelin. System suitability— Test for required detectability: To 1 mL of the sample so-lution add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of taltirelin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of taltirelin is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Taltirelin Tablets add V/2 mL of the mobile phase and exactly V/10 mL of the internal standard solu-tion, and sonicate for 10 minutes while occasional shaking. Then, add the mobile phase to make V mL so that each mL contains about 0.1 mg of taltirelin hydrate (C17H23N7O5.4H2 O), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of taltirelin hydrate (C17H23N7O5.4H2O) ＝ MS × QT/QS × V/500 × 1.178 MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of o-acetanisidide (1 in 2500). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Taltirelin Tablets is not less than 85z. Start the test with 1 tablet of Taltirelin Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 5.6 mg of taltirelin hydrate (C17H23N7O5.4H2O), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of taltirelin hydrate for assay (separately determine the water <2.48> in the same manner as Taltirelin Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the following conditions, and determine the peak areas, AT and AS, of taltirelin in each solution. Dissolution rate (z) with respect to the labeled amount of taltirelin hydrate (C17H23N7O5.4H2O) ＝ MS × AT/AS × V?/V × 1/C × 18 × 1.178 MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of taltirelin hydrate (C17H23N7O5.4H2O) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of taltirelin are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of taltirelin is not more than 2.0z.
Weigh accurately, and powder not less than 20 Tal-tirelin Tablets. Weigh accurately a portion of the powder, equivalent to about 5 mg of taltirelin hydrate (C17H23N7O5. 4H2O), add 25 mL of the mobile phase and exactly 5 mL of the internal standard solution, shake for 20 minutes, add the mobile phase to make 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 2 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of taltirelin hydrate for assay (separately determine the water <2.48> in the same manner as Taltirelin Hydrate), and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal stand-ard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01>, according to the following conditions, and calculate the ratios, QT and QS, of the peak area of taltirelin to that of the internal stand-ard. Amount (mg) of taltirelin hydrate (C17H23N7O5.4H2O) ＝ MS × QT/QS × 1/10 × 1.178 MS: Amount (mg) of taltirelin hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of o-acetanisidide (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 2.5 with phos-phoric acid, and add 1.7 g of sodium 1-octanesulfonate. To ‌850 mL of this solution add 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of taltirelin is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, taltirelin and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of taltirelin to that of the internal standard is not more than 1.0z.
Tamsulosin Hydrochloride Extended-release Tablets
Tamsulosin Hydrochloride Extended-release Tablets contain not less than 94.0z and not more than 106.0z of the labeled amount of tamsulosin hydro-chloride (C20H28N2O5S.HCl: 444.97). Method of preparation Prepare as directed under Tablets, with Tamsulosin Hydrochloride. Identification To an amount of powdered Tamsulosin Hy-drochloride Extended-release Tablets, equivalent to 1 mg of Tamsulosin Hydrochloride, add about 5 g of porcelain balls with about 5 mm in diameter, add 20 mL of 0.2 mol/L so-dium hydroxide TS, warm at 509C for 10 minutes, and shake vigorously for 15 minutes. Then, add 7 mL of acetonitrile, shake slightly, and centrifuge. Take the supernatant liquid, add 2.5 g of sodium chloride and 5 mL of ethyl acetate, shake vigorously for 5 minutes, and centrifuge. Take the su-pernatant liquid, evaporate to dryness at 509C in a water bath under reduced pressure, dissolve the residue with 20 mL of water, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 222 nm and 226 nm, and between 278 nm and 282 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Tamsulosin Hydrochloride Extended-release Tablets add about 5 g of porcelain balls with about 5 mm in diameter and 5 mL of water, and shake to disintegrate the tablet. Add 20 mL of a solution of sodium hydroxide (1 in 500), warm at 509C for 10 minutes, shake vigorously for 30 minutes, and add 10 mL of acetonitrile and 5 mL of 0.2 mol/L hydrochloric acid TS. To this solution add exactly 5 mL of the internal standard solution for every 0.1 mg of tamsulosin hydrochloride, add the mobile phase to make 50 mL, shake slightly, and centrifuge. Filter the supernatant liq-uid through a membrane filter with a pore size not exceeding 0.45 mm. To V mL of the filtrate add the mobile phase to make V? mL so that each mL contains about 2 mg of tam-sulosin hydrochloride (C20H28N2O5S.
To an amount of powdered Tamsulosin Hy-drochloride Extended-release Tablets, equivalent to 1 mg of Tamsulosin Hydrochloride, add about 5 g of porcelain balls with about 5 mm in diameter, add 20 mL of 0.2 mol/L so-dium hydroxide TS, warm at 509C for 10 minutes, and shake vigorously for 15 minutes. Then, add 7 mL of acetonitrile, shake slightly, and centrifuge. Take the supernatant liquid, add 2.5 g of sodium chloride and 5 mL of ethyl acetate, shake vigorously for 5 minutes, and centrifuge. Take the su-pernatant liquid, evaporate to dryness at 509C in a water bath under reduced pressure, dissolve the residue with 20 mL of water, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits maxima between 222 nm and 226 nm, and between 278 nm and 282 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Tamsulosin Hydrochloride Extended-release Tablets add about 5 g of porcelain balls with about 5 mm in diameter and 5 mL of water, and shake to disintegrate the tablet. Add 20 mL of a solution of sodium hydroxide (1 in 500), warm at 509C for 10 minutes, shake vigorously for 30 minutes, and add 10 mL of acetonitrile and 5 mL of 0.2 mol/L hydrochloric acid TS. To this solution add exactly 5 mL of the internal standard solution for every 0.1 mg of tamsulosin hydrochloride, add the mobile phase to make 50 mL, shake slightly, and centrifuge. Filter the supernatant liq-uid through a membrane filter with a pore size not exceeding 0.45 mm. To V mL of the filtrate add the mobile phase to make V? mL so that each mL contains about 2 mg of tam-sulosin hydrochloride (C20H28N2O5S.HCl), and use this solu-tion as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of tamsulosin hydrochloride (C20H28N2O5S.HCl) ＝ MS × QT/QS × V?/V × 1/100 MS: Amount (mg) of tamsulosin hydrochloride for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (1 in 25,000). Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Tam-sulosin Hydrochloride Extended-release Tablets, and pow-der. Weigh accurately a portion of the powder, equivalent to about 0.1 mg of tamsulosin hydrochloride (C20H28N2O5S. HCl), add about 5 g of porcelain balls with about 5 mm in diameter and 5 mL of water, shake, then add 20 mL of a so-lution of sodium hydroxide (1 in 500), warm at 509C for 10 minutes, and shake vigorously for 30 minutes. To this solu-tion add 10 mL of acetonitrile, 5 mL of 0.2 mol/L hydro-chloric acid TS and exactly 5 mL of the internal standard so-lution, then add 5 mL of the mobile phase, shake slightly, and centrifuge. Filter the supernatant liquid through a mem-brane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accu-rately about 10 mg of tamsulosin hydrochloride for assay, previously dried at 1059C for 2 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 2 mL of this so-lution, add exactly 10 mL of the internal standard solution, add the mobile phase to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tamsulosin to that of the internal standard. Amount (mg) of tamsulosin hydrochloride (C20H28N2O5S.HCl) ＝ MS × QT/QS × 1/100 MS: Amount (mg) of tamsulosin hydrochloride for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in the mobile phase (1 in 25,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.4 mL of perchloric acid and 1.5 g of sodium hydroxide in 950 mL of water, adjust to pH 2.0 with sodium hydroxide TS, and add water to make 1000 mL. To 700 mL of this solution add 300 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of tamsulosin is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and tamsulosin are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the ratio of the peak area of tamsulosin to that of the internal standard is not more than 1.0z.‌‌
Tazobactam and Piperacillin for Injection
注射用タゾバクタム･ピペラシリン
Tazobactam and Piperacillin for Injection is a preparation for injection which is dissolved before use. It contains not less than 93.
Tazobactam and Piperacillin for Injection oc-curs as white to pale yellowish white, masses or powder.
(1) Determine 1H spectrum of a solution of Tazobactam and Piperacillin for Injection in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropionate-d4 for nuclear mag-netic resonance spectroscopy as an internal reference com-pound: it exhibits a singlet signal A at around d 4.2 ppm, a multiplet signal B at d 7.3 – 7.5 ppm, a doublet signal C at around d 7.8 ppm and a doublet signal D at around d 8.1 ppm. The ratio of integrated intensity of these signals, A:B and C:D, is about 1:5 and about 1:1, respectively. (2) Tazobactam and Piperacillin for Injection responds to Qualitative Tests <1.09> (1) for sodium salt. pH <2.54> The pH of a solution of an amount of Tazobac-tam and Piperacillin for Injection, equivalent to 4.0 g (po-tency) of Piperacillin Hydrate, in 40 mL of water is 5.1 to 6.3.
(1) Clarity and color of solution—A solution of an amount of Tazobactam and Piperacillin for Injection, equivalent to 4.0 g (potency) of Piperacillin Hydrate, in 40 mL of water is clear and colorless. (2) Related substances—Keep the sample solution at 59C. Dissolve an amount of Tazobactam and Piperacillin for Injection, equivalent to 0.1 g (potency) of Piperacillin Hy-drate, in 100 mL of dissolving solution, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add dissolving solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 0.06 to piperacillin, obtained from the sample solution is not larger than 1.3 times the peak area of tazobactam from the stand-ard solution, the area of the peak, having the relative reten-tion time of about 0.05, about 0.07, about 0.19, about 0.45 and about 0.53, from the sample solution is not larger than 1/10 times the peak area of tazobactam from the standard solution, and the total area of the peaks, having the relative retention time of about 0.05, about 0.06, about 0.07, about 0.19, about 0.45 and about 0.53, from the sample solution is not larger than 1.5 times the peak area of tazobactam from the standard solution. Furthermore, the area of the peak, having the relative retention time of about 1.20 and about 1.36, from the sample solution is not larger than 1/5 times the peak area of piperacillin from the standard solution, the area of the peak, having the relative retention time of about 0.15 and about 0.63, from the sample solution is not larger than 3/10 times the peak area of piperacillin from the stand-ard solution, the area of the peak, having the relative reten-tion time of about 0.91 and about 1.53, from the sample so-lution is not larger than 2/5 times the peak area of piperacil-lin from the standard solution, the total area of the peaks eluted between the relative retention time of about 0.85 and about 0.87, from the sample solution is not larger than 1/2 times the peak area of piperacillin from the standard solu-tion, the total area of the peaks, having the relative retention time of about 0.85 and about 0.87, from the sample solution is not larger than 1.5 times the peak area of piperacillin from the standard solution, and the area of the peak other than tazobactam, piperacillin and the peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of piperacillin from the standard solution. The total area of the peaks other than tazobactam, piperacillin and the peaks, having the relative retention time of about 0.05, about 0.06, about 0.07, about 0.19, about 0.45 and about 0.53, from the sample solution is not larger than 4.0 times the peak area of piperacillin from the standard solu-tion. For the area of the peaks, having the relative retention time of about 0.05, about 0.06, about 0.07, about 0.15, about 0.19, about 0.45, about 0.53, about 0.63, about 0.68, about 0.79, about 0.91 and about 1.53, multiply their correc-tion factors 2.09, 0.70, 0.92, 0.42, 0.69, 0.56, 0.19, 1.37, 1,93, 1.64, 1.73 and 1.29, respectively, and for the total area of the peaks having the relative retention time of about 0.85 and about 0.87 and the total area of the peaks that are eluted between the peaks having the relative retention time of about 0.85 and about 0.87, multiply their correction factors, 1.79 and 2.50, respectively. Dissolving solution: To 950 mL of diluted 1 mol/L dipotassium hydrogen phosphate TS for buffer solution (1 in 100) adjusted to pH 6.5 with phosphoric acid, add 50 mL of acetonitrile. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase and flow rate: Pro-ceed as directed in the operating conditions in the Assay (1). Time span of measurement: For 36 minutes after injec- tion, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add dissolving solution to make exactly 20 mL. Confirm that the peak area of tazobactam obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, tazobactam and piperacillin are eluted in this order with the resolution between these peaks being not less than 50, and the number of theoretical plates and the symmetry factor of the peak of tazobactam are not less than 40,000 and not more than 1.5, respectively, and those of piperacillin are not less than 150,000 and not more than 1.5, respec-tively. Furthermore, when warm the sample solution at 409C for 60 minutes and proceed with 20 mL of this solution under the above conditions, the resolution between the two peaks, having the relative retention time of about 0.85 and about 0.87 to piperacillin, is not less than 2.9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviations of the peak area of tazobactam and piperacillin are not more than 2.0z, respec-tively. Water <2.48> Weigh accurately the mass of the content of 1 container of Tazobactam and Piperacillin for Injection, dis-solve in 20 mL of methanol for water determination, and perform the test with this solution according to the direct titration of Volumetric titration: not more than 0.6z. Per-form a blank determination in the same manner, and make any necessary correction. Bacterial endotoxins <4.01> Less than 0.07 EU/mg (po-tency) of Piperacillin Hydrate. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Tazobactam—Dissolve the contents of 10 con-tainers of Tazobactam and Piperacillin for Injection in a suitable amount of dissolving solution. Washout these empty containers with dissolving solution, combine the washings and the former solution, and add dissolving solution to make exactly V mL so that each mL contains about 5 mg (potency) of Tazobactam. Pipet 5 mL of this solution, add dissolving solution to make exactly 200 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 25 mg (potency) of Tazobactam RS, dissolve in 10 mL of acetonitrile, dilute with an amount of diluted 1 mol/L dipotassium hydrogen phosphate TS for buffer solution (1 in 100) adjusted to pH 6.5 with phosphoric acid to make ex-actly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions and determine the peak areas, AT and AS, of tazobactam in each solution. ‌Amount [g (potency)] of tazobactam (C10H12N4O5S) in 1 container of Tazobactam and Piperacillin for Injection ＝ MS × AT/AS × V/50,000 MS: Amount [mg (potency)] of Tazobactam RS taken Dissolving solution: To 950 mL of diluted 1 mol/L dipotassium hydrogen phosphate TS for buffer solution (1 in 100) adjusted to pH 6.5 with phosphoric acid, add 50 mL of acetonitrile. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: Dissolve 1.74 g of dipotassium hydrogen phosphate in 1000 mL of water, and adjust to pH 2.6 with phosphoric acid. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. solution. Amount [g (potency)] of piperacillin (C23H27N5O7S) in 1 container of Tazobactam and Piperacillin for Injection ＝ MS × AT/AS × V/12,500 MS: Amount [mg (potency)] of Piperacillin RS taken Dissolving solution: To 950 mL of diluted 1 mol/L dipotassium hydrogen phosphate TS for buffer solution (1 in 100) adjusted to pH 6.5 with phosphoric acid, add 50 mL of acetonitrile. Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: When the procedure is run with 20 mL of the solution for system suitability test obtained in the Assay (1) under the above operating conditions, tazobactam and piperacillin are eluted in this order with the resolution between these peaks being not less than 50, and the number of theoretical plates and the symmetry factor of the peak of piperacillin are not less than 100,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) tions, the relative standard deviations of the peak area of piperacillin is not more than 1.0z. 0 – 5 100 0 5 – 15 100 → 76 0 → 24 15 – 25 76 → 65 24 → 35 25 – 36 65 35 Flow rate: 1.5 mL per minute. System suitability— System performance: Dissolve 50 mg (potency) of piper-acillin hydrate in the standard solution to make 50 mL, and use this solution as the solution for system suitability test. When the procedure is run with 20 mL of the solution for sys-tem suitability test under the above operating conditions, tazobactam and piperacillin are eluted in this order with the resolution between these peaks being not less than 50, and the number of theoretical plates and the symmetry factor of the peak of tazobactam are not less than 25,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above condi-tions, the relative standard deviations of the peak area of tazobactam is not more than 1.0z. (2) Piperacillin—Dissolve the contents of 10 containers of Tazobactam and Piperacillin for Injection in a suitable amount of dissolving solution. Washout these empty con-tainers with dissolving solution, combine the washings and the former solution, and add dissolving solution to make ex-actly V mL so that each mL contains about 40 mg (potency) of Piperacillin Hydrate. Pipet 5 mL of this solution, add dis-solving solution to make exactly 200 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 50 mg (potency) of Piperacillin RS, dissolve in 2.5 mL of acetonitrile, dilute with an amount of diluted 1 mol/L dipotassium hydrogen phosphate TS for buffer solution (1 in 100) adjusted to pH 6.5 with phosphoric acid to make ex-actly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of piperacillin in each
Teceleukin for Injection (Genetical Recombination)
注射用テセロイキン（遺伝子組換え）
Teceleukin for Injection (Genetical Recombination) is a preparation for injection which is dissolved before use. It contains not less than 70.
Teceleukin for Injection (Genetical Recombi-nation) occurs as a white, light mass or powder.
Dissolve the content of 1 container of Teceleukin for Injection (Genetical Recombination) in 1 mL of sterilized purified water, dilute exactly with potency meas-uring medium for teceleukin to make the sample stock solu- ‌tion containing about 200 units of Teceleukin (Genetical Recombination) per mL. Proceed as directed in the Identifi-cation (1) under Teceleukin (Genetical Recombination). pH <2.54> Being specified separately when the drug is granted approval based on the Law.
Clarity and color of solution—Dissolve the content of 1 container of Teceleukin for Injection (Genetical Recom-bination) in 1 mL of water: the solution is clear and color-less. Loss on drying Transfer the content of the container of Teceleukin for Injection (Genetical Recombination) to a weighing bottle under the atmosphere not exceeding 10z relative humidity, and perform the test as directed in the Water content determination described in the Minimum Requirements for Biological Products: not more than 5z. Bacterial endotoxins <4.01> Less than 5 EU/350,000 units. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Calculate as |M － A| ＝ 0. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Dissolve the content of 1 container of Teceleukin for Injection (Genetical Recombination) in exactly 1 mL of steri-lized purified water, dilute exactly with culture medium for assay of teceleukin to make the sample solution containing a definite concentration of 10 to 50 units/mL (estimate). Pro-ceed as directed in the Assay under Teceleukin (Genetical Recombination), and calculate the amount (unit) of teceleu-kin in 1 container by the following formula. Amount (unit) of teceleukin in 1 container ＝ S × DT/DS × d × 1 S: Concentration of the standard solution (unit/mL) d: Dilution coefficient when sample solution prepared 1: Volume (mL) of the sample solution
Telmisartan and Amlodipine Besilate Tablets
テルミサルタン･アムロジピンベシル酸塩錠
Telmisartan and Amlodipine Besilate Tablets con-tain not less than 95.0z and not more than 105.0z of the labeled amount of telmisartan (C33H30N4O2: 514.62) and amlodipine besilate (C20H25ClN2O5. C6H6O3S: 567.05) Method of preparation Prepare as directed under Tablets, with Telmisartan and Amlodipine Besilate. Identification (1) Perform the test with 5 mL each of the sample solution and standard solution obtained in the Assay (1) as directed under Liquid Chromatography <2.01> accord- ing to the following conditions: the retention times of the peaks of telmisartan in the chromatograms obtained from the sample solution and the standard solution are the same, and both absorption spectra of these peaks exhibit similar in-tensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 270 nm; spectrum range of measurement: 210 – 400 nm). System suitability – System performance: Proceed as directed in the system suitability in the Assay (1). (2) Perform the test with 5 mL each of the sample solu-tion and standard solution obtained in the Assay (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions: the retention times of the peaks of amlodipine besilate in the chromatograms obtained from the sample solution and the standard solution are the same, and both spectra of these peaks in the chromatograms exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (2). Detector: A photodiode array detector (wavelength: 237 nm; spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (2). Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Telmisartan—To 1 tablet of Telmisartan and Am-lodipine Besilate Tablets add 4V/5 mL of the dissolving so-lution, disintegrate by sonicating, and add the dissolving so-lution to make exactly V mL so that each mL contains about 1.
(1) Perform the test with 5 mL each of the sample solution and standard solution obtained in the Assay (1) as directed under Liquid Chromatography <2.01> accord- ing to the following conditions: the retention times of the peaks of telmisartan in the chromatograms obtained from the sample solution and the standard solution are the same, and both absorption spectra of these peaks exhibit similar in-tensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (1). Detector: A photodiode array detector (wavelength: 270 nm; spectrum range of measurement: 210 – 400 nm). System suitability – System performance: Proceed as directed in the system suitability in the Assay (1). (2) Perform the test with 5 mL each of the sample solu-tion and standard solution obtained in the Assay (2) as di-rected under Liquid Chromatography <2.01> according to the following conditions: the retention times of the peaks of amlodipine besilate in the chromatograms obtained from the sample solution and the standard solution are the same, and both spectra of these peaks in the chromatograms exhibit similar intensities of absorption at the same wavelengths. Operating conditions— Column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay (2). Detector: A photodiode array detector (wavelength: 237 nm; spectrum range of measurement: 210 – 400 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay (2). Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Telmisartan—To 1 tablet of Telmisartan and Am-lodipine Besilate Tablets add 4V/5 mL of the dissolving so-lution, disintegrate by sonicating, and add the dissolving so-lution to make exactly V mL so that each mL contains about 1.6 mg of telmisartan (C33H30N4O2). Centrifuge this solu-tion, pipet 5 mL of the supernatant liquid, add the buffer so-lution to make exactly 25 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay (1). Amount (mg) of telmisartan (C33H30N4O2) ＝ MS × AT/AS × V/50 MS: Amount (mg) of telmisartan for assay taken Dissolving solution: Dissolve 2 g of ammonium dihydro- gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. Buffer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Amlodipine Besilate—To 1 tablet of Telmisartan and Amlodipine Besilate Tablets add 4V/5 mL of the dissolving solution, disintegrate by sonicating, and add the dissolving solution to make exactly V mL so that each mL contains about 0.138 mg of amlodipine besilate (C20H25ClN2O5. C6H6O3S). Centrifuge this solution, pipet 5 mL of the super-natant liquid, add the buffer solution to make exactly 25 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay (2). Amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × AT/AS × V/250 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Dissolving solution: Dissolve 2 g of ammonium dihydro-gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. Buffer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Dissolution <6.10> (1) Telmisartan—Being specified sepa-rately when the drug is granted approval based on the Law. (2) Amlodipine Besilate—Being specified separately when the drug is granted approval based on the Law.
(1) Telmisartan—Weigh accurately the mass of not less than 20 Telmisartan and Amlodipine Besilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 80 mg of telmisartan (C33H30N4O2), add 40 mL of the dissolving solution, disintegrate by sonicating, and add the dissolving solution to make exactly 50 mL. Cen-trifuge this solution, pipet 5 mL of the supernatant liquid, add the buffer solution to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of telmisartan for assay, previously dried at 1059C for 4 hours, add the dissolving solution to make ex-actly 50 mL, and use this solution as the telmisartan stand-ard stock solution. Pipet 5 mL of the telmisartan standard stock solution, add the buffer solution to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of telmisartan in each solution. pine Besilate RS (separately determine the water <2.48> in the same manner as Amlodipine Besilate) and add the dissolving solution to make exactly 100 mL. Pipet 20 mL of this solu-tion, add the dissolving solution to make exactly 50 mL, and use this solution as the amlodipine besilate standard stock solution. Pipet 5 mL of the amlodipine besilate standard stock solution, add the buffer solution to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography Amount (mg) of telmisartan (C H N O ) <2.01> according to the following conditions, and determine ＝ MS × AT/AS 33 30 4 2 the peak areas, AT and AS, of amlodipine in each solution. MS: Amount (mg) of telmisartan for assay taken Dissolving solution: Dissolve 2 g of ammonium dihydro- gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. Buffer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.5 with phosphoric acid. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 2.0 2.0 – 7.0 90 → 20 10 → 80 7.0 – 8.0 Flow rate: 0.8 mL per minute. System suitability— System performance: To each 5 mL of the telmisartan standard stock solution and the amlodipine besilate standard stock solution obtained in (2) add the buffer solution to make 25 mL. When the procedure is run with 5 mL of this so-lution under the above operating conditions, amlodipine and telmisartan are eluted in this order with the resolution be-tween these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of telmisartan is not more than 1.0z. (2) Amlodipine Besilate—Weigh accurately the mass of not less than 20 Telmisartan and Amlodipine Besilate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 6.9 mg of amlodipine besilate (C20H25ClN2O5.C6H6O3S), add 40 mL of the dissolving solu-tion, disintegrate by sonicating, and add the dissolving solu-tion to make exactly 50 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add the buffer solution to make exactly 25 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 35 mg of Amlodi- Amount (mg) of amlodipine besilate (C20H25ClN2O5.C6H6O3S) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of Amlodipine Besilate RS taken, cal-culated on the anhydrous basis Dissolving solution: Dissolve 2 g of ammonium dihydro-gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. Buffer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 237 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.5 with phosphoric acid. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 2.0 2.0 – 7.0 90 → 20 10 → 80 7.0 – 8.0 Flow rate: 0.8 mL per minute. System suitability— System performance: To 5 mL each of the telmisartan standard stock solution obtained in (1) and the amlodipine besilate standard stock solution add the buffer solution to make 25 mL. When the procedure is run with 5 mL of this so-lution under the above operating conditions, amlodipine and telmisartan are eluted in this order with the resolution be-tween these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of amlodipine is not more than 1.0z.
‌Telmisartan and Hydrochlorothiazide Tablets
テルミサルタン・ヒドロクロロチアジド錠
Related substances—To a quantity of powdered Telmisartan and Hydrochlorothiazide Tablets, equivalent to 12.5 mg of Hydrochlorothiazide, add 40 mL of the dissolv-ing solution, disperse by sonicating, add the dissolving solu-tion to make exactly 50 mL. Centrifuge this solution, and use the supernatant liquid as the sample solution. Pipet 1 mL of the sample solution, add the dissolving solution to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the area of the peak having the relative retention time of about 0.9 to hydrochlorothiazide, obtained from the sample solution is not larger than the peak area of hydrochlorothiazide from the standard solution. Dissolving solution: Dissolve 2 g of ammonium dihydro- 0 – 8 90 → 50 10 → 50 8 – 12 50 50 12 – 18 50 → 20 50 → 80 18 – 20 20 80 Flow rate: 1.0 mL per minute. System suitability— Test for required detectability: Pipet 5 mL of the standard solution and add the dissolving solution to make exactly 50 mL. Conˆrm that the peak area of hydrochlorothiazide ob-tained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of hydrochlorothiazide are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Telmisartan—To 1 tablet of Telmisartan and Hydro-chlorothiazide Tablets add 4V/5 mL of the dissolving solu-tion, disintegrate by sonicating, add the dissolving solution to make exactly V mL so that each mL contains about 1.6 mg of telmisartan (C33H30N4O2). Centrifuge this solution, pipet 5 mL of the supernatant liquid, add the buŠer solution to make exactly 25 mL, and use this solution as the sample solution. Proceed as directed in the Assay (1). Amount (mg) of telmisartan (C33H30N4O2) ＝ MS × AT/AS × V/50 MS: Amount (mg) of telmisartan for assay taken Dissolving solution: Dissolve 2 g of ammonium dihydro- gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. BuŠer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Hydrochlorothiazide—To 1 tablet of Telmisartan and Hydrochlorothiazide Tablets add 4V/5 mL of the dis-solving solution, disintegrate by sonicating, add the dissolv-ing solution to make exactly V mL so that each mL contains about 0.25 mg of hydrochlorothiazide (C7H8ClN3O4S2). Cen-trifuge this solution, pipet 5 mL of the supernatant liquid, add the buŠer solution to make exactly 25 mL, and use this solution as the sample solution. Proceed as directed in the Assay (2). Amount (mg) of hydrochlorothiazide (C H ClN O S ) area of telmisartan is not more than 2.0z. (2) Hydrochlorothiazide—When the test is performed at 75 revolutions per minute according to the Paddle method, using 900 mL of 2nd ‰uid for dissolution test as the dissolu-tion medium, the dissolution rate in 45 minutes of Telmisar-tan and Hydrochlorothiazide Tablets is not less than 80z. Start the test with 1 tablet of Telmisartan and Hydrochlo-rothiazide Tablets, withdraw not less than 20 mL of the me-dium at the speciˆed minute after starting the test, and ˆlter ＝ MS × AT/AS × V/50 7 8 3 4 2 through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard not less than 15 mL of the ˆrst ˆltrate, MS: Amount (mg) of Hydrochlorothiazide RS taken Dissolving solution: Dissolve 2 g of ammonium dihydro-gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. BuŠer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Dissolution <6.10> (1) Telmisartan—When the test is per-formed at 50 revolutions per minute according to the Paddle method, using 900 mL of 2nd ‰uid for dissolution test as the dissolution medium, the dissolution rates in 45 minutes of a telmisartan 40-mg and hydrochlorothiazide 12.5-mg tablet and a telmisartan 80-mg and hydrochlorothiazide 12.5-mg tablet are not less than 85z and not less than 80z, respec-tively. Start the test with 1 tablet of Telmisartan and Hydrochlo-rothiazide Tablets, withdraw not less than 20 mL of the me-dium at the speciˆed minute after starting the test, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.45 mm. Discard not less than 15 mL of the ˆrst ˆltrate, pipet V mL of the subsequent ˆltrate, add the dissolution medium to make exactly V? mL so that each mL contains about 44 mg of telmisartan (C33H30N4O2), and use this solu-tion as the sample solution. Separately, weigh accurately about 44 mg of telmisartan for assay, previously dried at 1059C for 4 hours, dissolve in 10 mL of a solution of meglu-mine in methanol (1 in 250), and add methanol to make ex-actly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of telmisar-tan in each solution. Dissolution rate (z) with respect to the labeled amount of telmisartan (C33H30N4O2) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of telmisartan for assay taken C: Labeled amount (mg) of telmisartan (C33H30N4O2) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of telmisartan are not less than 25,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak pipet V mL of the subsequent ˆltrate, add the dissolution medium to make exactly V? mL so that each mL contains about 14 mg of hydrochlorothiazide (C7H8ClN3O4S2), and use this solution as the sample solution. Separately, weigh accurately about 14 mg of Hydrochlorothiazide RS, previ-ously dried at 1059C for 2 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of hydro-chlorothiazide in each solution. Dissolution rate (z) with respect to the labeled amount of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Hydrochlorothiazide RS taken C: Labeled amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay (1). System suitability— System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of hydrochlorothiazide are not less than 1000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 2.0z.
(1) Telmisartan—Weigh accurately the mass of not less than 20 Telmisartan and Hydrochlorothiazide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 80 mg of telmisartan (C33H30N4O2), add 40 mL of the dissolving solution, sonicate, and add the dis-solving solution to make exactly 50 mL. Centrifuge this solu-tion, pipet 5 mL of the supernatant liquid, add the buŠer so-lution to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of telmisartan for assay, previously dried at 1059C for 4 hours, and dissolve in the dissolving solution to make exactly 50 mL. Pipet 5 mL of this solution, add the buŠer solution to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of telmisar-tan in each solution. Amount (mg) of telmisartan (C33H30N4O2) ＝ MS × AT/AS MS: Amount (mg) of telmisartan for assay taken ‌Dissolving solution: Dissolve 2 g of ammonium dihydro-gen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. To 1000 mL of this solution add 1000 mL of acetonitrile. BuŠer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 7.5 cm in length, packed with octylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 3.5 with phosphoric acid. Mobile phase B: Acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 2 2 – 7 90 → 20 10 → 80 7 – 8 Flow rate: 0.8 mL per minute. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of telmisartan are not less than 15,000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of telmisartan is not more than 1.0z. (2) Hydrochlorothiazide—Weigh accurately the mass of not less than 20 Telmisartan and Hydrochlorothiazide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 12.5 mg of hydrochlorothiazide (C7H8ClN3O4S2), add 40 mL of the dissolving solution, sonicate, and add the dissolving solution to make exactly 50 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add the buŠer solution to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 12.5 mg of Hydrochlorothiazide RS, previ-ously dried at 1059C for 2 hours, and dissolve in the dissolv-ing solution to make exactly 50 mL. Pipet 5 mL of this solu-tion, add the buŠer solution to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of hydrochlorothiazide in each solution. Amount (mg) of hydrochlorothiazide (C H ClN O S ) BuŠer solution: Dissolve 2 g of ammonium dihydrogen phosphate in 1000 mL of water, and adjust to pH 1.8 with phosphoric acid. Operating conditions— Proceed as directed in the operating conditions in (1). System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of hydrochlorothiazide are not less than 1500 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z.
Powder Telmisartan Tablets. To a portion of the powder, equivalent to 0.7 mg of Telmisartan, add 100 mL of methanol, shake well, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Determine the absorption spectrum of the filtrate as directed under Ultravi-olet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 226 nm and 230 nm and between 295 nm and 299 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test.
Weigh accurately the mass of not less than 20 Tel-misartan Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 80 mg of telmisartan (C33H30N4O2), add 80 mL of a mixture of water and metha-nol (1:1), shake thoroughly, and add a mixture of water and methanol (1:1) to make exactly 100 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add a mixture of water and metha-nol (1:1) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of telmisartan for assay, previously dried at 1059C for 4 hours, add 10 mL of a solution of meglumine in a mixture of water and methanol (1:1) (1 in 500), dissolve by shaking well, and add a mixture of water and methanol (1:1) to make ex-actly 25 mL. Pipet 5 mL of this solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of telmisartan in each solution. To 1 tablet of Telmisartan Tablets add 4V/5 mL of a mix-ture of water and methanol (1:1), disintegrate the tablet by sonicating, and add a mixture of water and methanol (1:1) to Amount (mg) of telmisartan (C ＝ MS × AT/AS × 4 33H30N4O2) make exactly V mL so that each mL contains about 0.8 mg of telmisartan (C33H30N4O2). Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, dis-card 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of telmisartan (C33H30N4O2) ＝ MS × AT/AS × V/25 MS: Amount (mg) of telmisartan for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Telmisartan Tablets is not less than 85z. Start the test with 1 tablet of Telmisartan Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 5 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 11 mg of telmisartan (C33H30N4O2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of telmisartan for assay, previously dried at 1059C for 4 hours, add 10 mL of a solution of meglumine in methanol (1 in 500), dissolve by sonicating, and add methanol to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 200 mL, and use this solution as the standard solu-tion. Determine the absorbances, AT and AS, at 296 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the dis-solution medium as the control. Dissolution rate (z) with respect to the labeled amount of telmisartan (C33H30N4O2) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of telmisartan for assay taken C: Labeled amount (mg) of telmisartan (C33H30N4O2) in 1 tablet MS: Amount (mg) of telmisartan for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 295 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 2 g of diammonium hydrogen-phosphate in 1000 mL of water, and adjust to pH 3.0 with diluted phosphoric acid (1 in 10). To 300 mL of this solution add 700 mL of methanol. Flow rate: Adjust so that the retention time of telmisartan is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of telmisartan are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of telmisartan is not more than 1.0z.
Temocapril Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled the area of the peak other than temocapril obtained from the sample solution is not larger than 1/5 times the peak area of temocapril from the standard solution, and the total area of amount of temocapril hydrochloride (C HCl: 513.07). 23H28N2O5S2. the peaks other than temocapril from the sample solution is not larger than 1/2 times the peak area of temocapril from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 234 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile (63:37). Flow rate: Adjust so that the retention time of temocapril is about 11 minutes. Time span of measurement: About 4 times as long as the retention time of temocapril, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add diluted acetonitrile (1 in 2) to make exactly 10 mL. Confirm that the peak area of temocapril obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of temocapril are not less than 7000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of temocapril is not more than 2.0z. Water <2.48> Not more than 1.0z (0.3 g, coulometric titration). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.8 g of Temocapril Hydro-chloride, dissolve in 80 mL of a mixture of acetic anhydrate and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 51.31 mg of C23H28N2O5S2.HCl Containers and storage Containers—Well-closed contain-ers. Method of preparation Prepare as directed under Tablets, with Temocapril Hydrochloride. Identification To an amount of powdered Temocapril Hy-drochloride Tablets, equivalent to 2.5 mg of Temocapril Hy-drochloride, add 25 mL of diluted acetonitrile (1 in 2), shake vigorously for 10 minutes, and centrifuge. To 5 mL of the supernatant liquid add diluted acetonitrile (1 in 2) to make 25 mL, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Temocapril Hydrochloride Tablets add ex-actly 20 mL of diluted acetonitrile (1 in 2), and sonicate for 10 minutes. Furthermore, shake for 10 minutes, and centri-fuge. Pipet V mL of the supernatant liquid equivalent to about 0.8 mg of temocapril hydrochloride (C23H28N2O5S 2.HCl), add exactly 2 mL of the internal standard solution, then add diluted acetonitrile (1 in 2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 40 mg of temocapril hydrochloride for assay (separately determine the water <2.48> in the same manner as Temocapril Hydrochloride), dissolve in diluted acetonitrile (1 in 2) to make exactly 200 mL. Pipet 4 mL of this solution, add exactly 2 mL of the internal standard solution, then add diluted acetonitrile (1 in 2) to make 20 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of temocapril to that of the internal standard. Amount (mg) of temocapril hydrochloride (C23H28N2O5S2.HCl) ＝ MS × QT/QS × 1/V × 2/5 MS: Amount (mg) of temocapril hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 3000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, temocapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of temocapril to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu- ‌tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Temocapril Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Temocapril Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.
To an amount of powdered Temocapril Hy-drochloride Tablets, equivalent to 2.5 mg of Temocapril Hy-drochloride, add 25 mL of diluted acetonitrile (1 in 2), shake vigorously for 10 minutes, and centrifuge. To 5 mL of the supernatant liquid add diluted acetonitrile (1 in 2) to make 25 mL, and determine the absorption spectrum of this solu-tion as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 232 nm and 236 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Temocapril Hydrochloride Tablets add ex-actly 20 mL of diluted acetonitrile (1 in 2), and sonicate for 10 minutes. Furthermore, shake for 10 minutes, and centri-fuge. Pipet V mL of the supernatant liquid equivalent to about 0.8 mg of temocapril hydrochloride (C23H28N2O5S 2.HCl), add exactly 2 mL of the internal standard solution, then add diluted acetonitrile (1 in 2) to make 20 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 40 mg of temocapril hydrochloride for assay (separately determine the water <2.48> in the same manner as Temocapril Hydrochloride), dissolve in diluted acetonitrile (1 in 2) to make exactly 200 mL. Pipet 4 mL of this solution, add exactly 2 mL of the internal standard solution, then add diluted acetonitrile (1 in 2) to make 20 mL, and use this solu-tion as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of temocapril to that of the internal standard. Amount (mg) of temocapril hydrochloride (C23H28N2O5S2.HCl) ＝ MS × QT/QS × 1/V × 2/5 MS: Amount (mg) of temocapril hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 3000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, temocapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of temocapril to that of the internal standard is not more than 1.0z. Dissolution <6.10> When the test is performed at 50 revolu- ‌tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Temocapril Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Temocapril Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 1.1 mg of temocapril hydro-chloride (C23H28N2O5S2.HCl), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of temocapril hydrochloride for assay (separately determine the water <2.48> in the same manner as Temocapril Hydro-chloride), and dissolve in diluted acetonitrile (1 in 2) to make exactly 50 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of temocapril in each solution. Dissolution rate (z) with respect to the labeled amount of temocapril hydrochloride (C23H28N2O5S2.HCl) ＝ MS × AT/AS × V?/V × 1/C × 9/2 MS: Amount (mg) of temocapril hydrochloride for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of temocapril hydrochloride (C23H28N2O5S2.HCl) in 1 tablet Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile (43:32). Flow rate: Adjust so that the retention time of temocapril is about 7 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of temocapril are not less than 9000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of temocapril is not more than 2.0z. uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of temocapril to that of the internal standard. Amount (mg) of temocapril hydrochloride (C23H28N2O5S2.HCl) ＝ MS × QT/QS × 1/5 MS: Amount (mg) of temocapril hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of propyl parahy-droxybenzoate in diluted acetonitrile (1 in 2) (1 in 3000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 234 nm). Column: A stainless steel column 6.0 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile (63:37). Flow rate: Adjust so that the retention time of temocapril is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, temocapril and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of temocapril to that of the internal standard is not more than 1.0z.
Weigh accurately about 0.8 g of Temocapril Hydro-chloride, dissolve in 80 mL of a mixture of acetic anhydrate and acetic acid (100) (7:3), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 51.31 mg of C23H28N2O5S2.HCl
(1) Take out the contents of Teprenone Capsules, to a quantity of the content, equivalent to 0.1 g of Teprenone, add 10 mL of ethanol (99.5), shake well, and centrifuge. To 2 mL of the supernatant liquid add 1 mL of a solution of phosphomolybdic acid n-hydrate in acetic acid (100) (1 in 100), heat in a water bath for 5 minutes, add 5-6 drops of sulfuric acid, and continue heating: a blue to bluish green color develops. (2) Take out the contents of Teprenone Capsules, to a quantity of the content, equivalent to 0.1 g of Teprenone, add 10 mL of ethanol (99.5), shake well, and centrifuge. To 2 mL of the supernatant liquid add 2 mL of 2,4-dinitrophenyl hydrazine TS, and shake: a yellow to orange-yellow precipitate is formed. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the following method: it meets the requirement of the Content uniformity test. Take out the contents of 1 capsule of Teprenone Capsules, add exactly 1 mL of the internal standard solution for each 10 mg of teprenone (C23H38O), and add ethyl acetate to make V mL so that each mL contains 1 mg of teprenone (C23H38O). Stand for 30 minutes with shaking occasionally, and filter. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of Teprenone RS, add exactly 5 mL of the internal standard solution, then add ethyl acetate to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay. the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the sum of the peak areas of mono-cis and all-trans isomer of teprenone, AT and AS, in each solution. Dissolution rate (z) with respect to the labeled amount of teprenone (C23H38O) ＝ MS × AT/AS × V?/V × 1/C × 180 MS: Amount (mg) of Teprenone RS taken C: Labeled amount (mg) of teprenone (C23H38O) in 1 cap-sule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of acetonitrile and water (87:13). Flow rate: Adjust so that the retention time of all-trans isomer of teprenone is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the mono-cis and the all-trans isomer of teprenone are eluted in this order with the resolution between these peaks being not less than 1.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the sum of the peak areas of the mono-cis and all-trans isomer of tepre-none is not more than 1.5z.
Take out the contents of not less than 20 Teprenone Capsules. Weigh accurately the total mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of teprenone (C23H38O), add exactly 5 mL of the internal standard solution, add ethyl acetate to make 50 mL. Stand for 30 minutes with shaking Amount (mg) of teprenone (C ＝ MS × QT/QS × V/50 23H 38O) occasionally, and filter. Discard the first 10 mL of the fil-trate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of Teprenone RS, MS: Amount (mg) of Teprenone RS taken Internal standard solution—A solution of di-n-butyl phtha-late in ethyl acetate (1 in 200). add exactly 5 mL of the internal standard solution, add ethyl acetate to make 50 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Teprenone. Dissolution <6.10> When the test is performed at 100 revo- lutions per minute according to the Paddle method using the Amount (mg) of teprenone (C23H38 ＝ M S × QT/QS sinker, using 900 mL of a solution of sodium lauryl sulfate in disodium hydrogen phosphate-citric acid buffer solution (pH 6.8) (1 in 20) as the dissolution medium, the dissolution rate in 60 minutes of Teprenone Capsules is not less than 70z. Start the test with 1 capsule of Teprenone Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of teprenone (C23H38 O), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Teprenone RS, and dissolve in ethanol (99.5) to make exactly 50 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 50 mL, and use this solution as the standard solution. Perform MS: Amount (mg) of Teprenone RS taken Internal standard solution—A solution of di-n-butyl phtha-late in ethyl acetate (1 in 200).
To quantity of Terbinafine Hydrochloride Cream, equivalent to 10 mg of Terbinafine Hydrochloride, dissolve in 20 mL of 2-propanol, and use this solution as the sample solution. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 20 mL of 2-propanol, and use this terbinafine are eluted in this order with the resolution be-tween these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of terbinafine is not more than 1.0z.
Terbinafine Hydrochloride Solution is a liquid for external use. It contains not less than 95.
To a volume of Terbinafine Hydrochloride Solution, equivalent to 10 mg of Terbinafine Hydrochloride, add methanol to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with the upper layer of a mixture of 80 volumes of hexane, 20 volumes of ethyl acetate and 1 volume of ammonia solution (28) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution shows the same Rf value with the spot from the standard solution. pH Being specified separately when the drug is granted ap-proval based on the Law.
Weigh accurately an amount of Terbinafine Hydro-chloride Cream, equivalent to about 10 mg of terbinafine hy-drochloride (C21H25N.HCl), dissolve in 2-propanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of terbinafine hy-drochloride for assay, previously dried at 1059C for 4 hours, and dissolve in 2-propanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of terbinafine in each solution. Amount (mg) of terbinafine hydrochloride (C21H25N.HCl) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of terbinafine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of tetramethylam-monium hydroxide (9 in 2000) adjusted to pH 8.0 with diluted phosphoric acid (1 in 25), acetonitrile and tetra-hydrofuran (2:2:1). Flow rate: Adjust so that the retention time of terbinafine is about 8.5 minutes. System suitability— System performance: Dissolve 40 mg of terbinafine hydro-chloride for assay and 3.5 mg of terphenyl in 200 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, terphenyl and Method of preparation Prepare as directed under Liquids and Solutions for Cutaneous Application, with Terbinafine Hydrochloride. Identification To a volume of Terbinafine Hydrochloride Solution, equivalent to 10 mg of Terbinafine Hydrochloride, add methanol to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with the upper layer of a mixture of 80 volumes of hexane, 20 volumes of ethyl acetate and 1 volume of ammonia solution (28) to a distance of about 15 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the prin-cipal spot obtained from the sample solution shows the same Rf value with the spot from the standard solution. pH Being specified separately when the drug is granted ap-proval based on the Law. Assay Weigh accurately an amount of Terbinafine Hydro-chloride Solution, equivalent to about 10 mg of terbinafine hydrochloride (C21H25N.HCl), add methanol to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of terbinafine hy-drochloride for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of terbinafine in each solution. Amount (mg) of terbinafine hydrochloride (C21H25N.HCl) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of terbinafine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). ‌Column temperature: A constant temperature of about 259C.‌ Mobile phase: A mixture of a solution of tetramethylam-monium hydroxide (9 in 2000) adjusted to pH 8.0 with diluted phosphoric acid (1 in 25), acetonitrile and tetra-hydrofuran (2:2:1). Flow rate: Adjust so that the retention time of terbinafine is about 8.5 minutes. System suitability— System performance: Dissolve 40 mg of terbinafine hydro-chloride for assay and 3.5 mg of terphenyl in 200 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, terphenyl and terbinafine are eluted in this order with the resolution be-tween these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of terbinafine is not more than 1.0z.
Terbinafine Hydrochloride Spray contains not less than 95.
To an amount of Terbinafine Hydrochloride Spray, equivalent to 10 mg of Terbinafine Hydrochloride, add methanol to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with the upper layer of a mixture of 80 volumes of hexane, 20 volumes of ethyl acetate and 1 volume of ammonia solution (28) to a distance of about 15 cm, and air-dry the plate. Examine the plate under ultraviolet light (main wavelength: 254 nm): the principal spot obtained from the sample solution shows the same Rf value with the spot from the standard solution. pH Being specified separately when the drug is granted ap-proval based on the Law.
Weigh accurately an amount of Terbinafine Hydro-chloride Spray, equivalent to about 10 mg of terbinafine hy-drochloride (C21H25N.HCl), dissolve in methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of terbinafine hy-drochloride for assay, previously dried at 1059C for 4 hours, dissolve in methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of terbinafine in each solution. Amount (mg) of terbinafine hydrochloride (C21H25N.HCl) ＝ MS × AT/AS × 1/4 MS: Amount (mg) of terbinafine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of tetramethylam-monium hydroxide (9 in 2000) adjusted to pH 8.0 with diluted phosphoric acid (1 in 25), acetonitrile and tetra-hydrofuran (2:2:1). Flow rate: Adjust so that the retention time of terbinafine is about 8.5 minutes. System suitability— System performance: Dissolve 40 mg of terbinafine hydro-chloride for assay and 3.5 mg of terphenyl in 200 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, terphenyl and terbinafine are eluted in this order with the resolution be-tween these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of terbinafine is not more than 1.0z.
Terbinafine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of terbinafine hydrochloride (C21H25N.HCl: 327.89). Method of preparation Prepare as directed under Tablets, with Terbinafine Hydrochloride. Identification To an amount of powdered Terbinafine Hydrochloride Tablets, equivalent to 10 mg of Terbinafine Hydrochloride, add 10 mL of methanol, shake thoroughly, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with the upper layer of a mixture of 80 volumes of hexane, 20 volumes of ethyl acetate and 1 volume of ammonia solution (28) to a distance of about 15 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ‌To 1 tablet of Terbinafine Hydrochloride Tablets add 40 mL of methanol, shake thoroughly until completely inte-grated, and add methanol to make exactly 50 mL. Centri-fuge this solution, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL so that each mL contains about 0.
To an amount of powdered Terbinafine Hydrochloride Tablets, equivalent to 10 mg of Terbinafine Hydrochloride, add 10 mL of methanol, shake thoroughly, centrifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of terbinafine hydrochloride for assay in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with the upper layer of a mixture of 80 volumes of hexane, 20 volumes of ethyl acetate and 1 volume of ammonia solution (28) to a distance of about 15 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the principal spot ob-tained from the sample solution and the spot from the stand-ard solution show the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. ‌To 1 tablet of Terbinafine Hydrochloride Tablets add 40 mL of methanol, shake thoroughly until completely inte-grated, and add methanol to make exactly 50 mL. Centri-fuge this solution, pipet V mL of the supernatant liquid, add methanol to make exactly V? mL so that each mL contains about 0.28 mg of terbinafine hydrochloride (C21H25N.HCl), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of terbinafine hydrochloride (C21H25N.HCl) ＝ MS × AT/AS × 5 MS: Amount (mg) of terbinafine hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 282 nm). Amount (mg) of terbinafine hydrochloride (C ＝ MS × AT/AS × V?/V × 1/2 21H 25N.HCl) Column: A stainless steel column 4.0 mm in inside diame-ter and 125 mm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame- MS: Amount (mg) of terbinafine hydrochloride for assay taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 30 minutes of Terbinafine Hydrochloride Tablets is not less than 75z. Start the test with 1 tablet of Terbinafine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 0.16 mg of ter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of tetramethylam-monium hydroxide (9 in 2000) adjusted to pH 8.0 with diluted phosphoric acid (1 in 25), acetonitrile and tetra-hydrofuran (2:2:1). Flow rate: Adjust so that the retention time of terbinafine is about 8.5 minutes. System suitability— System performance: Dissolve 40 mg of terbinafine hydro-chloride for assay and 3.5 mg of telphenyl in 200 mL of methanol. When the procedure is run with 10 mL of this so-lution under the above operating conditions, telphenyl and terbinafine are eluted in this order with the resolution be- terbinafine hydrochloride (C 21H 25N.HCl). Pipet 2 mL of this tween these peaks being not less than 6. System repeatability: When the test is repeated 6 times solution, add diluted acetic acid (100) (1 in 100) to make ex-actly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 16 mg of terbinafine hy-drochloride for assay, previously dried at 1059C for 4 hours, dissolve in diluted acetic acid (100) (1 in 100) to make exactly 100 mL. Pipet 5 mL of this solution, add 5 mL of the disso-lution medium, add diluted acetic acid (100) (1 in 100) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 283 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared by adding diluted acetic acid (100) (1 in 100) to 5 mL of the dissolution medium to make 50 mL, as the blank. Dissolution rate (z) with respect to the labeled amount of terbinafine hydrochloride (C21H25N.HCl) ＝ MS × AT/AS × V?/V × 1/C × 900 MS: Amount (mg) of terbinafine hydrochloride for assay taken C: Labeled amount (mg) of terbinafine hydrochloride (C21H25N.HCl) in 1 tablet
Weigh accurately the mass of not less than 20 Ter-binafine Hydrochloride Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.14 g of terbinafine hydrochloride (C21H25N.HCl), add 40 mL of methanol, shake thoroughly, then add methanol to make exactly 50 mL. Centrifuge, pipet 5 mL of the supernatant liquid, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 28 mg of terbinafine hydrochloride for assay, previ-ously dried at 1059C for 4 hours, dissolve in methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of terbinafine in each solution. with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of terbinafine is not more than 1.0z.
テストステロンエナント酸エステル注射液
Testosterone Enanthate Injection is an oily solution for injection. It contains not less than 90.
Testosterone Enanthate Injection is a clear, colorless or pale yellow oily liquid.
Measure a volume of Testosterone Enan-thate Injection, equivalent to 0.05 g of Testosterone Enan-thate, add 8 mL of petroleum ether, and extract with three 10-mL portions of diluted acetic acid (100) (7 in 10). Com-bine the extracts, wash with 10 mL of petroleum ether, add 0.5 mL of diluted sulfuric acid (7 in 10) to 0.1 mL of the ex-tract, and heat on a water bath for 5 minutes. Cool, and add 0.5 mL of iron (III) chloride-acetic acid TS: the color of the solution is blue. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> Perform the test ac-cording to Method 2: it meets the requirement. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure accurately a volume of Testosterone Enan-thate Injection, equivalent to about 25 mg of testosterone enanthate (C26H40O3), and dissolve in chloroform to make exactly 25 mL. Pipet 3 mL of this solution, add chloroform to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Testosterone Propionate RS, proceed in the same manner as for the sample solution, and use this solution as the standard solution. Pipet 5 mL each of the sample solution and stand-ard solution, add exactly 10 mL of isoniazid TS, add metha-nol to make exactly 20 mL, and allow to stand for 45 minutes. Determine the absorbances, AT and AS, of these so-lutions at 380 nm, respectively, as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution obtained by proceeding with 5 mL of chloroform as the blank. Amount (mg) of testosterone enanthate (C26H40O3) ＝ MS × AT/AS × 1.163 MS: Amount (mg) of Testosterone Propionate RS taken
テストステロンプロピオン酸エステル注射液
Testosterone Propionate Injection is an oily solution for injection. It contains not less than 92.
Testosterone Propionate Injection is a clear, colorless or pale yellow oily liquid.
Dissolve the residue obtained as directed in the procedure in the Assay in exactly 20 mL of methanol, and use this solution as the sample solution. Separately, dis-solve 1 mg of Testosterone Propionate RS in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of chloroform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. MS: Amount (mg) of Testosterone Propionate RS taken Internal standard solution—A solution of progesterone in methanol (9 in 100,000).
(i) Chromatographic tube A glass tube about 1 cm in inside diameter and about 18 cm in length, with a glass filter (G3) at the lower end. Chromatographic column To about 2 g of silica gel for liquid chromatography add 5 mL of dichloromethane, and mix gently. Transfer and wash into the chromatographic tube with the aid of dichloromethane, allow to elute the dichloromethane through the column, and put a filter paper on the upper end of the silica gel. Standard solution Weigh accurately about 10 mg of Testosterone Propionate RS, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution and methanol to make 20 mL. Sample stock solution To exactly a volume of Testosterone Propionate Injection, equivalent to about 20 mg of testosterone propionate (C22H32O3), add dichloro-methane to make exactly 20 mL. Procedure Transfer exactly 2 mL of the sample stock solution into the chromatographic column, and elute to the upper surface of the silica gel. Wash the inner surface of the chromatographic tube with 15 mL of dichlorometh-ane, elute to the upper surface of the silica gel, and discard the effluent. Elute 15 mL of a mixture of dichloromethane and methanol (39:1), discard the first 5 mL of the effluent, and collect the subsequent effluent. Wash the lower part of the column with a few amount of dichloromethane, combine the washings and the effluent, and evaporate the solvent under reduced pressure. Dissolve the residue so obtained with methanol to make exactly 20 mL. Pipet 5 mL of this so-lution, add exactly 5 mL of the internal standard solution and methanol to make 20 mL, and use this solution as the sample solution. Perform the test with 5 mL each of the sam-ple solution and standard solution as directed in the Assay under Testosterone Propionate. 107.5z of the labeled amount of testosterone propionate (C22H32O3: 344.49). Amount (mg) of testosterone propionate (C ＝ MS × QT/QS × 2 22H 32O3) Method of preparation Prepare as directed under Injec-tions, with Testosterone Propionate. Description Testosterone Propionate Injection is a clear, colorless or pale yellow oily liquid. Identification Dissolve the residue obtained as directed in the procedure in the Assay in exactly 20 mL of methanol, and use this solution as the sample solution. Separately, dis-solve 1 mg of Testosterone Propionate RS in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Devel-op the plate with a mixture of chloroform and diethylamine (19:1) to a distance of about 15 cm, and air-dry the plate. MS: Amount (mg) of Testosterone Propionate RS taken Internal standard solution—A solution of progesterone in methanol (9 in 100,000).
Thallium (201Tl) Chloride Injection is an aqueous in-jection It contains thallium-201 (201Tl) in the form of thal-lous chloride.
Thallium (201Tl) Chloride Injection is a clear, colorless liquid. Theophylline
(1) To a quantity of powdered Thiamazole Tablets, equivalent to 0.05 g of Thiamazole, add 20 mL of hot ethanol (95), shake for 15 minutes, filter, and evaporate the filtrate on a water bath to dryness. Dissolve the residue in 10 mL of water, filter if necessary, and use this solution as the sample solution. To 1 mL of the sample solution add 1 mL of sodium hydroxide TS, shake, and add 3 drops of so-dium pentacyanonitrosylferrate (III) TS: a yellow color de-velops, and it gradually changes to yellow-green to green. To this solution add 1 mL of acetic acid (31): it changes to blue. (2) With 2 mL of the sample solution obtained in (1), proceed as directed in the Identification (2) under Thiama-zole.
Weigh accurately and powder not less than 20 Thiamazole Tablets. Weigh accurately a quantity of the powder, equivalent to about 0.15 g of thiamazole (C4H6N2S), add 80 mL of water, shake for 15 minutes, add water to make exactly 100 mL, and centrifuge. Filter, dis-card the first 20 mL of the filtrate, pipet 50 mL of the subse-quent filtrate, add 1 mL of bromothymol blue TS, and if a blue color develops, neutralize with 0.1 mol/L hydrochloric acid VS until the color of the solution changes to green. To this solution add 4.5 mL of 0.1 mol/L sodium hydroxide VS from a burette, add 15 mL of 0.1 mol/L silver nitrate VS while stirring, and titrate <2.50> with 0.1 mol/L sodium hy- droxide VS. Continue the titration until a persistent blue-green color is produced, and determine the total volume of 0.1 mol/L sodium hydroxide VS consumed. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 11.42 mg of C4H6N2S
Thiamylal Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 93.
Thiamylal Sodium for Injection occurs as light yellow, crystals, powder or masses. It is hygroscopic. It is gradually decomposed by light.
(1) To 1.0 g of Thiamylal Sodium for In-jection add 20 mL of ethanol (95), shake vigorously, and filter. Dissolve the precipitate so obtained in 1 mL of water, and add 1 mL of barium chloride TS: a white precipitate is produced. Centrifuge this solution, take off the supernatant liquid, and to the precipitate add dilute hydrochloric acid dropwise: the precipitate dissolves with effervescence. (2) To 50 mg of Thiamylal Sodium for Injection add 100 mL of ethanol (95), shake vigorously, and filter. To 3 mL of the filtrate add ethanol (95) to make 200 mL. Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 236 nm and 240 nm, and between 287 nm and 291 nm. pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Thiamylal Sodium for Injection in 40 mL of water is between 10.5 and 11.5.
Related substances—To 0.10 g of Thiamylal So-dium for Injection add 10 mL of ethanol (95), shake vigor-ously, filter, and use the filtrate as the sample solution. Pro-ceed as directed in the Purity (3) under Thiamylal Sodium. Bacterial endotoxins <4.01> Less than 1.0 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require- ‌ment.‌ Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Open carefully 10 containers of Thiamylal Sodium for Injection, dissolve the contents with water, wash out the inside of each container with water, combine them, and add water to make exactly V mL so that each mL contains about 5 mg of thiamylal sodium (C12H17N2NaO2S). Pipet 5 mL of this solution, and add 0.5 mL of dilute hydrochloric acid and methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add exactly 10 mL of the internal standard solution and the mobile phase to make 200 mL, and use this solution as the sample solution. Proceed the test with the sample solu-tion as directed in the Assay under Thiamylal Sodium. Amount (mg) of thiamylal sodium (C12H17N2NaO2S) in 1 container ＝ MS × QT/QS × V/50 × 1.086 MS: Amount (mg) of Thiamylal RS taken Internal standard solution—A solution of phenyl benzoate in methanol (3 in 500).
注射用チオペンタールナトリウム
Thiopental Sodium for Injection is a preparation for injection which is dissolved before use. It contains not less than 93.
Thiopental Sodium for Injection is a light yel-low, powder or mass, and has a slight, characteristic odor. It is very soluble in water, and practically insoluble in de-hydrated diethyl ether. It is hygroscopic.
(1) Dissolve 0.1 g of Thiopental Sodium for Injection in 10 mL of water, and add 0.5 mL of barium chloride TS: a white precipitate is formed. Collect the pre-cipitate, and add dilute hydrochloric acid dropwise: the pre-cipitate dissolves with effervescence. (2) Proceed as directed in the Identification under Thiopental Sodium. pH <2.54> Dissolve 1.0 g of Thiopental Sodium for Injec-tion in 40 mL of water: the pH of this solution is between 10.2 and 11.2.
Proceed as directed in the Purity under Thiopental Sodium. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, 809C, 4 hours). Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Open carefully 10 containers of Thiopental Sodium for Injection, dissolve each content with water, wash each container with water, combine the washings with the former solution, and add water to make exactly 1000 mL. Pipet 10 mL of this solution, and add water to make exactly 100 mL. Measure exactly a volume (V mL) of this solution, equiva-lent to about 15 mg of thiopental sodium (C11H17N2NaO2S), and add water to make exactly 1000 mL. Pipet 10 mL of this solution, add 15 mL of diluted dilute sodium hydroxide TS (1 in 100), add water to make exactly 30 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 46 mg of thiopental for assay, previously dried at 1059 C for 3 hours, dissolve in 50 mL of dilute sodium hydroxide TS, and add water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and determine the absorbances, AT and AS, at 304 nm. ‌Amount (mg) of thiopental sodium (C11H17N2NaO2S) in each sample of Thiopental Sodium for Injection‌ ＝ MS × AT/AS × 300/V × 1.091 MS: Amount (mg) of thiopental sodium for assay taken
Tiapride Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of tiapride (C15H24N2O4S: 328.43). Method of preparation Prepare as directed under Tablets, with Tiapride Hydrochloride. Identification To a quantity of powdered Tiapride Hy-drochloride Tablets, equivalent to 10 mg of tiapride (C15H24N2O4S), add 100 mL of 0.1 mol/L hydrochloric acid TS, shake well, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 286 nm and 290 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Tiapride Hydrochloride Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, sonicate until the tablet is disintegrated, and add 4V/10 mL of methanol. To this solution add exactly V/10 mL of the internal standard solution, shake for 30 minutes, and add methanol to make V A solution of Tiapride Hydrochloride (1 in 20) re- sponds to Qualitative Tests <1.09> for chloride. mL so that each mL contains about 1 mg of tiapride (C15 H24 Purity (1) Heavy metals <1.
To a quantity of powdered Tiapride Hy-drochloride Tablets, equivalent to 10 mg of tiapride (C15H24N2O4S), add 100 mL of 0.1 mol/L hydrochloric acid TS, shake well, and filter. Determine the absorption spec-trum of the filtrate as directed under Ultraviolet-visible Spec-trophotometry <2.24>: it exhibits a maximum between 286 nm and 290 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Tiapride Hydrochloride Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, sonicate until the tablet is disintegrated, and add 4V/10 mL of methanol. To this solution add exactly V/10 mL of the internal standard solution, shake for 30 minutes, and add methanol to make V A solution of Tiapride Hydrochloride (1 in 20) re- sponds to Qualitative Tests <1.09> for chloride. mL so that each mL contains about 1 mg of tiapride (C15 H24
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tiapride Hydrochloride according to Method 1 and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.20 g of Tiapride Hy-drochloride in 10 mL of methanol and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add methanol to make exactly 100 mL. Pipet 1 mL of this solution, add methanol to make exactly 10 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot rapidly 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography under a stream of nitrogen. Develop the plate with a mixture of water, 1-butanol and acetic acid (100) (2:2:1) to a distance of about 10 cm, and air-dry, and then dry the plate at 809C for 30 minutes. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained N2O4S). Centrifuge this solution for 10 minutes, and use the supernatant liquid as the sample solution. Proceed as di-rected in the Assay. Amount (mg) of tiapride (C15H24N2O4S) ＝ MS × QT/QS × V/100 × 0.900 MS: Amount (mg) of tiapride hydrochloride for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 500). Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Tiapride Hydrochloride Tablets and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.1 g of tiapride (C15H24N2O4S), add about 10 mL of 0.1 mol/L hy-drochloric acid TS and 40 mL of methanol, add exactly 10 mL of the internal standard solution, shake for 30 minutes, and add methanol to make 100 mL. Centrifuge this solution ‌and use the supernatant liquid as the sample solution. Sepa-rately, weigh accurately about 0.11 g of tiapride chloride for assay, previously dried at 1059C for 2 hours, dissolve in 10 mL of 0.1 mol/L hydrochloric acid TS, add exactly 10 mL of the internal standard solution, add methanol to make 100 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of Tiapride to that of the internal standard. Amount (mg) of tiapride (C15H24N2O4S) ＝ MS × QT/QS × 0.900 MS: Amount (mg) of tiapride chloride for assay taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 11.2 g of sodium perchlorate in 800 mL of water, add 5 mL of diluted perchloric acid (17 in 2000). To 800 mL of this solution add 200 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of tiapride is about 8 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, tiapride and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of tiapride to that of the internal standard is not more than 1.0z.
Tiaramide Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of tiaramide (C15H18ClN3O3S: 355.84). Method of preparation Prepare as directed under Tablets, with Tiaramide Hydrochloride. Identification (1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 285 nm and 289 nm, and between 292 nm and 296 nm. (2) To a quantity of powdered Tiaramide Hydrochloride Tablets, equivalent to 0.1 g of tiaramide (C15H18ClN3O3S), add 10 mL of diluted ethanol (7 in 10), shake well, filter, and use the filtrate as the sample solution. Separately, dissolve 0.11 g of tiaramide hydrochloride for assay in 10 mL of di-luted ethanol (7 in 10), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop with a mixture of 1-butamol, water and acetic acid (100) (4:2:1) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly Dragendorff's TS for spraying followed by diluted nitric acid (1 in 50) on the plate: the principal spot obtained from the sample solution and the spot from the standard solution are yellow-red in color and have the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Tiaramide Hydrochloride Tablets add 3V/5 mL of 0.1 mol/L hydrochloric acid TS, shake for 60 minutes. Add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of tiaramide (C15H18ClN3O3S), and filter.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 285 nm and 289 nm, and between 292 nm and 296 nm. (2) To a quantity of powdered Tiaramide Hydrochloride Tablets, equivalent to 0.1 g of tiaramide (C15H18ClN3O3S), add 10 mL of diluted ethanol (7 in 10), shake well, filter, and use the filtrate as the sample solution. Separately, dissolve 0.11 g of tiaramide hydrochloride for assay in 10 mL of di-luted ethanol (7 in 10), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography, develop with a mixture of 1-butamol, water and acetic acid (100) (4:2:1) to a distance of about 10 cm, and dry the plate at 1009C for 30 minutes. Spray evenly Dragendorff's TS for spraying followed by diluted nitric acid (1 in 50) on the plate: the principal spot obtained from the sample solution and the spot from the standard solution are yellow-red in color and have the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Tiaramide Hydrochloride Tablets add 3V/5 mL of 0.1 mol/L hydrochloric acid TS, shake for 60 minutes. Add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of tiaramide (C15H18ClN3O3S), and filter. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 55 mg of tiaramide hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 294 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Start the test with 1 tablet of Tiaramide Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 56 mg of tiaramide (C15H18ClN3O3S), and use this solution as the sample solu-tion. Separately, weigh accurately about 15 mg of tiaramide hydrochloride for assay, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 25 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, at 294 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of tiaramide (C15H18ClN3O3S) ＝ MS × AT/AS × V?/V × 1/C × 360 × 0.907 MS: Amount (mg) of tiaramide hydrochloride for assay taken C: Labeled amount (mg) of tiaramide (C15H18ClN3O3S) in 1 tablet
Weigh accurately the mass of more than 20 Tiaramide Hydrochloride Tablets, and powder. Weigh accu-rately an amount of the powder, equivalent to about 0.1 g of tiaramide (C15H18ClN3O3S), add 60 mL of 0.1 mol/L hydro-chloric acid TS, shake for 30 minutes, add 0.1 mol/L hydro-chloric acid TS to make exactly 100 mL, and filter. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, add water to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 0.11 g of tiaramide hydrochloride for assay, previ-ously dried at 1059C for 3 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, of the sample solution and standard so-lution at 294 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of tiaramide (C15H18ClN3O3S) ＝ MS × AT/AS × 0.907 MS: Amount (mg) of tiaramide hydrochloride for assay taken
Ticlopidine Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ticlopidine Hydrochloride according to Method 4, and amount of ticlopidine hydrochloride (C HCl: 300.25). 14H14ClNS. perform the test (not more than 2 ppm). Related substances—Dissolve 0.5 g of Ticlopidine Hydrochloride in 20 mL of a solution of hydrochloric acid in methanol (1 in 20,000), and use this solution as the sample solution. To exactly 5 mL of the sample solution add a solu-tion of hydrochloric acid in methanol (1 in 20,000) to make exactly 200 mL, and use this solution as the standard solu-tion (1). Separately, pipet 1 mL of the sample solution, add a solution of hydrochloric acid in methanol (1 in 20,000) to make exactly 50 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution (1) on a plate of silica gel for thin-layer chromatography (Plate 1), and spot 10 mL each of the sample solution and standard solution (2) on another plate of silica gel for thin-layer chromatogra-phy (Plate 2). Develop the plates with an upper layer of a mixture of water, 1-butanol and acetic acid (100) (5:4:1) to a distance of about 15 cm, and air-dry the plates. Spray evenly a solution of ninhydrin in acetone (1 in 50) on Plate 1, and heat the plate at 1009C for 20 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution (1). Allow Plate 2 to stand in an iodine vapor for 30 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the stand-ard solution (2). Formaldehyde—Dissolve 0.80 g of Ticlopidine Hy- Method of preparation Prepare as directed under Tablets, with Ticlopidine Hydrochloride. Identification Determine the absorption spectrum of the sample solution obtained in the Uniformity of dosage units as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 212 nm and 216 nm, and between 231 nm and 235 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Ticlopidine Hydrochloride Tablets add 70 mL of water, thoroughly shake until the tablet is completely disintegrated, then add water to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make ex-actly V? mL so that each mL contains about 20 mg of ticlopi-dine hydrochloride (C14H14ClNS.
Determine the absorption spectrum of the sample solution obtained in the Uniformity of dosage units as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 212 nm and 216 nm, and between 231 nm and 235 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Ticlopidine Hydrochloride Tablets add 70 mL of water, thoroughly shake until the tablet is completely disintegrated, then add water to make exactly 100 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add water to make ex-actly V? mL so that each mL contains about 20 mg of ticlopi-dine hydrochloride (C14H14ClNS.HCl), and use this solution as the sample solution. Separately, weigh accurately about 25 mg of ticlopidine hydrochloride for assay (separately de-termine the water <2.48> in the same manner as Ticlopidine Hydrochloride), and dissolve in water to make exactly 50 mL. Pipet 2 mL of this solution, add water to make exactly 50 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 233 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. ‌Amount (mg) of ticlopidine hydrochloride (C14H14ClNS.HCl) ＝ MS × AT/AS × V?/V × 2/25 MS: Amount (mg) of ticlopidine hydrochloride for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 35 minutes of Ticlopidine Hydrochloride Tablets is not less than 85z. Start the test with 1 tablet of Ticlopidine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 11 mg of ticlopidine hydro-chloride (C14H14ClNS.HCl), and use this solution as the sam-ple solution. Separately, weigh accurately about 22 mg of ticlopidine hydrochloride for assay (separately determine the water <2.48> in the same manner as Ticlopidine Hydrochlo-ride), and dissolve in water to make exactly 200 mL. Pipet 2 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, at 233 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of ticlopidine hydrochloride (C14H14ClNS.HCl) ＝ MS × AT/AS × V?/V × 1/C × 45 MS: Amount (mg) of ticlopidine hydrochloride for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of ticlopidine hydrochloride (C14H14ClNS.HCl) in 1 tablet
To 20 tablets of Ticlopidine Hydrochloride Tablets, add 400 mL of a mixture of water and methanol (1:1), soni-cate until the tablets are completely disintegrated, and add the mixture of water and methanol (1:1) to make exactly 500 mL. Filter this solution through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, equivalent to about 20 mg of ticlopidine hydrochloride (C14H14 ClNS.HCl), add exactly 4 mL of the internal standard solu-tion, then add the mixture of water and methanol (1:1) to make 100 mL. To 2 mL of this solution add the mixture of water and methanol (1:1) to make 20 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 25 mg of ticlopidine hydrochloride for assay (separate-ly determine the water <2.48> in the same manner as Ticlopi-dine Hydrochloride), dissolve in a suitable amount of a mix-ture of water and methanol (1:1), add exactly 5 mL of the in-ternal standard solution, then add the mixture of water and methanol (1:1) to make 50 mL. Pipet 2 mL of this solution, add a mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of ticlopidine to that of the in-ternal standard. Amount (mg) of ticlopidine hydrochloride (C14H14ClNS.HCl) in 1 tablet ＝ MS × QT/QS × 1/V × 20 MS: Amount (mg) of ticlopidine hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in a mixture of water and methanol (1:1) (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 233 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and 0.05 mol/L phosphate buffer solution (pH 3.5) (7:3). Flow rate: Adjust so that the retention time of ticlopidine is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and ticlopidine are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ticlopidine to that of the internal standard is not more than 1.0z.
(1) To a quantity of powdered Tipepidine Hibenzate Tablets, equivalent to 44 mg of Tipepidine Hiben-zate, add 5 mL of water, shake for 1 minute, add 10 mL of sodium hydroxide TS, and extract with two 20-mL portions of chloroform. Combine the extracts, wash with 10 mL of water, and filter the chloroform layer. Evaporate the filtrate on a water bath to dryness, dissolve the residue in 0.2 mL of 1 mol/L hydrochloric acid TS and 2 mL of water, and add 5 dried in a desiccator (in vacuum, phosphorus (V) oxide, 609 C) for 3 hours, and dissolve in 80 mL of diluted ethanol (3 in 4) by warming occasionally. After cooling, add diluted ethanol (3 in 4) to make exactly 100 mL, then pipet 20 mL of this solution, add water to make exactly 900 mL, and use this solution as the standard solution. Determine the absor-bances, AT1 and AS1, at 286 nm, and AT2 and AS2, at 360 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of tipepidine hibenzate (C15H17NS2.C14H10O4) ＝ MS × (AT1 － AT2/AS1 － AS2) × 1/C × 20 MS: Amount (mg) of tipepidine hibenzate for assay taken C: Labeled amount (mg) of tipepidine hibenzate (C15H17NS2.C14H10O4) in 1 tablet
(1) Lead—Place 1.0 g of Titanium Oxide in a platinum crucible, add 10.0 g of potassium hydrogen sulfate, heat gently with caution at the beginning, then raise the tem-perature gradually, and heat strongly with occasional shak-ing until the contents fuse to yield a clear liquid. Cool, add Amount (mg) of tipepidine hibenzate (C H NS .C H O ) 30 mL of a solution of diammonium hydrogen citrate (9 in ＝ MS × QT/QS 15 17 2 14 10 4 20) and 50 mL of water, dissolve by heating on a water bath, cool, add water to make 100 mL, and use this solution as the MS: Amount (mg) of tipepidine hibenzate for assay taken Internal standard solution—A solution of dibucaine hydro-chloride in methanol (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of sodium lauryl sulfate in diluted phosphoric acid (1 in 1000) (1 in 500), acetonitrile and 2-propanol (3:2:1). Flow rate: Adjust the flow rate so that the retention time of tipepidine is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, tipepidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of tipepidine to that of the internal standard is not more than 1.0z.
Weigh accurately and powder not less than 20 Tipepidine Hibenzate Tablets. Weigh accurately a portion of the powder, equivalent to about 22 mg of tipepidine hiben-zate (C15H17NS2.C14H10O4), add 10 mL of diluted acetic acid (100) (1 in 2) and 30 mL of methanol, and warm for 10 minutes with occasional shaking. After cooling, add diluted methanol (1 in 2) to make exactly 50 mL, and filter. Discard the first 10 mL of the filtrate, pipet 5 mL of the subsequent ‌filtrate, add exactly 5 mL of the internal standard solution, then add diluted methanol (1 in 2) to make 25 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 22 mg of tipepidine hibenzate for assay, previ-ously dried in a desiccator (in vacuum, phosphorus (V) oxide, 609C) for 3 hours, dissolve in 10 mL of diluted acetic acid (100) (1 in 2) and 30 mL of methanol, and add diluted methanol (1 in 2) to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, then add diluted methanol (1 in 2) to make exactly 25 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of tipepidine to that of the inter-nal standard, respectively. When fused by heating with potassium hydrogen sulfate, with potassium hydroxide, or with potassium carbonate, it changes to soluble salts. Shake 1 g of Titanium Oxide with 10 mL of water: the mixture is neutral. Identification Heat 0.5 g of Titanium Oxide with 5 mL of sulfuric acid until white fumes are evolved, cool, add cau-tiously water to make 100 mL, and filter. To 5 mL of the fil-trate add 2 to 3 drops of hydrogen peroxide TS: a yellow-red color develops. Purity (1) Lead—Place 1.0 g of Titanium Oxide in a platinum crucible, add 10.0 g of potassium hydrogen sulfate, heat gently with caution at the beginning, then raise the tem-perature gradually, and heat strongly with occasional shak-ing until the contents fuse to yield a clear liquid. Cool, add Amount (mg) of tipepidine hibenzate (C H NS .C H O ) 30 mL of a solution of diammonium hydrogen citrate (9 in ＝ MS × QT/QS 15 17 2 14 10 4 20) and 50 mL of water, dissolve by heating on a water bath, cool, add water to make 100 mL, and use this solution as the MS: Amount (mg) of tipepidine hibenzate for assay taken Internal standard solution—A solution of dibucaine hydro-chloride in methanol (1 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of a solution of sodium lauryl sulfate in diluted phosphoric acid (1 in 1000) (1 in 500), acetonitrile and 2-propanol (3:2:1). Flow rate: Adjust the flow rate so that the retention time of tipepidine is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, tipepidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of tipepidine to that of the internal standard is not more than 1.0z.
Tobramycin Injection is an aqueous injection. It contains not less than 90.
Tobramycin Injection occurs as a colorless or very pale yellow, clear liquid.
To a volume of Tobramycin Injection, equivalent to 10 mg (potency) of Tobramycin, add water to make 1 mL, and use this solution as the sample solution. Separately, dissolve 10 mg (potency) of Tobramycin RS in 1 mL of water, and use this solution as the standard solution. Then, proceed as directed in the Identification (2) under Tobramycin. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH <2.54> 5.0 – 7.0 high and the low concentration sample solution, respec-tively.
Tolnaftate Solution contains not less than 90.
(1) Spot 1 drop of Tolnaftate Solution on filter paper. Spray hydrogen hexachloroplatinate (IV)-potas-sium iodide TS on the paper: a light yellow color develops in the spot. (2) To a volume of Tolnaftate Solution, equivalent to 0.02 g of Tolnaftate, add chloroform to make 10 mL, and use this solution as the sample solution. Separately, dissolve 0.02 g of Tolnaftate RS in 10 mL of chloroform, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with toluene to a distance of about 12 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot obtained from the sample solution and that from the stand-ard solution show the same Rf value.
Pipet a volume of Tolnaftate Solution, equivalent to about 20 mg of tolnaftate (C19H17NOS), add exactly 4 mL of the internal standard solution, then add chloroform to make 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 0.4 g of Tolnaftate RS, previ-ously dried in vacuum at a pressure not exceeding 0.67 kPa at 659C for 3 hours, and dissolve in chloroform to make ex-actly 100 mL. Pipet 5 mL of this solution, add exactly 4 mL of the internal standard solution, then add chloroform to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tolnaftate to that of the internal standard. Amount (mg) of tolnaftate (C19H17NOS) ＝ MS × QT/QS × 1/20 MS: Amount (mg) of Tolnaftate RS taken Internal standard solution—A solution of diphenyl phthalate Amount (mg) of tolnaftate (C 19H 17NOS) ＝ MS × AT/AS in chloroform (3 in 200). Operating conditions— MS: Amount (mg) of Tolnaftate RS taken
Tosufloxacin Tosilate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O: 594.56). Method of preparation Prepare as directed under Tablets, with Tosufloxacin Tosilate Hydrate. Identification To a quantity of powdered Tosufloxacin Tosilate Tablets, equivalent to 75 mg of Tosufloxacin Tosi-late Hydrate, add 200 mL of a mixture of methanol and so-dium hydroxide TS (49:1), shake well, and centrifuge. To 2 mL of the supernatant liquid add 100 mL of a mixture of methanol and sodium hydroxide TS (49:1). Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 260 nm and 264 nm, between 341 nm and 345 nm, and between 356 nm and 360 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Tosufloxacin Tosilate Tablets add V/10 mL of water and shake until the tablet is disintegrated. Add methanol to make exactly V mL so that each mL contains about 1.
To a quantity of powdered Tosufloxacin Tosilate Tablets, equivalent to 75 mg of Tosufloxacin Tosi-late Hydrate, add 200 mL of a mixture of methanol and so-dium hydroxide TS (49:1), shake well, and centrifuge. To 2 mL of the supernatant liquid add 100 mL of a mixture of methanol and sodium hydroxide TS (49:1). Determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 260 nm and 264 nm, between 341 nm and 345 nm, and between 356 nm and 360 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Tosufloxacin Tosilate Tablets add V/10 mL of water and shake until the tablet is disintegrated. Add methanol to make exactly V mL so that each mL contains about 1.5 mg of tosufloxacin tosilate hydrate (C19H15F3N4O3. C7H8O3S.H2O). Shake this solution for 10 minutes, and cen-trifuge. Pipet 4 mL of the supernatant liquid, add exactly 4 mL of the internal standard solution and methanol to make 100 mL, and use this solution as the sample solution. Pro-ceed as directed in the Assay. Amount (mg) of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O) ＝ MS × QT/QS × V/20 × 1.031 MS: Amount (mg) of Tosufloxacin Tosilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 800). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 90 minutes of Tosufloxacin Tosilate Tablets is not less than 65z. Start the test with 1 tablet of Tosufloxacin Tosilate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Dis-card not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) to make exactly V? mL so that each mL contains about 17 mg of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O), and use this solution as the sample solution. Separately, weigh accurately about 21 mg of Tosufloxacin Tosilate RS (separately determine the water <2.48> in the same manner as Tosufloxacin Tosilate Hydrate), and dissolve in N,N-dimethylformamide to make exactly 25 mL. Pipet 2 mL of this solution, add 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the blank, and determine the ab-sorbances, AT and AS, at 346 nm. Dissolution rate (z) with respect to the labeled amount of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O) ＝ MS × AT/AS × V?/V × 1/C × 72 × 1.031 MS: Amount (mg) of Tosufloxacin Tosilate RS taken, cal-culated on the anhydrous basis C: Labeled amount (mg) of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O) in 1 tablet
Weigh accurately the mass of not less than 20 Tosufloxacin Tosilate Tablets, and powder. Weigh accu-rately a portion of the powder, equivalent to about 0.15 g of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O), add 10 mL of water and methanol to make exactly 100 mL, shake for 10 minutes, and centrifuge. Pipet 4 mL of the su-pernatant liquid, add exactly 4 mL of the internal standard solution and methanol to make 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 30 mg of Tosufloxacin Tosilate RS (separately deter-mine the water <2.48> in the same manner as Tosufloxacin Tosilate Hydrate), add 2 mL of water, and dissolve in meth-anol to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 4 mL of the internal standard solution and meth-anol to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample so-lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tosufloxacin to that of the internal standard. Amount (mg) of tosufloxacin tosilate hydrate (C19H15F3N4O3.C7H8O3S.H2O) ＝ MS × QT/QS × 5 × 1.031 MS: Amount (mg) of Tosufloxacin Tosilate RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (1 in 800). Operating conditions— Proceed as directed in the operating conditions in the Assay under Tosufloxacin Tosilate Hydrate. System suitability— Proceed as directed in the system suitability in the Assay under Tosufloxacin Tosilate Hydrate.
Tranexamic Acid Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of tranexamic acid (C8H15NO2: 157.21). Method of preparation Prepare as directed under Cap-sules, with Tranexamic Acid. Identification Take an amount of powdered contents of Tranexamic Acid Capsules, equivalent to 0.5 g of Tranexamic Acid, add 50 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and heat for 3 minutes: a dark purple color develops. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the MS: Amount (mg) of Tranexamic Acid RS taken C: Labeled amount (mg) of tranexamic acid (C8H15NO2) in 1 capsule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 11.0 g of anhydrous sodium dihy-drogen phosphate in 500 mL of water, and add 10 mL of triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid, add water to make 600 mL, and add 400 mL of methanol. Flow rate: Adjust so that the retention time of tranexamic acid is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of tranexamic acid are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 2.0z. Assay Weigh accurately the mass of the contents of not less than 20 Tranexamic Acid Capsules, and powder. Weigh accurately an amount of the powder, equivalent to about 0.1 g of tranexamic acid (C8H15NO2), add 30 mL of water, shake well, and add water to make exactly 50 mL. Centri-fuge, filter the supernatant liquid through a membrane filter with pore size of not more than 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Separately, weigh accurately about 50 mg of Tranexamic Acid RS, previously dried at 1059C for 2 hours, dissolve in water to make exactly 25 mL, and use this solu-tion as the standard solution. Perform the test with exactly 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of tranexamic acid in each solution. dissolution rate in 15 minutes of Tranexamic Acid Capsules is not less than 80z. Start the test with 1 tablet of Tranexamic Acid Capsules, Amount (mg) of tranexamic acid (C8H ＝ MS × AT/AS × 2 15NO2) withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.
Take an amount of powdered contents of Tranexamic Acid Capsules, equivalent to 0.5 g of Tranexamic Acid, add 50 mL of water, shake well, and filter. To 5 mL of the filtrate add 1 mL of ninhydrin TS, and heat for 3 minutes: a dark purple color develops. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the MS: Amount (mg) of Tranexamic Acid RS taken C: Labeled amount (mg) of tranexamic acid (C8H15NO2) in 1 capsule Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 11.0 g of anhydrous sodium dihy-drogen phosphate in 500 mL of water, and add 10 mL of triethylamine and 1.4 g of sodium lauryl sulfate. Adjust the pH to 2.5 with phosphoric acid, add water to make 600 mL, and add 400 mL of methanol. Flow rate: Adjust so that the retention time of tranexamic acid is about 8 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of tranexamic acid are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 2.0z.
Weigh accurately the mass of the contents of not less than 20 Tranexamic Acid Capsules, and powder. Weigh accurately an amount of the powder, equivalent to about 0.1 g of tranexamic acid (C8H15NO2), add 30 mL of water, shake well, and add water to make exactly 50 mL. Centri-fuge, filter the supernatant liquid through a membrane filter with pore size of not more than 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sam-ple solution. Separately, weigh accurately about 50 mg of Tranexamic Acid RS, previously dried at 1059C for 2 hours, dissolve in water to make exactly 25 mL, and use this solu-tion as the standard solution. Perform the test with exactly 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of tranexamic acid in each solution. dissolution rate in 15 minutes of Tranexamic Acid Capsules is not less than 80z. Start the test with 1 tablet of Tranexamic Acid Capsules, Amount (mg) of tranexamic acid (C8H ＝ MS × AT/AS × 2 15NO2) withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.28 mg of tranexamic acid (C8H15NO2), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of Tranexamic Acid RS, previously dried at 1059C for 2 hours, dissolve in water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of tranexamic acid in each solution. Dissolution rate (z) with respect to the labeled amount of tranexamic acid (C8H15NO2) ＝ MS × AT/AS × V?/V × 1/C × 900 MS: Amount (mg) of Tranexamic Acid RS taken Operating conditions— Detector, column, and mobile phase: Proceed as directed in the operating conditions in the Assay under Tranexamic Acid. Column temperature: A constant temperature of about 359C. Flow rate: Adjust so that the retention time of tranexamic acid is about 16 minutes. System suitability— System performance: To 5 mL of the standard solution add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in 10,000) and water to make 50 mL. When the procedure is run with 30 mL of this solution under the above operating conditions, tranexamic acid and 4-(aminomethyl)benzoic acid are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times ‌with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 1.0z.‌
Tranexamic Acid Injection is an aqueous injection. It contains not less than 95.
Tranexamic Acid Injection is a clear and color-less liquid.
To a volume of Tranexamic Acid Injection, equivalent to 50 mg of Tranexamic Acid, add water to make 5 mL, add 1 mL of ninhydrin TS, and heat: a dark purple color develops. pH <2.54> 7.0 – 8.0 Bacterial endotoxins <4.01> Not more than 0.12 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. acid are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 1.0z.
To an amount of powdered Tranexamic Acid Tablets, equivalent to 0.5 g of Tranexamic Acid, add 50 mL of water, shake well, and filter. To 5 mL of the fil-trate add 1 mL of ninhydrin TS, and heat for 3 minutes: a dark purple color develops. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution Being specified separately when the drug is granted approval based on the Law.
Weigh accurately the mass of not less than 20 Tranexamic Acid Tablets, and powder. Weigh accurately a quantity of the powder, equivalent to about 5 g of tranexam-ic acid (C8H15NO2), add 150 mL of water, disintegrate the Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. Assay Pipet a volume of Tranexamic Acid Injection, equivalent to about 0.1 g of tranexamic acid (C8H15NO2), add water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Tranexamic Acid RS, previously dried at 1059C for 2 hours, dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with ex-actly 30 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of tranexamic acid in each solution. Amount (mg) of tranexamic acid (C8H15NO2) ＝ MS × AT/AS × 2 MS: Amount (mg) of Tranexamic Acid RS taken Operating conditions— Detector, column, and mobile phase: Proceed as directed in the operating conditions in the Assay under Tranexamic Acid. Column temperature: A constant temperature of about 359C. Flow rate: Adjust so that the retention time of tranexamic acid is about 16 minutes. System suitability— System performance: To 5 mL of the standard solution add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in 10,000) and water to make 50 mL. When the procedure is run with 30 mL of this solution under the above operating conditions, tranexamic acid and 4-(aminomethyl)benzoic tablets completely by sonicating, and add water to make ex-actly 200 mL. Centrifuge, pipet 4 mL of the supernatant liq-uid, and add water to make exactly 50 mL. Filter through a membrane filter with a pore size of not more than 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 50 mg of Tranexamic Acid RS, previously dried at 105 9C for 2 hours, dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 30 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of tranexamic acid in each solution. Amount (mg) of tranexamic acid (C8H15NO2) ＝ MS × AT/AS × 100 MS: Amount (mg) of Tranexamic Acid RS taken Operating conditions— Detector, column, and mobile phase: Proceed as directed in the operating conditions in the Assay under Tranexamic Acid. Column temperature: A constant temperature of about 359C. Flow rate: Adjust so that the retention time of tranexamic acid is about 16 minutes. System suitability— System performance: To 5 mL of the standard solution add 1 mL of a solution of 4-(aminomethyl)benzoic acid (1 in 10,000) and water to make 50 mL. When the procedure is run with 30 mL of this solution under the above operating conditions, tranexamic acid and 4-(aminomethyl)benzoic acid are eluted in this order with the resolution between these ‌peaks being not less than 3. System repeatability: When the test is repeated 6 times with 30 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of tranexamic acid is not more than 1.0z.
Tranilast Capsules contain not less than 95.0z and not more than 105.0z of the labeled amount of tranilast (C18H17NO5: 327.33). Method of preparation Prepare as directed under Cap-sules, with Tranilast. Identification To an amount of the content of Tranilast Capsules, equivalent to 0.1 g of Tranilast, add 180 mL of diethyl ether, shake thoroughly, filter, and evaporate the fil-trate to dryness on a water bath. Determine the absorption spectrum of a solution of the residue in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 333 nm and 337 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. Shake the contents and the empty capsule shell of 1 Tranilast Capsules with a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly V mL so that each mL contains about 0.
To an amount of the content of Tranilast Capsules, equivalent to 0.1 g of Tranilast, add 180 mL of diethyl ether, shake thoroughly, filter, and evaporate the fil-trate to dryness on a water bath. Determine the absorption spectrum of a solution of the residue in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>: it exhibits a maximum between 333 nm and 337 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Conduct this procedure using light-resistant vessels. Shake the contents and the empty capsule shell of 1 Tranilast Capsules with a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly V mL so that each mL contains about 0.5 mg of tranilast (C18H17NO5), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of tranilast (C18H17NO5) ＝ MS × QT/QS × V/50 MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of disodium hydrogen phosphate-citric acid buffer solution (pH 5.5) as the dissolution medium, the dissolution rate in 60 minutes of Tranilast Capsules is not less than 75z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with 1 capsule of Tranilast Capsules, withdraw not less than 20 mL of the me-dium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 2nd fluid for dis-solution test to make exactly V? mL so that each mL con-tains about 5.6 mg of tranilast (C18H17NO5), and use this so-lution as the sample solution. Separately, weigh accurately about 28 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in 2nd fluid for dissolution test to make exactly 100 mL. Pipet 5 mL of this solution, and add 2nd fluid for dissolution test to make exactly 50 mL. Then, pipet 5 mL of this solution, add 2nd fluid for dissolution test to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 332 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of tranilast (C18H17NO5) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of tranilast for assay taken C: Labeled amount (mg) of tranilast (C18H17NO5) in 1 capsule
Conduct this procedure using light-resistant vessels. Weigh accurately the mass of the contents of not less than 20 Tranilast Capsules, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of tranilast (C18H17NO5), shake with a suitable amount of a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and aceto-nitrile (7:3), then add the same mixture to make exactly 200 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and aceto-nitrile (7:3) to make 50 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in a mixture of 0.05 mol/L phosphate buffer so-lution (pH 7.0) and acetonitrile (7:3) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the inter-nal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tranilast to that of the internal standard. Amount (mg) of tranilast (C18H17NO5) ＝ MS × QT/QS × 4 MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). ‌Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of tranilast is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and tranilast are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of tranilast to that of the internal standard is not more than 1.0z.
Tranilast Fine Granules contain not less than 95.0z and not more than 105.0z of the labeled amount of tranilast (C18H17NO5: 327.33). Method of preparation Prepare as directed under Gran-ules, with Tranilast. Identification To an amount of Tranilast Fine Granules, equivalent to 0.1 g of Tranilast, add 180 mL of diethyl ether, shake thoroughly, filter, and evaporate the filtrate to dryness on a water bath. Determine the absorption spectrum of a so-lution of the residue in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 333 nm and 337 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Tranilast Fine Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. Shake the total content of 1 package of Tranilast Fine Granules with a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly V mL so that each mL contains about 0.
To an amount of Tranilast Fine Granules, equivalent to 0.1 g of Tranilast, add 180 mL of diethyl ether, shake thoroughly, filter, and evaporate the filtrate to dryness on a water bath. Determine the absorption spectrum of a so-lution of the residue in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 333 nm and 337 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Tranilast Fine Granules in single-dose packages meet the requirement of the Content uniformity test. Conduct this procedure using light-resistant vessels. Shake the total content of 1 package of Tranilast Fine Granules with a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly V mL so that each mL contains about 0.5 mg of tranilast (C18H17NO5), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make 50 mL, and use this solution as the sample so-lution. Then, proceed as directed in the Assay. Amount (mg) of tranilast (C18H17NO5) ＝ MS × QT/QS × V/50 MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citric acid buffer solu-tion (pH 5.5) as the dissolution medium, the dissolution rate in 30 minutes of Tranilast Fine Granules is not less than 75z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with an accurately weighed amount of Tranilast Fine Granules, equivalent to about 0.1 g of tranilast (C18H17NO5), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add 2nd fluid for dissolution test to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 28 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in 2nd fluid for dissolution test to make exactly 100 mL. Pipet 5 mL of this solution, and add 2nd fluid for dissolution test to make exactly 50 mL. Then, pipet 5 mL of this solution, add 2nd fluid for dis-solution test to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances, AT and A S, at 332 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of tranilast (C18H17NO5) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount (mg) of tranilast for assay taken MT: Amount (g) of Tranilast Fine Granules taken C: Labeled amount (mg) of tranilast (C18H17NO5) in 1 g
Conduct this procedure using light-resistant vessels. Powder Tranilast Fine Granules. Weigh accurately a portion of the powder, equivalent to about 0.1 g of tranilast (C18H17NO5), shake with a suitable amount of a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and aceto-nitrile (7:3), then add the same mixture to make exactly 200 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the fil-trate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and aceto-nitrile (7:3) to make 50 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in a mixture of 0.05 mol/L phosphate buffer so-lution (pH 7.0) and acetonitrile (7:3) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the inter-nal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tranilast to that of the internal standard. Amount (mg) of tranilast (C18H17NO5) ＝ MS × QT/QS × 4 MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌length: 255 nm).‌ Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of tranilast is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL ethanol (99.5) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in ethanol (99.5) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, then add ethanol (99.5) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of tranilast to that of the inter-nal standard. of the standard solution under the above operating condi-tions, the internal standard and tranilast are eluted in this order with the resolution between these peaks being not less Amount (mg) of tranilast (C ＝ MS × QT/QS × 1/5 18H17NO5) than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of tranilast to that of the internal standard is not more than 1.0z.
Tranilast for Syrup is a preparation for syrup, which is suspended before use. It contains not less than 95.
To an amount of Tranilast for Syrup, equivalent to 0.1 g of Tranilast, add 180 mL of diethyl ether, shake thoroughly, filter, and evaporate the filtrate to dryness on a water bath. Determine the absorption spectrum of a so-lution of the residue in methanol (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 333 nm and 337 nm. Uniformity of dosage units <6.02> Perform the test accord- ‌ing to the following method: Tranilast for Syrup in single-dose packages meet the requirement of the Content unifor-mity test. Conduct this procedure using light-resistant vessels. Shake the total content of 1 package of Tranilast for Syrup with a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3), then add the same mixture to make exactly V mL so that each mL contains about 0.5 mg of tranilast (C18H17NO5), and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet 10 mL of the subsequent filtrate, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make 50 mL, and use this solu- nitrile (7:3) to make 50 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 25 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly 50 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, then add a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of tranilast to that of the internal standard. tion as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of tranilast (C ＝ MS × QT/QS × 4 18H17NO5) Amount (mg) of tranilast (C18H17NO5) ＝ MS × QT/QS × V/50 MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of disodium hydrogen phosphate-citric acid buffer solu-tion (pH 5.5) as the dissolution medium, the dissolution rate in 60 minutes of Tranilast for Syrup is not less than 75z. Conduct this procedure without exposure to light, using light-resistant vessels. Start the test with an accurately weighed amount of Tranilast for Syrup, equivalent to about 0.1 g of tranilast (C18H17NO5), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet 5 mL of the subsequent filtrate, add 2nd fluid for dissolution test to make exactly 100 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 28 mg of tranilast for assay, previously dried at 1059C for 3 hours, and dissolve in 2nd fluid for dissolution test to make exactly 100 mL. Pipet 5 mL of this solution, and add 2nd fluid for dissolution test to make exactly 50 mL. Then, pipet 5 mL of this solution, add 2nd fluid for dissolution test to make exactly 25 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 332 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of tranilast (C18H17NO5) ＝ MS/MT × AT/AS × 1/C × 360 MS: Amount (mg) of tranilast for assay taken MT: Amount (g) of Tranilast for Syrup taken C: Labeled amount (mg) of tranilast (C18H17NO5) in 1 g
Conduct this procedure using light-resistant vessels. Powder Tranilast for Syrup. Weigh accurately the a portion of the powder, equivalent to about 0.1 g of tranilast (C18H17NO5), shake with a suitable amount of a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and aceto-nitrile (7:3), then add the same mixture to make exactly 200 mL, and filter through a membrane filter with a pore size MS: Amount (mg) of tranilast for assay taken Internal standard solution—Ethyl parahydroxybenzoate in a mixture of 0.05 mol/L phosphate buffer solution (pH 7.0) and acetonitrile (7:3) (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of tranilast is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and tranilast are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of tranilast to that of the internal standard is not more than 1.0z.
Tranilast Ophthalmic Solution is an aqueous oph-thalmic preparation. It contains not less than 95.
Tranilast Ophthalmic Solution occurs as a clear and pale yellow liquid.
When add 2 mL of dilute hydrochloric acid to a volume of Tranilast Ophthalmic Solution, equivalent to about 50 mg of Tranilast, a white precipitate is produced. Collect the precipitate by filtration, wash the precipitate with two 10-mL portions of water, and dry at 1059C for 3 hours. Dissolve 5 mg of the precipitate in methanol to make 100 mL. To 5 mL of this solution add methanol to make 50 mL, and determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 333 nm and 337 nm. Osmotic pressure ratio Being specified separately when the drug is granted approval based on the Law. pH Being specified separately when the drug is granted ap-proval based on the Law. Foreign insoluble matter <6.11> It meets the requirement. Insoluble particulate matter <6.08> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Conduct this procedure using light-resistant vessels. To exactly a volume of Tranilast Ophthalmic Solution, equivalent to about 5 mg of tranilast (C18H17NO5), add exactly 10 mL of the internal standard solution, then add MS: Amount (mg) of tranilast for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in ethanol (99.5) (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 255 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (3:2). Flow rate: Adjust so that the retention time of tranilast is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and tranilast are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of tranilast to that of the internal standard is not more than 1.0z.
Trichlormethiazide Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of trichlormethiazide (C8H8Cl3N3O4S2: 380.66). Method of preparation Prepare as directed under Tablets, with Trichlormethiazide. 0 – 10 100 0 10 – 20 100 → 0 0 → 100 Flow rate: 1.5 mL per minute. Time span of measurement: About 2.5 times as long as the retention time of trichlormethiazide, beginning after the sol-vent peak. System suitability— Test for required detectability: Dissolve 25 mg of Trichlor-methiazide in 50 mL of acetonitrile. To 1 mL of this solution add acetonitrile to make 50 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solu-tion for system suitability test, and add acetonitrile to make exactly 20 mL. Confirm that the peak area of trichlor-methiazide obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the solution for system suitability test. System performance: To 5 mL of the solution for system suitability test add 5 mL of water, and warm in a water bath of 609C for 30 minutes. When the procedure is run with 10 mL of this solution, after cooling, under the above operating conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide and trichlormethiazide are eluted in this order, the relative reten-tion time of 4-amino-6-chlorobenzene-1,3-disulfonamide to trichlormethiazide is about 0.3, and the number of theoreti-cal plates and the symmetry factor of the peak of trichlor-methiazide are not less than 5000 and not more than 1.2, respectively. System repeatability: When the test is repeated 3 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of trichlormethiazide is not more than 2.0z. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Trichlormethiazide Tablets add V/5 mL of diluted phosphoric acid (1 in 50), and disintegrate the tablet. Add 2V/5 mL of acetonitrile, shake vigorously for 15 minutes, add the mobile phase to make exactly V mL so that each mL contains about 40 mg of trichlormethiazide (C8H8Cl3N3O4S2).
To 10 Trichlormethiazide Tablets add V/10 mL of diluted phosphoric acid (1 in 50), and disintegrate the tablets. Add V/2 mL of acetonitrile, shake vigorously for 15 minutes, add the mobile phase to make exactly V mL so that each mL contains about 0.2 mg of trichlormethiazide (C8H8Cl3N3O4S2), and centrifuge. Pipet 5 mL of the superna-tant liquid, add the mobile phase to make exactly 25 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 4 mL of the filtrate, and ＝ MS × AT/AS × V/500 8 8 3 3 4 2 use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 20 mg of Trichlormethiazide MS: Amount (mg) of Trichlormethiazide RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Trichlormethiazide Tablets is not less than 75z. Start the test with 1 tablet of Trichlormethiazide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add diluted phosphoric acid (1 in 50) to make exactly V? mL so that each mL contains about 1.1 mg of trichlor-methiazide (C8H8Cl3N3O4S2), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Trichlormethiazide RS, previously dried at 1059C for 3 hours, and dissolve in acetonitrile to make exactly 200 mL. Pipet 2 mL of this solution, add diluted phosphoric acid (1 in 50) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 40 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, ATa and ASa, of trichlormethiazide obtained with the sample solution and standard solution, and the area, ATb, of the peak, hav-ing the relative retention time of about 0.3 to trichlor-methiazide, obtained with the sample solution. Dissolution rate (z) with respect to the labeled amount of trichlormethiazide (C8H8Cl3N3O4S2) ＝ MS × (ATa ＋ 0.95ATb)/ASa × V?/V × 1/C × 9/2 MS: Amount (mg) of Trichlormethiazide RS taken C: Labeled amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2) in 1 tablet RS, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of trichlormethiazide in each solu-tion. Amount (mg) of trichlormethiazide (C8H8Cl3N3O4S2) in 1 tablet ＝ MS × AT/AS × V/1000 MS: Amount (mg) of Trichlormethiazide RS taken Operating conditions— Proceed as directed in the operating conditions in the Assay under Trichlormethiazide. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of trichlormethiazide are not less than 5000 and not more than 1.2, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of trichlormethiazide is not more than 1.0z.
トリクロホスナトリウムシロップ
(1) Weigh a portion of Triclofos Sodium Syrup, equivalent to 0.25 g of Triclofos Sodium, add 40 mL of water, shake well, add 5 mL of diluted sulfuric acid (3 in 50), and extract with 25 mL of 3-methyl-1-butanol. Take 5 mL of the extract, evaporate on a water bath to dryness, and add 1 mL of diluted sulfuric acid (1 in 2) and 1 mL of a solu-tion of potassium permanganate (1 in 20) to the residue. Heat in a water bath for 5 minutes, add 7 mL of water, and then add a solution of oxalic acid dihydrate (1 in 20) until the color of the solution disappears. To 1 mL of this solution add 1 mL of pyridine and 1 mL of a solution of sodium hy-droxide (1 in 5), and heat in a water bath, while shaking, for 1 minute: a light red color develops in the pyridine layer. (2) Take 10 mL of the extract obtained in (1), evaporate on a water bath to dryness, add 1 g of anhydrous sodium carbonate to the residue, and heat for 10 minutes. After cooling, dissolve the residue in 40 mL of water, filter if nec-essary, and render the filtrate acidic with dilute nitric acid: the solution responds to Qualitative Tests <1.09> (2) for chlo-ride. The remainder of the filtrate responds to Qualitative Tests <1.09> (1) for chloride and to Qualitative Tests <1.09> for phosphate. pH <2.54> 6.0 – 6.5
Weigh accurately a portion of Triclofos Sodium Syrup, equivalent to 0.13 g of Triclofos Sodium, add 15 mL of water, 1 mL of sodium hydroxide TS and 15 mL of diethyl ether, shake for 1 minute, and separate the water layer. Wash the diethyl ether layer with 1 mL of water, and combine the washing with above water layer. To this solu-tion add 2.5 mL of diluted sulfuric acid (3 in 50), and extract with four 10-mL portions of 3-methyl-1-butanol. Combine the 3-methyl-1-butanol extracts, and add 3-methyl-1-butanol to make exactly 50 mL. Measure exactly 10 mL each of this solution, and dilute potassium hydroxide-ethanol TS, place in a glass ampule, fire-seal, mix, and heat at 1209C for 2 hours in an autoclave. After cooling, transfer the contents to a flask, add 20 mL of diluted nitric acid (63 in 500) and ex-actly 25 mL of 0.02 mol/L silver nitrate VS, shake well, and titrate <2.50> the excess silver nitrate with 0.02 mol/L ammo-nium thiocyanate VS (indicator: 2 to 3 drops of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.02 mol/L silver nitrate VS ＝ 1.676 mg of C2H3Cl3NaO4P
Trientine Hydrochloride Capsules contain not less than 90.0z and not more than 110.0z of the labeled amount of trientine hydrochloride (C6H18N4.2HCl: 219.16). Method of preparation Prepare as directed under Cap-sules, with Trientine Hydrochloride. Identification Take out the contents of Trientine Hydro-chloride Capsules, dry under reduced pressure not exceeding 0.67 kPa at 409C for 4 hours, and determine the infrared absorption spectrum as directed in the paste method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 3220 cm－1, 2120 cm－1, 1641 cm－1, 1620 cm－1, 1556 cm－1, 1502 cm－1 and 1116 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Trientine Hydrochloride Capsules is not less than 85z. Start the test with 1 capsule of Trientine Hydrochloride Capsules, withdraw not less than 25 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.
Take out the contents of Trientine Hydro-chloride Capsules, dry under reduced pressure not exceeding 0.67 kPa at 409C for 4 hours, and determine the infrared absorption spectrum as directed in the paste method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of about 3220 cm－1, 2120 cm－1, 1641 cm－1, 1620 cm－1, 1556 cm－1, 1502 cm－1 and 1116 cm－1. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Trientine Hydrochloride Capsules is not less than 85z. Start the test with 1 capsule of Trientine Hydrochloride Capsules, withdraw not less than 25 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.28 mg of trientine hydro-chloride (C6H18N4.2HCl), and use this solution as the sample solution. Separately, weigh accurately about 28 mg of trien-tine hydrochloride for assay, previously dried under reduced pressure not exceeding 0.67 kPa at 409C for 4 hours, dis-solve in water to make exactly 100 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample so-lution and standard solution separately, add exactly 5 mL of a mixture of disodium hydrogen phosphate-citric acid buffer solution (pH 8.2) and cupper (II) sulfate pentahydrate solu-tion (1 in 20) (4:1). Determine the absorbances, AT1 and AS1 at 580 nm, and AT2 and AS2 at 410 nm, of these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution obtained in the same manner with 10 mL of water as the blank. Dissolution rate (z) with respect to the labeled amount of trientine hydrochloride (C6H18N4.2HCl) ＝ Ms × (AT1 － AT2)/(AS1 － AS2) × V?/V × 1/C × 900 MS: Amount (mg) of trientine hydrochloride for assay taken C: Labeled amount (mg) of trientine hydrochloride (C6H18N4.2HCl) in 1 capsule
Take out the contents of not less than 20 Trientine Hydrochloride Capsules, weigh accurately the mass of the contents, and powder. Weigh accurately a portion of the powder, equivalent to about 0.25 g of trientine hydrochlo-ride (C6H18N4.2HCl), add 70 mL of methanol, dissolve by sonicating if necessary, and add methanol to make exactly 100 mL. Filter through a membrane filter with a pore size not exceeding 0.45 mm, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 0.25 g of trientine hydrochlo-ride for assay, previously dried under reduced pressure not exceeding 0.67 kPa at 409C for 4 hours, dissolve in methanol to make exactly 100 mL, and use this solution as the stand-ard solution. Pipet 5 mL each of the sample solution and standard solution separately, add exactly 10 mL of disodium hydrogen phosphate-citric acid buffer solution (pH 8.2) and exactly 1 mL of cupper (II) sulfate pentahydrate solution (1 in 20), and shake. Determine the absorbances, AT and AS, at 580 nm of these solutions, obtained with the sample solution and the standard solution, as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution obtained in the same manner with 5 mL of methanol as a blank. Amount (mg) of trientine hydrochloride (C6H18N4.2HCl) ＝ MS × AT/AS MS: Amount (mg) of trientine hydrochloride for assay taken
Containers—Tight containers. Storage—At 2 – 89C.
Trihexyphenidyl Hydrochloride Tablets contain not less than 93.0z and not more than 107.0z of the labeled amount of trihexyphenidyl hydrochloride (C20H31NO.HCl: 337.93). Method of preparation Prepare as directed under Tablets, with Trihexyphenidyl Hydrochloride. Identification (1) Weigh a quantity of powdered Trihexy-phenidyl Hydrochloride Tablets, equivalent to 0.1 g of at 408 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, respectively. Amount (mg) of trihexyphenidyl hydrochloride (C20H31NO.HCl) ＝ MS × AT/AS × V/1000 MS: Amount (mg) of Trihexyphenidyl Hydrochloride RS taken, calculated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medi-um, the dissolution rate in 30 minutes of Trihexyphenidyl Hydrochloride Tablets is not less than 70z. Start the test with 1 tablet of Trihexyphenidyl Hydrochlo-ride Tablets, withdraw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL ‌of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 2.
(1) Weigh a quantity of powdered Trihexy-phenidyl Hydrochloride Tablets, equivalent to 0.1 g of at 408 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, respectively. Amount (mg) of trihexyphenidyl hydrochloride (C20H31NO.HCl) ＝ MS × AT/AS × V/1000 MS: Amount (mg) of Trihexyphenidyl Hydrochloride RS taken, calculated on the dried basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medi-um, the dissolution rate in 30 minutes of Trihexyphenidyl Hydrochloride Tablets is not less than 70z. Start the test with 1 tablet of Trihexyphenidyl Hydrochlo-ride Tablets, withdraw not less than 30 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL ‌of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 2.2 mg of trihexyphenidyl hydrochloride (C20H31NO.HCl), and use this solution as the sample solution. Separately, weigh accu-rately about 10 mg of Trihexyphenidyl Hydrochloride RS, previously dried at 1059C for 3 hours, and dissolve in the dissolution medium to make exactly 100 mL. Pipet 2 mL of this solution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Pipet 20 mL each of the sample solution, the standard solution and the dissolution medium, add exactly 1 mL of diluted acetic acid (31) (1 in 10), and immediately add 5 mL of bromocresol green-sodium hydroxide-acetic acid-sodium acetate TS, and shake. Then, add exactly 10 mL each of dichloromethane, shake well, centrifuge, and take the dichloromethane layer. Determine the absorbances, AT, AS and AB, of these dichloromethane layers at 415 nm as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using dichloromethane as the blank. Dissolution rate (z) with respect to the labeled amount of trihexyphenidyl hydrochloride (C20H31NO.HCl) ＝ MS × (AT － AB)/(AS － AB) × V?/V × 1/C × 18 MS: Amount (mg) of Trihexyphenidyl Hydrochloride RS taken C: Labeled amount (mg) of trihexyphenidyl hydrochloride (C20H31NO.HCl) in 1 tablet
Weigh accurately and powder not less than 20 Tri-hexyphenidyl Hydrochloride Tablets. Weigh accurately a portion of the powder, equivalent to about 5 mg of trihexy-phenidyl hydrochloride (C20H31NO.HCl), dissolve in 2 mL of dilute hydrochloric acid and 60 mL of water by warming on a water bath for 10 minutes with occasional shaking. After cooling, add 2 mL of methanol and water to make ex-actly 100 mL, and use this solution as the sample solution. Dissolve about 50 mg of Trihexyphenidyl Hydrochloride RS (determine previously its loss on drying <2.41> under the same conditions as Trihexyphenidyl Hydrochloride), weighed accurately, in methanol, add methanol to make ex-actly 20 mL. Pipet 2 mL of this solution, add 2 mL of dilute hydrochloric acid and water to make exactly 100 mL, and use this solution as the standard solution. Pipet 10 mL each of the sample solution and standard solution into glass-stoppered centrifuge tubes, add exactly 10 mL each of bromocresol purple-dipotassium hydrogenphosphate-citric acid TS and 15 mL each of chloroform, stopper tightly, shake thoroughly, and centrifuge. Pipet 10 mL each of the chloroform layers, and add chloroform to make exactly 50 mL. Determine the absorbances, AT and AS, of the subse-quent solutions of the sample solution and standard solution at 408 nm as directed under Ultraviolet-visible Spectropho-tometry <2.24>, respectively. Amount (mg) of trihexyphenidyl hydrochloride (C20H31NO.HCl) ＝ MS × AT/AS × 1/10 MS: Amount (mg) of Trihexyphenidyl Hydrochloride RS taken, calculated on the dried basis
Trimetazidine Hydrochloride Tablets contain not less than 94.0z and not more than 106.0z of the labeled amount of trimetazidine hydrochloride (C14H22N2O3.2HCl: 339.26). Method of preparation Prepare as directed under Tablets, with Trimetazidine Hydrochloride. Identification Shake a quantity of powdered Trimetazidine Hydrochloride Tablets, equivalent to 10 mg of Trimetazidine cate for 10 minutes. Shake the solution for 10 minutes, and add the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly 20 mL. Centrifuge, pipet V mL of the supernatant liquid, equivalent to about 0.75 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl), add ex-actly 5 mL of the internal standard solution, add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of trimetazidine hydrochloride for assay (separately determine the water <2.48> in the same manner as Trimetazi-dine Hydrochloride), and dissolve in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly 200 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution and 0.1 mol/L hydro-chloric acid TS to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay. Amount (mg) of trimetazidine hydrochloride (C14H22N2O3.2HCl) ＝ MS × QT/QS × 1/2V MS: Amount (mg) of trimetazidine hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of parahy-droxybenzoic acid in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) (7 in 40,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Trimetazidine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Trimetazidine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 3.
Shake a quantity of powdered Trimetazidine Hydrochloride Tablets, equivalent to 10 mg of Trimetazidine cate for 10 minutes. Shake the solution for 10 minutes, and add the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly 20 mL. Centrifuge, pipet V mL of the supernatant liquid, equivalent to about 0.75 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl), add ex-actly 5 mL of the internal standard solution, add 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of trimetazidine hydrochloride for assay (separately determine the water <2.48> in the same manner as Trimetazi-dine Hydrochloride), and dissolve in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly 200 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution and 0.1 mol/L hydro-chloric acid TS to make 50 mL, and use this solution as the standard solution. Proceed as directed in the Assay. Amount (mg) of trimetazidine hydrochloride (C14H22N2O3.2HCl) ＝ MS × QT/QS × 1/2V MS: Amount (mg) of trimetazidine hydrochloride for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of parahy-droxybenzoic acid in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) (7 in 40,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of Trimetazidine Hydrochloride Tablets is not less than 80z. Start the test with 1 tablet of Trimetazidine Hydrochloride Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, and add water to make exactly V? mL so that each mL contains about 3.3 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl). Pipet 3 mL of this solu-tion, add exactly 3 mL of 0.1 mol/L hydrochloric acid TS, and use this solution as the sample solution. Separately, weigh accurately about 17 mg of trimetazidine hydrochloride for assay (separately determine the water <2.48> in the same manner as Trimetazidine Hydrochloride), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 50 mL. Pipet 5 mL of this so-lution, and add water to make exactly 25 mL. Pipet 3 mL of this solution, add exactly 3 mL of 0.1 mol/L hydrochloric acid TS, and use this solution as the standard solution. Per-form the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and de-termine the peak areas, AT and AS, of trimetazidine in each solution. ‌Dissolution rate (z) with respect to the labeled amount of trimetazidine hydrochloride (C14H22N2O3.2HCl) ＝ MS × AT/AS × V?/V × 1/C × 18 MS: Amount (mg) of trimetazidine hydrochloride for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of trimetazidine hydrochloride (C14H22N2O3.2HCl) in 1 tablet Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of trimetazidine are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of trimetazidine is not more than 1.5z.
Weigh accurately not less than 20 tablets of Trimetazidine Hydrochloride Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 3 mg of trimetazidine hydrochloride (C14H22N2O3.2HCl), add about 15 mL of a mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1), and sonicate for 10 minutes. Then shake for 10 minutes, add the mixture of 0.1 mol/L hydro-chloric acid TS and ethanol (99.5) (1:1) to make exactly 20 mL, and centrifuge. To exactly 5 mL of the supernatant liq-uid add exactly 5 mL of the internal standard solution and 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of trimetazidine hydrochloride for assay (sepa-rately determine the water <2.48> in the same manner as Trimetazidine Hydrochloride), and dissolve in the mixture of 0.1 mol/L hydrochloric acid TS and ethanol (99.5) (1:1) to make exactly 200 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution and 0.1 mol/L hydrochloric acid TS to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of trimetazidine to that of the internal standard. Amount (mg) of trimetazidine hydrochloride (C H N O .2HCl) ＝ M × Q /Q × 1/10 System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, trimetazidine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of trimetazidine to that of the internal stand-ard is not more than 1.0z.
Troxipide Fine Granules contain not less than 93.0z and not more than 107.0z of the labeled amount of troxipide (C15H22N2O4: 294.35). Method of preparation Prepare as directed under Gran-ules, with Troxipide. Identification To a quantity of Troxipide Fine Granules, equivalent to 20 mg of Troxipide, add 100 mL of 0.1 mol/L hydrochloric acid TS, stir, and filter. To 4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 50 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Troxipide Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Troxi-pide Fine Granules, add 80 mL of 0.1 mol/L hydrochloric acid TS, stir for 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of troxipide (C15H22N2O4).
To a quantity of Troxipide Fine Granules, equivalent to 20 mg of Troxipide, add 100 mL of 0.1 mol/L hydrochloric acid TS, stir, and filter. To 4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 50 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: Troxipide Fine Granules in single-dose packages meet the requirement of the Content uniformity test. To the total amount of the content of 1 package of Troxi-pide Fine Granules, add 80 mL of 0.1 mol/L hydrochloric acid TS, stir for 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of troxipide (C15H22N2O4). Centrifuge this solu-tion, pipet 2 mL of the supernatant liquid, add exactly 3 mL of the internal standard solution, and water to make 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of troxipide (C15H22N2O4) ＝ MS × QT/QS × V/25 MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 ‌mL of water as the dissolution medium, the dissolution rate in 60 minutes of Troxipide Fine Granules is not less than 85z. Weigh accurately an amount of Troxipide Fine Granules, equivalent to about 0.1 g of troxipide (C15H22N2O4), with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet 4 mL of the subsequent filtrate, add water to make exactly 20 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 20 mg of Troxipide RS, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 200 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 258 nm. Dissolution rate (z) with respect to the labeled amount of troxipide (C15H22N2O4) ＝ MS/MT × AT/AS × 1/C × 450 System suitability— System performance: When the procedure is run with 20 mL of standard solution under the above operating condi-tions, troxipide and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of troxipide to that of the internal standard is not more than 1.0z.
Troxipide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of troxi-pide (C H N O : 294.35). 15 22 2 4 MS: Amount (mg) of Troxipide RS taken MT: Amount (mg) of Troxipide Fine Granules taken C: Labeled amount (mg) of troxipide (C15H22N2O4) in 1 g Assay Weigh accurately an amount of Troxipide Fine Granules, equivalent to about 0.5 g of troxipide (C15H22N2O4), add 200 mL of 0.1 mol/L hydrochloric acid TS, stir for 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 250 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add 0.1 mol/L hydrochloric acid TS to make exactly 10 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, and water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Troxi-pide RS, previously dried at 1059C for 2 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 25 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, and water to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of troxipide to that of the internal standard. Amount (mg) of troxipide (C15H22N2O4) ＝ MS × QT/QS × 20 MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 258 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: To diluted phosphoric acid (1 in 500) add diethylamine to adjust the pH to 3.0. To 1500 mL of this so-lution add 100 mL of methanol and 50 mL of tetrahydrofu-ran. Flow rate: Adjust so that the retention time of troxipide is about 7 minutes. Method of preparation Prepare as directed under Tablets, with Troxipide. Identification Weigh accurately an amount of powdered Troxipide Tablets, equivalent to 0.1 g of Troxipide, add 250 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Troxipide Tablets add 90 mL of 0.1 mol/L hydrochloric acid TS, shake well to disintegrate, shake for another 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of troxipide (C15H22N2O4).
Weigh accurately an amount of powdered Troxipide Tablets, equivalent to 0.1 g of Troxipide, add 250 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Troxipide Tablets add 90 mL of 0.1 mol/L hydrochloric acid TS, shake well to disintegrate, shake for another 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of troxipide (C15H22N2O4). Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 3 mL of the inter-nal standard solution, add water to make 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of troxipide (C15H22N2O4) ＝ MS × QT/QS × V/25 MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Troxipide Tablets is not less than 70z. Start the test with 1 tablet of Troxipide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of troxipide (C15H22N2O4), and use this solution as the sample solution. Separately weigh accurately about 20 mg of Troxipide RS, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 200 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and ‌use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 258 nm. Dissolution rate (z) with respect to the labeled amount of troxipide (C15H22N2O4) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Troxipide RS taken C: Labeled amount (mg) of Troxipide (C15H22N2O4) in 1 tablet
Weigh accurately an amount of Troxipide Fine Granules, equivalent to about 0.5 g of troxipide (C15H22N2O4), add 200 mL of 0.1 mol/L hydrochloric acid TS, stir for 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly 250 mL. Centrifuge this solution, pipet 5 mL of the supernatant liquid, add 0.1 mol/L hydrochloric acid TS to make exactly 10 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, and water to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Troxi-pide RS, previously dried at 1059C for 2 hours, and dissolve in 0.1 mol/L hydrochloric acid TS to make exactly 25 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, and water to make 100 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of troxipide to that of the internal standard. Amount (mg) of troxipide (C15H22N2O4) ＝ MS × QT/QS × 20 MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 258 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: To diluted phosphoric acid (1 in 500) add diethylamine to adjust the pH to 3.0. To 1500 mL of this so-lution add 100 mL of methanol and 50 mL of tetrahydrofu-ran. Flow rate: Adjust so that the retention time of troxipide is about 7 minutes. Method of preparation Prepare as directed under Tablets, with Troxipide. Identification Weigh accurately an amount of powdered Troxipide Tablets, equivalent to 0.1 g of Troxipide, add 250 mL of 0.1 mol/L hydrochloric acid TS, shake, and filter. To 4 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 100 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maximum between 256 nm and 260 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Troxipide Tablets add 90 mL of 0.1 mol/L hydrochloric acid TS, shake well to disintegrate, shake for another 10 minutes, and add 0.1 mol/L hydrochloric acid TS to make exactly V mL so that each mL contains about 1 mg of troxipide (C15H22N2O4). Centrifuge this solution, pipet 2 mL of the supernatant liquid, add exactly 3 mL of the inter-nal standard solution, add water to make 100 mL, and use this solution as the sample solution. Proceed as directed in the Assay. Amount (mg) of troxipide (C15H22N2O4) ＝ MS × QT/QS × V/25 MS: Amount (mg) of Troxipide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in 0.1 mol/L hydrochloric acid TS (3 in 2000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Troxipide Tablets is not less than 70z. Start the test with 1 tablet of Troxipide Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.8 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of troxipide (C15H22N2O4), and use this solution as the sample solution. Separately weigh accurately about 20 mg of Troxipide RS, previously dried at 1059C for 2 hours, and dissolve in water to make exactly 200 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and ‌use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and de-termine the absorbances, AT and AS, at 258 nm. Dissolution rate (z) with respect to the labeled amount of troxipide (C15H22N2O4) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Troxipide RS taken C: Labeled amount (mg) of Troxipide (C15H22N2O4) in 1 tablet Assay Weigh accurately the mass of not less than 20 Troxi-pide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 1 g of troxipide (C H N O ), L-Tryptophan
Ubenimex Capsules contain not less than 93.0z and not more than 107.0z of the labeled amount of ubeni-mex (C16H24N2O4: 308.37). Method of preparation Prepare as directed under Cap-sules, with Ubenimex. Identification To a quantity of the contents of Ubenimex Capsules, equivalent to 25 mg of Ubenimex, add water to make 50 mL, shake well, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) visible Spectrophotometry <2.24>: it exhibits maxima be-tween 250 nm and 254 nm, between 255 nm and 259 nm, and 0 – 20 100 0 20 – 60 100 → 0 0 → 100 60 – 70 0 100 Flow rate: Adjust so that the retention time of ubenimex is about 14 minutes. Time span of measurement: About 5 times as long as the retention time of ubenimex, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of ubenimex obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ubenimex are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ubenimex is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Assay Weigh accurately about 0.5 g of Ubenimex, previ-ously dried, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.84 mg of C16H24N2O4 Containers and storage Containers—Tight containers. between 261 nm and 265 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 capsule of Ubenimex Capsules add 30 mL of a mix-ture of water and acetonitrile (7:3), shake well for 30 minutes, and add a mixture of water and acetonitrile (7:3) to make exactly 50 mL. Centrifuge this solution and filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 5 mL of the filtrate, pipet V mL of the subsequent filtrate, equivalent to about 3 mg of ubenimex (C16H24N2O4), add exactly 4 mL of the internal standard solution, add a mixture of water and acetonitrile (7:3) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of ubenimex for assay, previously dried at 809C for 4 hours under reduced pressure, and dissolve in a mixture of water and acetonitrile (7:3) to make exactly 100 mL. Pipet 15 mL of this solution, add exactly 4 mL of the internal standard solution, add a mixture of water and acetonitrile (7:3) to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ubenimex to that of the internal standard. Amount (mg) of ubenimex (C16H24N2O4) ＝ MS × QT/QS × 1/V × 15/2 MS: Amount (mg) of ubenimex for assay taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in a mixture of water and acetonitrile (7:3) (1 in 2000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, ubenimex and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ubenimex to that of the internal standard is not more than 2.0z. Dissolution <6.10> When the test is performed at 50 revolu- ‌tions per minute according to the Paddle method using the sinker, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Ubenimex Capsules is not less than 70z. Start the test with 1 capsule of Ubenimex Capsules, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add a mixture of water and acetonitrile (7:3) to make exactly V? mL so that each mL contains about 11 mg of ubenimex (C16H24N2O4), and use this solution as the sample solution.
To a quantity of the contents of Ubenimex Capsules, equivalent to 25 mg of Ubenimex, add water to make 50 mL, shake well, and filter. Determine the absorp-tion spectrum of the filtrate as directed under Ultraviolet- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) visible Spectrophotometry <2.24>: it exhibits maxima be-tween 250 nm and 254 nm, between 255 nm and 259 nm, and 0 – 20 100 0 20 – 60 100 → 0 0 → 100 60 – 70 0 100 Flow rate: Adjust so that the retention time of ubenimex is about 14 minutes. Time span of measurement: About 5 times as long as the retention time of ubenimex, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of ubenimex obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ubenimex are not less than 5000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ubenimex is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (0.5 g, in vacu-um, 809C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.5 g of Ubenimex, previ-ously dried, dissolve in 60 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determination in the same man-ner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 30.84 mg of C16H24N2O4
To a quantity of powdered Ursodeoxycholic Acid Granules, equivalent to 20 mg of Ursodeoxycholic Acid, add 10 mL of methanol, and shake for 20 minutes. Centrifuge this solution, pipet 4 mL of the supernatant liq-uid, and evaporate the methanol under reduced pressure. To the residue add 4 mL of acetone, disperse by sonication, cen-trifuge, and use the supernatant liquid as the sample solu-tion. Separately, dissolve 10 mg of ursodeoxycholic acid in 5 mL of acetone, and use this solution as the standard solu-tion. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL of the sam-ple solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of isooctane, ethanol (99.5), ethyl acetate and acetic acid (100) (10:6:3:1) to a distance of about 15 cm, and air-dry the plate. Dry the plate further at 1209C for 30 minutes, imme-diately splay evenly a solution of phosphomolybdic acid n-hydrate in ethanol (99.5) (1 in 5), and heat the plate at 1209C for 3 to 5 minutes: the principle spot obtained from the sam-ple solution and the spot from the standard solution show a blue color and the same Rf value. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medi-um, the dissolution rate in 15 minutes of Ursodeoxycholic Acid Granules is not less than 80z. Start the test with an accurately weigh amount of Ur-sodeoxycholic Acid Granules, equivalent to about 50 mg of ursodeoxycholic acid (C24H40O4), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, use the subsequent filtrate as the sample solu-tion. Separately, weigh accurately about 22 mg of ursodeox-ycholic acid for assay, previously dried at 1059C for 2 hours, and dissolve in acetonitrile to make exactly 100 mL. Pipet 5 ‌mL of this solution, add the dissolution medium to make ex-actly 20 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of ursodeoxycholic acid in each solution. Dissolution rate (z) with respect to the labeled amount of ursodeoxycholic acid (C24H40O4) ＝ MS/MT × AT/AS × 1/C × 225 MS: Amount (mg) of ursodeoxycholic acid for assay taken MT: Amount (g) of Ursodeoxycholic Acid Granules taken C: Labeled amount (mg) of ursodeoxycholic acid (C24H40O4) in 1 g Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and symmetry fac-tor of the peak of ursodeoxycholic acid are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ursodeoxycholic acid is not more than 2.0z.
Weigh accurately an amount of powdered Ursodeo-xycholic Acid Granules, equivalent to about 0.1 g of ursodeoxycholic acid (C24H40O4), add exactly 20 mL of the internal standard solution, shake for 10 minutes, and centri-fuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Separately, weigh accurately about 0.1 g of ursodeoxycholic acid for assay, previously dried at 1059C for 2 hours, dissolve in exactly 20 mL of the internal stand-ard solution, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ursodeoxycholic acid to that of the internal standard. Amount (mg) of ursodeoxycholic acid (C24H40O4) ＝ MS × QT/QS MS: Amount (mg) of ursodeoxycholic acid for assay taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (4 in 5) (7 in 200,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame- eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ursodeoxycholic acid to that of the internal standard is not more than 1.0z.
Ursodeoxycholic Acid Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of Ursodeoxycholic Acid (C24H40O4: 392.57). Method of preparation Prepare as directed under Tablets, with Ursodeoxycholic Acid. Identification To a quantity of powdered Ursodeoxycholic Acid Tablets, equivalent to 20 mg of Ursodeoxycholic Acid, add 10 mL of methanol, and shake for 20 minutes. Centri-fuge this solution, pipet 4 mL of the supernatant liquid, and evaporate the methanol under reduced pressure. To the residue add 4 mL of acetone, disperse by sonication, centri-fuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of ursodeoxycholic acid in 5 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of isooctane, ethanol (99.5), ethyl acetate and acetic acid (100) (10:6:3:1) to a distance of about 15 cm, and air-dry the plate. Dry the plate further at 1209C for 30 minutes, and im-mediately splay evenly a solution of phosphomolybdic acid n-hydrate in ethanol (95) (1 in 5) on the plate, and heat at 1209C for 3 to 5 minutes: the principal spot obtained from the sample solution and the spot obtained from the standard solution show a blue color and the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take 1 tablet of Ursodeoxycholic Acid Tablets and add exactly V mL of the internal standard solution so that each mL contains about 5 mg of ursodeoxycholic acid (C24H40O4), disperse by sonicating, then agitate to mix for 10 minutes and then centrifuge.
To a quantity of powdered Ursodeoxycholic Acid Tablets, equivalent to 20 mg of Ursodeoxycholic Acid, add 10 mL of methanol, and shake for 20 minutes. Centri-fuge this solution, pipet 4 mL of the supernatant liquid, and evaporate the methanol under reduced pressure. To the residue add 4 mL of acetone, disperse by sonication, centri-fuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of ursodeoxycholic acid in 5 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of isooctane, ethanol (99.5), ethyl acetate and acetic acid (100) (10:6:3:1) to a distance of about 15 cm, and air-dry the plate. Dry the plate further at 1209C for 30 minutes, and im-mediately splay evenly a solution of phosphomolybdic acid n-hydrate in ethanol (95) (1 in 5) on the plate, and heat at 1209C for 3 to 5 minutes: the principal spot obtained from the sample solution and the spot obtained from the standard solution show a blue color and the same Rf value. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. Take 1 tablet of Ursodeoxycholic Acid Tablets and add exactly V mL of the internal standard solution so that each mL contains about 5 mg of ursodeoxycholic acid (C24H40O4), disperse by sonicating, then agitate to mix for 10 minutes and then centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as the sample solution. Then, proceed as di-rected in the Assay. Amount (mg) of ursodeoxycholic acid (C H O ) ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ＝ MS × QT/QS × V/20 24 40 4 Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile for liquid chromatography (11:9). Flow rate: Adjust so that the retention time of ursodeoxy-cholic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ursodeoxycholic acid and the internal standard are MS: Amount (mg) of ursodeoxycholic acid for assay taken Internal standard solution—A solution of ethyl benzoate in diluted methanol (4 in 5) (7 in 200,000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution me-dium, the dissolution rates in 30 minutes of a 50-mg tablet and in 45 minutes of a 100-mg tablet are not less than 80z and not less than 70z, respectively. ‌Start the test with 1 tablet of Ursodeoxycholic Acid Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 10 mL of the first filtrate and pipet V mL of the subsequent filtrate. Add the dissolution medium to make exactly V? mL so that each mL contains about 56 mg of ursodeoxycholic acid (C24H40O4), and use the solution as the sample solution. Separately weigh accurately about 22 mg of ursodeoxycholic acid for assay, previously dried at 1059C for 2 hours, and dissolve in acetonitrile to make exactly 100 mL. Pipet 5 mL of this solution, add the dissolution medium to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 100 mL each of sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of ur-sodeoxycholic acid in each solution.‌ Dissolution rate (z) with respect to the labeled amount of ursodeoxycholic acid (C24H40O4) ＝ MS × AT/AS × V?/V × 1/C × 225 MS: Amount (mg) of ursodeoxycholic acid for assay taken C: Labeled amount (mg) of ursodeoxycholic acid in 1 tablet (C24H40O4) Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-dition, the number of theoretical plates and symmetry factor of the peak of ursodeoxycholic acid are not less than 3000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ursodeoxycholic acid is not more than 2.0z.
Weigh accurately the mass of not less than 20 Ur-sodeoxycholic Acid Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 0.1 g of ur-sodeoxycholic acid (C24H40O4), add exactly 20 mL of the in-ternal standard solution, shake for 10 minutes, and centri-fuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm, and use the filtrate as gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of diluted phosphoric acid (1 in 500) and acetonitrile for liquid chromatography (11:9). Flow rate: Adjust so that the retention time of ursodeoxy-cholic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution according to the above operating conditions, ursodeoxycholic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ursodeoxycholic acid to that of the internal standard is not more than 1.0z.
Valaciclovir Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of valaciclovir (C13H20N6O4: 324.34). Method of preparation Prepare as directed under Tablets, with Valaciclovir Hydrochloride. Identification Powder Valaciclovir Hydrochloride Tablets. To a portion of the powder, equivalent to 50 mg of valaciclovir (C13H20N6O4), add 90 mL of 0.1 mol/L hydro-chloric acid TS, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and to 2 mL of the subsequent filtrate add diluted phosphoric acid (1 in 1000) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 251 nm and 255 nm and a shoulder between 277 nm and 287 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Valaciclovir Hy-drochloride Tablets is not less than 75z. Start the test with 1 tablet of Valaciclovir Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 1 mL of the first filtrate, pipet V mL of the subsequent filtrate, add diluted phosphoric acid (1 in 1000) to make exactly V? mL so that each mL contains about 11 mg of valaciclovir (C13H20N6O4), and use this solution as the sample solution.
Powder Valaciclovir Hydrochloride Tablets. To a portion of the powder, equivalent to 50 mg of valaciclovir (C13H20N6O4), add 90 mL of 0.1 mol/L hydro-chloric acid TS, shake, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and to 2 mL of the subsequent filtrate add diluted phosphoric acid (1 in 1000) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhib-its a maximum between 251 nm and 255 nm and a shoulder between 277 nm and 287 nm. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the dissolution rate in 30 minutes of Valaciclovir Hy-drochloride Tablets is not less than 75z. Start the test with 1 tablet of Valaciclovir Hydrochloride Tablets, withdraw not less than 10 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Dis-card not less than 1 mL of the first filtrate, pipet V mL of the subsequent filtrate, add diluted phosphoric acid (1 in 1000) to make exactly V? mL so that each mL contains about 11 mg of valaciclovir (C13H20N6O4), and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Valaciclovir Hydrochloride RS (separately determine the water <2.48> and the residual solvent in the same man-ners as Valaciclovir Hydrochloride), and dissolve in diluted phosphoric acid (1 in 1000) to make exactly 250 mL. Pipet 5 mL of this solution, add diluted phosphoric acid (1 in 1000) to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 254 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using diluted phosphoric acid (1 in 1000) as the blank. Dissolution rate (z) with respect to the labeled amount of valaciclovir (C13H20N6O4) ＝ MS × AT/AS × V?/V × 1/C × 36 × 0.899 MS: Amount (mg) of Valaciclovir Hydrochloride RS taken, calculated on the anhydrous and residual sol-vent-free basis C: Labeled amount (mg) of valaciclovir (C13H20N6O4) in 1 tablet ‌Assay Weigh accurately the mass of not less than 20 Valaciclovir Hydrochloride Tablets, and powder. Weigh ac-curately a portion of the powder, equivalent to about 1 g of valaciclovir (C13H20N6O4), add 120 mL of 0.1 mol/L hydro-chloric acid TS, and sonicate for 10 minutes. Add 0.1 mol/L hydrochloric acid TS to make exactly 200 mL. Filter this so-lution through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, pipet 2 mL of the subsequent filtrate, add diluted phosphoric acid (1 in 1000) to make exactly 100 mL, and use this solu-tion as the sample solution. Separately, weigh accurately about 30 mg of Valaciclovir Hydrochloride RS (separately determine the water <2.48> and the residual solvent in the same manners as Valaciclovir Hydrochloride), dissolve in diluted phosphoric acid (1 in 1000) to make exactly 250 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of valaciclovir in each solution. Amount (mg) of valaciclovir (C13H20N6O4) ＝ MS × AT/AS × 40 × 0.899 MS: Amount (mg) of Valaciclovir Hydrochloride RS taken, calculated on the anhydrous and residual sol-vent-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with 18-crown ether-immobi-lized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 109C. Mobile phase: A mixture of diluted phosphoric acid (1 in 1000) and methanol (19:1). Flow rate: Adjust so that the retention time of valaciclovir is about 4.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of valaciclovir are not less than 600 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valaciclovir is not more than 1.0z.
Valsartan and Hydrochlorothiazide Tablets
バルサルタン・ヒドロクロロチアジド錠
Valsartan and Hydrochlorothiazide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of valsartan (C24H29N5O3: 435.52) and hydrochlorothiazide (C7H8ClN3O4S2: 297.74). Method of preparation Prepare as directed under Tablets, with Valsartan and Hydrochlorothiazide. Identiˆcation (1) To a quantity of powdered Valsartan and Hydrochlorothiazide Tablets, equivalent to 80 mg of Valsartan, add 5 mL of acetone, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 16 mg of valsartan in 1 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (15:5:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the two spots obtained from the sample solution and the spot from the standard solution show the same Rf value. (2) To a quantity of powdered Valsartan and Hydrochlo-rothiazide Tablets, equivalent to 6.25 mg of Hydrochloro-thiazide, add 5 mL of acetone, shake, centrifuge, and use the supernatant liquid as the sample solution. Separately, dis-solve 12.5 mg of hydrochlorothiazide in 10 mL of acetone, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with ‰uorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, hexane and acetic acid (100) (15:5:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): one of the two spots obtained from the sample solution and the spot from the standard solution show the same Rf value. Uniformity of dosage units <6.02> (1) Valsartan—Per-form the Mass variation test, or the Content uniformity test according to the following method: it meets the requirement. To 1 tablet of Valsartan and Hydrochlorothiazide Tablets add 10 mL of water, and shake until the tablet is disintegrat-ed. Add 10 mL of acetonitrile, shake thoroughly, and add a mixture of water and acetonitrile (1:1) to make exactly 50 mL. Centrifuge this solution, pipet V mL of the supernatant liquid, add a mixture of water and acetonitrile (1:1) to make exactly V? mL so that each mL contains about 0.
(1) Valsartan—Weigh accurately the mass of not less than 20 tablets of Valsartan and Hydrochlorothiazide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 80 mg of valsartan (C24H29N5O3), add 10 mL of water, and shake. Add 10 mL of acetonitrile, shake thoroughly, add a mixture of water and acetonitrile (1:1) to make exactly 50 mL, and centrifuge. Pipet 5 mL of the supernatant liquid, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this so-lution as the sample solution. Separately, weigh accurately about 40 mg of Valsartan RS (separately determine the water <2.48> and the residual solvent in the same manner as Valsar- tan), dissolve in a mixture of water and acetonitrile (1:1) to make exactly 25 mL, and use this solution as the valsartan standard stock solution. Pipet 5 mL of the valsartan stand-ard stock solution, add a mixture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, ‌of valsartan in each solution. Amount (mg) of valsartan (C24H29N5O3) ＝ MS × AT/AS × 2 MS: Amount (mg) of Valsartan RS taken, calculated on the anhydrous and residual solvent-free basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 271 nm). Column: A stainless steel column 3.0 mm in inside diame-ter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile and tri‰uoroacetic acid (900:100:1). Mobile phase B: A mixture of acetonitrile, water and tri‰uoroacetic acid (900:100:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. lowing conditions, and determine the peak areas, AT and AS, of hydrochlorothiazide in each solution. Amount (mg) of hydrochlorothiazide (C7H8ClN3O4S2) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of Hydrochlorothiazide RS taken Operating conditions— Proceed as directed in the operating conditions in (1). System suitability— System performance: Dissolve 1 mg of 4-amino-6-chlorobenzene-1,3-disulfonamide in a mixture of water and acetonitrile (1:1) to make 200 mL. To 1 mL of this solution, 5 mL of the valsartan standard stock solution in (1) and 5 mL of the hydrochlorothiazide standard stock solution add a mixture of water and acetonitrile (1:1) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide, hydrochlorothiazide and valsartan are eluted in this order with the resolution between the peaks of 4-amino-6-chlorobenzene-1,3-disulfonamide and hydrochloro-thiazide being not less than 1.5. System repeatability: When the test is repeated 6 times Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrochlorothiazide is not more than 1.0z. 0 – 25 90 → 10 10 → 90 Flow rate: Adjust so that the retention time of valsartan is about 16 minutes. System suitability— System performance: Dissolve 1 mg of 4-amino-6-chlorobenzene-1,3-disulfonamide in a mixture of water and acetonitrile (1:1) to make 200 mL. To 1 mL of this solution, 5 mL of the valsartan standard stock solution and 5 mL of the hydrochlorothiazide standard stock solution in (2) add a mixture of water and acetonitrile (1:1) to make 20 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, 4-amino-6-chlorobenzene-1,3-disulfonamide, hydrochlorothiazide and valsartan are eluted in this order with the resolution between the peaks of 4-amino-6-chlorobenzene-1,3-disulfonamide and hydrochloro-thiazide being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of valsartan is not more than 1.0z. (2) Hydrochlorothiazide—Weigh accurately the mass of not less than 20 tablets of Valsartan and Hydrochloro-thiazide Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 6.25 mg of hydrochloro-thiazide (C7H8ClN3O4S2), add 10 mL of water, and shake. Add 10 mL of acetonitrile, shake thoroughly, add a mixture of water and acetonitrile (1:1) to make exactly 50 mL, and centrifuge. Pipet 5 mL of the supernatant liquid, add a mix-ture of water and acetonitrile (1:1) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 12.5 mg of Hydrochlorothiazide RS, previously dried at 1059C for 2 hours, and dissolve in a mix-ture of water and acetonitrile (1:1) to make exactly 50 mL, and use this solution as the hydrochlorothiazide standard stock solution. Pipet 2.5 mL of the hydrochlorothiazide standard stock solution, add a mixture of water and aceto-nitrile (1:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- Vancomycin Hydrochloride
Valsartan Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of val-sartan (C24H29N5O3: 435.52). Method of preparation Prepare as directed under Tablets, with Valsartan. Identification Determine the absorption spectra of the sample solution and the standard solution in the range 220 to 350 nm, which are obtained in the Uniformity of dosage units, as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrums with each other: both spectra exhibit similar intensities of absorption at the same wavelengths. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Valsartan Tablets add V/10 mL of water, and shake until the tablet is disintegrated. Add V/2 mL of methanol, shake thoroughly, add methanol to make exactly V mL so that each mL contains about 0.
Determine the absorption spectra of the sample solution and the standard solution in the range 220 to 350 nm, which are obtained in the Uniformity of dosage units, as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrums with each other: both spectra exhibit similar intensities of absorption at the same wavelengths. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Valsartan Tablets add V/10 mL of water, and shake until the tablet is disintegrated. Add V/2 mL of methanol, shake thoroughly, add methanol to make exactly V mL so that each mL contains about 0.4 mg of valsartan (C24H29N5O3) for 20-mg tablet and 40-mg tablet, or contains about 0.8 mg of valsartan (C24H29N5O3) for 80-mg tablet and 160-mg tablet, and centrifuge. Pipet V? mL of the superna-tant liquid, equivalent to 0.8 mg of valsartan (C24H29N5O3), add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Valsartan RS (separately determine the water <2.48> and the residual solvent in the same manner as Valsar- tan), dissolve in 10 mL of water, and add methanol to make exactly 100 mL. Pipet 2 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 250 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of valsartan (C24H29N5O3) ＝ MS × AT/AS × V/V? × 1/50 MS: Amount (mg) of Valsartan RS taken, calculated on the anhydrous and residual solvent-free basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate of a 20-mg tablet, 40-mg tablet and 80-mg tablet in 30 minutes are not less than 75z, 75z and 80z, respectively, and of a 160-mg tablet in 45 minutes is not less than 75z. Start the test with 1 tablet of Valsartan Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.5 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add water to make exactly V? mL so that each mL con-tains about 22 mg of valsartan (C24H29N5O3), and use this so-lution as the sample solution. Separately, weigh accurately about 22 mg of Valsartan RS (separately determine the water <2.48> and the residual solvent in the same manners as Val- sartan), and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, at 250 nm of the sample solution and standard solution as directed under Ultraviolet- ‌visible Spectrophotometry <2.24>, using water as the control. Dissolution rate (z) with respect to the labeled amount of valsartan (C24H29N5O3) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Valsartan RS taken, calculated on the anhydrous and residual solvent-free basis C: Labeled amount (mg) of valsartan (C24H29N5O3) in 1 tablet
Weigh accurately the mass of not less than 20 Val-sartan Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 50 mg of valsartan (C24H29N5O3), add 60 mL of the mobile phase, shake thor-oughly, add the mobile phase to make exactly 100 mL, and centrifuge. Pipet 5 mL of the supernatant liquid, add exactly 3 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Valsartan RS (separately, determine the water <2.48> and the residual sol-vent in the same manner as Valsartan), and dissolve in the mobile phase to make exactly 100 mL. Pipet 5 mL of this so-lution, add exactly 3 mL of the internal standard solution, add the mobile phase to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of valsartan to that of the internal standard. Amount (mg) of valsartan (C24H29N5O3) ＝ MS × QT/QS MS: Amount (mg) of Valsartan RS taken, calculated on the anhydrous and residual solvent-free basis Internal standard solution—A solution of diclofenac sodium in the mobile phase (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 3 mm in inside diameter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, acetonitrile, and acetic acid (100) (500:500:1). Flow rate: Adjust so that the retention time of valsartan is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, valsartan and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of valsartan to that of the internal standard is not more than 1.0z.
Vancomycin Hydrochloride for Injection is a prepa-ration for injection which is dissolved before use. It contains not less than 90.
Vancomycin Hydrochloride for Injection oc-curs as white masses or a white powder.
(1) Dissolve an amount of Vancomycin Hydrochloride for Injection, equivalent to 5 mg (potency) of
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, and standard solu-tions—Proceed as directed in the Assay under Vancomycin Hydrochloride. Sample solutions—Weigh accurately the contents of not less than 10 Vancomycin Hydrochloride for Injection. Weigh accurately an amount of the content, equivalent to about 25 mg (potency) of Vancomycin Hydrochloride, and dissolve in water to make exactly 25 mL. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 4.5) to make solutions so that each mL contains 100 mg (potency) and 25 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Vasopressin Injection is an aqueous injection. It is a synthetic vasopressin consisting of 9 amino acid residues. It contains not less than 90.
Vasopressin Injection is a clear and colorless liquid. pH <2.54> 3.0 – 4.0
(1) Clarity and color of solution—Dissolve an amount of Vancomycin Hydrochloride for Injection, equiva-lent to 0.5 g (potency) of Vancomycin Hydrochloride, in 10 mL of water: the solution is clear and colorless to pale yel-low, and the absorbance of the solution, determined at 465 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.05. (2) Related substances—Dissolve an amount of Vanco-mycin Hydrochloride for Injection, equivalent to 0.1 g (po-tency) of Vancomycin Hydrochloride, in 10 mL of the mo-bile phase A, and use this solution as the sample solution. Proceed as directed in the Purity (2) under Vancomycin Hydrochloride. Water <2.48> Not more than 5.0z (0.1 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (3:1)). Bacterial endotoxins <4.01> Less than 0.25 EU/mg (po-tency). Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test. Foreign insoluble matter <6.06> Perform the test according to the Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement. [113-79-1] Vasopressin Injection is an aqueous injection. It is a synthetic vasopressin consisting of 9 amino acid residues. It contains not less than 90.0z and not more than 120.0z of the labeled units of vasopressin (C46H65N15O12S2). Method of preparation Prepare as directed under Injec-tions, with vasopressin. Description Vasopressin Injection is a clear and colorless liquid. pH <2.54> 3.0 – 4.0 Purity Related substances—To a suitable amount of Vasopressin Injection add diluted acetic acid (100) (1 in 400) so that each mL contains 20 Units of vasopressin (C46H65N15O12S2), and use this solution as the sample solu-tion. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area percentage method: the amount of the peak eluted before vasopressin is not more than 2.0z, and the total amount of the peaks other than vasopressin is not more than 10.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 6.6 g of diammonium hydrogen phosphate in 950 mL of water, adjust to pH 3.0 with phos-phoric acid, and add water to make 1000 mL. To 950 mL of this solution add 50 mL of acetonitrile. Mobile phase B: Dissolve 6.6 g of diammonium hydrogen ‌phosphate in 950 mL of water, adjust to pH 3.0 with phos-phoric acid, and add water to make 1000 mL. To 450 mL of this solution add 550 mL of acetonitrile. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 45 45 – 90 90 → 30 10 → 70 90 – 100 Flow rate: About 0.6 mL per minute. Time span of measurement: About 3 times as long as the retention time of vasopressin. System suitability— Test for required detectability: To 1 mL of the sample so-lution add diluted acetic acid (100) (1 in 400) to make 100 mL, and use this solution as the solution for system suitabil-ity test. Pipet 1 mL of the solution for system suitability test, and add diluted acetic acid (100) (1 in 400) to make exactly 10 mL. Confirm that the peak area of vasopressin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: When the procedure is run with 20 mL of the solution for system suitability test under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of vasopressin are not less than 17,500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of vasopressin is not more than 2.0z. Bacterial endotoxins <4.01> Less than 15 EU/ Unit. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Pipet V mL of Vasopressin Injection, equivalent to about 40 units of vasopressin, add diluted acetic acid (100) (1 in 400) to make exactly 25 mL, and use this solution as the sample solution. Separately, dissolve Vasopressin RS in diluted acetic acid (100) (1 in 400) so that each mL contains about 100 units of vasopressin, then dilute exactly with diluted acetic acid (100) (1 in 400) so that each mL contains about 1.6 units of vasopressin, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of vasopressin in each solution. Amount (Unit) of vasopressin in 1 mL of Vasopressin Injection ＝ MS × AT/AS × 25/V MS: Amount (unit) of vasopressin in 1 mL of the standard solution Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.6 g of diammonium hydrogen phosphate in 950 mL of water, adjust to pH 3.0 with phos-phoric acid, and add water to make 1000 mL. To 870 mL of this solution add 130 mL of acetonitrile. Flow rate: About 1 mL per minute. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of vasopressin are not less than 9500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of vasopressin is not more than 2.0z.
Verapamil Hydrochloride Injection is an aqueous injection. It contains not less than 93.
Verapamil Hydrochloride Injection is a clear, colorless liquid. Identiˆcation To 1 mL of the sample solution obtained in the Assay, add 0.02 mol/L hydrochloric acid TS to make 50 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm, and between 276 nm and 280 nm. pH Being speciˆed separately when the drug is granted ap-proval based on the Law. Bacterial endotoxins <4.01> Less than 12 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane ˆltration method: it meets the requirement.
Pipet a volume of Verapamil Hydrochloride Injec-tion, equivalent to about 10 mg of verapamil hydrochloride (C27H38N2O4.HCl), add 0.02 mol/L hydrochloric acid TS to make exactly 10 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of ver-apamil hydrochloride for assay, previously dried at 1059C for 2 hours, dissolve in 0.02 mol/L hydrochloric acid TS to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of ver-apamil in each solution. Amount (mg) of verapamil hydrochloride (C27H38N2O4.HCl) ＝ MS × AT/AS × 1/5 MS: Amount (mg) of verapamil hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 279 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water and perchlo-ric acid (550:450:1). Flow rate: Adjust so that the retention time of verapamil is about 5 minutes. ‌System Suitability—‌ System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of verapamil are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of verapamil is not more than 1.0z.
Verapamil Hydrochloride Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of verapamil hydrochloride (C H N O .HCl: 1059C for 2 hours, dissolve in a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of verapamil in each solution. Amount (mg) of verapamil hydrochloride (C27H38N2O4.HCl) in 1 tablet ＝ MS × AT/AS × V/500 MS: Amount (mg) of verapamil hydrochloride for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol, water and perchlo- 491.06). 27 38 2 4 ric acid (550:450:1). Flow rate: Adjust so that the retention time of verapamil Method of preparation Prepare as directed under Tablets, with Verapamil Hydrochloride. Identification To 2.5 mL of the sample solution obtained in the Assay add the mixture of methanol and 0.1 mol/L hy-drochloric acid TS (3:1) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 228 nm and 232 nm, and between 277 nm and 281 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Verapamil Hydrochloride Tablets add 7V/10 mL of a mixture of methanol and 0.1 mol/L hydro-chloric acid TS (3:1), and sonicate until the tablet is disin-tegrated. After cooling, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly V mL so that each mL contains about 0.
To 2.5 mL of the sample solution obtained in the Assay add the mixture of methanol and 0.1 mol/L hy-drochloric acid TS (3:1) to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits maxima between 228 nm and 232 nm, and between 277 nm and 281 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Verapamil Hydrochloride Tablets add 7V/10 mL of a mixture of methanol and 0.1 mol/L hydro-chloric acid TS (3:1), and sonicate until the tablet is disin-tegrated. After cooling, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly V mL so that each mL contains about 0.8 mg of verapamil hydrochlo-ride (C27H38N2O4.HCl). Centrifuge this solution, and use the supernatant liquid as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of verapamil hydrochloride (C27H38N2O4.HCl) ＝ MS × AT/AS × V/50 MS: Amount (mg) of verapamil hydrochloride for assay taken Disintegration <6.09> It meets the requirement.
To 25 Verapamil Hydrochloride Tablets, add 7V/10 mL a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1), and sonicate until the tablets are disintegrated. Fur-ther, sonicate for about 5 minutes. After cooling, add a mix-ture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly V mL so that each mL contains about 2 mg of verapamil hydrochloride (C27H38N2O4.HCl). Centrifuge this solution, pipet 10 mL of the supernatant liquid, add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (3:1) to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of verapamil hydrochloride for assay, previously dried at is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of verapamil are not less than 2000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of verapamil is not more than 1.0z.
Vinblastine Sulfate for Injection is a preparation for injection, which is dissolved before use. It contains not less than 90.
Vinblastine Sulfate for Injection occurs as white to pale yellow, light masses or powder. It is freely soluble in water. The pH of a solution (1 in 1000) is 3.5 – 5.0.
Proceed as directed in the Identification (1) under Vinblastine Sulfate. ‌Purity Related substances—Dissolve 4 mg of Vinblastine Sulfate for Injection in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 25 mL, and use this solution as the standard solution. Perform the test with exactly 200 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than vinblastine obtained from the sample solution is not larger than 1/2 times the peak area of vinblastine from the stand-ard solution, and the total area of the peaks other than vin-blastine from the sample solution is not larger than 2 times the peak area of vinblastine from the standard solution. Operating conditions— Perform as directed in the operating conditions in Purity (2) under Vinblastine Sulfate. System suitability— Perform as directed in the system suitability in Purity (2) under Vinblastine Sulfate. Bacterial endotoxins <4.01> Less than 10 EU/mg. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Dissolve 1 Vinblastine Sulfate for Injection in water to make exactly V mL so that each mL contains about 0.4 mg of vinblastine sulfate (C46H58N4O9.H2SO4), and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of Vinblastine Sulfate RS (previously determine the loss on drying under the same conditions as Vinblastine Sulfate), dissolve in water to make exactly 25 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Vinblastine Sulfate. Amount (mg) of vinblastine sulfate (C46H58N4O9.H2SO4) ＝ MS × AT/AS × 25/V MS: Amount (mg) of Vinblastine Sulfate RS taken, calcu-lated on the dried basis Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take an amount of Vinblastine Sulfate for Injection, equivalent to 0.10 g of vinblastine sulfate (C46H58N4O9.H2SO4), dissolve each content with a suitable amount of water, transfer into a 100-mL volumetric flask, wash each container with water, transfer the washings into the volumetric flask, and add water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Vinblastine Sulfate RS (previously determine the loss on drying under the same con-ditions as Vinblastine Sulfate), dissolve in water to make ex-actly 25 mL, and use this solution as the standard solution. Proceed as directed in the Assay under Vinblastine Sulfate. and colored containers may be used. Storage—Light-resistant, at 2 to 89C. Vincristine Sulfate
Thiamine Nitrate occurs as white, crystals or crystalline powder. It is odorless or a slight, characteristic odor. It is sparingly soluble in water, and very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 1939C (with decomposition).
To a portion of Thiamine Chloride Hydro-chloride Powder, equivalent to 0.02 g of Thiamine Chloride Hydrochloride, add 50 mL of water and 10 mL of dilute ace-tic acid, shake, and filter. Proceed with 5 mL of the filtrate 3-(4-Amino-2-methylpyrimidin-5-ylmethyl)-5-(2-hydroxyethyl)-4-methylthiazolium nitrate [532-43-4] Thiamine Nitrate, when dried, contains not less than 98.0z and not more than 102.0z of thiamine nitrate (C12H17N5O4S). Description Thiamine Nitrate occurs as white, crystals or crystalline powder. It is odorless or a slight, characteristic odor. It is sparingly soluble in water, and very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 1939C (with decomposition). Identification (1) Take 2-mL portions of a solution of Thiamine Nitrate (1 in 500), and add 2 to 3 drops of iodine TS: a red-brown precipitate or turbidity is produced. Upon ‌further addition of 1 mL of 2,4,6-trinitrophenol TS, a yellow precipitate or turbidity is produced. To 1 mL of a solution of Thiamine Nitrate (1 in 500) add 1 mL of lead (II) acetate TS and 1 mL of a solution of sodium hydroxide (1 in 10), and warm: the color of the solu-tion changes through yellow to brown, and on standing, a black-brown precipitate is produced. To 5 mL of a solution of Thiamine Nitrate (1 in 500) add 2.5 mL of sodium hydroxide TS and 0.5 mL of potas-sium hexacyanoferrate (III) TS. Then add 5 mL of 2-methyl-1-propanol, shake the mixture vigorously for 2 minutes, allow to stand, and examine under ultraviolet light (main wavelength: 365 nm): the 2-methyl-1-propanol layer shows a blue-purple fluorescence. This fluorescence disappears when the mixture is acidified, but reappears when it is again made alkaline. A solution of Thiamine Nitrate (1 in 50) responds to Qualitative Tests <1.09> (1) and (2) for nitrate. pH <2.54> Dissolve 1.0 g of Thiamine Nitrate in 100 mL of water: the pH of this solution is between 6.5 and 8.0.
(1) Chloride <1.03>—Perform the test with 0.20 g of Thiamine Nitrate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.053z). Sulfate <1.14>—Dissolve 1.5 g of Thiamine Nitrate in 30 mL of water and 2 mL of dilute hydrochloric acid, and add water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS and 2 mL of dilute hydrochloric acid, and add water to make 50 mL (not more than 0.011z). Heavy metals <1.07>—Dissolve 1.0 g of Thiamine Nitrate in 30 mL of water by warming, cool, and add 12 mL of 6 mol/L acetic acid TS and water to make 50 mL. Per-form the test with this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 1.0z (0.5 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.2z (1 g).
Weigh accurately about 0.1 g each of Thiamine Nitrate, previously dried, and Thiamine Chloride Hydro-chloride RS (separately determine the water <2.48> in the same manner as Thiamine Chloride Hydrochloride), and dis-solve them in the mobile phase to make exactly 50 mL. To 10 mL each of the solutions, accurately measured, add exactly 5 mL each of the internal standard solution, add the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions and calculate the ratios, QT and QS, of the peak area of thiamine to that of the internal standard. ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 1.1 g of sodium l-octanesulfonate in 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mL of this solution add 400 mL of a mixture of methanol and acetonitrile (3:2). Flow rate: Adjust so that the retention time of thiamine is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, thiamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of thiamine to that of the internal standard is not more than 1.0z.
Cyanocobalamin Injection is a clear, light red to red liquid.
Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ul-traviolet-visible Spectrophotometry <2.24>: it exhibits maxi-ma between 277 nm and 279 nm, between 360 nm, and 362 nm and between 548 nm and 552 nm. Determine the absor-bances, A1 and A2, of this solution at the wavelengths of maximum absorption between 360 nm and 362 nm, and be-tween 548 nm and 552 nm, respectively: the ratio A2/A1 is not less than 0.29 and not more than 0.32. Bacterial endotoxins <4.01> Less than 0.30 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Cyanocobalamin Injection, equivalent to about 2 mg of cyanocobalamin (C H CoN O P), add water to make exactly 100 mL, and MS: Amount (mg) of Cyanocobalamin RS taken, calcu-lated on the dried basis
リボフラビンリン酸エステルナトリウム注射液
Riboflavin Sodium Phosphate Injection is a clear, yellow to orange-yellow liquid. pH: 5.0 – 7.0
(1) To a measured volume of Riboflavin Sodium Phosphate Injection, equivalent to 1 mg of Riboflavin, add water to make 100 mL, and proceed with this solution as directed in the Identification (1) and (2) under Riboflavin Sodium Phosphate. To a measured volume of Riboflavin Sodium Phos-phate Injection, equivalent to 0.05 g of Riboflavin, and evaporate on a water bath to dryness. Proceed with this residue as directed in the Identification (4) under Riboflavin Sodium Phosphate. Bacterial endotoxins <4.01> Less than 10 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Membrane filtration method: it meets the requirement.
Conduct this procedure without exposure to light, using light-resistant vessels. To an accurately measured volume of Riboflavin Sodium Phosphate Injection, equiva-lent to about 15 mg of riboflavin (C17H20N4O6), add diluted acetic acid (100) (1 in 500) to make exactly 1000 mL, and use this solution as the sample solution. Proceed as directed in the Assay under Riboflavin Sodium Phosphate. Amount (mg) of Riboflavin (C17H20N4O6) ＝ MS × (AT － AT?)/(AS － AS?) MS: Amount (mg) of Riboflavin RS taken
Shake a portion of Riboflavin Powder, equivalent to 1 mg of Riboflavin, with 100 mL of water, filter, and proceed with the filtrate as directed in the Identifi-cation (1) and (2) under Riboflavin.
Rancidity—Riboflavin Powder is free from any un-pleasant or rancid odor or taste. Dissolution <6.10> When the test is performed at 75 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 60 minutes of Riboflavin Powder is not less than 80z. Conduct this procedure without exposure to light. Start the test with an accurately weighed amount of Riboflavin Powder, equivalent to about 5 mg of riboflavin (C17H20N4O6), withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 22 mg of Riboflavin RS, pre-viously dried at 1059C for 2 hours, dissolve in water by warming and add water to make exactly 200 mL after cool-ing. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. De-termine the absorbances, AT and AS, at 445 nm of the sam-ple solution and standard solution as directed under Ultravi- ‌olet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of riboflavin (C17H20N4O6) ＝ MS/MT × AT/AS × 1/C × 45/2 MS: Amount (mg) of Riboflavin RS taken MT: Amount (g) of Riboflavin Powder taken C: Labeled amount (mg) of riboflavin (C17H20N4O6) in 1 g
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately Riboflavin Powder equivalent to about 15 mg of riboflavin (C17H20N4O6), add 800 mL of diluted acetic acid (100) (1 in 400), and extract by warming for 30 minutes with occasional shaking. Cool, dilute with water to make exactly 1000 mL, and filter through a glass filter (G4). Use this filtrate as the sample solution, and proceed as directed in the Assay under Riboflavin. Amount (mg) of riboflavin (C17H20N4O6) ＝ MS × (AT － AT?)/(AS － AS?) MS: Amount (mg) of Riboflavin RS taken
Ascorbic Acid Injection occurs as a clear, col-orless liquid.
(1) Measure a volume of Ascorbic Acid Injection, equivalent to 0.5 g of Ascorbic Acid, and add water to make 25 mL. Proceed with 5 mL each of the solu-tion as directed in the Identification (1) under Ascorbic Acid. Measure a volume of Ascorbic Acid Injection, equivalent to 5 mg of Ascorbic Acid. Add a solution of metaphosphoric acid (1 in 50) to make 5 mL, and proceed JP XVIII Official Monographs / Ascorbic Acid and Calcium Pantothenate Tablets 485 with this solution as directed in the Identification (2) under Ascorbic Acid. Ascorbic Acid Injection responds to Qualitative Tests for sodium salt. pH <2.54> 5.6 – 7.4 Bacterial endotoxins <4.01> Less than 0.15 EU/mg. ‌Extractable volume <6.05> It meets requirement.‌ Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. der, equivalent to 0.5 g of Ascorbic Acid, add 30 mL of water, shake for 1 minute, and filter. Proceed with 5 mL each of the filtrate as directed in the Identification (1) under Ascorbic Acid. Weigh a portion of Ascorbic Acid Powder, equiva-lent to about 0.01 g of Ascorbic Acid, add 10 mL of a solu-tion of metaphosphoric acid (1 in 50), shake for 1 minute, and filter. Proceed with 5 mL of the filtrate as directed in the Identification (2) under Ascorbic Acid.
Rancidity—Ascorbic Acid Powder is free from any unpleasant or rancid odor and taste.
Weigh accurately a portion of Ascorbic Acid Pow-der, equivalent to about 0.1 g of L-ascorbic acid (C H O ), 6 8 6 Sterility <4.06> Perform the test according to the Mem- brane filtration method: it meets the requirement. Assay Measure exactly a volume of Ascorbic Acid Injec-tion, equivalent to about 0.1 g of L-ascorbic acid (C6H8O6), previously diluted with metaphosphoric acid-acetic acid TS, if necessary, and add metaphosphoric acid-acetic acid TS to make exactly 200 mL. Measure exactly 2 mL of the solution, and shake with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL of hydrogen peroxide TS. Titrate <2.50> with 2,6-dichloroindophenol sodium TS for titration until a light red color persists for 5 seconds. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 2, 6-dichlorophenol-indophenol sodium TS for titration ＝ A mg of C6H8O6 A is decided by the following standardization of 2,6-dichloroindophenol sodium TS for titration. 2,6-Dichlorophenol-indophenol sodium TS for titration: Preparation—Dissolve 42 mg of sodium hydrogen carbon-ate in 50 mL of water, add 0.5 g of 2,6-dichloroindophenol sodium dihydrate and water to make 200 mL, and filter. Pre-pare before use. Standardization—Weigh accurately about 50 mg of Ascor-bic Acid RS, previously dried in a desiccator (silica gel) for 24 hours, and dissolve in metaphosphoric acid-acetic acid TS to make exactly 100 mL. Pipet 2 mL of this solution, shake with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL of hydrogen peroxide TS, and titrate <2.50> with 2,6-dichloroindophenol sodium TS for titration until a light red color persists for 5 seconds. Perform a blank determination extract with several successive portions of metaphosphoric acid-acetic acid TS, combine the extracts, and filter. Wash the residue with metaphosphoric acid-acetic acid TS. Com-bine the filtrates and washings, and add metaphosphoric acid-acetic acid to make exactly 200 mL. Pipet 2 mL of the solution, and shake with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL of hydrogen peroxide TS. Titrate <2.50> with 2,6-dichloroindophenol sodium TS for titration until a light red color persists for 5 seconds. Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 2,6-dichlorophenol-indophenol sodium TS for titration ＝ A mg of C6H8O6 A is decided by the following standardization of 2,6-dichloroindophenol sodium TS for titration. 2,6-Dichlorophenol-indophenol sodium TS for titration: Preparation—Dissolve 42 mg of sodium hydrogen carbon-ate in 50 mL of water, add 0.05 g of 2,6-dichloroindophenol sodium dihydrate and water to make 200 mL, and filter. Pre-pare before use. Standardization—Weigh accurately about 50 mg of Ascor-bic Acid RS, previously dried in a desiccator (silica gel) for 24 hours, and dissolve in metaphosphoric acid-acetic acid TS to make exactly 100 mL. Pipet 2 mL of this solution, shake with 8 mL of metaphosphoric acid-acetic acid TS and 2 mL of hydrogen peroxide TS, and titrate <2.50> with 2,6-dichloroindophenol sodium TS for titration until a light red color persists for 5 seconds. Perform a blank determination in the same manner, and make any necessary correction. Calculate the quantity (A mg) of L-ascorbic acid (C H O ) in the same manner, and make any necessary correction. Calculate the quantity (A mg) of L-ascorbic acid (C H O ) equivalent to 1 mL of this test solution. 6 8 6 equivalent to 1 mL of this test solution. 6 8 6
(1) Weigh a portion of Ascorbic Acid Pow- Ascorbic Acid and Calcium Pantothenate Tablets
Voglibose Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of vogli- uid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Proceed as di-rected in the Assay. Amount (g) of voglibose (C10H21NO7) ＝ MS × AT/AS × V/500 MS: Amount (mg) of voglibose for assay taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 30 minutes of Voglibose Tablets is not less than 85z. Start the test with 1 tablet of Voglibose Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent fil-trate, add the mobile phase to make exactly V? mL so that each mL contains about 0.
Shake vigorously an amount of powdered Voglibose Tablets, equivalent to 5 mg of Voglibose, with 40 mL of water, and centrifuge. Transfer the supernatant liquid to a chromatographic column [prepared by pouring 1.0 mL of strongly acidic ion-exchange resin (H type) for column chromatography (100 to 200 mm in particle diameter) into a chromatographic column 8 mm in inside diameter and 130 mm in height], and allow to flow at a rate of about 5 mL per minute. Then wash the column with 200 mL of water, and allow to flow with 10 mL of diluted ammonia TS (1 in 4) at a rate of about 5 mL per minute. Filter the effluent solution 2 times through a membrane filter with a pore size not exceed-ing 0.22 mm. Evaporate the filtrate to dryness at 509C under reduced pressure, dissolve the residue with 0.5 mL of a mix-ture of water and methanol (1:1), and use this solution as the sample solution. Separately, dissolve 20 mg of voglibose for assay in 2 mL of the mixture of water and methanol (1:1), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 20 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ace-tone, ammonia water (28) and water (5:3:1) to a distance of about 12 cm, air-dry the plate, and allow to stand in iodine vapors: the principal spot obtained from the sample solution and the spot from the standard solution show a yellow-brown color, and the same Rf value. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Voglibose Tablets add exactly V mL of the mobile phase so that the solution contains about 40 mg of voglibose (C10H21NO7) per mL, disintegrate the tablet com-pletely by shaking, and centrifuge. Filter the supernatant liq- of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of voglibose in each solution. Dissolution rate (z) with respect to the labeled amount of voglibose (C10H21NO7) ＝ MS × AT/AS × V?/V × 1/C × 9/20 MS: Amount (mg) of voglibose for assay taken C: Labeled amount (mg) of voglibose (C10H21NO7) in 1 tablet Operating conditions— Apparatus, detector, column, column temperature, reac-tion coil, cooling coil, reaction reagent, reaction tempera-ture, and flow rate of reaction reagent: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 1.56 g of sodium dihydrogen phos-phate dihydrate in 500 mL of water. To this solution add a suitable amount of a solution, prepared by dissolving 3.58 g of disodium hydrogen phosphate decahydrate in 500 mL of water, to adjust to pH 6.5. To 500 mL of this solution add 500 mL of acetonitrile. Cooling temperature: A constant temperature of about 259C. Flow rate of mobile phase: Adjust so that the retention time of voglibose is about 6 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of voglibose are not less than 2000 and not more than1.5, respectively. System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voglibose is not more than 3.0z. ‌Assay To 20 tables of Voglibose Tablets add 80 mL of the mobile phase, and completely disintegrate by shaking. To an exact volume of the solution, equivalent to about 4 mg of voglibose (C10H21NO7), add the mobile phase to make ex-actly 100 mL, and centrifuge. Filter the supernatant liquid through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 1 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 20 mg of voglibose for assay (previously de-termine the water <2.48> in the same manner as Voglibose), and dissolve in the mobile phase to make exactly 25 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of voglibose in each solution. Amount (mg) of voglibose (C10H21NO7) ＝ MS × AT/AS × 1/500 MS: Amount of voglibose for assay taken, calculated on area of voglibose is not more than 2.0z.
Voriconazole for Injection is a preparation for in-jection which is dissolved before use. It contains not less than 93.
Voriconazole for Injection is white, masses or powder.
To 5 mL of the sample solution obtained in the Assay add the mobile phase in the Assay to make 25 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. pH Being specified separately when the drug is granted ap-proval based on the Law.
(1) Related substances—Dissolve the content of 1 container of Voriconazole for Injection in water so that each mL contains about 10 mg of voriconazole (C16H14F3N5O). To 5 mL of this solution add the mobile phase to make 100 mL, and use this solution as the sample solution. Pipet 5 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak, having the relative retention time of about 0.26 to voriconazole, ob-tained from the sample solution is not larger than 2.5 times the peak area of voriconazole from the standard solution, the area of the peak, having the relative retention time of about 0.32, from the sample solution is not larger than the peak area of voriconazole from the standard solution, the area of the peak, having the relative retention time of about 0.5, from the sample solution is not larger than 2 times the peak area of voriconazole from the standard solution, and the area of the peak other than voriconazole, the peak hav-ing the relative retention time of about 0.61 and the peaks mentioned above from the sample solution is not larger than the peak area of voriconazole from the standard solution. Furthermore, the total area of the peaks other than voriconazole and the peak having the relative retention time of about 0.61 from the sample solution is not larger than 7 times the peak area of voriconazole from the standard solu-tion. For the areas of the peaks, having the relative retention times of about 0.26, about 0.32 and about 0.5, multiply their correction factors, 0.7, 0.7 and 1.2, respectively. Operating conditions— Detector, column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 1.3 times as long as the retention time of voriconazole. System suitability— System performance: Suspend 0.1 g of voriconazole in 10 mL of a solution of sodium hydroxide (1 in 25), add the mo-bile phase to make 20 mL, and allow to stand for 30 minutes. To 1 mL of this solution add the mobile phase to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, the resolu-tion between the peaks, having the relative retention times about 0.26 and about 0.32 to voriconazole, is not less than 1.5. System repeatability: To 5 mL of the standard solution add the mobile phase to make 10 mL. When the test is repeated 6 times with 20 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of voriconazole is not more than 5.0z. (2) Enantiomer—Dissolve the content of 1 container of Voriconazole for Injection in the mobile phase so that each mL contains about 1 mg of voriconazole (C16H14F3N5O). To 5 mL of this solution add the mobile phase to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add the mobile phase to make ex-actly 100 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 10 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the area of the peak of the enan-tiomer, having the relative retention time of about 1.3 to voriconazole, obtained from the sample solution is not larger than 4 times the peak area of voriconazole from the standard solution. Operating conditions— Proceed as directed in the operating conditions in the Purity (3) under Voriconazole. System suitability— Proceed as directed in the system suitability in the Purity (3) under Voriconazole. Bacterial endotoxins <4.01> Less than 1.5 EU/mg. Uniformity of dosage units <6.02> It meets the requirement of the Mass variation test (T: 106.0z). Foreign insoluble matter <6.06> Perform the test according to Method 2: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Take 10 containers of Voriconazole for Injection, dissolve the contents of each in the mobile phase, combine the solutions, and add the mobile phase to make exactly 1000 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 50 mg of Voriconazole RS (separately determine the water <2.48> in the same manner as Voriconazole), and dissolve in the mo-bile phase to make exactly 50 mL. Pipet 5 mL of this solu-tion, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of voriconazole in each solution. ‌Amount (mg) of voriconazole (C16H14F3N5O) in 1 container of Voriconazole for Injection ＝ MS × AT/AS × 4 MS: Amount (mg) of Voriconazole RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 256 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.9 g of ammonium formate in 1000 mL of water, and adjust to pH 4.0 with formic acid. To 550 mL of this solution add 300 mL of methanol and 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of voricona-zole is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of voriconazole are not less than 5000 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voriconazole is not more than 1.0z.
Voriconazole Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of voriconazole (C16H14F3N5O: 349.31). Method of preparation Prepare as directed under Tablets, with Voriconazole. Identification To 5 mL of the sample solution obtained in the Assay add the mobile phase in the Assay to make 25 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Voriconazole Tablets add small amount of water to disintegrate the tablet, add V/2 mL of the mobile phase, stir for 20 minutes, and add the mobile phase to make exactly V mL so that each mL contains about 1 mg of voriconazole (C16H14F3N5O).
To 5 mL of the sample solution obtained in the Assay add the mobile phase in the Assay to make 25 mL. Determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 254 nm and 258 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Voriconazole Tablets add small amount of water to disintegrate the tablet, add V/2 mL of the mobile phase, stir for 20 minutes, and add the mobile phase to make exactly V mL so that each mL contains about 1 mg of voriconazole (C16H14F3N5O). Centrifuge, pipet 5 mL of the supernatant liquid, add the mobile phase to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of voriconazole (C16H14F3N5O) ＝ MS × AT/AS × V/20 MS: Amount (mg) of Voriconazole RS taken, calculated on the anhydrous basis Dissolution <6.10> When the test is performed at 50 revolu- tions per minute according to the Paddle method, using 900 mL of 1st fluid for dissolution test as the dissolution me-dium, the value Q in 30 minutes of Voriconazole Tablets is 80z. Start the test with 1 tablet of Voriconazole Tablets, with-draw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 22 mg of voriconazole (C16H14F3N5O), and use this solution as the sample solution. Separately, weigh accurately about 18 mg of Voriconazole RS (separately determine the water <2.48> in the same man-ner as Voriconazole), dissolve in 2 mL of methanol, and add the dissolution medium to make exactly 200 mL. Pipet 10 mL of this solution, add the dissolution medium to make ex-actly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, at 256 nm of the sample solution and standard solution as directed under Ul-traviolet-visible Spectrophotometry <2.24>, using the dissolu-tion medium as the blank. Dissolution rate (z) with respect to the labeled amount of voriconazole (C16H14F3N5O) ＝ MS × AT/AS × V?/V × 1/C × 90 MS: Amount (mg) of Voriconazole RS taken, calculated on the anhydrous basis C: Labeled amount (mg) of voriconazole (C16H14F3N5O) in 1 tablet
Weigh accurately the mass of not less than 20 Voriconazole Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 50 mg of voricona-zole (C16H14F3N5O), add the mobile phase, stir, and add the mobile phase to make exactly 50 mL. Centrifuge, pipet 5 mL of the supernatant liquid, add the mobile phase to make ex-actly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of Voriconazole RS (separately determine the water <2.48> in the same man-ner as Voriconazole), and dissolve in the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of voriconazole in each solution. Amount (mg) of voriconazole (C16H14F3N5O) ＝ MS × AT/AS × 5/2 MS: Amount (mg) of Voriconazole RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 256 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 1.9 g of ammonium formate in 1000 mL of water, and adjust to pH 4.0 with formic acid. To 550 mL of this solution add 300 mL of methanol and 150 mL of acetonitrile. Flow rate: Adjust so that the retention time of voricona-zole is about 9 minutes. ‌System suitability—‌ System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of voriconazole are not less than 5000 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of voriconazole is not more than 1.0z.
Warfarin Potassium Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of warfarin potassium (C19H15KO4: 346.42). Method of preparation Prepare as directed under Tablets, with Warfarin Potassium. Identification (1) Determine the absorption spectrum of the solution T2 obtained in the Assay, using 0.02 mol/L po-tassium hydroxide TS as the blank, as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 306 nm and 310 nm, and a minimum between 258 nm and 262 nm. Separately, determine the absorption spectrum of the solution T1 obtained in the Assay, using 0.02 mol/L hydrochloric acid TS as the blank, as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 281 nm and 285 nm and between 303 nm and 307 nm, and a minimum between 243 nm and 247 nm. (2) Weigh a quantity of Warfarin Potassium Tablets, equivalent to 0.01 g of Warfarin Potassium, add 10 mL of acetone, shake, and filter. Heat the filtrate on a water bath to evaporate the acetone. To the residue add 10 mL of diethyl ether and 2 mL of dilute hydrochloric acid, and shake: the aqueous layer responds to Qualitative Tests <1.09> for potassium salt. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Powder 1 tablet of Warfarin Potassium Tablets, add 40 mL of water, and shake vigorously for 30 minutes. Add water to make exactly V mL of this solution containing about 20 mg of warfarin potassium (C19H15KO4) per ml. Filter this solution, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 40 mg of Warfarin Potassium MS: Amount (mg) of Warfarin Potassium RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 0.5-mg, 1-mg and 2-mg tablet and in 30 minutes of 5-mg tablet of Warfarin Potassium Tablets are not less than 80z. Start the test with 1 tablet of Warfarin Potassium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.
(1) Determine the absorption spectrum of the solution T2 obtained in the Assay, using 0.02 mol/L po-tassium hydroxide TS as the blank, as directed under Ultra-violet-visible Spectrophotometry <2.24>: it exhibits a maxi-mum between 306 nm and 310 nm, and a minimum between 258 nm and 262 nm. Separately, determine the absorption spectrum of the solution T1 obtained in the Assay, using 0.02 mol/L hydrochloric acid TS as the blank, as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 281 nm and 285 nm and between 303 nm and 307 nm, and a minimum between 243 nm and 247 nm. (2) Weigh a quantity of Warfarin Potassium Tablets, equivalent to 0.01 g of Warfarin Potassium, add 10 mL of acetone, shake, and filter. Heat the filtrate on a water bath to evaporate the acetone. To the residue add 10 mL of diethyl ether and 2 mL of dilute hydrochloric acid, and shake: the aqueous layer responds to Qualitative Tests <1.09> for potassium salt. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. Powder 1 tablet of Warfarin Potassium Tablets, add 40 mL of water, and shake vigorously for 30 minutes. Add water to make exactly V mL of this solution containing about 20 mg of warfarin potassium (C19H15KO4) per ml. Filter this solution, discard the first 5 mL of the filtrate, and use the subsequent filtrate as the sample solution. Sepa-rately, weigh accurately about 40 mg of Warfarin Potassium MS: Amount (mg) of Warfarin Potassium RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rates in 15 minutes of 0.5-mg, 1-mg and 2-mg tablet and in 30 minutes of 5-mg tablet of Warfarin Potassium Tablets are not less than 80z. Start the test with 1 tablet of Warfarin Potassium Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 0.56 mg of warfarin potassium (C19H15KO4), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Warfarin Potassium RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Pipet 5 mL of this solu-tion, add water to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 100 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of warfarin in each solution. Dissolution rate (z) with respect to the labeled amount of warfarin potassium (C19H15KO4) ＝ MS × AT/AS × V?/V × 1/C × 9/4 MS: Amount (mg) of Warfarin Potassium RS taken C: Labeled amount (mg) of warfarin potassium (C19H15KO4) in 1 tablet Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 283 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol, water and phos-phoric acid (700:300:1). Flow rate: Adjust so that the retention time of warfarin is about 6 minutes. System suitability— System performance: When the procedure is run with 100 mL of the standard solution under the above conditions, the number of theoretical plates and the symmetry factor of the ‌peak of warfarin are not less than 2000 and not more than 2.0, respectively.‌‌ System repeatability: When the test is repeated 6 times with 100 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of warfarin is not more than 2.0z.
Weigh accurately and powder not less than 20 Warfarin Potassium Tablets. Weigh accurately a portion of the powder, equivalent to about 4 mg of warfarin potassium (C19H15KO4), add 80 mL of water, shake vigorously for 15 minutes, and add water to make exactly 100 mL. Filter this solution, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. Separately, weigh accurately about 80 mg of Warfarin Potassium RS, previ-ously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the stand-ard solution. Pipet 10 mL each of the sample solution and standard solution, add 0.02 mol/L hydrochloric acid TS to make exactly 20 mL, and use these solutions as the solution T1 and the solution S1, respectively. Separately, pipet 10 mL each of the sample solution and standard solution, add 0.02 mol/L potassium hydroxide TS to make exactly 20 mL, and use these solutions as the solution T2 and the solution S2, respectively. Determine the absorbances, AT and AS, of the solution T1 and the solution S1 at 272 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the solu-tion T2 and the solution S2 as the blank, respectively. Amount (mg) of warfarin potassium (C19H15KO4) ＝ MS × AT/AS × 1/20 MS: Amount (mg) of Warfarin Potassium RS taken
Water for Injection is a clear and colorless liquid, having no odor.
Total organic carbon <2.59>—Not more than 0.50 mg/L. Conductivity <2.51> When the test is performed according to the following method, the conductivity (259C) is not more than 2.1 mS・cm－1. Transfer a suitable amount of Water for Injection to a beaker, and stir the water specimen. Adjust the temperature to 25 ± 19C, and begin agitating the water specimen vigorously, while observing its conductivity periodically. When the change in conductivity becomes not greater than 0.1 mS・cm－1 per 5 minutes, adopt the observed value as the conductivity of the water specimen. Bacterial endotoxins <4.01> Less than 0.25 EU/mL. Sterile Water for Injection in Containers
Weak Opium Alkaloids and Scopolamine Injection
弱アヘンアルカロイド・スコポラミン注射液
Weak Opium Alkaloids and Scopolamine Injection is an aqueous injection. It contains not less than 0.
Weak Opium Alkaloids and Scopolamine In-jection is a clear, colorless or light brown liquid. It is affected by light. pH: 2.5 – 3.5
(1) To 1 mL of Weak Opium Alkaloids and Scopolamine Injection add 1 mL of ethanol (99.5), mix, and use this solution as the sample solution. Proceed with the sample solution as directed in the Identification (1) under Opium Alkaloids Hydrochlorides. (2) To 2 mL of Weak Opium Alkaloids and Scopolamine Injection add 2 mL of ammonia TS, extract with 10 mL of diethyl ether, and filter the diethyl ether layer. Evaporate the filtrate on a water bath to dryness, add 1 mL of ethanol (99.5) to the residue, and heat to dissolve. Allow to stand this solution in an ice water for 30 minutes with occasional shaking. After crystals are formed, use the supernatant liq-uid as the sample solution. Separately, dissolve 0.03 g of Scopolamine Hydrobromide RS in 100 mL of water, proceed with 2 mL of this solution in the same manner as for the sample solution, and use a solution so obtained as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of methanol and ammonia water (28) (200:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly Dragendorff's TS for spraying on the plate: a spot with an Rf value of about 0.7 among the several spots from the sample solution and an orange colored spot from the standard solution show the same color tone, and have the same Rf value (scopolamine). Extractable volume <6.05> It meets the requirements.
(1) Morphine—Pipet 2 mL of Weak Opium Alkaloids and Scopolamine Injection, add exactly 10 mL of the internal standard solution and water to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of morphine hydrochloride hydrate for assay, dissolve in exactly 10 mL of the internal standard solution, add water to make 50 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of morphine to that of the internal standard. Amount (mg) of morphine (C17H19NO3) ＝ MS × QT × QS × 0.887 MS: Amount (mg) of morphine hydrochloride hydrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of etilefrin hydro-chloride (1 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.0 g of sodium lauryl sulfate in 500 mL of diluted phosphoric acid (1 in 1000), and adjust the pH to 3.0 with sodium hydroxide TS. To 240 mL of this solution add 70 mL of tetrahydrofuran, and mix. Flow rate: Adjust so that the retention time of morphine is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, morphine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of morphine to that of the internal standard is not more than 2.0z. (2) Scopolamine hydrobromide hydrate—Pipet 4 mL of Weak Opium Alkaloids and Scopolamine Injection, and add exactly 2 mL of the internal standard solution. To this solu-tion add 10 mL of diluted dilute hydrochloric acid (1 in 10), and shake with two 10-mL portions of dichloromethane. Remove the dichloromethane layer, to the water layer add 2 mL of ammonia TS, add immediately 20 mL of dichloro-methane, shake vigorously, filter the dichloromethane ex-tract through a filter paper on which 5 g of anhydrous sodi-um sulfate is placed, and evaporate the filtrate to dryness under reduced pressure. To the residue add 0.5 mL of 1,2-dichloroethane and 0.5 mL of bis-trimethyl silyl acetamide, stopper tightly, warm in a water bath at 609C for 15 minutes, and use this solution as the sample solution. Separately, weigh accurately about 60 mg of Scoporamine Hydrobromide RS (separately determine the loss on drying <2.41> under the same conditions as Scopolamine Hydro- bromide Hydrate), and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution. Proceed with this solution in the same manner as for the sample solution, and use so obtained solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the follow-ing conditions, and calculate the ratios, QT and QS, of the peak area of scopolamine to that of the internal standard. Amount (mg) of scopolamine hydrobromide hydrate (C17H21NO4.HBr.3H2O) ＝ MS × QT/QS × 1/50 × 1.141 MS: Amount (mg) of Scopolamine Hydrobromide RS taken, calculated on the dried basis Internal standard solution—A solution of homatropine hydrobromide (1 in 4000). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1.5 m in length, packed with 180 to 250 mm siliceous earth for gas chromatography coated in 1 to 3z with 50z phenyl-methyl silicone polymer for gas chromatography. Column temperature: A constant temperature of about 2109C. Carrier gas: Nitrogen or helium. Flow rate: Adjust so that the retention time of scopola-mine is about 8 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, the internal standard and scopolamine are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 5 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of scopolamine to that of the internal standard is not more than 2.0z.
White Ointment is white in color. It has a slight, characteristic odor.
Xylitol Injection is a clear, colorless liquid. It has a sweet taste.
Measure a volume of Xylitol Injection, equivalent to 0.1 g of Xylitol, add water to make 10 mL, and use this solution as the sample solution. Separately, dissolve 0.1 g of xylitol in 10 mL of water, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethanol (95), ammonia solution (28) and water (25:4:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly silver nitrate-ammonia TS, and dry the plate at 1059C for 15 minutes: the spots from the sample solution and standard solution show a black-brown color and the same Rf value. pH <2.54> 4.5 – 7.5 Bacterial endotoxins <4.01> Less than 0.50 EU/mL. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Measure exactly a volume of Xylitol Injection, equivalent to about 5 g of xylitol (C5H12O5), and add water to make exactly 250 mL. Measure exactly 10 mL of this solu-tion, and add water to make exactly 100 mL. Then, pipet 10 mL of this solution into an iodine flask, and proceed as di-rected in the Assay under Xylitol. Each mL of 0.1 mol/L sodium thiosulfate VS ＝ 1.902 mg of C5H12O5
Zaltoprofen Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 44 mg of zaltoprofen (C17H14O3S), and use this solution as the sample solution.
Powder a suitable amount of Zaltoprofen Tablets. To a portion of the powder, equivalent to 80 mg of Zaltoprofen, add 30 mL of ethanol (99.5), shake well, and centrifuge. To 1 mL of the supernatant liquid add ethanol (99.5) to make 20 mL. To 2 mL of this solution add ethanol (99.5) to make 25 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm and between 329 nm and 333 nm, and a shoulder between 238 nm and 248 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Zaltoprofen Tablets add 4 mL of water, and shake to disintegrate. Add a suitable amount of ethanol (95), shake, then add ethanol (95) to make exactly V mL so that each mL contains about 4 mg of zaltoprofen (C17H14O3S), and centrifuge. Pipet 2 mL of the supernatant liquid, add ex-actly 10 mL of the internal standard solution and ethanol (95) to make 50 mL, and use this solution as the sample solu-tion. Proceed as directed in the Assay. Amount (mg) of zaltoprofen (C17H14O3S) ＝ MS × QT/QS × V/20 MS: Amount (mg) of zaltoprofen for assay taken Internal standard solution—A solution of benzyl benzoate in acetonitrile (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Zaltoprofen Tablets is not less than 75z. Start the test with 1 tablet of Zaltoprofen Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent ethanol (95) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of zaltoprofen for assay, previously dried at 1059C for 4 hours, add 4 mL of water and ethanol (95) to make exactly 20 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and ethanol (95) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of zaltoprofen to that of the internal standard. Amount (mg) of zaltoprofen (C17H14O3S) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of zaltoprofen for assay taken Internal standard solution—A solution of benzyl benzoate in acetonitrile (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (300:200:1). Flow rate: Adjust so that the retention time of zaltoprofen is about 4 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, zaltoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of zaltoprofen to that of the internal standard is not more than 1.0z. ‌Containers and storage Containers—Tight containers. Zidovudine
To 10 tablets of Zaltoprofen Tablets add 40 mL of water, shake to disintegrate, then add a suitable amount of ethanol (95), shake, add ethanol (95) to make exactly 200 mL, and centrifuge. Pipet an amount of the supernatant zaltoprofen (C 17H 14O3S: 298.36). liquid, equivalent to about 8 mg of zaltoprofen (C17H14O3S), add exactly 10 mL of the internal standard solution and Method of preparation Prepare as directed under Tablets, with Zaltoprofen. Identification Powder a suitable amount of Zaltoprofen Tablets. To a portion of the powder, equivalent to 80 mg of Zaltoprofen, add 30 mL of ethanol (99.5), shake well, and centrifuge. To 1 mL of the supernatant liquid add ethanol (99.5) to make 20 mL. To 2 mL of this solution add ethanol (99.5) to make 25 mL, and determine the absorption spec-trum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima between 227 nm and 231 nm and between 329 nm and 333 nm, and a shoulder between 238 nm and 248 nm. Uniformity of dosage units <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Zaltoprofen Tablets add 4 mL of water, and shake to disintegrate. Add a suitable amount of ethanol (95), shake, then add ethanol (95) to make exactly V mL so that each mL contains about 4 mg of zaltoprofen (C17H14O3S), and centrifuge. Pipet 2 mL of the supernatant liquid, add ex-actly 10 mL of the internal standard solution and ethanol (95) to make 50 mL, and use this solution as the sample solu-tion. Proceed as directed in the Assay. Amount (mg) of zaltoprofen (C17H14O3S) ＝ MS × QT/QS × V/20 MS: Amount (mg) of zaltoprofen for assay taken Internal standard solution—A solution of benzyl benzoate in acetonitrile (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 2nd fluid for dissolution test as the dissolution medium, the dissolution rate in 30 minutes of Zaltoprofen Tablets is not less than 75z. Start the test with 1 tablet of Zaltoprofen Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent ethanol (95) to make 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 80 mg of zaltoprofen for assay, previously dried at 1059C for 4 hours, add 4 mL of water and ethanol (95) to make exactly 20 mL. Pipet 2 mL of this solution, add exactly 10 mL of the internal standard solution and ethanol (95) to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of zaltoprofen to that of the internal standard. Amount (mg) of zaltoprofen (C17H14O3S) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of zaltoprofen for assay taken Internal standard solution—A solution of benzyl benzoate in acetonitrile (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of acetonitrile, water and acetic acid (100) (300:200:1). Flow rate: Adjust so that the retention time of zaltoprofen is about 4 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, zaltoprofen and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of zaltoprofen to that of the internal standard is not more than 1.0z. ‌Containers and storage Containers—Tight containers. Zidovudine
Zinc Oxide Ointment contains not less than 18.
Zinc Oxide Ointment is white in color. ‌Identification Place 1 g of Zinc Oxide Ointment in a cruci-ble, melt by warming, heat gradually raising the temperature until the mass is thoroughly charred, and then ignite it strongly: a yellow color is produced, and disappears on cool-ing. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the filtrate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is formed (zinc oxide).‌
Calcium, magnesium and other foreign inorganic matters—Place 2.0 g of Zinc Oxide Ointment in a crucible, melt by warming, and heat gradually raising the tempera- filter. Boil the residue on a filter paper with 10 mL of water, cool, and add 1 drop of iodine TS: a dark blue-purple color is produced (starch).
Zinc Oxide Starch Powder occurs as a white powder.
(1) Place 1 g of Zinc Oxide Starch Powder in a crucible, heat gradually, raising the temperature until it is charred, and then ignite strongly: a yellow color develops, and disappears on cooling. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the filtrate add 2 to 3 drops of potassium hexa-cyanoferrate (II) TS: a white precipitate is formed (zinc oxide). (2) Shake throughly 1 g of Oxide Starch Powder with 10 mL of water and 5 mL of dilute hydrochloric acid, and Description Zinc Sulfate Hydrate occurs as colorless crys-tals or white crystalline powder. It is very soluble in water, and very slightly soluble in ethanol (99.5). It effloresces in dry air. Identification (1) A solution of Zinc Sulfate Hydrate (1 in 20) responds to Qualitative Tests <1.09> for zinc salt. (2) A solution of Zinc Sulfate Hydrate (1 in 20) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Zinc Sulfate Hydrate in 20 mL of water: the pH of the solution is between 4.4 and 6.0.
(1) Clarity and color of solution—Dissolve 0.25 g of Zinc Sulfate Hydrate in 5 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Dissolve 1.0 g of Zinc Sulfate Hydrate in 10 mL of water contained in a Nessler tube. Add 20 mL of potassium cyanide TS, and mix well. Add 2 drops of sodium sulfide TS, and allow the mixture to stand for 5 minutes. Observe vertically against a white background, the color of the solution is not more intense than the following control solution. Control solution: To 1.0 mL of Standard Lead Solution add 10 mL of water and 20 mL of potassium cyanide TS, and mix well. Add 2 drops of sodium sulfide TS (not more than 10 ppm). Alkali earth metals and alkali metals—Dissolve 2.0 g of Zinc Sulfate Hydrate in 150 mL of water, add a suitable amount of ammonium sulfide TS to complete the precipita-tion, and add water to make exactly 200 mL. Shake well, and filter through a dry filter paper. Discard the first 20 mL of the filtrate, take exactly 100 mL of the subsequent filtrate, evaporate to dryness, and ignite as directed under Residue on Ignition <2.44>: the mass of the residue is not more than 5.0 mg. Arsenic <1.11>—Prepare the test solution with 1.0 g of Zinc Sulfate Hydrate according to Method 1, and per-form the test (not more than 2 ppm). Loss on drying <2.41> Not less than 35.5z and not more than 38.5z (1 g, 1059C, 3 hours).
Weigh accurately about 0.3 g of Zinc Sulfate Hy-drate, and dissolve in water to make exactly 100 mL. Meas-ure exactly 25 mL of this solution, add 100 mL of water and 2 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.01 mol/L disodium dihydoro-gen ethylenediamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). ‌Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS‌ ＝ 2.876 mg of ZnSO4.7H2O
Zinc Sulfate Ophthalmic Solution contains not less than 0.
Zinc Sulfate Ophthalmic Solution is a clear, colorless liquid.
(1) Zinc Sulfate Ophthalmic Solution re-sponds to Qualitative Tests <1.09> for zinc salt. Zinc Sulfate Ophthalmic Solution responds to Quali-tative Tests <1.09> for borate. Zinc Sulfate Ophthalmic Solution responds to Quali-tative Tests <1.09> for chloride.
Pipet 25 mL of Zinc Sulfate Ophthalmic Solution, add 100 mL of water and 2 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.01 mol/L disodium dihydrogen ethylenediamine tetraace-tate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.876 mg of ZnSO4.7H2O
Zolpidem Tartrate Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of zolpidem tartrate [(C19H21N3O)2.C4H6O6: 764.87]. Method of preparation Prepare as directed under Tablets, with Zolpidem Tartrate. Identification To 1 tablet of Zolpidem Tartrate Tablets add 100 mL of 0.1 mol/L hydrochloric acid TS, shake for 30 minutes, and filter. Discard the first 20 mL of the filtrate, to a volume of the subsequent filtrate, equivalent to 1 mg of Zolpidem Tartrate, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 235 nm and 239 nm and between 292 nm and 296 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Zolpidem Tartrate Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disintegrate the tablet by shaking for 15 minutes. Add 2V/5 mL of methanol, then add exactly V/10 mL of the internal standard solution, shake for 15 minutes, and add methanol to make V mL so that each mL contains about 0.
To 1 tablet of Zolpidem Tartrate Tablets add 100 mL of 0.1 mol/L hydrochloric acid TS, shake for 30 minutes, and filter. Discard the first 20 mL of the filtrate, to a volume of the subsequent filtrate, equivalent to 1 mg of Zolpidem Tartrate, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>: it exhibits maxima between 235 nm and 239 nm and between 292 nm and 296 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirements of the Content uniformity test. To 1 tablet of Zolpidem Tartrate Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disintegrate the tablet by shaking for 15 minutes. Add 2V/5 mL of methanol, then add exactly V/10 mL of the internal standard solution, shake for 15 minutes, and add methanol to make V mL so that each mL contains about 0.1 mg of zolpidem tartrate [(C19H21N3O)2.C4H6O6]. Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 25 mg of zolpidem tartrate for assay (separately determine the water <2.48> in the same manner as Zolpidem Tartrate), and dissolve in 25 mL of 0.1 mol/L hy-drochloric acid TS, add exactly 25 mL of the internal stand-ard solution, then add methanol to make 250 mL, and use this solution as the standard solution. Then, proceed as di-rected in the Assay. Amount (mg) of zolpidem tartrate [(C19H21N3O)2.C4H6O6] ＝ MS × QT/QS × V/250 MS: Amount (mg) of zolpidem tartrate for assay taken, calculated on the anhydrous basis Internal standard solution—A solution of benzyl parahy-droxybenzoate in methanol (1 in 1000). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 15 minutes of Zolpidem Tartrate Tablets is not less than 80z. Start the test with 1 tablet of Zolpidem Tartrate Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add 2nd fluid for dissolution test to make exactly V? mL so that each mL contains about 2.8 mg of zolpidem tartrate [(C19H21N3O)2.C4H6O6], and use this solution as the sample solution. Separately, weigh accurately about 22 mg of zolpidem tartrate for assay (separately determine the water <2.48> in the same manner as Zolpidem Tartrate), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 200 mL. Pipet 25 mL of this solution, add 2nd fluid for dissolution test to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 242 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using diluted 2nd fluid for dissolution test (1 in 2) as the blank. Dissolution rate (z) with respect to the labeled amount of zolpidem tartrate [(C19H21N3O)2.C4H6O6] ＝ MS × AT/AS × V?/V × 1/C × 45/4 MS: Amount (mg) of zolpidem tartrate for assay taken, calculated on the anhydrous basis C: Labeled amount (mg) of zolpidem tartrate [(C19H21N3O)2.C4H6O6] in 1 tablet
To 20 Zolpidem Tartrate Tablets add V/10 mL of 0.1 mol/L hydrochloric acid TS, and disintegrate the tablet by shaking for 15 minutes. Add 2V/5 mL of methanol, then add exactly V/10 mL of the internal standard solution, shake for 15 minutes, and add methanol to make V mL so that each mL contains about 1 mg of zolpidem tartrate [(C19H21N3O)2.C4H6O6]. Centrifuge this solution, add to 1 ‌mL of the supernatant liquid add a mixture of methanol and 0.1 mol/L hydrochloric acid TS (9:1) to make 10 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of zolpidem tartrate for assay (separately determine the water <2.48> in the same manner as Zolpidem Tartrate), and dissolve in 25 mL of 0.1 mol/L hy-drochloric acid TS, add exactly 2.5 mL of the internal stand-ard solution, then add methanol to make 250 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01>, and calculate the ratios, QT and QS, of the peak area of zolpidem to that of the internal standard. Amount (mg) of zolpidem tartrate [(C19H21N3O)2.C4H6O6] in 1 tablet of Zolpidem Tartrate Tablets ＝ MS × QT/QS × V/500 Zonisamide
Zonisamide Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of zonisamide (C8H8N2O3S: 212.23). Method of preparation Prepare as directed under Tablets, with Zonisamide. Identification To 5 mL of the sample solution obtained in the Assay add 5 mL of methanol. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 237 nm and 241 nm, between 243 nm and 247 nm, and between 282 nm and 286 nm. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Zonisamide Tablets add V/25 mL of water, disintegrate completely by sonicating, add 7V/10 mL of methanol, and shake for 15 minutes. Add methanol to make exactly V mL so that each mL contains about 0.
To 5 mL of the sample solution obtained in the Assay add 5 mL of methanol. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits maxima be-tween 237 nm and 241 nm, between 243 nm and 247 nm, and between 282 nm and 286 nm. Uniformity of dosage unit <6.02> Perform the Mass varia-tion test, or the Content uniformity test according to the fol-lowing method: it meets the requirement. To 1 tablet of Zonisamide Tablets add V/25 mL of water, disintegrate completely by sonicating, add 7V/10 mL of methanol, and shake for 15 minutes. Add methanol to make exactly V mL so that each mL contains about 0.5 mg of zonisamide (C8H8N2O3S). Centrifuge this solution, pipet 3 mL of the supernatant liquid, add methanol to make exactly 50 mL, and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of zonisamide (C H N O S) test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac- ＝ MS × AT/AS × V/75 8 8 2 3 cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of zonisamide to that of the in-ternal standard. Amount (mg) of zonisamide (C8H8N2O3S) ＝ MS × QT/QS × 2 MS: Amount (mg) of Zonisamide RS taken Internal standard solution—A solution of 4-aminoacetophe-none in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 239 nm). Column: A stainless steel column 5 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water and tetrahydrofuran (5:1). Flow rate: Adjust so that the retention time of zonisamide is about 11 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and zonisamide are eluted in this order with the resolution between these peaks being not MS: Amount (mg) of Zonisamide RS taken Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of water as the dissolution medium, the dissolution rate in 45 minutes of 25-mg tablet is not less than 75z, and those in 10 minutes and 45 minutes of 100-mg tablet are not more than 65z and not less than 70z, respectively. Start the test with 1 tablet of Zonisamide Tablets. In the case of 25-mg tablets, withdraw not less than 20 mL of the medium at the specified minutes after starting the test. In the case of 100-mg tablets, withdraw exactly 20 mL of the me-dium at the specified minutes after starting the test, and supply exactly 20 mL of water warmed to 37 ± 0.59C imme-diately after withdrawing of the medium every time. Filter these media through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add water to make exactly V? mL so that each mL contains about 22 mg of zonisamide (C8H8N2O3S), and use this solution as the sample solution. Separately, weigh accurately about 22 mg of Zonisamide RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 50 mL, and use this so-lution as the standard solution. Determine the absorbances, AT(n) and AS, of the sample solution and standard solution at 285 nm as directed under Ultraviolet-visible Spectropho- ‌tometry <2.24>. Dissolution rate (z) with respect to the labeled amount of zonisamide (C H N O S) on the nth medium withdrawing tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. (n ＝ 1, 2) 8 8 2 3 (2) Determine the infrared absorption spectrum of Zopiclone as directed in the potassium bromide disk method S ＋ ∑ ＝ M ×AT(n) n－1 ØAT(i ) × 1 »× V? × 1 × 90 under Infrared Spectrophotometry <2.25>, and compare the  AS i＝1 AS 45  V C spectrum with the Reference Spectrum: both spectra exhibit MS: Amount (mg) of Zonisamide RS taken C: Labeled amount (mg) of zonisamide (C8H8N2O3S) in 1 tablet
(1) Heavy metals <1.07>—Proceed with 2.0 g of Zopiclone according to Method 2, and perform the test. Pre-pare the control solution with 4.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 40 mg of Zopiclone in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au- ＝ MS × AT/AS × 2 8 8 2 3 tomatic integration method: the peak areas of related sub-stance A, having the relative retention time of about 0.1 to MS: Amount (mg) of Zonisamide RS taken
Weigh accurately the mass of not less than 20 Zonisamide Tablets, and powder. Weigh accurately a por-tion of the powder, equivalent to about 75 mg of zonisamide (C H N O S), and moisten with 2 mL of water. Add 70 mL similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Zopiclone in 21 times its mass of 2-propanol, heat under a reflux condenser for 15 minutes, and gradually cool to 59C or below. Maintain the temperature for more than 2 hours, filter this solution, wash the residue with 2-propanol, dry, and perform the test using the residue. 8 8 2 3 of methanol, shake for 15 minutes, and add methanol to make exactly 100 mL. Centrifuge this solution, pipet 2 mL of the supernatant liquid, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 38 mg of Zonisamide RS, previously dried at 1059C for 3 hours, dissolve in 1 mL of water and methanol to make exactly 50 mL. Pipet 2 mL of this solu-tion, add methanol to make exactly 50 mL, and use this solu-tion as the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution at 284 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of zonisamide (C H N O S) Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Zopiclone according to Method 2, and perform the test. Pre-pare the control solution with 4.0 mL of Standard Lead So-lution (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 40 mg of Zopiclone in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au- ＝ MS × AT/AS × 2 8 8 2 3 tomatic integration method: the peak areas of related sub-stance A, having the relative retention time of about 0.1 to MS: Amount (mg) of Zonisamide RS taken
Zopiclone Tablets contain not less than 95.0z and not more than 105.0z of the labeled amount of zopiclone (C17H17ClN6O3: 388.81). Method of preparation Prepare as directed under Tablets, with Zopiclone. Identification To a quantity of powdered Zopiclone Tablets, equivalent to 30 mg of Zopiclone, add 60 mL of 0.1 mol/L hydrochloric acid TS, shake thoroughly, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and filter. To 2 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 214 nm and 218 nm, and between 302 nm and 306 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Zopiclone Tablets add the mobile phase, sonicate while occasional shaking to disintegrate the tablet, add the mobile phase to make exactly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/10 mL of the inter-nal standard solution, add the mobile phase to make V? mL so that each mL contains about 0.
To a quantity of powdered Zopiclone Tablets, equivalent to 30 mg of Zopiclone, add 60 mL of 0.1 mol/L hydrochloric acid TS, shake thoroughly, add 0.1 mol/L hydrochloric acid TS to make 100 mL, and filter. To 2 mL of the filtrate add 0.1 mol/L hydrochloric acid TS to make 50 mL. Determine the absorption spectrum of this so-lution as directed under Ultraviolet-visible Spectrophotome-try <2.24>: it exhibits maxima between 214 nm and 218 nm, and between 302 nm and 306 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Zopiclone Tablets add the mobile phase, sonicate while occasional shaking to disintegrate the tablet, add the mobile phase to make exactly 50 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/10 mL of the inter-nal standard solution, add the mobile phase to make V? mL so that each mL contains about 0.1 mg of zopiclone (C17H17ClN6O3), and use this solution as the sample solution. Then, proceed as directed in the Assay. Amount (mg) of zopiclone (C17H17ClN6O3) ＝ MS × QT/QS × V?/V × 1/10 MS: Amount (mg) of zopiclone for assay taken Internal standard solution—A solution of salicylic acid in the mobile phase (1 in 800). Dissolution <6.10> When the test is performed at 50 revolu-tions per minute according to the Paddle method, using 900 mL of 0.05 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) as the dissolution medium, the dissolution rate in 30 minutes of Zopiclone Tablets is not less than 80z. Start the test with 1 tablet of Zopiclone Tablets, withdraw not less than 20 mL of the medium at the specified minute after starting the test, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard not less than 10 mL of the first filtrate, pipet V mL of the subsequent filtrate, add the dissolution medium to make exactly V? mL so that each mL contains about 8.3 mg of zopiclone (C17H17ClN6O3), and use this solution as the sample solution. Separately, weigh accurately about 21 mg of zopiclone for assay (separately determine the loss on drying <2.41> in the same conditions as Zopiclone), and dissolve in the dissolu-tion medium to make exactly 100 mL. Pipet 4 mL of this so-lution, add the dissolution medium to make exactly 100 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sample solution and stand-ard solution at 304 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Dissolution rate (z) with respect to the labeled amount of zopiclone (C17H17ClN6O3) ＝ MS × AT/AS × V?/V × 1/C × 36 MS: Amount (mg) of zopiclone for assay taken, calculated on the dried basis C: Labeled amount (mg) of zopiclone (C17H17ClN6O3) in 1 tablet
To 20 tablets of Zopiclone Tablets add the mobile phase, sonicate while occasional shaking to disintegrate the tablets, add the mobile phase to make exactly 500 mL, and filter through a membrane filter with a pore size not exceeding 0.45 mm. Discard the first 10 mL of the filtrate, pipet V mL of the subsequent filtrate, add exactly V?/10 mL of the internal standard solution, add the mobile phase to make V? mL so that each mL contains about 0.1 mg of zopiclone (C17H17ClN6O3), and use this solution as the sam-ple solution. Separately, weigh accurately about 50 mg of zopiclone for assay (separately determine the loss on drying <2.41> in the same conditions as Zopiclone), and dissolve in the mobile phase to make exactly 20 mL. Pipet 4 mL of this solution, add exactly 10 mL of the internal standard solu-tion, add the mobile phase to make 100 mL, and use this so-lution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of zopiclone to that of the internal standard. Amount (mg) of zopiclone (C17H17ClN6O3) in 1 tablet of Zopiclone Tablet ＝ MS × QT/QS × V?/V × 1/20 MS: Amount (mg) of zopiclone for assay taken, calculated on the dried basis Internal standard solution—A solution of salicylic acid in the mobile phase (1 in 800). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 304 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: To 378 mL of diluted acetic acid (100) (57 in 5000), add 222 mL of a solution of sodium acetate trihy-drate (17 in 625), and add 400 mL of acetonitrile. Flow rate: Adjust so that the retention time of zopiclone is about 9.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and zopiclone are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of zopiclone to that of the internal standard is not more than 1.0z.
Japanese Ministry of Health, Labour and Welfare
Japanese Pharmacopoeia XVIII - Official Publication
Chemical structure image from official JP XVIII monograph
Chemical Structure - Dihydroergotoxine Mesilate
化学構造 - ジヒドロエルゴトキシンメシル酸塩
Image 2 - Insulin Aspart (Genetical Recombination)
画像2 - インスリン アスパルト(遺伝子組換え)
Image 3 - Insulin Aspart (Genetical Recombination)
画像3 - インスリン アスパルト(遺伝子組換え)
Image 4 - Insulin Aspart (Genetical Recombination)
画像4 - インスリン アスパルト(遺伝子組換え)
Chemical Structure - Insulin Aspart (Genetical Recombination)
化学構造 - インスリン アスパルト(遺伝子組換え)
Image 2 - Insulin Glargine (Genetical Recombination)
画像2 - インスリン グラルギン（遺伝子組換え）
Image 3 - Insulin Glargine (Genetical Recombination)
画像3 - インスリン グラルギン（遺伝子組換え）
Image 4 - Insulin Glargine (Genetical Recombination)
画像4 - インスリン グラルギン（遺伝子組換え）
Image 5 - Insulin Glargine (Genetical Recombination)
画像5 - インスリン グラルギン（遺伝子組換え）
Chemical Structure - Insulin Glargine (Genetical Recombination)
化学構造 - インスリン グラルギン（遺伝子組換え）
化学構造 - インスリン ヒト（遺伝子組換え）
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アレンドロン酸ナトリウム水和物
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Chemical Structure - Alprostadil Alfadex
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Chemical Structure - Aprindine Hydrochloride
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化学構造 - アスポキシシリン水和物
画像2 - アトルバスタチンカルシウム水和物
アトルバスタチンカルシウム水和物
画像3 - アトルバスタチンカルシウム水和物
画像4 - アトルバスタチンカルシウム水和物
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Chemical Structure - Benoxinate Hydrochloride
化学構造 - オキシブプロカイン塩酸塩
Chemical Structure - Benserazide Hydrochloride
Chemical Structure - Benzethonium Chloride
Chemical Structure - Benzylpenicillin Benzathine Hydrate
化学構造 - ベンジルペニシリンベンザチン水和物
Chemical Structure - Bepotastine Besilate
化学構造 - ベポタスチンベシル酸塩
Chemical Structure - Berberine Chloride Hydrate
化学構造 - ベルベリン塩化物水和物
Chemical Structure - Betahistine Mesilate
化学構造 - ベタヒスチンメシル酸塩
Chemical Structure - Betamethasone Dipropionate
化学構造 - ベタメタゾンジプロピオン酸エステル
Chemical Structure - Betamethasone Sodium Phosphate
化学構造 - ベタメタゾンリン酸エステルナトリウム
ベタメタゾンリン酸エステルナトリウム
Chemical Structure - Betamethasone Valerate
化学構造 - ベタメタゾン吉草酸エステル
Chemical Structure - Betaxolol Hydrochloride
Chemical Structure - Bethanechol Chloride
Chemical Structure - Biperiden Hydrochloride
Chemical Structure - Bisoprolol Fumarate
化学構造 - ビソプロロールフマル酸塩
Chemical Structure - Bleomycin Hydrochloride
Chemical Structure - Bromfenac Sodium Hydrate
化学構造 - ブロムフェナクナトリウム水和物
ブロムフェナクナトリウム水和物
Chemical Structure - Bromhexine Hydrochloride
Chemical Structure - Bromocriptine Mesilate
化学構造 - ブロモクリプチンメシル酸塩
Chemical Structure - Bucumolol Hydrochloride
Chemical Structure - Bufetolol Hydrochloride
Chemical Structure - Buformin Hydrochloride
Chemical Structure - Bunazosin Hydrochloride
Chemical Structure - Bupivacaine Hydrochloride Hydrate
化学構造 - ブピバカイン塩酸塩水和物
Chemical Structure - Bupranolol Hydrochloride
Chemical Structure - Buprenorphine Hydrochloride
化学構造 - ブプレノルフィン塩酸塩
Chemical Structure - Butyl Parahydroxybenzoate
化学構造 - パラオキシ安息香酸ブチル
化学構造 - カモスタットメシル酸塩
Chemical Structure - Capsicum and Salicylic Acid Spirit
化学構造 - トウガラシ・サリチル酸精
Chemical Structure - Carbazochrome Sodium Sulfonate Hydrate
化学構造 - カルバゾクロムスルホン酸ナトリウム水和物
カルバゾクロムスルホン酸ナトリウム水和物
Chemical Structure - Carteolol Hydrochloride
化学構造 - セファロチンナトリウム
Chemical Structure - Cefatrizine Propylene Glycolate
化学構造 - セファトリジンプロピレングリコール
化学構造 - セファゾリンナトリウム
Chemical Structure - Cefazolin Sodium Hydrate
化学構造 - セファゾリンナトリウム水和物
Chemical Structure - Cefbuperazone Sodium
化学構造 - セフブペラゾンナトリウム
Chemical Structure - Cefcapene Pivoxil Hydrochloride Hydrate
化学構造 - セフカペン ピボキシル塩酸塩水和物
化学構造 - セフジトレン ピボキシル
Image 2 - Cefepime Dihydrochloride Hydrate
Image 3 - Cefepime Dihydrochloride Hydrate
Image 4 - Cefepime Dihydrochloride Hydrate
Chemical Structure - Cefepime Dihydrochloride Hydrate
化学構造 - セフェピム塩酸塩水和物
Chemical Structure - Cefmenoxime Hydrochloride
化学構造 - セフメタゾールナトリウム
Chemical Structure - Cefminox Sodium Hydrate
化学構造 - セフミノクスナトリウム水和物
化学構造 - セフォジジムナトリウム
Chemical Structure - Cefoperazone Sodium
化学構造 - セフォペラゾンナトリウム
化学構造 - セフォタキシムナトリウム
Chemical Structure - Cefotiam Hexetil Hydrochloride
化学構造 - セフォチアム ヘキセチル塩酸塩
Chemical Structure - Cefotiam Hydrochloride
Chemical Structure - Cefozopran Hydrochloride
化学構造 - セフピラミドナトリウム
画像2 - セフポドキシム プロキセチル
画像3 - セフポドキシム プロキセチル
画像4 - セフポドキシム プロキセチル
Chemical Structure - Cefpodoxime Proxetil
化学構造 - セフポドキシム プロキセチル
Chemical Structure - Cefroxadine Hydrate
化学構造 - セフスロジンナトリウム
Chemical Structure - Ceftazidime Hydrate
化学構造 - セフチゾキシムナトリウム
Chemical Structure - Ceftriaxone Sodium Hydrate
化学構造 - セフトリアキソンナトリウム水和物
セフトリアキソンナトリウム水和物
化学構造 - セフロキシム アキセチル
Chemical Structure - Cellulose Acetate Phthalate
Image 2 - Celmoleukin (Genetical Recombination)
画像2 - セルモロイキン（遺伝子組換え）
Image 3 - Celmoleukin (Genetical Recombination)
画像3 - セルモロイキン（遺伝子組換え）
Image 4 - Celmoleukin (Genetical Recombination)
画像4 - セルモロイキン（遺伝子組換え）
Chemical Structure - Celmoleukin (Genetical Recombination)
化学構造 - セルモロイキン（遺伝子組換え）
Chemical Structure - Cetirizine Hydrochloride
Chemical Structure - Cetotiamine Hydrochloride Hydrate
化学構造 - セトチアミン塩酸塩水和物
Chemical Structure - Cetraxate Hydrochloride
Chemical Structure - Chenodeoxycholic Acid
Chemical Structure - Chloramphenicol Palmitate
化学構造 - クロラムフェニコールパルミチン酸エステル
Chemical Structure - Chloramphenicol Sodium Succinate
化学構造 - クロラムフェニコールコハク酸エステルナトリウム
クロラムフェニコールコハク酸エステルナトリウム
Chemical Structure - Chlorhexidine Hydrochloride
化学構造 - クロルヘキシジン塩酸塩
Chemical Structure - Chlormadinone Acetate
化学構造 - クロルマジノン酢酸エステル
Chemical Structure - Chlorphenesin Carbamate
化学構造 - クロルフェネシンカルバミン酸エステル
Chemical Structure - Chlorpheniramine Maleate
化学構造 - クロルフェニラミンマレイン酸塩
Chemical Structure - Chlorpromazine Hydrochloride
化学構造 - クロルプロマジン塩酸塩
Chemical Structure - Chorionic Gonadotrophin
化学構造 - ヒト絨毛性性腺刺激ホルモン
Chemical Structure - Cibenzoline Succinate
化学構造 - シベンゾリンコハク酸塩
化学構造 - シラスタチンナトリウム
Chemical Structure - Cinchocaine Hydrochloride
Chemical Structure - Ciprofloxacin Hydrochloride Hydrate
化学構造 - シプロフロキサシン塩酸塩水和物
Chemical Structure - Citric Acid Hydrate
化学構造 - クレボプリドリンゴ酸塩
Chemical Structure - Clemastine Fumarate
化学構造 - クレマスチンフマル酸塩
Chemical Structure - Clindamycin Hydrochloride
化学構造 - クリンダマイシン塩酸塩
Chemical Structure - Clindamycin Phosphate
化学構造 - クリンダマイシンリン酸エステル
Chemical Structure - Clobetasol Propionate
化学構造 - クロベタゾールプロピオン酸エステル
Chemical Structure - Clocapramine Hydrochloride Hydrate
化学構造 - クロカプラミン塩酸塩水和物
Chemical Structure - Clofedanol Hydrochloride
化学構造 - クロフェダノール塩酸塩
化学構造 - クロミフェンクエン酸塩
Chemical Structure - Clomipramine Hydrochloride
Chemical Structure - Clonidine Hydrochloride
Chemical Structure - Cloperastine Fendizoate
化学構造 - クロペラスチンフェンジゾ酸塩
Chemical Structure - Cloperastine Hydrochloride
Chemical Structure - Clopidogrel Sulfate
Chemical Structure - Clorazepate Dipotassium
化学構造 - クロラゼプ酸二カリウム
Chemical Structure - Cloxacillin Sodium Hydrate
化学構造 - クロキサシリンナトリウム水和物
クロキサシリンナトリウム水和物
Chemical Structure - Codeine Phosphate Hydrate
化学構造 - コデインリン酸塩水和物
Chemical Structure - Colistin Sodium Methanesulfonate
化学構造 - コリスチンメタンスルホン酸ナトリウム
コリスチンメタンスルホン酸ナトリウム
Chemical Structure - Compound Acrinol and Zinc Oxide Oil
化学構造 - 複方アクリノール・チンク油
Chemical Structure - Compound Iodine Glycerin
化学構造 - 複方ヨード・グリセリン
Chemical Structure - ‌Compound Methyl Salicylate Spirit‌
化学構造 - 複方サリチル酸メチル精
Chemical Structure - Compound Salicylic Acid Spirit
Chemical Structure - Concentrated Glycerol
Chemical Structure - Containers and storage Containers—Hermetic containers. Storage—Light-resistant.
Chemical Structure - Containers and storage Containers—Well-closed contain-ers.
化学構造 - コルチゾン酢酸エステル
Chemical Structure - Croconazole Hydrochloride
Chemical Structure - Cyclopentolate Hydrochloride
化学構造 - シクロペントラート塩酸塩
Chemical Structure - Cyclophosphamide Hydrate
化学構造 - シクロホスファミド水和物
Chemical Structure - Cyproheptadine Hydrochloride Hydrate
化学構造 - シプロヘプタジン塩酸塩水和物
Chemical Structure - d-Chlorpheniramine Maleate
化学構造 - d-クロルフェニラミンマレイン酸塩
Chemical Structure - Daiokanzoto Extract
Chemical Structure - Dantrolene Sodium Hydrate
化学構造 - ダントロレンナトリウム水和物
Chemical Structure - Daunorubicin Hydrochloride
Chemical Structure - Deferoxamine Mesilate
化学構造 - デフェロキサミンメシル酸塩
Chemical Structure - Demethylchlortetracycline Hydrochloride
化学構造 - デメチルクロルテトラサイクリン塩酸塩
Chemical Structure - Dental Iodine Glycerin
化学構造 - 歯科用ヨード・グリセリン
Chemical Structure - Dental Paraformaldehyde Paste
化学構造 - 歯科用パラホルムパスタ
Chemical Structure - Dental Phenol with Camphor
化学構造 - 歯科用フェノール・カンフル
Chemical Structure - ‌Dental Triozinc Paste‌
化学構造 - 歯科用トリオジンクパスタ
Chemical Structure - Dextromethorphan Hydrobromide Hydrate
化学構造 - デキストロメトルファン臭化水素酸塩水和物
化学構造 - ジクロフェナクナトリウム
Chemical Structure - Dicloxacillin Sodium Hydrate
化学構造 - ジクロキサシリンナトリウム水和物
ジクロキサシリンナトリウム水和物
Chemical Structure - Diethylcarbamazine Citrate
化学構造 - ジエチルカルバマジンクエン酸塩
Chemical Structure - Difenidol Hydrochloride
Chemical Structure - Diflorasone Diacetate
化学構造 - ジフロラゾン酢酸エステル
Chemical Structure - Diflucortolone Valerate
化学構造 - ジフルコルトロン吉草酸エステル
Chemical Structure - Dihydrocodeine Phosphate
化学構造 - ジヒドロコデインリン酸塩
Chemical Structure - Dihydroergotamine Mesilate
化学構造 - ジヒドロエルゴタミンメシル酸塩
Chemical Structure - ‌Dilazep Hydrochloride Hydrate
Chemical Structure - Diltiazem Hydrochloride
Chemical Structure - Dilute Iodine Tincture
Chemical Structure - Dimemorfan Phosphate
化学構造 - ジメモルファンリン酸塩
Chemical Structure - Diphenhydramine Hydrochloride
化学構造 - ジフェンヒドラミン塩酸塩
Chemical Structure - Diphenhydramine, Phenol and Zinc Oxide Liniment
化学構造 - ジフェンヒドラミン・フェノール・亜鉛華リニメント
Diphenhydramine, Phenol and Zinc Oxide Liniment
ジフェンヒドラミン・フェノール・亜鉛華リニメント
Chemical Structure - Disodium Edetate Hydrate
化学構造 - エデト酸ナトリウム水和物
Chemical Structure - ‌dl-Methylephedrine Hydrochloride
化学構造 - dl-メチルエフェドリン塩酸塩
Chemical Structure - Dobutamine Hydrochloride
Chemical Structure - Donepezil Hydrochloride
Chemical Structure - Dopamine Hydrochloride
Chemical Structure - Doxapram Hydrochloride Hydrate
化学構造 - ドキサプラム塩酸塩水和物
化学構造 - ドキサゾシンメシル酸塩
Chemical Structure - Doxorubicin Hydrochloride
Chemical Structure - Doxycycline Hydrochloride Hydrate
化学構造 - ドキシサイクリン塩酸塩水和物
Chemical Structure - Ecabet Sodium Hydrate
化学構造 - エカベトナトリウム水和物
化学構造 - エコチオパートヨウ化物
Chemical Structure - Edrophonium Chloride
Chemical Structure - Emedastine Fumarate
化学構造 - エメダスチンフマル酸塩
化学構造 - エナラプリルマレイン酸塩
化学構造 - エンビオマイシン硫酸塩
Chemical Structure - Eperisone Hydrochloride
Chemical Structure - Ephedrine Hydrochloride
Chemical Structure - Epirubicin Hydrochloride
画像2 - エポエチン アルファ（遺伝子組換え）
画像3 - エポエチン アルファ（遺伝子組換え）
画像4 - エポエチン アルファ（遺伝子組換え）
画像5 - エポエチン アルファ（遺伝子組換え）
画像6 - エポエチン アルファ（遺伝子組換え）
画像7 - エポエチン アルファ（遺伝子組換え）
化学構造 - エポエチン アルファ（遺伝子組換え）
化学構造 - エポエチン ベータ（遺伝子組換え）
Chemical Structure - Ergometrine Maleate
化学構造 - エルゴメトリンマレイン酸塩
Chemical Structure - Ergotamine Tartrate
Chemical Structure - Erythromycin Ethylsuccinate
化学構造 - エリスロマイシンエチルコハク酸エステル
Chemical Structure - Erythromycin Lactobionate
化学構造 - エリスロマイシンラクトビオン酸塩
Chemical Structure - Erythromycin Stearate
化学構造 - エリスロマイシンステアリン酸塩
化学構造 - エストラジオール安息香酸エステル
Chemical Structure - Ethacridine Lactate
Chemical Structure - Ethambutol Hydrochloride
化学構造 - エチニルエストラジオール
Chemical Structure - Ethyl L-Cysteine Hydrochloride
化学構造 - L-エチルシステイン塩酸塩
Chemical Structure - Ethyl Parahydroxybenzoate
化学構造 - パラオキシ安息香酸エチル
Chemical Structure - Ethylmorphine Hydrochloride Hydrate
化学構造 - エチルモルヒネ塩酸塩水和物
Chemical Structure - Etidronate Disodium
化学構造 - エチドロン酸二ナトリウム
Chemical Structure - Etilefrine Hydrochloride
画像2 - ファロペネムナトリウム水和物
画像3 - ファロペネムナトリウム水和物
Chemical Structure - Faropenem Sodium Hydrate
化学構造 - ファロペネムナトリウム水和物
化学構造 - フェンタニルクエン酸塩
Chemical Structure - Fexofenadine Hydrochloride
化学構造 - フェキソフェナジン塩酸塩
画像2 - フィルグラスチム（遺伝子組換え）
画像3 - フィルグラスチム（遺伝子組換え）
画像4 - フィルグラスチム（遺伝子組換え）
画像5 - フィルグラスチム（遺伝子組換え）
画像6 - フィルグラスチム（遺伝子組換え）
画像7 - フィルグラスチム（遺伝子組換え）
化学構造 - フィルグラスチム（遺伝子組換え）
Chemical Structure - Flavin Adenine Dinucleotide Sodium
化学構造 - フラビンアデニンジヌクレオチドナトリウム
フラビンアデニンジヌクレオチドナトリウム
Chemical Structure - Flavoxate Hydrochloride
化学構造 - フロモキセフナトリウム
Chemical Structure - Fludrocortisone Acetate
化学構造 - フルドロコルチゾン酢酸エステル
Chemical Structure - Fluocinolone Acetonide
化学構造 - フルオシノロンアセトニド
化学構造 - フルオレセインナトリウム
Chemical Structure - Fluphenazine Enanthate
化学構造 - フルフェナジンエナント酸エステル
Chemical Structure - Flurazepam Hydrochloride
Chemical Structure - Fluvoxamine Maleate
化学構造 - フルボキサミンマレイン酸塩
Chemical Structure - Foeniculated Ammonia Spirit
化学構造 - アンモニア・ウイキョウ精
Chemical Structure - Formoterol Fumarate Hydrate
化学構造 - ホルモテロールフマル酸塩水和物
Chemical Structure - Fosfomycin Calcium Hydrate
化学構造 - ホスホマイシンカルシウム水和物
ホスホマイシンカルシウム水和物
化学構造 - ホスホマイシンナトリウム
Chemical Structure - Fursultiamine Hydrochloride
化学構造 - フルスルチアミン塩酸塩
化学構造 - ガベキサートメシル酸塩
Chemical Structure - Gati‰oxacin Hydrate
化学構造 - ガチフロキサシン水和物
Chemical Structure - Glacial Acetic Acid
Chemical Structure - Glucagon (Genetical Recombination)
化学構造 - グルカゴン(遺伝子組換え)
Chemical Structure - Gonadorelin Acetate
Chemical Structure - Goshajinkigan Extract
Chemical Structure - Guanethidine Sulfate
Chemical Structure - Hachimijiogan Extract
Chemical Structure - Homatropine Hydrobromide
化学構造 - ホマトロピン臭化水素酸塩
Chemical Structure - Homochlorcyclizine Hydrochloride
化学構造 - ホモクロルシクリジン塩酸塩
Chemical Structure - Human Menopausal Gonadotrophin
化学構造 - ヒト下垂体性性腺刺激ホルモン
Chemical Structure - Hydralazine Hydrochloride
Chemical Structure - Hydrochloric Acid Lemonade
Chemical Structure - Hydrochlorothiazide
Chemical Structure - Hydrocortisone Acetate
化学構造 - ヒドロコルチゾン酢酸エステル
Chemical Structure - Hydrocortisone Butyrate
化学構造 - ヒドロコルチゾン酪酸エステル
Chemical Structure - Hydrocortisone Sodium Phosphate
化学構造 - ヒドロコルチゾンリン酸エステルナトリウム
ヒドロコルチゾンリン酸エステルナトリウム
Chemical Structure - Hydrocortisone Sodium Succinate
化学構造 - ヒドロコルチゾンコハク酸エステルナトリウム
ヒドロコルチゾンコハク酸エステルナトリウム
Chemical Structure - Hydrocortisone Succinate
化学構造 - ヒドロコルチゾンコハク酸エステル
Chemical Structure - Hydrocotarnine Hydrochloride Hydrate
化学構造 - ヒドロコタルニン塩酸塩水和物
Chemical Structure - Hydrophilic Petrolatum
Chemical Structure - Hydroxocobalamin Acetate
化学構造 - ヒドロキソコバラミン酢酸塩
Chemical Structure - Hydroxyethylcellulose
化学構造 - ヒドロキシエチルセルロース
Chemical Structure - Hydroxyzine Hydrochloride
Chemical Structure - Hydroxyzine Pamoate
化学構造 - ヒドロキシジンパモ酸塩
Chemical Structure - Hypromellose Phthalate
化学構造 - ヒプロメロースフタル酸エステル
化学構造 - イブプロフェンピコノール
Chemical Structure - Idarubicin Hydrochloride
Chemical Structure - Ifenprodil Tartrate
化学構造 - イフェンプロジル酒石酸塩
画像10 - ジヒドロコデインリン酸塩散 10
Chemical Structure - Imidapril Hydrochloride
Chemical Structure - Imipramine Hydrochloride
Chemical Structure - Indenolol Hydrochloride
Chemical Structure - Iodine, Salicylic Acid and Phenol Spirit
化学構造 - ヨード・サリチル酸・フェノール精
Chemical Structure - Ipratropium Bromide Hydrate
化学構造 - イプラトロピウム臭化物水和物
Image 2 - Irinotecan Hydrochloride Hydrate
Image 3 - Irinotecan Hydrochloride Hydrate
Image 4 - Irinotecan Hydrochloride Hydrate
Image 5 - Irinotecan Hydrochloride Hydrate
Chemical Structure - Irinotecan Hydrochloride Hydrate
化学構造 - イリノテカン塩酸塩水和物
Chemical Structure - Irsogladine Maleate
化学構造 - イルソグラジンマレイン酸塩
Chemical Structure - ‌Isosorbide Dinitrate
Chemical Structure - Isosorbide Mononitrate 70%/Lactose 30%
化学構造 - 70%一硝酸イソソルビド乳糖末
Chemical Structure - Isoxsuprine Hydrochloride
Chemical Structure - Josamycin Propionate
化学構造 - ジョサマイシンプロピオン酸エステル
Chemical Structure - Kainic Acid Hydrate
Chemical Structure - Kakkontokasenkyushin'i Extract
Chemical Structure - Kanamycin Monosulfate
Chemical Structure - Keishibukuryogan Extract
Chemical Structure - Ketamine Hydrochloride
化学構造 - ケトチフェンフマル酸塩
Chemical Structure - Kitasamycin Acetate
化学構造 - キタサマイシン酢酸エステル
Chemical Structure - Kitasamycin Tartrate
化学構造 - キタサマイシン酒石酸塩
Chemical Structure - L-Arginine Hydrochloride
Chemical Structure - L-Cysteine Hydrochloride Hydrate
化学構造 - L-システイン塩酸塩水和物
Chemical Structure - L-Histidine Hydrochloride Hydrate
化学構造 - L-ヒスチジン塩酸塩水和物
Chemical Structure - l-Isoprenaline Hydrochloride
Chemical Structure - L-Lysine Hydrochloride
Chemical Structure - Labetalol Hydrochloride
化学構造 - ラタモキセフナトリウム
Chemical Structure - Lenampicillin Hydrochloride
Image 2 - Lenograstim (Genetical Recombination)
画像2 - レノグラスチム（遺伝子組換え）
Image 3 - Lenograstim (Genetical Recombination)
画像3 - レノグラスチム（遺伝子組換え）
Image 4 - Lenograstim (Genetical Recombination)
画像4 - レノグラスチム（遺伝子組換え）
Image 5 - Lenograstim (Genetical Recombination)
画像5 - レノグラスチム（遺伝子組換え）
Image 6 - Lenograstim (Genetical Recombination)
画像6 - レノグラスチム（遺伝子組換え）
Image 7 - Lenograstim (Genetical Recombination)
画像7 - レノグラスチム（遺伝子組換え）
Image 8 - Lenograstim (Genetical Recombination)
画像8 - レノグラスチム（遺伝子組換え）
Chemical Structure - Lenograstim (Genetical Recombination)
化学構造 - レノグラスチム（遺伝子組換え）
Chemical Structure - Leuprorelin Acetate
化学構造 - リュープロレリン酢酸塩
Chemical Structure - Levallorphan Tartrate
化学構造 - レバロルファン酒石酸塩
Chemical Structure - Levofloxacin Hydrate
化学構造 - レボフロキサシン水和物
Chemical Structure - Levomepromazine Maleate
化学構造 - レボメプロマジンマレイン酸塩
Chemical Structure - Levothyroxine Sodium Hydrate
化学構造 - レボチロキシンナトリウム水和物
レボチロキシンナトリウム水和物
化学構造 - リマプロスト アルファデクス
Chemical Structure - Lincomycin Hydrochloride Hydrate
化学構造 - リンコマイシン塩酸塩水和物
Chemical Structure - Liothyronine Sodium
化学構造 - リオチロニンナトリウム
化学構造 - ロベンザリットナトリウム
Chemical Structure - Loxoprofen Sodium Hydrate
化学構造 - ロキソプロフェンナトリウム水和物
ロキソプロフェンナトリウム水和物
Chemical Structure - Lysozyme Hydrochloride
Chemical Structure - ‌Magnesium Sulfate Mixture‌
Chemical Structure - Manidipine Hydrochloride
Chemical Structure - Maprotiline Hydrochloride
Chemical Structure - Meclofenoxate Hydrochloride
化学構造 - メクロフェノキサート塩酸塩
Chemical Structure - Medroxyprogesterone Acetate
化学構造 - メドロキシプロゲステロン酢酸エステル
Chemical Structure - Mefloquine Hydrochloride
Chemical Structure - ‌Mepenzolate Bromide
Chemical Structure - Mepivacaine Hydrochloride
Chemical Structure - Mercaptopurine Hydrate
化学構造 - メルカプトプリン水和物
化学構造 - モンテルカストナトリウム
化学構造 - メテノロン酢酸エステル
Chemical Structure - ‌Metenolone Enanthate‌
化学構造 - メテノロンエナント酸エステル
Chemical Structure - Metformin Hydrochloride
Chemical Structure - Methamphetamine Hydrochloride
化学構造 - メタンフェタミン塩酸塩
Chemical Structure - Methyl Parahydroxybenzoate
化学構造 - パラオキシ安息香酸メチル
Chemical Structure - Methylbenactyzium Bromide
化学構造 - メチルベナクチジウム臭化物
Chemical Structure - Methylergometrine Maleate
化学構造 - メチルエルゴメトリンマレイン酸塩
Chemical Structure - Methylprednisolone Succinate
化学構造 - メチルプレドニゾロンコハク酸エステル
Chemical Structure - Metoprolol Tartrate
化学構造 - メトプロロール酒石酸塩
Chemical Structure - Mexiletine Hydrochloride
Chemical Structure - ‌Miconazole Nitrate
Chemical Structure - Microcrystalline Cellulose
Chemical Structure - Micronomicin Sulfate
化学構造 - ミクロノマイシン硫酸塩
Chemical Structure - Midecamycin Acetate
化学構造 - ミデカマイシン酢酸エステル
Chemical Structure - Minocycline Hydrochloride
Chemical Structure - Mitiglinide Calcium Hydrate
化学構造 - ミチグリニドカルシウム水和物
Chemical Structure - Morphine Hydrochloride Hydrate
Chemical Structure - Morphine Sulfate Hydrate
画像2 - モサプリドクエン酸塩水和物
画像3 - モサプリドクエン酸塩水和物
画像4 - モサプリドクエン酸塩水和物
Chemical Structure - ‌Mosapride Citrate Hydrate
化学構造 - モサプリドクエン酸塩水和物
Chemical Structure - Mukoi-Daikenchuto Extract
化学構造 - 無コウイ大建中湯エキス
画像2 - ムピロシンカルシウム水和物
画像3 - ムピロシンカルシウム水和物
Chemical Structure - Mupirocin Calcium Hydrate
化学構造 - ムピロシンカルシウム水和物
Chemical Structure - Nafamostat Mesilate
化学構造 - ナファモスタットメシル酸塩
Chemical Structure - Naloxone Hydrochloride
Chemical Structure - Naphazoline Hydrochloride
Chemical Structure - Naphazoline Nitrate
Image 2 - Nartograstim (Genetical Recombination)
画像2 - ナルトグラスチム（遺伝子組換え）
Chemical Structure - Nartograstim (Genetical Recombination)
化学構造 - ナルトグラスチム（遺伝子組換え）
Chemical Structure - Natural Aluminum Silicate
化学構造 - 天然ケイ酸アルミニウム
化学構造 - フラジオマイシン硫酸塩
Chemical Structure - Neostigmine Methylsulfate
化学構造 - ネオスチグミンメチル硫酸塩
Chemical Structure - ‌Nicardipine Hydrochloride
Chemical Structure - Nortriptyline Hydrochloride
化学構造 - ノルトリプチリン塩酸塩
Chemical Structure - Noscapine Hydrochloride Hydrate
化学構造 - ノスカピン塩酸塩水和物
Chemical Structure - Nux Vomica Tincture
Chemical Structure - Olmesartan Medoxomil
化学構造 - オルメサルタン メドキソミル
Chemical Structure - Olopatadine Hydrochloride
Chemical Structure - Orange Peel Tincture
Chemical Structure - ‌Orciprenaline Sulfate‌
化学構造 - オルシプレナリン硫酸塩
Chemical Structure - Oxprenolol Hydrochloride
化学構造 - オクスプレノロール塩酸塩
Image 2 - Oxycodone Hydrochloride Hydrate
Chemical Structure - Oxycodone Hydrochloride Hydrate
化学構造 - オキシコドン塩酸塩水和物
Image 2 - ‌Oxytetracycline Hydrochloride
画像2 - オキシテトラサイクリン塩酸塩
Chemical Structure - ‌Oxytetracycline Hydrochloride
化学構造 - オキシテトラサイクリン塩酸塩
Chemical Structure - Pancuronium Bromide
Chemical Structure - Papaverine Hydrochloride
化学構造 - パルナパリンナトリウム
Image 2 - Paroxetine Hydrochloride Hydrate
Image 3 - Paroxetine Hydrochloride Hydrate
Image 4 - Paroxetine Hydrochloride Hydrate
Chemical Structure - Paroxetine Hydrochloride Hydrate
化学構造 - パロキセチン塩酸塩水和物
Chemical Structure - Pas-calcium Hydrate
化学構造 - パラアミノサリチル酸カルシウム水和物
パラアミノサリチル酸カルシウム水和物
Chemical Structure - ‌Pazufloxacin Mesilate
化学構造 - パズフロキサシンメシル酸塩
Chemical Structure - Pemirolast Potassium
Chemical Structure - Penicillin G Potassium
化学構造 - ベンジルペニシリンカリウム
Chemical Structure - Pentobarbital Calcium
化学構造 - ペントバルビタールカルシウム
Chemical Structure - Pentoxyverine Citrate
化学構造 - ペントキシベリンクエン酸塩
Chemical Structure - Perphenazine Maleate
化学構造 - ペルフェナジンマレイン酸塩
Chemical Structure - Pethidine Hydrochloride
Chemical Structure - Phenethicillin Potassium
化学構造 - フェネチシリンカリウム
Chemical Structure - ‌Phenol and Zinc Oxide Liniment‌
化学構造 - フェノール・亜鉛華リニメント
Chemical Structure - Phenolsulfonphthalein
化学構造 - フェノールスルホンフタレイン
Chemical Structure - ‌Phenylephrine Hydrochloride‌
Chemical Structure - Pilocarpine Hydrochloride
Chemical Structure - Pilsicainide Hydrochloride Hydrate
化学構造 - ピルシカイニド塩酸塩水和物
Chemical Structure - Pioglitazone Hydrochloride
Chemical Structure - Pipemidic Acid Hydrate
Chemical Structure - Piperacillin Hydrate
Chemical Structure - Piperacillin Sodium
化学構造 - ピペラシリンナトリウム
化学構造 - ピペラジンアジピン酸塩
Chemical Structure - ‌Piperazine Phosphate Hydrate‌
化学構造 - ピペラジンリン酸塩水和物
Image 2 - ‌Pirenzepine Hydrochloride Hydrate
Chemical Structure - ‌Pirenzepine Hydrochloride Hydrate
化学構造 - ピレンゼピン塩酸塩水和物
画像2 - ピタバスタチンカルシウム水和物
ピタバスタチンカルシウム水和物
画像3 - ピタバスタチンカルシウム水和物
Chemical Structure - Pitavastatin Calcium Hydrate
化学構造 - ピタバスタチンカルシウム水和物
Chemical Structure - Pivmecillinam Hydrochloride
Chemical Structure - ‌Polymixin B Sulfate
Chemical Structure - ‌Potassium Guaiacolsulfonate‌
化学構造 - グアヤコールスルホン酸カリウム
グアヤコールスルホン酸カリウム
Chemical Structure - ‌Prasterone Sodium Sulfate Hydrate‌
化学構造 - プラステロン硫酸エステルナトリウム水和物
プラステロン硫酸エステルナトリウム水和物
化学構造 - プラバスタチンナトリウム
Chemical Structure - Prazosin Hydrochloride
Chemical Structure - Prednisolone Acetate
化学構造 - プレドニゾロン酢酸エステル
Chemical Structure - ‌Prednisolone Sodium Phosphate
化学構造 - プレドニゾロンリン酸エステルナトリウム
プレドニゾロンリン酸エステルナトリウム
Chemical Structure - Prednisolone Succinate
化学構造 - プレドニゾロンコハク酸エステル
Chemical Structure - Procainamide Hydrochloride
化学構造 - プロカインアミド塩酸塩
Chemical Structure - ‌Procaine Hydrochloride‌
Chemical Structure - Procarbazine Hydrochloride
Chemical Structure - Procaterol Hydrochloride Hydrate
化学構造 - プロカテロール塩酸塩水和物
Chemical Structure - ‌Prochlorperazine Maleate‌
化学構造 - プロクロルペラジンマレイン酸塩
Chemical Structure - ‌Promethazine Hydrochloride‌
Chemical Structure - Propafenone Hydrochloride
Chemical Structure - Propantheline Bromide
Chemical Structure - Propiverine Hydrochloride
Chemical Structure - Propranolol Hydrochloride
化学構造 - プロプラノロール塩酸塩
Chemical Structure - Propyl Parahydroxybenzoate
化学構造 - パラオキシ安息香酸プロピル
化学構造 - イソプロピルアンチピリン
Chemical Structure - Protirelin Tartrate Hydrate
化学構造 - プロチレリン酒石酸塩水和物
Chemical Structure - Purified Dehydrocholic Acid
Chemical Structure - Purified Sodium Hyaluronate
化学構造 - 精製ヒアルロン酸ナトリウム
Chemical Structure - Pyridostigmine Bromide
化学構造 - ピリドスチグミン臭化物
Chemical Structure - Pyridoxal Phosphate Hydrate
化学構造 - ピリドキサールリン酸エステル水和物
Chemical Structure - Quetiapine Fumarate
化学構造 - クエチアピンフマル酸塩
Chemical Structure - Quinapril Hydrochloride
Chemical Structure - ‌Quinidine Sulfate Hydrate
Chemical Structure - ‌Quinine Ethyl Carbonate‌
化学構造 - キニーネエチル炭酸エステル
Chemical Structure - Quinine Hydrochloride Hydrate
Chemical Structure - ‌Quinine Sulfate Hydrate
Chemical Structure - ‌Rabeprazole Sodium‌
化学構造 - ラベプラゾールナトリウム
Chemical Structure - Ranitidine Hydrochloride
Chemical Structure - Ribostamycin Sulfate
化学構造 - リボスタマイシン硫酸塩
Chemical Structure - Rilmazafone Hydrochloride Hydrate
化学構造 - リルマザホン塩酸塩水和物
Chemical Structure - Ritodrine Hydrochloride
Chemical Structure - Rosuvastatin Calcium
化学構造 - ロスバスタチンカルシウム
Chemical Structure - Roxatidine Acetate Hydrochloride
化学構造 - ロキサチジン酢酸エステル塩酸塩
画像2 - サッカリンナトリウム水和物
画像3 - サッカリンナトリウム水和物
画像4 - サッカリンナトリウム水和物
Chemical Structure - Saccharin Sodium Hydrate
化学構造 - サッカリンナトリウム水和物
Chemical Structure - Salicylic Acid Adhesive Plaster
Chemical Structure - ‌Salicylic Acid Spirit‌
Chemical Structure - Sarpogrelate Hydrochloride
化学構造 - サルポグレラート塩酸塩
Chemical Structure - Scopolamine Butylbromide
化学構造 - ブチルスコポラミン臭化物
Chemical Structure - Scopolamine Hydrobromide Hydrate
化学構造 - スコポラミン臭化水素酸塩水和物
Chemical Structure - Shakuyakukanzoto Extract
Chemical Structure - Shoseiryuto Extract
画像2 - シタグリプチンリン酸塩水和物
Chemical Structure - Sitagliptin Phosphate Hydrate
化学構造 - シタグリプチンリン酸塩水和物
Chemical Structure - Sivelestat Sodium Hydrate
化学構造 - シベレスタットナトリウム水和物
シベレスタットナトリウム水和物
Chemical Structure - Spectinomycin Hydrochloride Hydrate
化学構造 - スペクチノマイシン塩酸塩水和物
Chemical Structure - ‌Spiramycin Acetate
化学構造 - スピラマイシン酢酸エステル
Chemical Structure - ‌Streptomycin Sulfate
化学構造 - ストレプトマイシン硫酸塩
化学構造 - スクラルファート水和物
化学構造 - スルバクタムナトリウム
Chemical Structure - Sulbenicillin Sodium
化学構造 - スルベニシリンナトリウム
Chemical Structure - Sulfadiazine Silver
化学構造 - スルファメトキサゾール
Chemical Structure - Sulfamonomethoxine Hydrate
化学構造 - スルファモノメトキシン水和物
化学構造 - サラゾスルファピリジン
Chemical Structure - Sulfobromophthalein Sodium
化学構造 - スルホブロモフタレインナトリウム
スルホブロモフタレインナトリウム
Chemical Structure - Sulfur and Camphor Lotion
化学構造 - イオウ・カンフルローション
Chemical Structure - ‌Sulpyrine Hydrate‌
Chemical Structure - Sultamicillin Tosilate Hydrate
化学構造 - スルタミシリントシル酸塩水和物
Chemical Structure - Suxamethonium Chloride Hydrate
化学構造 - スキサメトニウム塩化物水和物
Chemical Structure - Talampicillin Hydrochloride
化学構造 - タモキシフェンクエン酸塩
Chemical Structure - ‌Tamsulosin Hydrochloride
Image 2 - Teceleukin (Genetical Recombination)
Image 3 - Teceleukin (Genetical Recombination)
Image 4 - Teceleukin (Genetical Recombination)
Image 5 - Teceleukin (Genetical Recombination)
Image 6 - Teceleukin (Genetical Recombination)
Chemical Structure - Teceleukin (Genetical Recombination)
化学構造 - テセロイキン（遺伝子組換え）
Chemical Structure - Temocapril Hydrochloride
Chemical Structure - ‌Terbinafine Hydrochloride
Chemical Structure - Terbutaline Sulfate
Chemical Structure - Testosterone Enanthate
化学構造 - テストステロンエナント酸エステル
Chemical Structure - Testosterone Propionate
化学構造 - テストステロンプロピオン酸エステル
Chemical Structure - Tetracaine Hydrochloride
Chemical Structure - Tetracycline Hydrochloride
化学構造 - テトラサイクリン塩酸塩
化学構造 - チアミラールナトリウム
化学構造 - チオペンタールナトリウム
Chemical Structure - Thioridazine Hydrochloride
Chemical Structure - Tiapride Hydrochloride
Chemical Structure - Tiaramide Hydrochloride
Chemical Structure - ‌Ticlopidine Hydrochloride‌
Chemical Structure - Timepidium Bromide Hydrate
化学構造 - チメピジウム臭化物水和物
化学構造 - チモロールマレイン酸塩
Chemical Structure - Tipepidine Hibenzate
化学構造 - チペピジンヒベンズ酸塩
Chemical Structure - Todralazine Hydrochloride Hydrate
化学構造 - トドララジン塩酸塩水和物
Chemical Structure - Tokishakuyakusan Extract
Chemical Structure - Tolperisone Hydrochloride
Chemical Structure - Tosufloxacin Tosilate Hydrate
化学構造 - トスフロキサシントシル酸塩水和物
Chemical Structure - ‌Tramadol Hydrochloride
Chemical Structure - Triamcinolone Acetonide
化学構造 - トリアムシノロンアセトニド
化学構造 - トリクロホスナトリウム
Chemical Structure - Trientine Hydrochloride
Chemical Structure - Trihexyphenidyl Hydrochloride
化学構造 - トリヘキシフェニジル塩酸塩
Chemical Structure - Trimebutine Maleate
化学構造 - トリメブチンマレイン酸塩
Chemical Structure - Trimetazidine Hydrochloride
Chemical Structure - ‌Trimetoquinol Hydrochloride Hydrate
化学構造 - トリメトキノール塩酸塩水和物
Chemical Structure - ‌Tulobuterol Hydrochloride‌
Chemical Structure - Ursodeoxycholic Acid
化学構造 - ウルソデオキシコール酸
Chemical Structure - Valaciclovir Hydrochloride
Chemical Structure - Vancomycin Hydrochloride
Chemical Structure - Verapamil Hydrochloride
Chemical Structure - Vinblastine Sulfate
Chemical Structure - Vincristine Sulfate
Chemical Structure - ‌10 Codeine Phosphate Powder
Chemical Structure - ‌10 dl-Methylephedrine Hydrochloride Powder
化学構造 - dl-メチルエフェドリン塩酸塩散 10
Chemical Structure - Acetylcholine Chloride for Injection
化学構造 - 注射用アセチルコリン塩化物
Chemical Structure - Acrinol and Zinc Oxide Ointment
化学構造 - アクリノール・亜鉛華軟膏
Chemical Structure - Alprostadil Injection
化学構造 - アルプロスタジル注射液
Chemical Structure - Bepotastine Besilate Tablets
化学構造 - ベポタスチンベシル酸塩錠
画像2 - カンデサルタン シレキセチル錠
画像3 - カンデサルタン シレキセチル錠
画像4 - カンデサルタン シレキセチル錠
画像5 - カンデサルタン シレキセチル錠
Chemical Structure - Candesartan Cilexetil Tablets
化学構造 - カンデサルタン シレキセチル錠
Chemical Structure - Compound Diastase and Sodium Bicarbonate Powder
化学構造 - 複方ジアスターゼ・重曹散
Chemical Structure - Compound Oxycodone and Atropine Injection
化学構造 - 複方オキシコドン・アトロピン注射液
Chemical Structure - ‌Compound Oxycodone Injection
化学構造 - 複方オキシコドン注射液
Chemical Structure - Compound Phellodendron Powder for Cataplasm
化学構造 - パップ用複方オウバク散
Chemical Structure - Compound Scopolia Extract and Diastase Powder
化学構造 - 複方ロートエキス・ジアスターゼ散
Chemical Structure - Compound Thianthol and Salicylic Acid Solution
化学構造 - 複方チアントール・サリチル酸液
Chemical Structure - Dextran 40 Injection
Chemical Structure - Diagnostic Sodium Citrate Solution
化学構造 - 診断用クエン酸ナトリウム液
Chemical Structure - Diastase and Sodium Bicarbonate Powder
Chemical Structure - Diluted Opium Powder
Chemical Structure - Diphenhydramine and Bromovalerylurea Powder
化学構造 - ジフェンヒドラミン・バレリル尿素散
Chemical Structure - Doripenem for Injection
Image 2 - Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
画像2 - ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Image 3 - Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
画像3 - ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Image 4 - Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
画像4 - ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Image 5 - Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
画像5 - ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Chemical Structure - Dorzolamide Hydrochloride and Timolol Maleate Ophthalmic Solution
化学構造 - ドルゾラミド塩酸塩･チモロールマレイン酸塩点眼液
Chemical Structure - Epinephrine Solution
画像2 - シロップ用ファロペネムナトリウム
画像3 - シロップ用ファロペネムナトリウム
Chemical Structure - Faropenem Sodium for Syrup
化学構造 - シロップ用ファロペネムナトリウム
Chemical Structure - Faropenem Sodium Tablets
化学構造 - ファロペネムナトリウム錠
Image 2 - Gati‰oxacin Ophthalmic Solution
Image 3 - Gati‰oxacin Ophthalmic Solution
Chemical Structure - Gati‰oxacin Ophthalmic Solution
化学構造 - ガチフロキサシン点眼液
Chemical Structure - Gentian and Sodium Bicarbonate Powder
Chemical Structure - Glycerin and Potash Solution
Chemical Structure - Hydrocortisone and Diphenhydramine Ointment
化学構造 - ヒドロコルチゾン・ジフェンヒドラミン軟膏
化学構造 - ヨウ化ヒプル酸ナトリウム (131I) 注射液
ヨウ化ヒプル酸ナトリウム (131I) 注射液
Image 2 - Irinotecan Hydrochloride Injection
Image 3 - Irinotecan Hydrochloride Injection
Chemical Structure - Irinotecan Hydrochloride Injection
化学構造 - イリノテカン塩酸塩注射液
Chemical Structure - Isotonic Sodium Chloride Solution
Chemical Structure - Kainic Acid and Santonin Powder
化学構造 - カイニン酸・サントニン散
Chemical Structure - L-Arginine Hydrochloride Injection
化学構造 - L-アルギニン塩酸塩注射液
画像2 - イソロイシン・ロイシン・バリン顆粒
化学構造 - イソロイシン・ロイシン・バリン顆粒
Chemical Structure - Lansoprazole Delayed-release Orally Disintegrating Tablets
化学構造 - ランソプラゾール腸溶性口腔内崩壊錠
Image 2 - Meglumine Iotalamate Injection
画像2 - イオタラム酸メグルミン注射液
Image 3 - Meglumine Iotalamate Injection
画像3 - イオタラム酸メグルミン注射液
Chemical Structure - Meglumine Iotalamate Injection
化学構造 - イオタラム酸メグルミン注射液
Chemical Structure - Meglumine Sodium Amidotrizoate Injection
化学構造 - アミドトリゾ酸ナトリウムメグルミン注射液
Chemical Structure - Morphine and Atropine Injection
化学構造 - モルヒネ・アトロピン注射液
Chemical Structure - ‌Mosapride Citrate Powder
化学構造 - モサプリドクエン酸塩散
Chemical Structure - Mosapride Citrate Tablets
化学構造 - モサプリドクエン酸塩錠
Chemical Structure - Naphazoline and Chlorpheniramine Solution
化学構造 - ナファゾリン・クロルフェニラミン液
Chemical Structure - Nartograstim for Injection (Genetical Recombination)
化学構造 - 注射用ナルトグラスチム（遺伝子組換え）
Chemical Structure - Opium Alkaloids and Atropine Injection
化学構造 - アヘンアルカロイド・アトロピン注射液
Chemical Structure - Opium Alkaloids and Scopolamine Injection
化学構造 - アヘンアルカロイド・スコポラミン注射液
Chemical Structure - Opium Alkaloids Hydrochlorides Injection
化学構造 - アヘンアルカロイド塩酸塩注射液
Chemical Structure - Opium Ipecac Powder
Chemical Structure - Panipenem and Betamipron for Injection
化学構造 - 注射用パニペネム･ベタミプロン
Image 2 - Pemirolast Potassium Ophthalmic Solution
画像2 - ペミロラストカリウム点眼液
Image 3 - Pemirolast Potassium Ophthalmic Solution
画像3 - ペミロラストカリウム点眼液
Chemical Structure - Pemirolast Potassium Ophthalmic Solution
化学構造 - ペミロラストカリウム点眼液
Chemical Structure - Phellodendron, Albumin Tannate and Bismuth Subnitrate Powder
化学構造 - オウバク・タンナルビン・ビスマス散
Chemical Structure - Phenolsulfonphthalein Injection
化学構造 - フェノールスルホンフタレイン注射液
Chemical Structure - Phenytoin Sodium for Injection
化学構造 - 注射用フェニトインナトリウム
Chemical Structure - Pitavastatin Calcium Orally Disintegrating Tablets
化学構造 - ピタバスタチンカルシウム口腔内崩壊錠
Chemical Structure - Pitavastatin Calcium Tablets
化学構造 - ピタバスタチンカルシウム錠
Chemical Structure - Polyethylene Glycol Ointment
Chemical Structure - Prednisolone Sodium Succinate for Injection
化学構造 - 注射用プレドニゾロンコハク酸エステルナトリウム
Image 2 - Purified Sodium Hyaluronate Injection
画像2 - 精製ヒアルロン酸ナトリウム注射液
Image 3 - Purified Sodium Hyaluronate Injection
画像3 - 精製ヒアルロン酸ナトリウム注射液
Chemical Structure - Purified Sodium Hyaluronate Injection
化学構造 - 精製ヒアルロン酸ナトリウム注射液
Image 2 - Purified Sodium Hyaluronate Ophthalmic Solution
画像2 - 精製ヒアルロン酸ナトリウム点眼液
Image 3 - Purified Sodium Hyaluronate Ophthalmic Solution
画像3 - 精製ヒアルロン酸ナトリウム点眼液
Image 4 - Purified Sodium Hyaluronate Ophthalmic Solution
画像4 - 精製ヒアルロン酸ナトリウム点眼液
Chemical Structure - Purified Sodium Hyaluronate Ophthalmic Solution
化学構造 - 精製ヒアルロン酸ナトリウム点眼液
Chemical Structure - Salicylated Alum Powder
化学構造 - サリチル・ミョウバン散
Chemical Structure - Saponated Cresol Solution
Chemical Structure - Scopolia Extract and Carbon Powder
化学構造 - ロートエキス・カーボン散
Chemical Structure - Scopolia Extract and Ethyl Aminobenzoate Powder
化学構造 - ロートエキス・アネスタミン散
Chemical Structure - Scopolia Extract Powder
Chemical Structure - ‌Silver Nitrate Ophthalmic Solution‌‌
Chemical Structure - Silver Protein Solution
Chemical Structure - Sodium Citrate Injection for Transfusion
化学構造 - 輸血用クエン酸ナトリウム注射液
Chemical Structure - Sodium Iotalamate Injection
化学構造 - イオタラム酸ナトリウム注射液
Chemical Structure - Sodium L-Lactate Ringer's Solution
化学構造 - L-乳酸ナトリウムリンゲル液
Chemical Structure - Sulfur, Salicylic Acid and Thianthol Ointment
化学構造 - イオウ・サリチル酸・チアントール軟膏
Chemical Structure - Swertia and Sodium Bicarbonate Powder
Image 2 - Tazobactam and Piperacillin for Injection
画像2 - 注射用タゾバクタム･ピペラシリン
Image 3 - Tazobactam and Piperacillin for Injection
画像3 - 注射用タゾバクタム･ピペラシリン
Chemical Structure - Tazobactam and Piperacillin for Injection
化学構造 - 注射用タゾバクタム･ピペラシリン
Image 2 - ‌Telmisartan and Hydrochlorothiazide Tablets
画像2 - テルミサルタン・ヒドロクロロチアジド錠
Image 3 - ‌Telmisartan and Hydrochlorothiazide Tablets
画像3 - テルミサルタン・ヒドロクロロチアジド錠
Chemical Structure - ‌Telmisartan and Hydrochlorothiazide Tablets
化学構造 - テルミサルタン・ヒドロクロロチアジド錠
Chemical Structure - Tolbutamide Tablets
Chemical Structure - Trichlormethiazide Tablets
化学構造 - トリクロルメチアジド錠
Image 2 - Valsartan and Hydrochlorothiazide Tablets
画像2 - バルサルタン・ヒドロクロロチアジド錠
Image 3 - Valsartan and Hydrochlorothiazide Tablets
画像3 - バルサルタン・ヒドロクロロチアジド錠
Chemical Structure - Valsartan and Hydrochlorothiazide Tablets
化学構造 - バルサルタン・ヒドロクロロチアジド錠
Chemical Structure - Vasopressin Injection
Chemical Structure - Weak Opium Alkaloids and Scopolamine Injection
化学構造 - 弱アヘンアルカロイド・スコポラミン注射液
Chemical Structure - Zinc Oxide Ointment
Chemical Structure - Zinc Oxide Starch Powder
Chemical Structure - Zinc Sulfate Ophthalmic Solution
Apparatus - Alcohol Number Determination
Illustration from JP XVIII General Tests, Processes and Apparatus
Test method illustration from JP XVIII General Tests
Diagram 10 - Size-exclusion chromatography
Diagram 11 - Size-exclusion chromatography
Diagram 12 - Size-exclusion chromatography
Diagram 13 - Size-exclusion chromatography
Diagram 14 - Size-exclusion chromatography
Diagram 15 - Size-exclusion chromatography
Diagram 16 - Size-exclusion chromatography
Diagram 17 - Size-exclusion chromatography
Diagram 2 - Size-exclusion chromatography
Diagram 3 - Size-exclusion chromatography
Diagram 4 - Size-exclusion chromatography
Diagram 5 - Size-exclusion chromatography
Diagram 6 - Size-exclusion chromatography
Diagram 7 - Size-exclusion chromatography
Diagram 8 - Size-exclusion chromatography
Diagram 9 - Size-exclusion chromatography
Apparatus - Size-exclusion chromatography
Chemical Structure - Aluminum Acetylsalicylate
化学構造 - アスピリンアルミニウム
Chemical Structure - Calcium Folinate Calcium Leucovorin
化学構造 - ホリナートカルシウム水和物
Chemical Structure - Calcium Gluconate Hydrate
化学構造 - グルコン酸カルシウム水和物
Chemical Structure - Calcium Lactate Hydrate
Chemical Structure - Calcium Levofolinate Hydrate
化学構造 - レボホリナートカルシウム水和物
レボホリナートカルシウム水和物
Chemical Structure - Calcium Pantothenate
化学構造 - パントテン酸カルシウム
Chemical Structure - Calcium Polystyrene Sulfonate
化学構造 - ポリスチレンスルホン酸カルシウム
ポリスチレンスルホン酸カルシウム
Chemical Structure - Calcium Sodium Edetate Hydrate
化学構造 - エデト酸カルシウムナトリウム水和物
エデト酸カルシウムナトリウム水和物
Chemical Structure - Potassium Canrenoate
Chemical Structure - Potassium Clavulanate
Chemical Structure - Sodium Acetate Hydrate
Chemical Structure - Sodium Aurothiomalate
化学構造 - 金チオリンゴ酸ナトリウム
Chemical Structure - Sodium Bicarbonate and Bitter Tincture Mixture
Sodium Bicarbonate and Bitter Tincture Mixture
Chemical Structure - Sodium Citrate Hydrate
化学構造 - クエン酸ナトリウム水和物
Chemical Structure - Sodium Cromoglicate
化学構造 - クロモグリク酸ナトリウム
Chemical Structure - Sodium Picosulfate Hydrate
化学構造 - ピコスルファートナトリウム水和物
ピコスルファートナトリウム水和物
Chemical Structure - Sodium Risedronate Hydrate
化学構造 - リセドロン酸ナトリウム水和物
化学構造 - レチノール酢酸エステル
Chemical Structure - Vitamin A Palmitate
化学構造 - レチノールパルミチン酸エステル
レチノールパルミチン酸エステル
Chemical Structure - Vitamin B1 Hydrochloride
Chemical Structure - Vitamin B2 Butyrate
化学構造 - リボフラビン酪酸エステル
Chemical Structure - Vitamin B2 Phosphate Ester
化学構造 - リボフラビンリン酸エステルナトリウム
リボフラビンリン酸エステルナトリウム
化学構造 - エルゴカルシフェロール
化学構造 - トコフェロール酢酸エステル
Chemical Structure - Vitamin E Calcium Succinate
化学構造 - トコフェロールコハク酸エステルカルシウム
トコフェロールコハク酸エステルカルシウム
Chemical Structure - Vitamin E Nicotinate
化学構造 - トコフェロールニコチン酸エステル
トコフェロールニコチン酸エステル
Method of preparation Prepare by one of the following
Acrinol and Zinc Oxide Oil contains not less than 44.
Acrinol and Zinc Oxide Oil is a yellow-white, slimy substance. Separation of a part of its ingredients oc-curs on prolonged standing.
(1) Shake well 1 g of Acrinol and Zinc Oxide Oil with 10 mL of diethyl ether, 2 mL of acetic acid (100) and 10 mL of water, and separate the water layer. Shake the layer with 5 mL of hydrochloric acid and 2 to 3 drops of sodium nitrite TS, and allow to stand: a dark red color is produced (acrinol). Place 1 g of Acrinol and Zinc Oxide Oil in a crucible, melt by warming, heat, gradually raising the temperature until the mass is thoroughly charred, and then ignite strongly: a yellow color is produced, and disappears on cool-ing. To the residue add 10 mL of water and 5 mL of dilute ‌hydrochloric acid, filter after thorough shaking, and to the filtrate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is formed (zinc oxide).‌ Shake well 0.2 g of Acrinol and Zinc Oxide Oil with 20 mL of ethanol (95) and 1 mL of acetic acid (100), centri-fuge, filter, and use the filtrate as the sample solution. Sepa-rately, dissolve 5 mg of acrinol in 50 mL of ethanol (95) and 2.5 mL of acetic acid (100), and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 2-propanol and acetic acid (100) (9:1) to a distance of about 10 cm, and air-dry the plate. Ex-amine under ultraviolet light (main wavelength: 365 nm): the spots from the sample solution and standard solution exhibit a blue fluorescence and show the same Rf value.
Transfer about 0.8 g of well-mixed Acrinol and Zinc Oxide Oil, accurately weighed, to a crucible, heat, gradually raising the temperature until the mass is throughly charred, then strongly heat until the residue becomes yellow. After cooling, dissolve the residue by addition of 1 mL of water and 1.5 mL of hydrochloric acid, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add 80 mL of water, then add sodium hydroxide solution (1 in 50) until slightly precipitates appear, and add 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7). Titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.069 mg of ZnO
Alendronate Sodium Hydrate contains not less than 99.
Alendronate Sodium Hydrate occurs as a white crystalline powder. It is sparingly soluble in water, and practically insoluble in ethanol (99.5). It dissolves in 0.1 mol/L trisodium citrate TS. Melting point: about 2529C (with decomposition, after drying).
(1) To 5 mL of a solution of Alendronate Sodium Hydrate (1 in 50) add 1 mL of ninhydrin TS, and heat: a blue-purple color develops. Determine the infrared absorption spectrum of Alen-dronate Sodium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Alendronate Sodium RS: both spectra ex-hibit similar intensities of absorption at the same wave num-bers. To 0.1 g of Alendronate Sodium Hydrate add 10 mL of a mixture of nitric acid and perchloric acid (1:1). Heat to concentrate to about 1 mL, add about 10 mL of water while hot, and neutralize with a solution of sodium hydroxide (2 in 5): the solution responds to Qualitative Tests <1.09> for phosphate. A solution of Alendronate Sodium Hydrate (1 in 100) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> The pH of a solution of 1.0 g of Alendronate Sodium Hydrate in 100 mL of freshly boiled and cooled water is between 4.0 and 5.0.
(1) Heavy metals <1.07>—Put 1.0 g of Alen-dronate Sodium Hydrate in a Kjeldahl flask, add 9 mL of a mixture of nitric acid and sulfuric acid (5:4), and heat until the solution becomes brown. After cooling, add 9 mL of a mixture of nitric acid and sulfuric acid (5:4), and heat again until the color changes from colorless to brown. After cool-ing, add 2 mL of nitric acid, strongly heat until brown fumes are no longer evolved, and heat until large amounts of white fumes are evolved. After cooling, add carefully 5 mL of water and 1 mL of hydrogen peroxide (30), heat until white fumes are no longer evolved, and continue heating for more 5 minutes. After cooling, if any yellow color remains, add 2 mL of nitric acid, and repeat the same procedure. After cooling, transfer the solution in the Kjeldahl flask to a beaker, wash out the inside of the flask with 5 mL of water, and add the washing to the beaker. Adjust to pH 3 – 5 with ammonia solution (28), transfer to a Nessler tube, add water to make 50 mL, and perform the test with this solution as the test solution. Prepare the control solution in the same proce-dure using the same amount of the reagents used for the preparation of the sample solution, add 1.0 mL of Standard Lead Solution and add water to make 50 mL (not more than 10 ppm). (2) Related substances—Dissolve 15 mg of Alendronate Sodium Hydrate in 25 mL of 0.1 mol/L trisodium citrate TS, and use this solution as the sample stock solution. Pipet 5 mL of the sample stock solution, and add 0.1 mol/L triso-dium citrate TS to make exactly 50 mL. Pipet 1 mL of this solution, add 0.1 mol/L trisodium citrate TS to make exactly 100 mL, and use this solution as the standard stock solution. To exactly 5 mL each of the sample stock solution and standard stock solution, add exactly 5 mL each of a solution of sodium tetraborate decahydrate (19 in 1000), acetonitrile and a solution of 9-fluorenylmethyl chloroformate in aceto-nitrile (1 in 250), shake for 45 seconds, and allow to stand for 30 minutes at room temperature. Then, add 20 mL of dichloromethane to them, shake for 60 seconds, centrifuge, and use the supernatant liquids so obtained as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: each peak area other than alendronic acid obtained from the sample solution is not larger than the peak area of alendronic acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 459C. Mobile phase A: Dissolve 2.94 g of trisodium citrate dihy-drate and 1.42 g of anhydrous disodium hydrogen phosphate in 900 mL of water, adjust to pH 8.0 with phosphoric acid, and add water to make 1000 mL. To 850 mL of this solution add 150 mL of acetonitrile for liquid chromatography. Mobile phase B: Dissolve 2.94 g of trisodium citrate dihy-drate and 1.42 g of anhydrous disodium hydrogen phosphate in 900 mL of water, adjust to pH 8.0 with phosphoric acid, and add water to make 1000 mL. To 300 mL of this solution add 700 mL of acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 15 100 → 50 0 → 50 15 – 25 50 → 0 50 → 100 Flow rate: About 1.8 mL per minute. Time span of measurement: About 5 times as long as the retention time of alendronic acid, beginning after the solvent peak. System suitability— System performance: Dissolve 15 mg of Alendronate So-dium Hydrate and 2 mg of 4-aminobutylic acid in 100 mL of 0.1 mol/L trisodium citrate TS. To 5 mL of this solution add 5 mL each of a solution of sodium tetraborate decahydrate (19 in 1000), acetonitrile and a solution of 9-fluorenylmethyl chloroformate in acetonitrile (1 in 250), then, proceed in the same manner as the sample solution. When the procedure is run with 20 mL of this solution under the above operating conditions, alendronic acid and 4-aminobutylic acid are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of alendronic acid is not more than 2.0z. Loss on drying <2.41> 16.1 – 17.1z (1 g, 1409C, 3 hours).
Weigh accurately about 10 mg each of Alendronate Sodium Hydrate and Alendronate Sodium RS (separately determine the loss on drying <2.41> in the same conditions as Alendronate Sodium Hydrate), dissolve in 0.1 mol/L triso-dium citrate TS to make exactly 100 mL, and use these solu-tions as the sample stock solution and the standard stock so- ‌lution, respectively. To exactly 5 mL each of these solutions add exactly 5 mL each of a solution of sodium tetraborate decahydrate (19 in 1000) and a solution of 9-fluorenylmethyl chloroformate in acetonitrile (1 in 2000), shake for 30 seconds, and allow to stand for 25 minutes. Then, add 25 mL of dichloromethane, shake for 60 seconds, centrifuge, and use the supernatant liquids so obtained as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of alendronic acid in each solu-tion. Amount (mg) of alendronate sodium (C4H12NNaO7P2) ＝ MS × AT/AS MS: Amount (mg) of Alendronate Sodium RS taken, cal-culated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.1 mm in inside diame-ter and 25 cm in length, packed with styrene-divinylbenzene copolymer for liquid chromatography (10 mm in particle di-ameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 14.7 g of trisodium citrate dihy-drate and 7.1 g of anhydrous disodium hydrogen phosphate in 900 mL of water, adjust to pH 8.0 with phosphoric acid, and add water to make 1000 mL. To 700 mL of this solution add 250 mL of acetonitrile for liquid chromatography and 50 mL of methanol. Flow rate: Adjust so that the retention time of alendronic acid is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of alendronic acid are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of alendronic acid is not more than 1.0z.
Aspirin Aluminum occurs as a white crystal-line powder. It is odorless or has a slight, acetic odor. It is practically insoluble in water, in methanol, in ethanol (95) and in diethyl ether. It dissolves, with decomposition, in sodium hydroxide TS and in sodium carbonate TS.
(1) Dissolve 0.1 g of Aspirin Aluminum in 10 mL of sodium hydroxide TS by heating, if necessary. Neutralize 2 mL of the solution with hydrochloric acid, and add 1 to 2 drops of iron (III) chloride TS: a red-purple color develops. Determine the absorption spectrum of the sample so-lution obtained in the Assay (1) as directed under Ultravio-let-visible Spectrophotometry <2.24>: it exhibits a maximum between 277 nm and 279 nm. Place 2 g of Aspirin Aluminum in a platinum cruci-ble, and ignite until charred. To the residue add 1 g of anhy-drous sodium carbonate, and ignite for 20 minutes. After cooling, to the residue add 15 mL of dilute hydrochloric acid, shake, and filter: the filtrate responds to Qualitative Tests <1.09> for aluminum salt.
(1) Salicylate—Using AT2 and AS2 obtained in the Assay (1), calculate the amount of salicylate [as salicylic acid (C7H6O3: 138.12)] by the following equation: salicylate con-tent is not more than 7.5z, calculated on the anhydrous basis. Amount (mg) of salicylic acid (C7H6O3) ＝ MS × AT2/AS2 × 1/4 MS: Amount (mg) of salicylic acid for assay taken Heavy metals <1.07>—Place 2.0 g of Aspirin Alumi-num in a porcelain crucible, cover the crucible loosely, and ignite at a low temperature until charred. After cooling, add 2 mL of nitric acid and 1 mL of sulfuric acid to the content of the crucible, heat gently the crucible until white fumes are evolved, and continue the heating until white fumes are no longer evolved, then ignite between 5009C and 6009C until the carbon is incinerated. When the incineration is not com-pleted, add 2 mL of nitric acid and 1 mL of sulfuric acid, and heat gently in the same manner, then ignite between 5009C and 6009C to incinerate completely. After cooling, add 2 mL of hydrochloric acid, and proceed as directed in Method 2, and perform the test. Prepare the control solution by using the same quantities of the same reagents as directed for the preparation of the test solution, and add 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Dissolve 1.0 g of Aspirin Aluminum in 15 mL of sodium hydroxide TS. To this solution add 1 drop of phenolphthalein TS, and with stirring, add dropwise hydrochloric acid until the red color of the solution disap-pears. Then add 2 mL of hydrochloric acid, cool with occa-sional shaking for 10 minutes, and filter with a glass filter (G3). Wash the residue with two 5 mL portions of 1 mol/L hydrochloric acid TS, and combine the filtrate and the wash-ings. Use this solution as the test solution, and perform the test (not more than 2 ppm). Water <2.48> Not more than 4.0z (0.15 g, direct titration).
(1) Aspirin—Weigh accurately about 0.1 g of Aspi-rin Aluminum, add 40 mL of sodium fluoride TS, and shake for 5 minutes. Allow the solution to stand for 10 minutes with frequent shaking. Extract the solution with six 20-mL portions of chloroform. Combine all chloroform extracts, and add chloroform to make exactly 200 mL. Measure ex-actly 10 mL of this solution, add chloroform to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 90 mg of salicylic acid for assay, previously dried in a desiccator (silica gel) for 3 hours, and dissolve in chloroform to make exactly 200 mL. Meas-ure exactly 5 mL of this solution, add chloroform to make exactly 200 mL, and use this solution as the standard solu-tion (1). Then weigh accurately about 90 mg of Aspirin RS, previously dried in a desiccator (silica gel) for 5 hours, and dissolve in chloroform to make exactly 200 mL. Measure ex-actly 10 mL of this solution, add chloroform to make exactly 100 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Ultra-violet-visible Spectrophotometry <2.24>. Determine the ab-sorbances, AT1 and AS1, of the sample solution and standard ‌490 solution (1) at 278 nm, and absorbances, AT2 and AS2, of these solution, at 308 nm, respectively. Then determine the absorbance AS3 of the standard solution (2) at 278 nm. Amount (mg) of aspirin (C9H8O4)   A － AT2 × AS1  (2) Determine the infrared absorption spectrum of Aspoxicillin Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or spec-trum of Aspoxicillin RS: both spectra exhibit similar intensi- ties of absorption at the same wave numbers. D T1 S ＝ M ×  AS3 AS2   Optical rotation <2.49> [a]20: ＋170 – ＋1859(0.2 g calcu-lated on the anhydrous bases, water, 20 mL, 100 mm). MS: Amount (mg) of Aspirin RS taken (2) Aluminum—Weigh accurately about 0.4 g of Aspirin Aluminum, and dissolve in 10 mL of sodium hydroxide TS. Add dropwise 1 mol/L hydrochloric acid TS to adjust the solution to a pH of about 1, add 20 mL of acetic acid-ammonium acetate buffer solution (pH 3.0) and 0.5 mL of Cu-PAN TS, and heat. While boiling, titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraace-tate VS until the color of the solution changes from red to yellow and persists for 1 minute. Perform a blank determi-nation in the same manner, and make any necessary correc-tion. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.349 mg of Al
Aluminum Monostearate is mainly aluminum com- test (not more than 5 ppm). Acid-consuming capacity Weigh accurately about 0.2 g of pounds of stearic acid (C acid (C16H32O2: 256.42). 18H 36O2: 284.48) and palmitic Dried Aluminum Hydroxide Gel, and transfer to a glass-stoppered flask. Add exactly 100 mL of 0.1 mol/L hydro-chloric acid VS, stopper the flask, shake at 37 ± 29C for 1 hour, and filter. Measure exactly 50 mL of the filtrate, and titrate <2.50> while thoroughly stirring, the excess hydrochlo-ric acid with 0.1 mol/L sodium hydroxide VS until the pH of the solution becomes to 3.5. The volume of 0.1 mol/L hy-drochloric acid VS consumed is not less than 250 mL per g of Dried Aluminum Hydroxide Gel. Aluminum Monostearate, when dried, contains not less than 7.
Aluminum Monostearate occurs as a white to yellow-white powder. It is odorless or has a faint, character-istic odor. It is practically insoluble in water, in ethanol (95) and in diethyl ether. ‌JP XVIII Official Monographs / Dried Aluminum Potassium Sulfate 441
(1) Heat 3 g of Aluminum Monostearate with 30 mL of hydrochloric acid in a water bath with occa-sional shaking for 10 minutes. After cooling, shake the mix-ture vigorously with 50 mL of water and 30 mL of diethyl ether for 3 minutes, and allow to stand. To the separated aqueous layer add sodium hydroxide TS until the solution becomes slightly turbid, and filter: the filtrate responds to Qualitative Tests <1.09> for aluminum salt. (2) Wash the diethyl ether layer separated in (1) with two 20-mL portions of water, and evaporate the diethyl ether layer on a water bath: the residue melts <1.13> at above 549C. Acid value for fatty acid <1.13> 193 – 210. Weigh accu-rately about 1 g of fatty acid obtained in the Identification (2), transfer a 250-mL glass-stoppered flask, add 100 mL of a mixture of diethyl ether and ethanol (95) (2:1), warm to dissolve, add several drops of phenolphthalein TS, and proceed as directed under Acid Value.
(1) Free fatty acid—Mix 1.0 g of Aluminum Monostearate with about 50 mL of a mixture of neutralized ethanol and diethyl ether (1:1), filter through dry filter paper, wash the vessel and the filter paper with a small amount of a mixture of neutralized ethanol and diethyl ether (1:1), combine the filtrate and the washings, and add 2.1 mL of 0.1 mol/L potassium hydroxide VS: a red color develops. Water-soluble salts—Heat 2.0 g of Aluminum Monostearate with 80 mL of water in a loosely stoppered conical flask on a water bath for 30 minutes with occasional shaking. After cooling, filter through dry filter paper, wash the residue with a small amount of water, combine the wash-ings with the filtrate, add water to make 100 mL, evaporate 50 mL of this solution on a water bath, and heat strongly at 6009C: the mass of the residue is not more than 10.0 mg. Heavy metals <1.07>—Heat 1.0 g of Aluminum Monostearate over a small flame with caution at the begin-ning, and continue the heating, gradually raising the temper-ature, to ash. After cooling, add 10 mL of diluted hydro-chloric acid (1 in 2), evaporate on a water bath, and boil the residue with 20 mL of water for 1 minute. Cool, filter, wash the residue with water, combine the filtrate and the wash-ings, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Evaporate 10 mL of diluted hydrochloric acid (1 in 2) on a water bath to dryness, add 2 mL of dilute acetic acid and 5.0 mL of Standard Lead Solution, dilute with water to make 50 mL, and use this solution as the control solution (not more than 50 ppm). Arsenic <1.11>—Mix 1.0 g of Aluminum Monostearate with 2 g of magnesium nitrate hexahydrate, ignite over a small flame, moisten the residue after cooling with 0.5 mL of nitric acid, and heat. Heat again the residue with 10 mL of dilute sulfuric acid until white fumes evolve, add water to make 5 mL, and perform the test with this solu-tion as the test solution (not more than 2 ppm). Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 3 hours).
Weigh accurately about 1 g of Aluminum Monostearate, previously dried, ignite gently to ash, and cool. Add dropwise 0.5 mL of nitric acid, evaporate on a water bath by heating, and then heat strongly between 9009C and 11009C to a constant mass. After cooling, weigh rapidly the ignited residue, and designate the mass as aluminum oxide (Al O : 101.96). Amount (mg) of aluminum (Al) ＝ amount (mg) of aluminum oxide (Al2O3) × 0.529
Ampicillin Sodium contains not less than 850 mg (potency) and not more than 950 mg (potency) per mg, calculated on the anhydrous basis.
Ampicillin Sodium occurs as white to light yellow-white, crystals or crystalline powder. It is very soluble in water, and sparingly soluble in ethanol (99.5).
(1) Determine the infrared absorption spec-trum of Ampicillin Sodium, previously dried in a desiccator (reduced pressure not exceeding 0.67 kPa, 609C) for 3 hours, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wave numbers. (2) Ampicillin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋246 – ＋2729(1 g calculated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Ampicillin Sodium in 10 mL of water is between 8.0 and 10.0.
(1) Clarity and color of solution—Dissolve 0.25 g (potency) of Ampicillin Sodium in 0.75 mL of water: the so-lution is clear, and its absorbance at 400 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.40. Heavy metals <1.07>—Proceed with 1.0 g of Ampicil- lin Sodium according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ampicillin Sodium according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Ampicillin Sodium in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks other than ampicillin obtained from the sample so-lution is not larger than the peak area of ampicillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 10 times as long as the retention time of ampicillin. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of ampicillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg of Ampicillin RS in a suitable amount of the mobile phase, add 5 mL of a solution of guaifenesin in the mobile phase (1 in 200) and the mobile phase to make 50 mL, and use this solution as the solution for system suitability test. When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, ampicillin and guaifenesin are eluted in this order with the resolution between these peaks being not less than 35. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ampicillin is not more than 1.0z. Water <2.48> Not more than 2.0z (0.2 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Ampicillin Sodium and Ampicillin RS, equivalent to about 50 mg (potency), dissolve them in a suitable amount of the mobile phase, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ampicillin to that of the internal standard. Amount [mg (potency)] of ampicillin (C16H19N3O4S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Ampicillin RS taken Internal standard solution—A solution of guaifenesin in the mobile phase (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.94 g of diammonium hydrogen phosphate in 850 mL of water, add 100 mL of acetonitrile, adjust to pH 5.0 with phosphoric acid, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of ampicillin is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ampicillin and guaifenesin are eluted in this order with the resolution between these peaks being not less than 35. System repeatability: When the test is repeated 6 times ‌JP XVIII Official Monographs / Ampicillin Sodium and Sulbactam Sodium for Injection 471 ‌with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of ampicillin to that of the internal standard is not more than 1.0z.
Anhydrous Dibasic Calcium Phosphate contains not less than 97.
Anhydrous Dibasic Calcium Phosphate oc-curs as white, crystalline powder or granules. It is practically insoluble in water and in ethanol (99.5). It dissolves in dilute hydrochloric acid and in dilute nitric acid.◆ Identiˆcation (1) Dissolve 0.1 g of Anhydrous Dibasic Calcium Phosphate in 10 mL of 2 mol/L hydrochloric acid TS by warming, add 2.5 mL of ammonia TS dropwise with shaking, and add 5 mL of ammonium oxalate TS: a white precipitate is produced. (2) Dissolve 0.1 g of Anhydrous Dibasic Calcium Phos-phate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-monium heptamolybdate TS after warming at 709C for 1 to 2 minutes: a yellow precipitate is produced.
(1) Acid-insoluble substances—Dissolve 5.0 g of Anhydrous Dibasic Calcium Phosphate in 40 mL of water and 10 mL of hydrochloric acid, and boil gently for 5 minutes. After cooling, collect the insoluble substance using a ˆlter paper for quantitative analysis. Wash with water until no more turbidity of the washings is produced when silver nitrate TS is added. Ignite to incinerate the residue and the ˆlter paper at 600 ± 509C: the mass is not more than 10 mg (not more than 0.2z). Chloride—To 0.20 g of Anhydrous Dibasic Calcium Phosphate add 20 mL of water and 13 mL of dilute nitric acid, dissolve by warming, if necessary, add water to make 100 mL, and ˆlter, if necessary. Put 50 mL of this solution in a Nessler tube, and use this as the sample solution. Separate-ly, transfer 0.70 mL of 0.01 mol/L hydrochloric acid VS to another Nessler tube, add 6 mL of dilute nitric acid and water to make 50 mL, and use this solution as the control so-lution. Add 1 mL of silver nitrate TS to the sample solution and control solution, mix well, and allow to stand for 5 minutes protecting from light. Compare the opalescence de- ‌JP XVIII Official Monographs / Dibasic Calcium Phosphate Hydrate 605 veloped in both solutions against a black background by viewing downward or transversely. The opalescence deve-loped in the sample solution is not more than that of the con-trol solution (not more than 0.25z). Sulfate—Dissolve 0.50 g of Anhydrous Dibasic Cal-cium Phosphate in 5 mL of water and 5 mL of dilute hydro-chloric acid, add water to make 100 mL, and ˆlter, if neces-sary. Put 20 mL of this solution in a Nessler tube, add 1 mL of dilute hydrochloric acid, and add water to make 50 mL, and use this as the sample solution. Transfer 1.0 mL of 0.005 mol/L sulfuric acid VS to another Nessler tube, add 1 mL of dilute hydrochloride acid and water to make 50 mL, and use this solution as the control solution. Add 2 mL of barium chloride TS to the sample solution and control solution, mix well, and allow to stand for 10 minutes. Compare the white turbidity produced in both solutions against a black back-ground by viewing downward or transversely. The turbidity produced in the sample solution is not thicker than that of the control solution (not more than 0.48z). Carbonate—Shake 1.0 g of Anhydrous Dibasic Cal- Dibasic Calcium Phosphate Hydrate
Atorvastatin Calcium Hydrate contains not less than 98.
Atorvastatin Calcium Hydrate occurs as a white to pale yellow-white crystalline powder. It is very soluble in methanol, freely soluble in dimethyl-sulfoxide, and very slightly soluble in water and in ethanol (99.5). It gradually turns yellowish white on exposure to light. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Atorvastatin Calcium Hydrate in methanol (1 in Mobile phase A: Dissolve 10.5 g of citric acid monohy-drate in 900 mL of water, adjust to pH 5.0 with ammonia so-lution (28), and add water to make 1000 mL. To 400 mL of this solution add 100 mL of acetonitrile and 100 mL of tetra-hydrofuran. Mobile phase B: A mixture of acetonitrile and tetra-hydrofuran (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 62,500) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) ence Spectrum or the spectrum of a solution of Atorvastatin Calcium RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ator-vastatin Calcium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Atorvastatin Calcium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, recrystallize the sample and the reference standard according to the method otherwise specified, filter and dry the crystals, and perform the test with the crystals. A gruel-like liquid of Atorvastatin Calcium Hydrate prepared by adding a small amount of dilute hydrochloric acid responds to the Qualitative Tests <1.09> (1) for calcium salt. A solution of Atorvastatin Calcium Hydrate in a mix-ture of methanol and water (7:3) (1 in 250) is also responds to Qualitative Tests <1.09> (3) for calcium salt. D Optical rotation <2.49> [a]25: －7 – －109(0.2 g, calculated on the anhydrous basis, dimethylsulfoxide, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Atorvastatin Calcium Hydrate according to Method 4, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 20 mg of Atorvastatin Calcium Hydrate in 20 mL of a mixture of water and aceto-nitrile (1:1), and use this solution as the sample solution. 0 – 40 93 7 40 – 80 93 → 60 7 → 40 Flow rate: Adjust so that the retention time of atorvasta-tin is about 16 minutes. Time span of measurement: About 5 times as long as the retention time of atorvastatin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of water and acetonitrile (1:1) to make exactly 100 mL. Confirm that the peak area of ator-vastatin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of atorvastatin are not less than 8000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of atorvastatin is not more than 2.0z. Water <2.48> 3.5 – 5.5z (50 mg, coulometric titration).
Weigh accurately about 20 mg each of Atorvastatin Calcium Hydrate and Atorvastatin Calcium RS (separately determine the water <2.48> in the same manner as Atorvasta-tin Calcium Hydrate), dissolve each in an adequate amount ‌JP XVIII Official Monographs / Atorvastatin Calcium Tablets 493 of a mixture of water and acetonitrile (1:1), add exactly 10 mL of the internal standard solution, then add a mixture of water and acetonitrile (1:1) to make 50 mL, and use these so-lutions as the sample solution and the standard solution, re-spectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of atorvastatin to that of the internal standard. Amount (mg) of atorvastatin calcium (C66H68CaF2N4O10) ＝ MS × QT/QS MS: Amount (mg) of Atorvastatin Calcium RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of butyl para-hydroxybenzoate in a mixture of water and acetonitrile (1:1) (1 in 1500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 10.5 g of citric acid monohydrate in 900 mL of water, adjust to pH 4.0 with ammonia solution photometry <2.24>: it exhibits a maximum between 244 nm and 248 nm. Uniformity of dosage units <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 tablet of Atorvastatin Calcium Tablets add 3V/5 mL of a mixture of water and methanol (1:1), and disintegrate the tablet by shaking. Add exactly V/10 mL of the internal standard solution, and add a mixture of water and methanol (1:1) to make V mL so that each mL contains about 0.1 mg of atorvastatin calcium hydrate (C66H68CaF2N4O10.3H2O). Centrifuge this solution, and use the supernatant liquid as the sample solution. Separately, weigh accurately about 22 mg of Atorvastatin Calcium RS (separately determine the water <2.48> in the same manner as Atorvastatin Calcium Hydrate), and dissolve in a mixture of water and methanol (1:1) to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, then add a mixture of water and methanol (1:1) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of atorvastatin to that of the internal standard. Amount (mg) of atorvastatin calcium hydrate (C H CaF N O .3H O) (28), and add water to make 1000 mL. To 530 mL of this 66 68 2 4 10 2 solution add 270 mL of acetonitrile and 200 mL of tetra-hydrofuran. Flow rate: Adjust so that the retention time of atorvasta-tin is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and atorvastatin are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of atorvastatin to that of the internal standard is not more than 1.0z.
Beraprost Sodium, when dried, contains not less than 98.
Beraprost Sodium occurs as a white powder. It is very soluble in methanol, and freely soluble in water and in ethanol (99.5). It is hygroscopic. Flow rate: Adjust so that the retention time of the second peak of beraprost is about 23 minutes. Time span of measurement: For 80 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add methanol to make 20 mL. To 1 mL of this solu- ‌532 tion add methanol to make 20 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add methanol to make exactly 10 mL. Confirm that the total area of the two peaks of beraprost obtained with 15 mL of this solution is equivalent to 14 to 26z of that with 15 mL of the solution for system suitability test. Beraprost Sodium Tablets
(1) Determine the absorption spectrum of a solution of Beraprost Sodium in methanol (3 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of previ-ously dried Beraprost Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Beraprost Sodium in methanol (1 in 1000) responds to Qualitative Tests <1.09> (1) for sodium salt.
Related substances—Dissolve 20 mg of Beraprost Sodium in 2 mL of methanol, and use this solution as the sample solution. Perform the test with 15 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calculate their amounts by the area percentage method: the amount of the peak having the relative retention time of about 0.5 to the second eluting principal peak of beraprost and the two adjacent peaks at the relative retention time of about 1.7 and another two adjacent peaks at the relative retention time of about 2.0 are not more than 0.2z, respectively, the amount of the peak at the relative retention time of about 1.2 is not more than 0.3z, the amount of the peak, other than the two peaks of beraprost and the peaks mentioned above, is less than 0.1z, and the total amount of the peaks, other than the two peaks of beraprost, is not more than 1.5z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 285 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (4 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase A: A mixture of water, acetonitrile, metha-nol and acetic acid (100) (640:330:30:1). Mobile phase B: A mixture of acetonitrile, water and acetic acid (100) (900:100:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. C24H29NaO5: 420.47 Monosodium (1RS,2RS,3aSR,8bSR )-2,3,3a,8b-tetrahydro-2-hydroxy-1-[(1E,3SR,4RS )-3-hydroxy-4-methyloct-1-en-6-yn-1-yl]-1H- Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) cyclopenta[b]benzofuran-5-butanoate 0 – 30 Monosodium (1RS,2RS,3aSR,8bSR )-2,3,3a,8b-tetrahydro-2-hydroxy- 30 – 45 100 → 56 0 → 44 1-[(1E,3SR,4SR )-3-hydroxy-4-methyloct-1-en-6-yn-1-yl]-1H- 45 – 60 cyclopenta[b]benzofuran-5-butanoate 60 – 70 56 → 0 44 → 100 [88475-69-8] 70 – 80 Beraprost Sodium, when dried, contains not less than 98.5z and not more than 101.0z of beraprost sodium (C24H29NaO5). Description Beraprost Sodium occurs as a white powder. It is very soluble in methanol, and freely soluble in water and in ethanol (99.5). It is hygroscopic. Flow rate: Adjust so that the retention time of the second peak of beraprost is about 23 minutes. Time span of measurement: For 80 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add methanol to make 20 mL. To 1 mL of this solu- ‌532 tion add methanol to make 20 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add methanol to make exactly 10 mL. Confirm that the total area of the two peaks of beraprost obtained with 15 mL of this solution is equivalent to 14 to 26z of that with 15 mL of the solution for system suitability test. Beraprost Sodium Tablets
Betamethasone Sodium Phosphate contains not less than 97.
Betamethasone Sodium Phosphate occurs as white to pale yellow-white, crystalline powder or masses. It is odorless. It is freely soluble in water, sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. It is hygroscopic. Melting point: about 2139C (with decomposition).
(1) Dissolve 2 mg of Betamethasone So-dium Phosphate in 2 mL of sulfuric acid: a brown color de-velops, and gradually changes to blackish brown. Prepare the test solution with 0.01 g of Betametha-sone Sodium Phosphate as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as an absorbing liquid: the test solution responds to Qualitative Tests <1.09> (2) for fluoride. Take 40 mg of Betamethasone Sodium Phosphate in a platinum crucible, and carbonize by heating. After cool-ing, add 5 drops of nitric acid, and incinerate by heating. To the residue add 10 mL of diluted nitric acid (1 in 50), and boil for several minutes. After cooling, filter if necessary, and use this solution as the sample solution. The sample so-lution responds to Qualitative Tests <1.09> (2) for phosphate. The sample solution neutralized with ammonia TS responds to Qualitative Tests <1.09> for sodium salt, and to Qualita-tive Tests <1.09> (1) and (3) for phosphate. Determine the infrared absorption spectrum of Beta-methasone Sodium Phosphate, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Betamethasone Sodium Phosphate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋99 – ＋1059(0.1 g calcu-lated on the anhydrous basis, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.10 g of Betamethasone Sodium Phos-phate in 20 mL of water: the pH of this solution is between 7.5 and 9.0.
(1) Clarity and color of solution—Dissolve 0.25 g of Betamethasone Sodium Phosphate in 10 mL of water: the solution is clear and colorless. Free phosphoric acid—Weigh accurately about 20 mg of Betamethasone Sodium Phosphate, dissolve in 20 mL of water, and use this solution as the sample solution. Sepa-rately, pipet 4 mL of Standard Phosphoric Acid Solution, add 20 mL of water, and use this solution as the standard so-lution. To each of the sample solution and the standard solu-tion add exactly 7 mL of dilute sulfuric acid, exactly 2 mL of hexaammonium heptamolybdate-sulfuric acid TS and ex-actly 2 mL of p-methylaminophenol sulfate TS, shake well, and allow to stand at 20 ± 19C for 15 minutes. To each add water to make exactly 50 mL, and allow to stand at 20 ± 19C for 15 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 20 mL of water in the same manner as the blank. Determine the absorbances, AT and AS, of each solution from the sample solution and standard solution at 730 nm: the amount of free phosphoric acid is not more than 0.5z. Amount (z) of free phosphoric acid (H3PO4) ＝ AT/AS × 1/M × 10.32 M: Amount (mg) of Betamethasone Sodium Phosphate taken, calculated on the anhydrous basis Betamethasone—Dissolve 20 mg of Betamethasone Sodium Phosphate in exactly 2 mL of methanol, and use this solution as the sample solution. Separately, dissolve 20 mg of Betamethasone RS in exactly 10 mL of methanol. Pipet 1 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a freshly prepared mixture of 1-butanol, water and acetic anhydride (3:1:1) to a distance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot from the sample solution ‌542 corresponding to the spot obtained from the standard solu-tion is not more intense than the spot from the standard so-lution. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, back titration).
Weigh accurately about 20 mg each of Betametha-sone Sodium Phosphate and Betamethasone Sodium Phos-phate RS (separately, determine the water <2.48> in the same manner as Betamethasone Sodium Phosphate), and dissolve each in methanol to make exactly 20 mL. Pipet 5 mL each of these solutions, and exactly 5 mL of the internal standard solution, then add methanol to make 50 mL, and use these solutions as the sample solution and standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of betamethasone phosphate to that of the internal standard. Amount (mg) of betamethasone sodium phosphate (C22H28FNa2O8P) ＝ MS × QT/QS MS: Amount (mg) of Betamethasone Sodium Phosphate RS taken, calculated on the anhydrous basis Internal standard solution—A solution of butyl parahy-droxybenzoate in methanol (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.6 g of tetra-n-butylammonium bromide, 3.2 g of disodium hydrogen phosphate dodecahy-drate and 6.9 g of potassium dihydrogen phosphate in 1000 mL of water, and add 1500 mL of methanol. Flow rate: Adjust so that the retention time of betametha-sone phosphate is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, betamethasone phosphate and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of betamethasone phosphate to that of the internal standard is not more than 1.0z.
Bromfenac Sodium Hydrate contains not less than 97.
Bromfenac Sodium Hydrate occurs as a yellow to orange crystalline powder. It is freely soluble in water, soluble in methanol, and slightly soluble in ethanol (99.5). It dissolves in a solution of sodium hydrogen carbonate (21 in 2500). Identiˆcation (1) Dissolve 10 mg of Bromfenac Sodium Hydrate in 500 mL of a solution of sodium hydrogen car-bonate (21 in 2500). Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Bromfenac Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Bromfenac Sodium Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Bromfenac Sodium RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. A solution of Bromfenac Sodium Hydrate (1 in 20) responds to Qualitative Tests <1.09> (1) for sodium salt. pH <2.54> Dissolve 1.0 g of Bromfenac Sodium Hydrate in 20 mL of water: the pH of the solution is between 8.4 and 10.2.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Bromfenac Sodium Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 50 mg of Bromfenac Sodium Hydrate in 100 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than bromfenac obtained from the sample so-lution is not larger than 1/10 times the peak area of brom-fenac from the standard solution, and the total area of the peaks other than bromfenac from the sample solution is not larger than the peak area of bromfenac from the standard solution. Operating conditions— Detector, column and column temperature: Proceed as di-rected in the operating conditions in the Assay. Mobile phase: Dissolve 3.85 g of ammonium acetate in 1000 mL of water, and adjust to pH 4.0 with acetic acid (100). To 570 mL of this solution add 430 mL of acetonitrile. Flow rate: Adjust so that the retention time of bromfenac is about 8 minutes. Time span of measurement: About 3 times as long as the retention time of bromfenac, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, and add methanol to make exactly 20 mL. Conˆrm that the peak area of bromfenac obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con- ditions, the number of theoretical plates and the symmetry factor of the peak of bromfenac are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bromfenac is not more than 2.0z. Water <2.48> 6.9 – 8.5z (0.15 g, volumetric titration, direct titration. Use a solution of imidazole for water deter-mination in methanol for water determination (1 in 80) instead of methanol for water determination).
Weigh accurately about 30 mg each of Bromfenac Sodium Hydrate and Bromfenac Sodium RS (separately de-termine the water <2.48> in the same manner as Bromfenac Sodium Hydrate), dissolve each in methanol to make exactly 50 mL. Pipet 5 mL each of these solutions, add the mobile phase to make them exactly 100 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 20 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of brom-fenac in each solution. Amount (mg) of bromfenac sodium (C15H11BrNNaO3) ＝ MS × AT/AS MS: Amount (mg) of Bromfenac Sodium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 266 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 3.85 g of ammonium acetate in 1000 mL of water. To 600 mL of this solution add 250 mL of methanol and 150 mL of tetrahydrofuran. Flow rate: Adjust so that the retention time of bromfenac is about 9 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of bromfenac are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of bromfenac is not more than 1.0z.
Caffeine and Sodium Benzoate occurs as a white powder. It is odorless, and has a slightly bitter taste. It is freely soluble in water, soluble in acetic acid (100) and in acetic anhydride, sparingly soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Dissolve 1 g of Caffeine and Sodium Benzoate in 10 mL of water in a separator, add 1 drop of phenolphthalein TS, and add carefully 0.01 mol/L sodium hydroxide VS dropwise until a faint red color develops. Ex-tract with three 20-mL portions of chloroform by thorough shaking, and separate the chloroform layer from the water layer. [Use the water layer for test (2).] Filter the combined chloroform extracts, evaporate the filtrate to dryness on a water bath, and proceed the following tests with the residue: To 2 mL of a solution of the residue (1 in 500) add tannic acid TS dropwise: a white precipitate, which dissolves upon the dropwise addition of tannic acid TS, is produced. To 0.01 g of the residue add 10 drops of hydrogen peroxide TS and 1 drop of hydrochloric acid, evaporate to dryness on a water bath: the residue acquires a yellow-red color. Invert the residue over a vessel containing 2 to 3 drops of ammonia TS: the color turns red-purple, and disappears upon the addition of 2 to 3 drops of sodium hydroxide TS. Dissolve 0.01 g of the residue in water to make 50 mL. To 5 mL of this solution add 3 mL of diluted acetic acid (31) (3 in 100) and 5 mL of a solution of pyridine (1 in 10), mix, add 2 mL of diluted sodium hypochlorite TS (1 in 5), and allow to stand for 1 minute. Add 2 mL of sodium thio-sulfate TS and 5 mL of sodium hydroxide TS to the solution: a yellow color develops. To 5 mL of the water layer obtained in (1) add 5 mL of water: the solution responds to the Qualitative Tests <1.09> (2) for benzoate. Heat Caffeine and Sodium Benzoate: white fumes are evolved. Ignite furthermore, and to the residue add hy-drochloric acid: bubbles are produced, and the solution re-sponds to Qualitative Tests <1.09> (1) for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Caffeine and Sodium Benzoate in 5 mL of water: the so-lution is clear and colorless. Alkalinity—Dissolve 1.0 g of Caffeine and Sodium Benzoate in 20 mL of water, and add 1 or 2 drops of phenol-phthalein TS: no red color develops. Chloride <1.03>—Dissolve 0.5 g of Caffeine and So- dium Benzoate in 10 mL of water, and add 30 mL of ethanol (95), 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 0.70 mL of 0.01 mol/L hydro-chloric acid VS, 30 mL of ethanol (95) and water to make 50 mL (not more than 0.050z). Chlorinated compounds—Dissolve 1.0 g of Caffeine and Sodium Benzoate in 40 mL of water, add 10 mL of dilute sulfuric acid, and extract with two 20-mL portions of diethyl ether. Allow the combined diethyl ether extracts to evaporate at room temperature to dryness. Place this residue and 0.7 g of calcium carbonate in a crucible, mix with a small amount of water, and dry. Ignite at about 6009C, dis-solve the residue in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washings, and add water to make 50 mL. To this so-lution add 0.5 mL of silver nitrate TS: the solution is not more turbid than the following control solution to which 0.5 mL of silver nitrate TS has been added. Control solution: Dissolve 0.7 g of calcium carbonate in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washings, and add 1.2 mL of 0.01 mol/L hydrochloric acid VS and water to make 50 mL. Heavy metals <1.07>—Dissolve 2.0 g of Caffeine and Sodium Benzoate in 47 mL of water, add slowly, with vigorous stirring, 3 mL of dilute hydrochloric acid, and filter. Discard the first 5 mL of the filtrate, neutralize the subsequent 25 mL of the filtrate with ammonia TS, and add 2 mL of dilute acetic acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution by adding 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Caffeine and Sodium Benzoate according to Method 1, and perform the test (not more than 2 ppm). Phthalic acid—To 0.10 g of Caffeine and Sodium Benzoate add 1 mL of water and 1 mL of resorcinol-sulfuric acid TS, and heat the mixture in an oil bath heated at a tem-perature between 1209C and 1259C to evaporate the water, then heat the residue for further 90 minutes, cool, and dis- ‌JP XVIII Official Monographs / Calcitonin Salmon 591 solve in 5 mL of water. To 1 mL of the solution add 10 mL of a solution of sodium hydroxide (43 in 500), shake, then examine under light at a wavelength between 470 nm and 490 nm: the green fluorescence of the solution is not more in-tense than that of the following control solution. Control solution: Dissolve 61 mg of potassium hydrogen phthalate in water to make exactly 1000 mL. Pipet exactly 1 mL of the solution, add 1 mL of resorcinol-sulfuric acid TS, and proceed as directed above. Readily carbonizable substances <1.15>—Proceed with 0.5 g of Caffeine and Sodium Benzoate, and perform the test: the solution is not more colored than Matching Fluid A. Loss on drying <2.41> Not more than 3.0z (2 g, 809C, 4 hours).
(1) Sodium benzoate—Weigh accurately about 0.2 g of Caffeine and Sodium Benzoate, previously dried, dissolve by warming in 50 mL of a mixture of acetic anhy-dride and acetic acid (100) (6:1), cool, and titrate <2.50> with 0.1 mol/L perchloric acid-dioxane VS to the first equiva-lence point (potentiometric titration). Perform a blank deter-mination in the same manner, and make any nccessary cor-rection. Each mL of 0.1 mol/L perchloric acid-dioxane VS ＝ 14.41 mg of C7H5NaO2 (2) Caffeine—Continue the titration <2.50> in (1) with 0.1 mol/L perchloric acid-dioxane VS from the first equiva-lence point to the second equivalence point (potentiometric titration). Each mL of 0.1 mol/L perchloric acid-dioxane VS ＝ 19.42 mg of C8H10N4O2
Weigh accurately about 0.4 g of Calcium Chloride Hydrate, and dissolve in water to make exactly 200 mL. Measure exactly 20 mL of this solution, add 40 mL of water, 2 mL of 8 mol/L potassium hydroxide TS and 0.1 g of NN indicator, and titrate <2.50> immediately with 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.940 mg of CaCl2.2H2O
Calcium Folinate Hydrate occurs as a white to light yellow crystalline powder. It is sparingly soluble in water, and practically insoluble in methanol and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Calcium Folinate Hydrate (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Calcium Folinate Hydrate RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cal-cium Folinate Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Calcium Folinate Hydrate (1 in 100) re-
(1) Clarity and color of solution—To 1.25 g of Calcium Folinate Hydrate add 50 mL of freshly boiled and cooled water, and warm to 409C, if necessary, to dissolve: the solution is clear, and the absorbance at 420 nm of it, de-termined as directed under Ultraviolet-visible Spectropho-tometry <2.24>, is not more than 0.25. Heavy metals <1.07>—Proceed with 0.40 g of Cal- cium Folinate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 50 ppm). Related substances—Dissolve 10 mg of Calcium Folinate Hydrate in 25 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than folinate obtained from the sample solution is not larger than the peak area of folinate from the standard solution, and the total area of the peaks other than folinate from the sample solution is not larger than 5 times the peak area of folinate from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of folinate, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution, and add water to make exactly 50 mL. Confirm that the peak area of folinate obtained with 20 mL of this so-lution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of folinate is not more than 2.0z. Water <2.48> Not less than 7.0z and not more than 17.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately about 10 mg each of Calcium Folinate Hydrate and Calcium Folinate Hydrate RS (separately determine the water <2.48> in the same manner as Calcium Folinate Hydrate), dissolve in water to make them exactly 25 mL. Pipet 5 mL each of these solutions, add the mobile phase to make exactly 25 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with exactly 20 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of folinate in each solution. sponds to Qualitative Tests <1.09> (2) and (3) for calcium salt. Optical rotation <2.49> [a]20: ＋14 – ＋199(0.1 g calculated Amount (mg) of calcium folinate (C ＝ MS × AT/AS 20H21CaN7O7) D on the anhydrous basis, water, 10 mL, 100 mm). pH <2.54> To 1.25 g of Calcium Folinate Hydrate add 50 mL of freshly boiled and cooled water, and warm to 409C, if necessary, to dissolve: the pH of this solution is between 6.8 and 8.0. MS: Amount (mg) of Calcium Folinate Hydrate RS taken, calculated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- ‌598‌ ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: A mixture of disodium hydrogen phosphate dodecahydrate solution (287 in 100,000), methanol and tetrabutylammonium hydroxide TS (385:110:4), adjusted to pH7.5 with phosphoric acid. Flow rate: Adjust so that the retention time of folinate is about 10 minutes. System suitability— System performance: Dissolve 10 mg each of Calcium Folinate Hydrate and folic acid in 100 mL of the mobile phase. When the procedure is run with 20 mL of this solution under the above operating conditions, folinate and folic acid are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of folinate is not more than 1.0z.
Calcium Gluconate Hydrate, when dried, contains not less than 99.
Calcium Gluconate Hydrate occurs as a white, crystalline powder or granules. It is soluble in water, and practically insoluble in ethanol (99.5).
(1) To separately 10 mg each of Calcium Gluconate Hydrate and calcium gluconate for thin-layer chromatography add 1 mL of water, dissolve by warming, and use these solutions as the sample solution and the stand-ard solution, respectively. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. De-velop the plate with a mixture of ethanol (95), water, ammo-nia solution (28) and ethyl acetate (5:3:1:1) to a distance of about 10 cm, air-dry the plate, and heat the plate at 1109C for 20 minutes. After cooling, spray evenly hexaammonium heptamolybdate-cerium (IV) sulfate TS on the plate, air-dry, and heat at 1109C for 10 minutes: the spots with the sample solution and the standard solution are the same in the Rf value and color tone. A solution of Calcium Gluconate Hydrate (1 in 40) responds to Qualitative Tests <1.09> (1), (2) and (3) for cal-cium salt. D Optical rotation <2.49> [a]20: ＋6 – ＋119 (after drying, 0.5 g, water, warming, after cooling, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Calcium Gluconate Hydrate in 20 mL of water by warming: the pH of the solution is be-tween 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Calcium Gluconate Hydrate in 50 mL of water by warm-ing: the solution is clear and colorless. Chloride <1.03>—Take 0.40 g of Calcium Gluconate Hydrate, and perform the test. Prepare the control solution with 0.80 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.071z). Sulfate <1.14>—Take 1.0 g of Calcium Gluconate Hydrate, and perform the test. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Dissolve 1.0 g of Calcium Gluconate Hydrate in 30 mL of water and 2 mL of dilute acetic acid by warming, cool, and add water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution, 2 mL of dilute acetic acid, and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Dissolve 0.6 g of Calcium Gluconate Hydrate in 5 mL of water by warming, add 5 mL of dilute sulfuric acid and 1 mL of bromine TS, and concentrate on a water bath to 5 mL. Perform the test with this solution as the test solution (not more than 3.3 ppm). Sucrose and reducing sugars—To 0.5 g of Calcium Gluconate Hydrate add 10 mL of water and 2 mL of dilute hydrochloric acid, and boil the solution for 2 minutes. After cooling, add 5 mL of sodium carbonate TS, allow to stand for 5 minutes, add water to make 20 mL, and filter. To 5 mL of the filtrate add 2 mL of Fehling's TS, and boil for 1 minute: no orange-yellow to red precipitate is formed imme-diately. Loss on drying <2.41> Not more than 1.0z (1 g, 809C, 2 hours).
Weigh accurately about 0.4 g of Calcium Gluconate Hydrate, previously dried, dissolve in 100 mL of water, add 2 mL of 8 mol/L potassium hydroxide TS and 0.1 g of NN indicator, and titrate <2.50> immediately with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 22.42 mg of C12H22CaO14.H2O
Calcium Hydroxide contains not less than 90.
Calcium Hydroxide occurs as a white powder. It has a slightly bitter taste. It is slightly soluble in water, very slightly soluble in boil-ing water, and practically insoluble in ethanol (95) and in diethyl ether. It dissolves in dilute acetic acid, in dilute hydrochloric acid ‌JP XVIII Official Monographs / Calcium Lactate Hydrate 599 and in dilute nitric acid. It absorbs carbon dioxide from air.
(1) Mix Calcium Hydroxide with 3 to 4 times its mass of water: the mixture is slushy and is alkaline. Dissolve 1 g of Calcium Hydroxide in 30 mL of dilute acetic acid, and boil. After cooling, neutralize with ammonia TS: the solution responds to Qualitative tests <1.09> (2) and for calcium salt.
(1) Acid-insoluble substances—To 5 g of Calcium Hydroxide add 100 mL of water, add hydrochloric acid dropwise with strring until the solution becomes acidic, and further add 1 mL of hydrochloric acid. Boil this solution for 5 minutes, cool, and filter through a tared glass filter (G4). Wash the residue with boiling water until the last washing exhibits no turbidity upon addition of silver nitrate TS, and dry at 1059C to constant mass: the mass is not more than 25 mg. Heavy metals <1.07>—Dissolve 1.0 g of Calcium Hy- droxide in 10 mL of dilute hydrochloric acid, evaporate on a water bath to dryness, dissolve the residue in 40 mL of water, and filter. To 20 mL of the filtrate add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 5 mL of dilute hydrochloric acid on a water bath to dryness, and add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 40 ppm). Magnesium and alkali metals—Dissolve 1.0 g of Cal-cium Hydroxide in a mixture of 20 mL of water and 10 mL of dilute hydrochloric acid, boil, neutralize with ammonia TS, and precipitate calcium oxalate completely by adding dropwise ammonium oxalate TS. Heat the mixture on a water bath for 1 hour, cool, dilute with water to 100 mL, shake, and filter. To 50 mL of the filtrate add 0.5 mL of sul-furic acid, evaporate to dryness, and ignite at 6009C to con-stant mass: the mass of the residue does not exceed 24 mg. Arsenic <1.11>—Dissolve 0.5 g of Calcium Hydroxide in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 4 ppm).
Weigh accurately about 1 g of Calcium Hydroxide, dissolve by adding 10 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Measure exactly 10 mL of this solution, add 90 mL of water and 1.5 mL of 8 mol/L potassium hydroxide TS, shake, allow to stand for 3 to 5 minutes, and then add 0.1 g of NN indicator. Titrate <2.50> immediately with 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS, until the red-purple color of the so-lution changes to blue. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 3.705 mg of Ca(OH)2
Calcium Lactate Hydrate, when dried, contains not less than 97.
Calcium Lactate Hydrate occurs as white, powder or granules. It is odorless, and has a slightly acid taste. A 1 g portion of it dissolves gradually in 20 mL of water, and it is slightly soluble in ethanol (95), and practically in-soluble in diethyl ether. It is partly efflorescent at ordinary temperature, and yields the anhydride at 1209C.
A solution of Calcium Lactate Hydrate (1 in 20) responds to Qualitative Tests <1.09> for calcium salt and for lactate.
(1) Clarity of solution—Dissolve 1.0 g of Calcium Lactate Hydrate in 20 mL of water by warming: the solution is clear. Acidity or alkalinity—To the solution obtained in (1) add 2 drops of phenolphthalein TS: no red color is pro-duced. Then add 0.50 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Heavy metals <1.07>—Dissolve 1.0 g of Calcium Lac- tate Hydrate in 30 mL of water and 5 mL of dilute acetic acid by warming, cool, add water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution from 2.0 mL of Standard Lead Solution and 2 mL of dilute acetic acid, and dilute with water to 50 mL (not more than 20 ppm). Magnesium or alkali metals—Dissolve 1.0 g of Cal-cium Lactate Hydrate in 40 mL of water, add 0.5 g of am-monium chloride, boil, then add 20 mL of ammonium oxa-late TS. Heat the mixture on a water bath for 1 hour, cool, dilute with water to 100 mL, and filter. To 50 mL of the fil-trate add 0.5 mL of sulfuric acid, evaporate to dryness, and ignite between 4509C and 5509C to constant mass: the mass of the residue is not more than 5 mg. Arsenic <1.11>—Dissolve 0.5 g of Calcium Lactate Hydrate in 2 mL of water and 3 mL of hydrochloric acid, and perform the test with this solution as the test solution (not more than 4 ppm). Volatile fatty acid—Warm 1.0 g of Calcium Lactate Hydrate with 2 mL of sulfuric acid: an odor of acetic acid or butyric acid is not perceptible. Loss on drying <2.41> 25.0 – 30.0z (1 g, 809C, 1 hour at first, then 1209C, 4 hours).
Weigh accurately about 0.5 g of Calcium Lactate Hydrate, previously dried, add water, dissolve by heating on a water bath, cool, and add water to make exactly 100 mL. Pipet 20 mL of this solution, then 80 mL of water and 1.5 mL of 8 mol/L potassium hydroxide TS, and allow to stand for 3 to 5 minutes. Add 0.1 g of NN indicator, and titrate ‌600 <2.50> immediately with 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solu-tion changes from red to blue. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.364 mg of C6H10CaO6
Calcium Levofolinate Hydrate contains not less than 97.
Calcium Levofolinate Hydrate occurs as a white to light yellow crystalline powder. It is sparingly soluble in water, and practically insoluble in methanol and in ethanol (99.5). It is hygroscopic. D Optical rotation [a]25: －10 – －159(0.25 g calculated on the anhydrous and residual solvent-free basis, 0.2 mol/L tris buffer solution (pH 8.1), 25 mL, 100 mm).
(1) Determine the absorption spectrum of a solution of Calcium Levofolinate Hydrate (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cal-cium Levofolinate Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Calcium Levofolinate Hydrate (1 in 200) responds to Qualitative Tests <1.09> (2) and (3) for cal-cium salt. pH <2.54> To 0.4 g of Calcium Levofolinate Hydrate add 50 mL of freshly boiled and cooled water, and warm to 409C, if necessary, to dissolve: the pH of the solution is be-tween 7.0 and 8.5.
(1) Clarity and color of solution—To 0.4 g of Cal-cium Levofolinate Hydrate add 50 mL of water, and warm to 409C, if necessary, to dissolve: the solution is clear, and its absorbance at 420 nm determined as directed under Ultra-violet-visible Spectrophotometry <2.24> is not more than 0.25. (2) Chloride—To 0.300 g of Calcium Levofolinate Hy- automatic integration method: the area of the peak other than levofolinate obtained from the sample solution is not larger than the peak area of levofolinate from the standard solution, and the total area of the peaks other than levofolinate from the sample solution is not larger than 5 times the peak area of levofolinate from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of levofolinate, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add water to make exactly 25 mL. Con-firm that the peak area of levofolinate obtained with 20 mL of this solution is equivalent to 14 to 26z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of levofolinate are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of levofolinate is not more than 2.0z. Diastereomer—Dissolve 50 mg of Calcium Levofo-linate Hydrate in 100 mL of water, and use this solution as the sample solution. Perform the test with 10 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late their amounts by the area percentage method: the amount of the peak of the diastereomer, having the relative retention time of about 2.0 to levofolinate, is not more than 0.3z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 286 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with human albumin chemically bonded silica gel for liquid chromatography (5 mm in particle ‌JP XVIII Official Monographs / Calcium Oxide 601 diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 3.4 g of sodium dihydrogen phos-phate dihydrate in 870 mL of water, adjust to pH 4.9 with sodium hydroxide TS or phosphoric acid, and add 110 mL of 2-propanol and 20 mL of acetonitrile. Flow rate: Adjust so that the retention time of levofolinate is about 16 minutes. System suitability— Test for required detectability: Dissolve 10 mg of Calcium Folinate RS in water to make 50 mL. To 1 mL of this solu-tion add the sample solution to make 20 mL, and use this so-lution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add water to make exactly 10 mL. Confirm that the peak area of the di-astereomer obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the solution for system suitability test under the above operating conditions, levofolinate and the diastereomer are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of the diastereomer is not more than 2.0z. Water <2.48> 12.0 – 17.0z (0.2 g, volumetric titration, direct titration ).
Weigh accurately about 10 mg each of Calcium Levofolinate Hydrate and Calcium Folinate RS (separately determine the water <2.48> in the same manner as Calcium Folinate Hydrate), and dissolve each in water to make ex-actly 25 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak area, AT, of levofolinate with the sample solution, and the peak area, AS, of folinate with the standard solution. Amount (mg) of calcium levofolinate (C20H21CaN7O7) ＝ MS × AT/AS MS: Amount (mg) of Calcium Folinate RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Adjust the pH of a mixture of diluted 0.05 mol/L disodium hydrogen phosphate TS (4 in 25), methanol and tetrabutylammonium hydroxide TS (385:110:4) to 7.5 with phosphoric acid. Flow rate: Adjust so that the retention time of folinate is about 10 minutes. System suitability— System performance: Dissolve 10 mg of folic acid in 50 mL of the mobile phase. To 5 mL of this solution add 5 mL of the standard solution. When the procedure is run with 20 mL of this solution under the above operating conditions, folinate and folic acid are eluted in this order with the reso-lution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of folinate is not more than 1.0z.
Calcium Oxide, when incinerated, contains not less than 98.
Calcium Oxide occurs as hard, white masses, containing a powder. It is odorless. It is very slightly soluble in boiling water, and practically insoluble in ethanol (95). One gram of Calcium Oxide dissolves almost completely in 2500 mL of water. It slowly absorbs moisture and carbon dioxide from air.
(1) Moisten Calcium Oxide with water: heat is generated and a white powder is obtained. Mix the powder with about 5 times its mass of water: the mixture is alkaline. (2) Dissolve 1 g of Calcium Oxide in 20 mL of water by adding a few drops of acetic acid (31): the solution responds to Qualitative Tests <1.09> for calcium salt.
(1) Acid-insoluble substances—Disintegrate 5.0 g of Calcium Oxide with a small amount of water, add 100 mL of water, add dropwise hydrochloric acid with stirring until the solution becomes acidic, and further add 1 mL of hydro-chloric acid. Boil the solution for 5 minutes, cool, filter through a glass filter (G4), wash the residue with boiling water until no turbidity is produced when silver nitrate TS is added to the last washing, and dry at 1059C to constant mass: the mass of the residue is not more than 10.0 mg. Carbonate—Disintegrate 1.0 g of Calcium Oxide with a small amount of water, mix thoroughly with 50 mL of water, allow to stand for a while, remove most of the super-natant milky liquid by decantation, and add an excess of dilute hydrochloric acid to the residue: no vigorous efferves-cence is produced. Magnesium and alkali metals—Dissolve 1.0 g of Cal-cium Oxide in 75 mL of water by adding dropwise hydro-chloric acid, and further add 1 mL of hydrochloric acid. Boil for 1 to 2 minutes, neutralize with ammonia TS, add drop-wise an excess of hot ammonium oxalate TS, heat the mix-ture on a water bath for 2 hours, cool, add water to make 200 mL, mix thoroughly, and filter. Evaporate 50 mL of the filtrate with 0.5 mL of sulfuric acid to dryness, and heat the residue strongly at 6009C to constant mass: the mass of the residue is not more than 15 mg. Loss on ignition <2.43> Not more than 10.0z (1 g, 9009C, constant mass).
Weigh accurately about 0.7 g of Calcium Oxide, pre-viously incinerated at 9009C to constant mass and cooled in a desiccator (silica gel), and dissolve in 50 mL of water and 8 ‌602 mL of diluted hydrochloric acid (1 in 3) by heating. Cool, and add water to make exactly 250 mL. Pipet 10 mL of the solution, add 50 mL of water, 2 mL of 8 mol/L potassium hydroxide TS and 0.1 g of NN indicator, and titrate <2.50> with 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS, until the red-purple color of the solution changes to blue. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.122 mg of CaO
Calcium Pantothenate contains not less than 98.
Calcium Pantothenate occurs as a white pow-der. It is freely soluble in water, and practically insoluble in ethanol (99.5). The pH of a solution prepared by dissolving 1.0 g of Cal-cium Pantothenate in 20 mL of water is between 7.0 and 9.0. It is hygroscopic. It shows crystal polymorphism.
(1) Determine the infrared absorption spec-trum of previously dried Calcium Pantothenate as directed in the potassium bromide disk method under Infrared Spec-trophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of dried Calcium Pantothenate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve the sample and the Reference Standard separately in water, evaporate water, dry the residues in vacuum for 24 hours using silica gel as a desiccant, and perform the test using these residues. A solution of Calcium Pantothenate (1 in 10) re-sponds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. D Optical rotation <2.49> [a]20: ＋25.0 – ＋28.59(1 g calcu-lated on the dried basis, water, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 1.0 g of Calcium Pantothenate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand- lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak, having the relative retention time of about 0.6 to pantothenic acid ob-tained from the sample solution is not larger than 1.2 times the peak area of pantothenic acid from the standard solu-tion, the area of the peak, having the relative retention time of about 0.8 is not larger than the peak area of pantothenic acid from the standard solution, the area of the peak, having the relative retention time of about 1.5 is not larger than 3/5 times the peak area of pantothenic acid from the standard solution, and the area of the peak other than pantothenic acid and the peaks mentioned above is not larger than 3/10 times the peak area of pantothenic acid from the standard solution. Additionally, the total area of the peaks other than pantothenic acid from the sample solution is not larger than 2.4 times the peak area of pantothenic acid from the stand-ard solution. For the areas of the peaks, having the relative retention time of about 0.6 and about 0.8 to pantothenic acid, multiply their correction factors, 19 and 13, respec-tively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of pantothenic acid, beginning after the sol-vent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add water to make exactly 10 mL. Con-firm that the peak area of pantothenic acid obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pantothenic acid are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pantothenic acid is not more than 2.0z. Alkaloids—Dissolve 50 mg of Calcium Pantothenate in 5 mL of water, add 0.5 mL of hexaammonium heptamo-lybdate TS and 0.5 mL of a solution of phosphoric acid (1 in 10): no white turbidity is produced. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 4 hours).
Weigh accurately about 20 mg each of Calcium Pan-tothenate and Calcium Pantothenate RS (separately deter-mine the loss on drying <2.41> in the same conditions as Cal-cium Pantothenate), dissolve each in water to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of pantothenic acid in each solution. ard Lead Solution (not more than 20 ppm). (2) Related substances—Dissolve 0.30 g of Calcium Pan-tothenate in 20 mL of water, and use this solution as the Amount (mg) of calcium pantothenate (C ＝ MS × AT/AS 18H 32CaN2O 10) sample solution. Pipet 1 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol- MS: Amount (mg) of Calcium Pantothenate RS taken, calculated on the dried basis Operating conditions— Detector: An ultraviolet absorption photometer (wave- ‌JP XVIII Official Monographs / Calcium Paraaminosalicylate Hydrate 603 length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 0.81 g of sodium 1-heptanesul-fonate and 1.36 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust to pH 2.1 with phos-phoric acid. To 980 mL of this solution add 10 mL of aceto-nitrile and 10 mL of methanol. Flow rate: Adjust so that the retention time of pantothenic acid is about 17 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pantothenic acid are not less than 10,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pantothenic acid is not more than 1.0z.
Calcium Polystyrene Sulfonate is a cation exchange resin prepared as the calcium form of the sulfonated styrene divinylbenzene copolymer. When dried, it contains not less than 7.
Calcium Polystyrene Sulfonate occurs as a pale yellow-white to light yellow powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether.
(1) Determine the infrared absorption spec-trum of Calcium Polystyrene Sulfonate, previously dried, as directed in the potassium bromide disk method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. (2) Mix 0.5 g of Calcium Polystyrene Sulfonate with 10 mL of dilute hydrochloric acid, filter, and neutralize the fil-trate with ammonia TS: the solution responds to Qualitative Tests <1.09> for calcium salt.
(1) Ammonium—Place 1.0 g of Calcium Polysty-rene Sulfonate in a flask, add 5 mL of sodium hydroxide TS, cover the flask with a watch glass having a moistened strip of red litmus paper on the underside, and boil for 15 minutes: the gas evolved does not change the red litmus paper to blue (not more than 5 ppm). Heavy metals <1.07>—Proceed with 2.0 g of Calcium Polystyrene Sulfonate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Calcium Polystyrene Sulfonate according to Method 3, and perform the test (not more than 2 ppm). Styrene—To 10.0 g of Calcium Polystyrene Sulfonate add 10 mL of acetone, shake for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of styrene in acetone to make exactly 100 mL. Pipet 1 mL of this solution, dilute with acetone to make exactly 100 mL, and use this solution as the standard solu-tion. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Gas Chro-matography <2.02> according to the following conditions. Determine the peak heights, HT and HS, of styrene in each solution: HT is not larger than HS. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A stainless steel column 3 mm in inside diameter and 2 m in length, having polyethylene glycol 20 M coated at the ratio of 15z on siliceous earth for gas chromatography (150 to 180 mm in particle diameter). Column temperature: A constant temperature of about 909C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of styrene is about 9 minutes. System suitability— System performance: Mix 10 mg of styrene with 1000 mL of acetone. When the procedure is run with 5 mL of this solu-tion under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of styrene are not less than 800 and 0.8 to 1.2, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak heights of styrene is not more than 5z. Sodium—Pipet 2 mL of the 50-mL solution obtained in the Assay (1), add 0.02 mol/L hydrochloric acid TS to make exactly 500 mL, and use this solution as the sample so-lution. Separately, weigh accurately 0.2542 g of sodium chlo-ride, previously dried at 1309C for 2 hours, and dissolve in 0.02 mol/L hydrochloric acid TS to make exactly 1000 mL. Pipet a suitable volume of this solution, and dilute with 0.02 mol/L hydrochloric acid TS to make a solution containing 1 to 3 mg of sodium (Na: 22.99) per mL, and use these solu-tions as the standard solutions. Perform the test with the sample solution and standard solutions according to Atomic Absorption Spectrophotometry <2.23> under the following conditions, and determine the amount of sodium in the sam-ple solution using the calibration curve obtained from the standard solutions: the amount of sodium is not more than 1z. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A sodium hollow-cathode lamp. Wavelength: 589.0 nm. Loss on drying <2.41> Not more than 10.0z (1 g, in vacu-um, 809C, 5 hours). Microparticles (i) Apparatus: Use an apparatus as shown in the illustration. (ii) Procedure: Weigh accurately about 5.5 g of Calcium Polystyrene Sulfonate, previously dried, add 300 mL of water of 259C, and mix for 5 minutes. Transfer this turbid solution to the sedimentation tube J, keeping a temperature at 259C, add water of 259C to 2 mm below the mark F of 20 cm of the sedimentation tube J, and then insert the pipet. Open the two-way stopcock C, exhaust air, add exactly water from the vent-hole D to the mark F of 20 cm, and close the two-way stopcock C. Shake the apparatus well ver-tically and horizontally, disperse Calcium Polystyrene Sul-fonate in water, and then open the two-way stopcock, and allow to stand at 25 ± 19C for 5 hours and 15 minutes. Then, draw exactly the meniscus of the turbid solution in sedimentation tube J up to the mark of pipet bulb A by suc-tion, open the two-way stopcock C to the outlet of pipet H, and transfer exactly measured 20 mL of the turbid solution to a weighing bottle. Repeat the procedure, and combine exactly measured 20 mL of the turbid solution. Evaporate 20 mL of this turbid solution on a water bath to dryness, dry to constant mass at 1059C, and weigh the residue as MS (g). Pipet 20 mL of used water, and weigh the residue in the same manner as MB (g). Calculate the difference mi (g) between MS and MB, and calculate the amount of microparticles (S ) by the following equation: the amount of microparticles is ‌608 Fig. Andreasen pipet not more than 0.1z. S (z) ＝ (mi × V )/(20 × MT) × 100 MT: Amount (g) of Calcium Polystyrene Sulfonate taken V: Actual volume (mL) to the mark of 20 cm at which the suction part of pipet is inserted
(1) Calcium—Weigh accurately about 1 g of Cal- Potassium exchange capacity—Pipet 50 mL of Stand-ard Potassium Stock Solution into a glass-stoppered flask containing about 1 g of dried Calcium Polystyrene Sul-fonate, accurately weighed, stir for 120 minutes, filter, and discard the first 20 mL of the filtrate. Pipet 5 mL of the sub-sequent filtrate, and add 0.02 mol/L hydrochloric acid TS to make exactly 100 mL. Pipet 10 mL of this solution, add 0.02 mol/L hydrochloric acid TS to make exactly 1000 mL, and use this solution as the sample solution. Separately, measure exactly a suitable volume of Standard Potassium Stock Solu-tion, dilute with 0.02 mol/L hydrochloric acid TS to make solutions containing 0.5 to 2.5 mg of potassium (K: 39.10) per mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solu-tions as directed under Atomic Absorption Spectrophotome-try <2.23> according to the following conditions, and deter-mine the amount, Y (mg), of potassium in 1000 mL of the sample solution, using the calibration curve obtained from the standard solutions. The exchange quantity for potassium per g of dried Calcium Polystyrene Sulfonate is 53 to 71 mg, calculating by the following equation. Exchange quantity (mg) for potassium (K) per g of dried Calcium Polystyrene Sulfonate ＝ (X － 100 Y )/M X: The amount (mg) of potassium in 50 mL of Standard Potassium Stock Solution before exchange M: The amount (g) of dried Calcium Polystyrene Sul-fonate taken Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A potassium hollow-cathode lamp. Wavelength: 766.5 nm.
Calcium Sodium Edetate Hydrate contains not less than 98.
Calcium Sodium Edetate Hydrate occurs as JP XVIII Official Monographs / Calcium Stearate 609 white, powder or particles. It is freely soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (99.5). It is hygroscopic.◆ ‌Identification (1) Dissolve 2 g of Calcium Sodium Edetate Hydrate in 10 mL of water, add 6 mL of a solution of lead (II) nitrate (33 in 1000), shake, and add 3 mL of po-tassium iodide TS: no yellow precipitate is formed. Make this solution alkaline by the addition of diluted ammonia so-lution (28) (7 in 50), and add 3 mL of ammonium oxalate TS: a white precipitate is formed. (2) Determine the infrared absorption spectrum of Cal- cium Sodium Edetate Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ A solution of Calcium Sodium Edetate Hydrate (1 in 20) responds to Qualitative Tests <1.09> (2) for sodium salt. pH <2.54> The pH of a solution of 2.0 g of Calcium So-dium Edetate Hydrate in 10 mL of water is 6.5 to 8.0.
◆(1) Clarity and color of solution—Dissolve 0.25 g of Calcium Sodium Edetate Hydrate in 10 mL of water: the solution is clear and colorless.◆ (2) Chloride <1.03>—Dissolve 0.70 g of Calcium Sodium Edetate Hydrate in water to make 20 mL. To this solution add 30 mL of dilute nitric acid, allow to stand for 30 minutes, and filter. To 10 mL of the filtrate add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.10z). (3) Heavy metals <1.07>—Proceed with 1.0 g of Cal-cium Sodium Edetate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ Disodium edetate—Dissolve 1.00 g of Calcium So-dium Edetate Hydrate in 50 mL of water, add 5 mL of am-monia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.01 mol/L magnesium chloride VS until the color of the solution changes from blue to red-purple (in-dicator: 40 mg of eriochrome black T-sodium chloride indi-cator): the amount of 0.01 mol/L magnesium chloride VS consumed is not more than 3.0 mL (not more than 1.0z). (5) Nitrilotriacetic acid—Conduct this procedure using light-resistant vessels. Dissolve 0.100 g of Calcium Sodium Edetate Hydrate in diluting solution to make exactly 25 mL, and use this solution as the sample solution. Separately, dis-solve 40.0 mg of nitrilotriacetic acid in diluting solution to make exactly 100 mL. Pipet 1 mL of this solution, add 0.1 mL of the sample solution, then add diluting solution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak areas, AT and AS, of nitrilotriacetic acid in each solution: AT is not larger than AS (not more than 0.1z). Diluting solution: Dissolve 10.0 g of iron (III) sulfate n-hydrate in 20 mL of 0.5 mol/L sulfuric acid TS and 780 mL of water, adjust to pH 2.0 with sodium hydroxide TS, and add water to make 1000 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 273 nm). Column: A stainless steel column 4.6 mm in inside diame- ter and 10 cm in length, packed with graphite carbon for liq-uid chromatography (mean pore size: 25 nm, specific sur-face: 120 m2/g, 5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 50.0 mg of iron (III) sulfate n-hydrate in 50 mL of 0.5 mol/L sulfuric acid TS, add 750 mL of water, adjust to pH 1.5 with 0.5 mol/L sulfuric acid TS or sodium hydroxide TS, and add 20 mL of ethylene glycol and water to make 1000 mL. Flow rate: 1.0 mL per minute (the retention time of nitrilotriacetic acid is about 5 minutes). System suitability— Test for required detectability: When perform the test with 20 mL of the standard solution under the above operating conditions, the SN ratio of the peak of nitrilotriacetic acid is not less than 50. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, nitrilotriacetic acid and edetic acid are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of nitrilotriacetic acid is not more than 1.0z.◆ Water <2.48> 5.0 – 13.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately about 0.5 g of Calcium Sodium Edetate Hydrate, and dissolve in water to make exactly 200 mL. Pipet 20 mL of this solution, add 80 mL of water, adjust to pH 2 – 3 with dilute nitric acid, and titrate <2.50> with 0.01 mol/L bismuth nitrate VS until the color of the so-lution changes from yellow to red (indicator: 2 drops of xylenol orange TS). Each mL of 0.01 mol/L bismuth nitrate VS ＝ 3.743 mg of C10H12CaN2Na2O8 ◆Containers and storage Containers—Tight containers.◆ Calcium Stearate
Calcium Stearate mainly consists of calcium salts of stearic acid (C18H36O2: 284.48) and palmitic acid (C16H32O2: 256.42). Calcium Stearate, when dried, contains not less than 6.
Calcium Stearate occurs as a white, light, bulky powder. It feels smooth when touched, and is adhesive to the skin. It is odorless or has a faint, characteristic odor. It is practically insoluble in water, in ethanol (95) and in diethyl ether.
(1) Shake vigorously 3 g of Calcium Stearate with 20 mL of diluted hydrochloric acid (1 in 2) and 30 mL of diethyl ether for 3 minutes, and allow to stand: the separated aqueous layer responds to Qualitative Tests <1.09> (1), (2) and (4) for calcium salt. (2) Wash the diethyl ether layer obtained in (1) with 20 mL and 10 mL of dilute hydrochloric acid and 20 mL of water successively, and evaporate the diethyl ether on a water bath: the residue melts <1.13> at a temperature not ‌610 below 549C.
(1) Heavy metals <1.07>—Heat gently 1.0 g of Calcium Stearate with caution at the beginning, and heat further, gradually raising the temperature, to incineration. After cooling, add 2 mL of hydrochloric acid, evaporate on a water bath to dryness, warm the residue with 20 mL of water and 2 mL of dilute acetic acid for 2 minutes, cool, filter, and wash the residue with 15 mL of water. Combine the filtrate and the washings, add water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution by evaporating 2 mL of hydrochlo-ric acid on a water bath to dryness and by adding 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). (2) Arsenic <1.11>—To 1.0 g of Calcium Stearate add 5 mL of diluted hydrochloric acid (1 in 2) and 20 mL of chlo-roform, shake vigorously for 3 minutes, allow to stand, and separate the water layer. Perform the test with the water layer as the test solution (not more than 2 ppm). Loss on drying <2.41> Not more than 4.0z (1 g, 1059C, 3 hours).
Weigh accurately about 0.5 g of Calcium Stearate, previously dried, heat gently with caution at first, and then ignite gradually to ash. Cool, add 10 mL of dilute hydro-chloric acid to the residue, warm for 10 minutes on a water bath, and transfer the contents to a flask with the aid of 10-mL, 10-mL, and 5-mL portions of hot water. Add so-dium hydroxide TS until the solution becomes slightly turbid, and then add 25 mL of 0.05 mol/L disodium dihy-drogen ethylenediamine tetraacetate VS, 10 mL of ammonia-ammonium chloride buffer solution (pH 10.7), 4 drops of eriochrome black T TS and 5 drops of methyl yellow TS, and titrate <2.50> rapidly the excess disodium dihydrogen ethylenediamine tetraacetate with 0.05 mol/L magnesium chloride VS, until the green color of the solution disappears and a red color develops. Perform a blank determination in the same manner. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.004 mg of Ca
Carbazochrome Sodium Sulfonate Hydrate contains not less than 98.
Carbazochrome Sodium Sulfonate Hydrate occurs as orange-yellow, crystals or crystalline powder. It is sparingly soluble in water, very slightly soluble in methanol and in ethanol (95), and practically insoluble in diethyl ether. A solution of Carbazochrome Sodium Sulfonate Hydrate (1 in 100) shows no optical rotation. Melting point: about 2109C (with decomposition).
(1) Determine the absorption spectrum of a solution of Carbazochrome Sodium Sulfonate Hydrate (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer- ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Car-bazochrome Sodium Sulfonate Hydrate as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Carbazochrome Sodium Sulfonate Hy-drate (1 in 100) responds to Qualitative Tests <1.09> (1) for sodium salt. pH <2.54> Dissolve 0.8 g of Carbazochrome Sodium Sul-fonate Hydrate in 50 mL of water by warming, and cool: the pH of this solution is between 5.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Carbazochrome Sodium Sulfonate Hydrate in 50 mL of water by warming, and allow to cool: the solution is clear. Perform the test with this solution as directed under Ultravi-olet-visible Spectrophotometry <2.24>: the absorbance at 590 nm is not more than 0.070. Heavy metals <1.07>—Proceed with 1.0 g of Car-bazochrome Sodium Sulfonate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 50 mg of Carbazo-chrome Sodium Sulfonate Hydrate in 100 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine each peak area of these solutions by the automatic integration method: the total area of the peaks other than carbazochrome sul-fonate obtained from the sample solution is not larger than the peak area of carbazochrome sulfonate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 360 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.2 g of ammonium dihydrogen phosphate in 1000 mL of water, and filter through a mem-brane filter (0.4 mm in pore size) if necessary. To 925 mL of this solution add 75 mL of ethanol (95), shake, and adjust the pH to 3 with phosphoric acid. Flow rate: Adjust so that the retention time of car-bazochrome sulfonate is about 7 minutes. Time span of measurement: About 3 times as long as the retention time of carbazochrome sulfonate, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 20 mL. Confirm that the peak area of carbazochrome sulfonate obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 10 mg each of Carbazo-chrome Sodium Sulfonate Hydrate and carbazochrome in 100 mL of water by warming. When the procedure is run with 10 mL of this solution under the above operating condi- ‌JP XVIII Official Monographs / Carbidopa Hydrate 625 tions, carbazochrome sulfonate and carbazochrome are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carbazochrome sulfonate is not more than 2.0z. Water <2.48> 13.0 – 16.0z (0.3 g, volumetric titration, direct titration).
Weigh accurately about 0.25 g of Carbazochrome Sodium Sulfonate Hydrate, dissolve in 50 mL of water, apply to a chromatographic column, 10 mm in diameter, previously prepared with 20 mL of strongly acidic ion exchange resin for column chromatography (type H), and allow to flow at a rate of 4 mL per minute. Wash the column with 150 mL of water, combine the washing and the former effluent solution, and titrate <2.50> with 0.05 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sodium hydroxide VS ＝ 16.11 mg of C10H11N4NaO5S
Carmellose Calcium occurs as a white to yel-lowish white powder. It is practically insoluble in ethanol (95) and in diethyl ether. It swells with water to form a suspension. The pH of a suspension, obtained by shaking 1.0 g of Car-mellose Calcium with 100 mL of water, is between 4.5 and 6.0. It is hygroscopic.◆
(1) Shake thoroughly 0.1 g of Carmellose Calcium with 10 mL of water, followed by 2 mL of sodium hydroxide TS, allow to stand for 10 minutes, and use this so-lution as the sample solution. To 1 mL of the sample solu-tion add water to make 5 mL. To 1 drop of this solution add 0.5 mL of chromotropic acid TS, and heat in a water bath for 10 minutes: a red-purple color develops. Shake 5 mL of the sample solution obtained in (1) with 10 mL of acetone: a white, flocculent precipitate is produced. Shake 5 mL of the sample solution obtained in (1) with 1 mL of iron (III) chloride TS: a brown, flocculent precipitate is produced. Ignite 1 g of Carmellose Calcium to ash, dissolve the residue in 10 mL of water and 6 mL of acetic acid (31), and filter, if necessary. Boil the filtrate, cool, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> (1) and (3) for calcium salt.
(1) Alkalinity—Shake thoroughly 1.0 g of Carmel-lose Calcium with 50 mL of freshly boiled and cooled water, and add 2 drops of phenolphthalein TS: no red color de-velops. Chloride <1.03>—Shake thoroughly 0.80 g of Car- mellose Calcium with 50 mL of water, add 10 mL of sodium hydroxide TS to dissolved, add water to make 100 mL, and use this solution as the sample solution. Heat 20 mL of the sample solution with 10 mL of 2 mol/L nitric acid TS on a water bath until a flocculent precipitate is produced. After cooling, centrifuge, and take out the supernatant liquid. Wash the precipitate with three 10-mL portions of water by centrifuging each time, combine the supernatant and the washings, and add water to make 100 mL. Take 25 mL of this solution, and add 1 mL of nitric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.36z). Sulfate <1.14>—Heat 10 mL of the sample solution obtained in (2) with 1 mL of hydrochloric acid in a water bath until a flocculent precipitate is produced. Cool, centri-fuge, and take out the supernatant liquid. Wash the precipi-tate with three 10-mL portions of water by centrifuging each time, combine the supernatant and the washings, and add water to make 100 mL. Perform the test with 25 mL this so-lution as the test solution. Prepare the control solution with 0.42 mL of 0.005 mol/L sulfuric acid VS. To the test solu-tion and the control solution add 1 mL of 3 mol/L hydro-chloric acid TS and 3 mL of barium chloride TS, then add water to make 50 mL, and mix. Allow to stand for 10 minutes, and compare the turbidity of these solutions: the turbidity obtained with the test solution is not more than that obtained with the control solution (not more than 1.0z). ‌632 (4) Heavy metals <1.07>—Proceed with 1.0 g of Car-mellose Calcium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 4 hours). Residue on ignition <2.44> 10 – 20z (after drying 1 g). ◆Containers and storage Containers—Tight containers.◆ Carmellose Sodium Carboxymethylcellulose Sodium
Carmellose Sodium occurs as a white to yel-lowish white, powder or granules. It has no taste. It is practically insoluble in methanol, in ethanol (95), in acetic acid (100) and in diethyl ether. It forms a viscid solution in water and in warm water. It is hygroscopic.
(1) Dissolve 0.2 g of Carmellose Sodium in 20 mL of warm water with stirring, cool, and use this solu-tion as the sample solution. To 1 mL of the sample solution add water to make 5 mL. To 1 drop of this solution add 0.5 mL of concentrated chromotropic acid TS, and heat in a water bath for 10 minutes: a red-purple color develops. To 10 mL of the sample solution obtained in test (1) add 1 mL of copper (II) sulfate TS: a blue flocculent precipi-tate is produced. To 3 g of Carmellose Sodium add 20 mL of methanol and 2 mL of dilute hydrochloric acid, boil gently on a water bath for 5 minutes, and filter. Evaporate the filtrate to dry-ness, and add 20 mL of water to the residue: the solution re-sponds to Qualitative Tests <1.09> for sodium salt. pH <2.54> Add 1.0 g of Carmellose Sodium in small por-tions to 100 mL of warm water with stirring, dissolve, and cool: the pH of this solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Firmly attach a glass plate of good quality 2 mm in thickness, to the bottom of a glass column 250 mm in height, 25 mm in inner diameter and 2 mm in thickness. This is used as an outer tube. Simi-larly prepare an inner tube by attaching a glass plate of good quality 2 mm in thickness to the bottom of a glass column 300 mm in height, 15 mm in inner diameter and 2 mm in thickness. Dissolve 1.0 g of Carmellose Sodium in 100 mL of water, pour this solution into the outer tube, and place on a piece of white paper on which 15 parallel black lines 1 mm in width and 1 mm in interval are drawn. Moving the inner tube up and down and observing from the upper part, deter-mine the height of the solution up to the lower edge of the inner tube when the distinction of the lines becomes impos-sible. Repeat the operation 3 times, and calculate the mean value: it is larger than that calculated from the similar opera-tion, using the following control solution. Control solution: To 5.50 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid, 5 mL of ethanol (95) and water to make 50 mL. Add 2 mL of barium chloride TS, mix well, and allow to stand for 10 minutes. Shake well this solution before use. Chloride <1.03>—Dissolve 0.5 g of Carmellose So- dium in 50 mL of water, and use this solution as the sample solution. Shake 10 mL of the sample solution with 10 mL of dilute nitric acid, heat to produce a flocculent precipitate in a water bath, cool, and centrifuge. Separate the supernatant liquid, wash the precipitate with three 10-mL portions of water, centrifuging each time, combine the supernatant liq-uid with the washings, and dilute with water to 200 mL. Per-form the test using 50 mL of this solution as the test solu-tion. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.640z). Sulfate <1.14>—Add 1 mL of hydrochloric acid to 10 mL of the sample solution obtained in (2), shake well, heat to produce a flocculent precipitate in a water bath, cool, and centrifuge. Separate the supernatant liquid, wash the precipi-tate with three 10-mL portions of water, centrifuging each time, combine the washings with the supernatant liquid men-tioned above, and dilute to 50 mL with water. Take 10 mL of this solution, dilute with water to 50 mL, and perform the test using this solution as the test solution. Prepare the con-trol solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.960z). Silicate—Weigh accurately about 1 g of Carmellose Sodium, ignite in a platinum dish, add 20 mL of dilute hy-drochloric acid, cover with a watch glass, and boil gently for 30 minutes. Remove the watch glass, and evaporate on a water bath to dryness with the aid of a current of air. Con-tinue heating for further 1 hour, add 10 mL of hot water, stir well, and filter through a filter paper for quantitative analy-sis. Wash the residue with hot water, dry together with the filter paper after no turbidity is produced on the addition of silver nitrate TS to the last washing, and then ignite to con-stant mass: the mass of the residue is not more than 0.5z. Heavy metals <1.07>—Proceed with 1.0 g of Carmel- lose Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Carmellose Sodium add 20 mL of nitric acid, heat gently until it becomes fluid, cool, add 5 mL of sulfuric acid, and heat until white fumes are evolved. Add, if necessary, 5 mL of nitric acid after cooling, and heat again. Repeat this operation until the solution becomes colorless or slightly yellow. After cooling, add 15 mL of a saturated solution of ammonium oxalate monohy-drate, and heat until white fumes are evolved again, cool, and dilute with water to 25 mL. Take 5 mL of this solution as the test solution, and perform the test. The solution has no more color than the following color standard. Color standard: Without using Carmellose Sodium, proceed in the same manner, then transfer 5 mL of this solu-tion to a generator bottle, add exactly 2 mL of Standard Arsenic Solution, and proceed as directed for the test with the test solution (not more than 10 ppm). Starch—Add 2 drops of iodine TS to 10 mL of the sample solution obtained in (2): no blue color develops. Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 4 hours).
Weigh accurately about 0.5 g of Carmellose So-dium, previously dried, add 80 mL of acetic acid (100), con-nect with a reflux condenser, and heat in an oil bath main-tained at 1309C for 2 hours. Cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per- ‌JP XVIII Official Monographs / Croscarmellose Sodium 633 form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 2.299 mg of Na
Carumonam Sodium contains not less than 850 mg (potency) and not more than 920 mg (potency) per mg, calculated on the anhydrous basis.
Carumonam Sodium occurs as a white to yel-lowish white, crystals or crystalline powder. It is freely soluble in water, soluble in formamide, very slightly soluble in methanol, and practically insoluble in ethanol (99.5) and in acetic acid (100).
(1) Determine the absorption spectrum of a solution of Carumonam Sodium (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Carumonam Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Carumonam Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Carumonam Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Carumonam Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a doublet signal A at around d 5.5 ppm, and a singlet signal B at around d 7.0 ppm. The ratio of the integrated intensity of these signals, A:B, is about 1:1. Carumonam Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋18.5 – ＋21.09(0.1 g calcu-lated on the anhydrous basis, water, 10 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Carumonam Sodium in 10 mL of water is between 5.0 and 6.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Carumonam Sodium in 5 mL of water: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 2.0 g of Carumonam Sodium according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Stand-ard Lead Solution (not more than 15 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Carumonam Sodium according to Method 4, and perform the test (not more than 1 ppm). Related substance 1—Weigh accurately about 0.1 g of Carumonam Sodium, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Carumonam Sodium RS, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, and add the mobile phase to make exactly 25 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of the related substances by the following equa-tion: the amount of the related substance having the relative retention time of about 0.7 to carumonam is not more than 4.0z, and each amount of the related substances other than the related substance having the relative retention time of about 0.7 is not more than 1.0z. Amount (z) of related substance ＝ MS/MT × AT/AS MS: Amount (g) of Carumonam Sodium RS taken MT: Amount (g) of Carumonam Sodium taken AS: Peak area of carumonam from the standard solution AT: Each peak area other than carumonam from the sam-ple solution Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of carumonam. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add the mobile phase to make ex-actly 50 mL. Confirm that the peak area of carumonam ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 40 mg of Carumonam So-dium in 20 mL of the mobile phase. To 5 mL of this solution add 5 mL of a solution of resorcinol in the mobile phase (9 in 1000) and the mobile phase to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, resorcinol and carumonam are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carumonam is not more than 2.0z. Related substance 2—Weigh accurately about 0.1 g of Carumonam Sodium, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 0.1 g of Carumonam Sodium RS, and dissolve in the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, and add the mobile phase to make exactly 25 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of the related substances by the following equa-tion: the amount of each related substance is not more than 1.0z. Amount (z) of related substance ＝ MS/MT × AT/AS MS: Amount (g) of Carumonam Sodium RS taken MT: Amount (g) of Carumonam Sodium taken AS: Peak area of carumonam from the standard solution AT: Each area of the peaks appeared after the peak of carumonam from the sample solution Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 10,000), methanol and acetic acid (100) (74:25:1). Flow rate: Dissolve 0.01 g of phthalic acid in the mobile phase to make 100 mL. Adjust so that the retention time of phthalic acid is about 6.5 minutes when the procedure is run with 10 mL of this solution. Time span of measurement: About 10 times as long as the retention time of carumonam. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add the mobile phase to make ex-actly 50 mL. Confirm that the peak area of carumonam ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 40 mg of Carumonam So-dium in 20 mL of the mobile phase. To 5 mL of this solution add 5 mL of a solution of resorcinol in the mobile phase (9 in 1000) and the mobile phase to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, resorcinol and carumonam are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of carumonam is not more than 2.0z. ‌JP XVIII Official Monographs / Carvedilol 637 Total amount of related substances—The total of the amounts of the related substances obtained in the Related substance 1 and the Related substance 2 is not more than 6.0z. Water <2.48> Not more than 2.0z (0.2 g, volumetric titra-tion, direct titration; Use a mixture of formamide for water determination and methanol for water determination (3:1) instead of methanol for water determination).
Weigh accurately an amount of Carumonam So-dium and Carumonam Sodium RS, equivalent to about 40 mg (potency), and dissolve each in the mobile phase to make exactly 20 mL. Measure exactly 5 mL each of these solu-tions, add exactly 5 mL of the internal standard solution and the mobile phase to make 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of carumonam to that of the internal standard. Amount [ mg (potency)] of carumonam (C12H14N6O10S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Carumonam Sodium RS taken Internal standard solution—A solution of resorcinol in the mobile phase (9 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 10,000), methanol and acetic acid (100) (97:2:1). Flow rate: Adjust so that the retention time of carumon-am is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and carumonam are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of carumonam to that of the internal stand-ard is not more than 1.0z.
Cefalotin Sodium contains not less than 920 mg (potency) and not more than 980 mg (potency) per mg, calculated on the anhydrous basis.
Cefalotin Sodium occurs as white to light yel-low-white, crystals or crystalline powder. It is freely soluble in water, slightly soluble in methanol, very slightly soluble in ethanol (95), and practically insoluble in acetonitrile.
(1) Determine the absorption spectrum of a solution of Cefalotin Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefalotin Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefalotin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefalotin Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefalotin Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic JP XVIII Official Monographs / Cefalotin Sodium for Injection 653 ‌Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a singlet signal A at around d 2.1 ppm, a singlet or sharp mul-tiplet signal B at around d 3.9 ppm, and a multiplet signal C at around d 7.0 ppm. The ratio of the integrated intensity of these signals, A:B:C, is about 3:2:2. Cefalotin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]25: ＋124 – ＋1349(5 g, water, 100 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Cefalotin Sodium in 10 mL of water is between 4.5 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefalotin Sodium in 10 mL of water: the solution is clear. The absorbance of this solution at 450 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.20. Heavy metals <1.07>—Proceed with 1.0 g of Cefalo- tin Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefalotin Sodium according to Method 3, and perform the test (not more than 2 ppm). Related substances—Pipet 1 mL of the standard solu-tion obtained in the Assay, add the mobile phase to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solu-tion obtained in the Assay and the standard solution pre-pared here as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks other than cefalotin from the sample solution is not larger than the peak area of cefalotin from the standard solution, and the total area of the peaks other than cefalotin obtained from the sample solution is not larger than 3 times the peak area of cefalotin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of cefalotin. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cefalotin ob-tained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Heat the standard solution in a water bath of 909C for 10 minutes, and cool. Measure ex-actly 2.5 mL of this solution, and add the mobile phase to make exactly 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak of cefalotin and the peak, hav-ing the relative retention time of about 0.5 to cefalotin, is not less than 9, and the symmetry factor of the peak of cefalotin is not more than 1.8. System repeatability: When the test is repeated 3 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefalotin is not more than 2.0z. Water <2.48> Not more than 1.0z (0.5 g, volumetric titra- tion, back titration).
Weigh accurately an amount of Cefalotin Sodium and Cefalotin Sodium RS, equivalent to about 25 mg (po-tency), and dissolve each in the mobile phase to make exactly 25 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of cefalotin in each solution. Amount [ mg (potency)] of cefalotin (C16H16N2O6S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefalotin Sodium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 17 g of sodium acetate trihydrate in 790 mL of water, and add 0.6 mL of acetic acid (100). If necessary adjust the pH to 5.9 ± 0.1 with diluted sodium hydroxide TS (1 in 10) or acetic acid (100). To this solution add 150 mL of acetonitrile and 70 mL of ethanol (95). Flow rate: Adjust so that the retention time of cefalotin is about 12 minutes. System suitability— System performance: Heat the standard solution in a water bath of 909C for 10 minutes, and cool. Measure ex-actly 2.5 mL of this solution, and add the mobile phase to make exactly 100 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the resolution between the peak of cefalotin and the peak, hav-ing the relative retention time of about 0.5 to cefalotin is not less than 9, and the symmetry factor of the peak of cefalotin is not more than 1.8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefalotin is not more than 1.0z.
Cefazolin Sodium Hydrate contains not less than 920 mg (potency) and not more than 975 mg (potency) per mg, calculated on the anhydrous basis.
Cefazolin Sodium Hydrate occurs as white to pale yellowish white crystals. It is freely soluble in water, sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether.
(1) Determine the absorption spectrum of a solution of Cefazolin Sodium Hydrate (1 in 50,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefazolin Sodium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefazolin Sodium Hydrate in heavy water for nuclear magnetic reso-nance spectroscopy (1 in 10), using sodium 3-trimethylsilyl- more than 1.0z, the amount of the peak other than cefazo-lin and the peak mentioned above is not more than 0.5z, and the total amount of the peaks other than cefazolin is not more than 2.0z. For the area of the peak, having the rela-tive retention time of about 0.2 to cefazolin, multiply the correction factor 1.43. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cefazolin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Confirm that the peak area of cefazolin obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suitability test. System performance: Dissolve 20 mg of Cefazolin Sodium Hydrate in 20 mL of a solution of p-acetanisidide in 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000). When the procedure is run with 5 mL of this solution under the above operating conditions, cefazolin and p-acetanisidide are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation ‌JP XVIII Official Monographs / Cefbuperazone Sodium 659 of the peak area of cefazolin is not more than 2.0z. Water <2.48> Not less than 13.7z and not more than 16.0z (0.1 g, volumetric titration, direct titration. Use a mixture of formamide for water determination and metha-nol for water determination (2:1) instead of methanol for water determination). Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency).
(1) Clarity and color of solution—Dissolve 1.0 g of Cefazolin Sodium Hydrate in 10 mL of water: the solu-tion is clear, and when determine the absorbance at 400 nm of this solution as directed under Ultraviolet-visible Spectro-photometry <2.24>, it is not more than 0.15. Heavy metals <1.07>—Proceed with 2.0 g of Cefazo- lin Sodium Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 of Cefazolin So-dium Hydrate in 20 mL of 0.1 mol/L phosphate buffer solu-tion (pH 7.0) and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method, and calculate the amount of them by the area percentage method: the amount of the peak hav- 14 13 8 4 3 2 ing the relative retention time of about 0.2 to cefazolin is not Monosodium (6R,7R)-3-(5-methyl-1,3,4-thiadiazol-2-ylsulfanylmethyl)-8-oxo-7-[2-(1H-tetrazol- yl)acetylamino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate pentahydrate [115850-11-8] Cefazolin Sodium Hydrate contains not less than 920 mg (potency) and not more than 975 mg (potency) per mg, calculated on the anhydrous basis. The po-tency of Cefazolin Sodium Hydrate is expressed as mass (potency) of cefazolin (C14H14N8O4S3: 454.51). Description Cefazolin Sodium Hydrate occurs as white to pale yellowish white crystals. It is freely soluble in water, sparingly soluble in methanol, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. Identification (1) Determine the absorption spectrum of a solution of Cefazolin Sodium Hydrate (1 in 50,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cefazolin Sodium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefazolin Sodium Hydrate in heavy water for nuclear magnetic reso-nance spectroscopy (1 in 10), using sodium 3-trimethylsilyl- more than 1.0z, the amount of the peak other than cefazo-lin and the peak mentioned above is not more than 0.5z, and the total amount of the peaks other than cefazolin is not more than 2.0z. For the area of the peak, having the rela-tive retention time of about 0.2 to cefazolin, multiply the correction factor 1.43. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cefazolin, beginning after the solvent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 100 mL, and use this solution as the solution for sys-tem suitability test. Pipet 1 mL of the solution for system suitability test, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 10 mL. Confirm that the peak area of cefazolin obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for system suitability test. System performance: Dissolve 20 mg of Cefazolin Sodium Hydrate in 20 mL of a solution of p-acetanisidide in 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000). When the procedure is run with 5 mL of this solution under the above operating conditions, cefazolin and p-acetanisidide are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation ‌JP XVIII Official Monographs / Cefbuperazone Sodium 659 of the peak area of cefazolin is not more than 2.0z. Water <2.48> Not less than 13.7z and not more than 16.0z (0.1 g, volumetric titration, direct titration. Use a mixture of formamide for water determination and metha-nol for water determination (2:1) instead of methanol for water determination). Bacterial endotoxins <4.01> Less than 0.10 EU/mg (po-tency).
Weigh accurately an amount of Cefazolin Sodium Hydrate and Cefazolin RS, equivalent to about 20 mg (po-tency), dissolve in exactly 20 mL of the internal standard so-lution, and use these solutions as the sample solution and standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of cefazolin to that of the internal standard. Amount [ mg (potency)] of cefazolin (C14H14N8O4S3) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefazolin RS taken Internal standard solution—A solution of p-acetanisidide in 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.27 g of disodium hydrogen phos-phate dodecahydrate and 0.47 g of citric acid monohydrate in water to make 935 mL. To this solution, add 65 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefazolin is about 8 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cefazolin and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of cefazolin is not more than 1.0z.
Cefazolin Sodium contains not less than 900 mg (potency) and not more than 975 mg (potency) per mg, calculated on the anhydrous basis.
Cefazolin Sodium occurs as a white to light yellow-white, crystals or crystalline powder. It is freely soluble in water and in formamide, slightly soluble in methanol, and practically insoluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Cefazolin Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Cefazolin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefazolin Sodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10), using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an inter-nal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits singlet signals, A and B, at around d 2.7 ppm and at around d 9.3 ppm, re-spectively. The ratio of integrated intensity of these signals, A:B, is about 3:1. Cefazolin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －19 – －239(2.5 g calculated as the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefazolin Sodium in 10 mL of water: pH of the solution is between 4.8 and 6.3.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefazolin Sodium in 10 mL of water: the solution is clear and colorless to pale yellow, and its absorbance at 400 nm determined as directed under Ultraviolet-visible Spectropho-tometry <2.24> is not more than 0.35. The test should be per-formed within 10 minutes after preparing of the solution. Heavy metals <1.07>—Proceed with 2.0 g of Cefazo-lin Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Cefazolin Sodium according to Method 3, and perform the test. When prepare the test solution, add 1.5 mL of hydrogen peroxide (30) after addition of 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 50), and then ignite (not more than 1 ppm). Related substances—Dissolve 0.10 g of Cefazolin So-dium in 20 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and use this solution as the sample solution. Prepare the sample solution before use. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late the amount of the peaks by the area percentage method: the amount of the peak, having the relative retention time of about 0.2 to cefazolin and the amount of the peak other than cefazolin and the peak mentioned above are not more than 1.5z, respectively. The total amount of the peaks other than cefazolin is not more than 2.5z. For the area of the peak, having the relative retention time of about 0.2 to the cefazo-lin, multiply the correction factor, 1.43. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cefazolin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Dissolve about 80 mg of Cefazolin RS in 0.1 mol/L phosphate buffer solution (pH 7.0) to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 20 mL. Confirm that the peak area of cefazolin obtained with 5 mL of this solution is equivalent to 3 to 7z of that with 5 mL of the solution for system suitability test. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak areas of cefazolin is not more than 1.0z. Water <2.48> Not more than 2.5z (1.0 g, volumetric titra-tion, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination).
Weigh accurately an amount of Cefazolin Sodium ‌JP XVIII Official Monographs / Cefazolin Sodium for Injection 657 and Cefazolin RS, equivalent to about 20 mg (potency), dis-solve each in the internal standard solution to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefazolin to that of the internal standard. drug is granted approval based on the Law. pH <2.54> The pH of a solution prepared by dissolving an amount of Cefazolin Sodium for Injection, equivalent to 1.0 g (potency) of Cefazolin Sodium, in 10 mL of water is 4.5 to 6.5. Purity (1) Clarity and color of solution—Conduct this procedure within 10 minutes after the preparation of the solutions. A solution prepared by dissolving an amount of Amount [mg (potency)] of cefazolin (C H N O S ) Cefazolin Sodium for Injection, equivalent to 1.0 g (po- ＝ MS × QT/QS × 1000 14 14 8 4 3 tency) of Cefazolin Sodium, in 10 mL of water is clear, and the absorbance of this solution at 400 nm, determined as di- MS: Amount [mg (potency)] of Cefazolin RS taken Internal standard solution—A solution of p-acetanisidide in 0.1 mol/L phosphate buffer solution (pH 7.0) (11 in 20,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.27 g of disodium hydrogen phos-phate dodecahydrate and 0.47 g of citric acid monohydrate in water to make 935 mL, and add 65 mL of acetonitrile. Flow rate: Adjust so that the retention time of cefazolin is about 8 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cefazolin and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of cefazolin to that of the internal standard is not more than 1.0z.
Cefbuperazone Sodium contains not less than 870 mg (potency) per mg, calculated on the anhydrous basis.
Cefbuperazone Sodium occurs as white to light yellow-white, powder or masses. It is very soluble in water, freely soluble in methanol and in pyridine, sparingly soluble in ethanol (95), and very slightly soluble in acetonitrile.
(1) Determine the absorption spectrum of a solution of Cefbuperazone Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Dissolve 0.1 g of Cefbuperazone Sodium in 0.5 mL of deuterated pyridine for nuclear magnetic resonance spec-troscopy and 1 drop of heavy water for nuclear magnetic resonance spectroscopy, and determine the 1H spectrum of this solution as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a triplet signal A at around d 1.1 ppm, and two doublet signals, B and C, at around d 1.6 ppm and at around d 5.1 ppm, respectively. The ratio of the inte-grated intensity of each signal, A:B:C, is about 3:3:1. Cefbuperazone Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋48 – ＋569(0.4 g calculated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefbuperazone Sodium in 4 mL of water: the pH of the solution is between 4.0 and 6.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefbuperazone Sodium in 4 mL of water: the solution is clear and light yellow. Heavy metals <1.07>—Proceed with 2.0 g of Cef- buperazone Sodium according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefbuperazone Sodium according to Method 4, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Cefbupera- ‌660 zone Sodium in 100 mL of the mobile phase, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method. Calculate the percentages of each peak area of related substances ob-tained from the sample solution against 50 times of the peak area of cefbuperazone from the standard solution; the amount of related substance I having the relative retention time of about 0.2 to cefbuperazone is not more than 2.0z, the amount of related substance II having the relative reten-tion time of about 0.6 is not more than 4.5z and the amount of related substance III having the relative retention time of about 1.6 is not more than 1.0z, and the total amount of these related substances is not more than 6.0z. For the peak areas of the related substances I and III, multi-ply their correction factors, 0.72 and 0.69, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of cefbuperazone. System suitability— Test for required detectability: Measure exactly 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cefbuperazone obtained with 25 mL of this solution is equivalent to 7 to 13z of that with 25 mL of the standard solution. System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefbuperazone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefbuperazone is not more than 2.0z. Water <2.48> Not more than 1.0z (3 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Cefbuperazone So-dium and Cefbuperazone RS, equivalent to about 0.1 g (po-tency), and dissolve each in the mobile phase to make exactly 100 mL. Measure exactly 10 mL each of these solutions, add exactly 10 mL of the internal standard solution and the mobile phase to make 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefbuperazone to that of the internal standard. Amount [ mg (potency)] of cefbuperazone (C22H29N9O9S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefbuperazone RS taken Internal standard solution—A solution of acetanilide in the mobile phase (1 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 2.0 g of tetra-n-propylammonium bromide in 1000 mL of a mixture of water, acetonitrile and acetic acid-sodium acetate buffer solution (pH 5.0) (83:13:4). Flow rate: Adjust so that the retention time of cefbupera-zone is about 16 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefbuperazone are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefbuperazone to that of the internal standard is not more than 1.0z.
Cefmetazole Sodium contains not less than 860 mg (potency) and not more than 965 mg (potency) per mg, calculated on the anhydrous basis.
Cefmetazole Sodium occurs as a white to light yellow-white, powder or mass. It is very soluble in water, freely soluble in methanol, slightly soluble in ethanol (95), and very slightly soluble in tetrahydrofuran. It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Cefmetazole Sodium (1 in 40,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cef-metazole Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefmeta-zole Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits singlet signals, A, B and C, at around d 3.6 ppm, at around d 4.1 ppm and at around d 5.2 ppm, respectively. The ratio of integrated intensity of each signal, A:B:C, is about 3:3:1. Cefmetazole Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋73 – ＋859(0.25 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefmetazole Sodium in 10 mL of water: the pH of the solution is between 4.2 and 6.2.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefmetazole Sodium in 10 mL of water: the solution is clear, and has no more color than the following control solution. Control solution: To a mixture of exactly 0.5 mL of Cobalt (II) Chloride CS and exactly 5 mL of Iron (III) Chloride CS add water to make exactly 50 mL. To exactly 15 mL of this solution add water to make exactly 20 mL. Heavy metals <1.07>—Proceed with 1.0 g of Cef-metazole Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefmetazole Sodium according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.50 g of Cefmetazole Sodium in 10 mL of water, and use this solution as the sam-ple solution. Pipet 4 mL, 2 mL, 1 mL, 0.5 mL and 0.25 mL of the sample solution, add water to them to make exactly 100 mL, and use these solutions as the standard solutions (1), (2), (3), (4) and (5), respectively. Separately, dissolve 0.10 g of 1-methyl-1H-tetrazole-5-thiol in water to make ex-actly 100 mL, and use this solution as the standard solution (6). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 1 mL each of the sample solution and standard solutions (1) to (6) on a plate of silica gel for thin-layer chromatography. Develop with a mixture of 1-butanol, water and acetic acid (100) (4:1:1) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the spot obtained from the sample solution corresponding to the spot from the standard solution (6) is not more intense than the spot from the stand-ard solution (6), and the spots other than this spot and other than the principal spot are not more intense than the spot from the standard solution (1). Furthermore, the total amount of the spots other than the principal spot from the sample solution, calculated by the comparison with the spots from the standard solutions (1), (2), (3), (4) and (5), is not more than 8.0z. Water <2.48> Not more than 1.0z (1 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Cefmetazole So-dium and Cefmetazole RS, equivalent to about 50 mg (po-tency), and dissolve each in the mobile phase to make exactly 25 mL. Pipet 1 mL each of these solutions, add exactly 10 mL of the internal standard solution, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with 10 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefmeta-zole to that of the internal standard. Amount [mg (potency)] of cefmetazole (C15H17N7O5S3) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefmetazole RS taken Internal standard solution—A solution of methyl para-hydroxybenzoate in the mobile phase (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 214 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.75 g of ammonium dihydrogen-phosphate in 700 mL of water, add 280 mL of methanol, 20 mL of tetrahydrofuran and 3.2 mL of 40z tetrabutylammo-nium hydroxide TS, and adjust to pH 4.5 with phosphoric acid. Flow rate: Adjust so that the retention time of cefmetazole is about 8 minutes. ‌678‌ System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefmetazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefmetazole to that of the internal standard is not more than 1.0z.
Cefminox Sodium Hydrate contains not less than 900 mg (potency) and not more than 970 mg (potency) per mg, calculated on the anhydrous basis.
Cefminox Sodium Hydrate occurs as a white to light yellow-white crystalline powder. It is freely soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Cefminox Sodium Hydrate (1 in 50,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefminox Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wave- ‌JP XVIII Official Monographs / Cefodizime Sodium 679 lengths. Determine the infrared absorption spectrum of Cef-minox Sodium Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefminox Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefmi-nox Sodium Hydrate in heavy water for nuclear magnetic resonance spectroscopy (1 in 30) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropanesulfonate for nuclear magnetic reso-nance spectroscopy as an internal reference compound: it exhibits a multiplet signal, A, at around d 3.2 ppm, a singlet signal, B, at around d 3.5 ppm, a singlet signal, C, at around d 4.0 ppm, and a singlet signal, D, at around d 5.1 ppm. The ratio of integrated intensity of each signal, A:B:C:D, is about 2:3:3:1. Cefminox Sodium Hydrate responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋62 – ＋729(50 mg, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.70 g of Cefminox Sodium Hydrate in 10 mL of water: the pH of the solution is between 4.5 and 6.0.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Cefminox Sodium Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Cefminox Sodium Hydrate according to Method 3, and perform the test (not more than 1 ppm). Water <2.48> Not less than 18.0z and not more than 20.0z (0.1 g, volumetric titration, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Escherichia coli NIHJ Culture medium—Use the medium iii in 3) under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 6.5 to 6.6 after sterilization. Standard solution—Weigh accurately an amount of Cefminox Sodium RS, equivalent to about 40 mg (potency), dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.05 mol/L phosphate buffer solution (pH 7.0) to make solutions so that each mL contains 40 mg (potency) and 20 mg (potency), and use these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively. Sample solution—Weigh accurately an amount of Cefminox Sodium Hydrate equivalent to about 40 mg (po-tency), dissolve in 0.05 mol/L phosphate buffer solution (pH 7.0) to make exactly 50 mL. Take exactly a suitable amount of this solution, add 0.05 mol/L phosphate buffer solution (pH 7.0) to make solutions so that each mL contains 40 mg (potency) and 20 mg (potency), and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively. Procedure—Incubate between 329C and 359C.
Cefodizime Sodium contains not less than 890 mg (potency) per mg, calculated on the anhydrous and ethanol-free basis.
Cefodizime Sodium occurs as a white to light yellow-white crystalline powder. It is very soluble in water, and practically insoluble in acetonitrile and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Cefodizime Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefodizime Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Cefodizime Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cefodizime Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cefodi-zime Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits singlet signals, A, B and C, at around d 2.3 ppm, at around d 4.0 ppm, and at around d 7.0 ppm. The ratio of the inte-grated intensity of these signals, A:B:C, is about 3:3:1. Cefodizime Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －56 – －629(0.2 g calculated on the anhydrous and ethanol-free basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefodizime Sodium in 10 mL of water: the pH of the solution is between 5.5 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefodizime Sodium in 10 mL of water: the solution is clear and pale yellow to light yellow. Heavy metals <1.07>—Weigh 1.0 g of Cefodizime Sodium in a crucible, cover loosely, and carbonize by gentle heating. After cooling, add 2 mL of sulfuric acid, heat gradually until the white fumes are no longer evolved, and ignite between 5009C and 6009C. Proceed according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefodizime Sodium according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 30 mg of Cefodizime Sodium in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peaks other than cefodizime obtained from the sample solution is not larger than the peak area of cefodizime from the stand-ard solution, and the total area of the peaks other than cefodizime from the sample solution is not larger than 3 times the peak area of cefodizime from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of ethanol is about 3 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ethanol and the internal standard are eluted in this order with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ethanol to that of the internal standard is not more than 2.0z. Water <2.48> Not more than 4.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Cefodizime Sodium and Cefodizime Sodium RS, equivalent to about 50 mg (po-tency), add exactly 10 mL of the internal standard solution to dissolve, add water to make 100 mL, and use these solu-tions as the sample solution and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of cefodizime to that of the internal standard. Time span of measurement: About 4 times as long as the retention time of cefodizime, beginning after the solvent peak. Amount [ mg (potency)] of cefodizime (C ＝ MS × QT/QS × 1000 20H20N6O7S4) System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mobile phase to make ex-actly 20 mL. Confirm that the peak area of cefodizime ob-tained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the standard solution. Ethanol—Weigh accurately about 1 g of Cefodizime Sodium, and dissolve in water to make exactly 10 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal stand-ard solution, and use this solution as the sample solution. Separately, weigh accurately about 2 g of ethanol for gas chromatography, and add water to make exactly 1000 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal standard solution, and use this solution as the standard solu-tion. Perform the test with 10 mL each of the sample solution and standard solution as directed under Gas Chromatogra-phy <2.02> according to the following conditions, and calcu-late the ratios, QT and QS, of the peak area of ethanol to that of the internal standard: the amount of ethanol is not more than 2.0z. Amount (z) of ethanol ＝ MS/MT × QT/QS MS: Amount (g) of ethanol for gas chromatography taken MT: Amount (g) of Cefodizime Sodium taken Internal standard solution—A solution of 1-propanol (1 in 400). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3.2 mm in inside diameter and 3 m in length, packed with tetrafluoroethylene polymer for gas chromatography (180 – 250 mm in particle diameter) coated in 15z with polyethylene glycol 20 M. Column temperature: A constant temperature of about 1009C. MS: Amount [mg (potency)] of Cefodizime Sodium RS taken Internal standard solution—A solution of anhydrous caffeine (3 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.80 g of potassium dihydrogen phosphate and 0.20 g of anhydrous disodium hydrogen phosphate in a suitable amount of water, and add 80 mL of acetonitrile and water to make 1000 mL. Flow rate: Adjust so that the retention time of cefodizime is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, cefodizime and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefodizime to that of the internal stand-ard is not more than 2.0z.
Cefoperazone Sodium contains not less than 871 mg (potency) and not more than 986 mg (potency) per mg, calculated on the anhydrous basis.
Cefoperazone Sodium occurs as a white to yel-lowish white crystalline powder. It is very soluble in water, soluble in methanol, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Cefoperazone Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the 1H spectrum of a solution of Cefoper-azone Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a triplet signal A at around d 1.2 ppm, a doublet signal B at around d 6.8 ppm, and a doublet signal C at around d 7.3 ppm. The ratio of integrated intensity of these signals, A:B:C, is about 3:2:2. Cefoperazone Sodium responds to the Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －15 – －259(1 g, water, 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefoperazone Sodium in 4 mL of water: the pH of the solution is between 4.5 and 6.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefoperazone Sodium in 10 mL of water: the solution is clear, and its absorbance at 400 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.18. Heavy metals <1.07>—Proceed with 1.0 g of Cefoper- azone Sodium according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefoperazone Sodium according to Method 4, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.1 g of Cefoperazone Sodium in 100 mL of water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add water to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 25 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method. Calculate the percentages of each peak area obtained from the sample solution against 50 times of the peak area of cefoperazone from the standard solution: the related substance I with the retention time of about 8 minutes is not more than 5.0z, the related substance II with that of about 17 minutes is not more than 1.5z, and the total amount of all related substances is not more than 7.0 z. For the peak areas of the related substances I and II, multiply their correction factors, 0.90 and 0.75, respectively. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cefoperazone, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 20 mL. Confirm that the peak area of cefoperazone obtained with 25 mL of this solution is equivalent to 3.5 to 6.5z of that with 25 mL of the standard solution. System performance: When the procedure is run with 25 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cefoperazone are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 25 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefoperazone is not more than 2.0z. Water <2.48> Not more than 1.0z (3 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Cefoperazone So-dium equivalent to about 0.1 g (potency), and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accu-rately an amount of Cefoperazone RS equivalent to about 20 mg (potency), dissolve in 1 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and add water to make exactly 20 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, and use this solution as the stand-ard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of cefoperazone to that of the internal standard. Amount [mg (potency)] of cefoperazone (C25H27N9O8S2) ＝ MS × QT/QS × 5000 MS: Amount [mg (potency)] of Cefoperazone RS taken Internal standard solution—A solution of acetanilide in a mixture of water and acetonitrile (43:7) (3 in 8000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- ‌682 ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 57 mL of acetic acid (100) add 139 mL of triethylamine and water to make 1000 mL. To 20 mL of this solution add 835 mL of water, 140 mL of acetonitrile and 5 mL of dilute acetic acid. Flow rate: Adjust so that the retention time of cefopera-zone is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and cefoperazone are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of cefoperazone to that of the internal standard is not more than 1.0z.
Cefotaxime Sodium contains not less than 916 mg (potency) and not more than 978 mg (potency) per mg, calculated on the dried basis.
Cefotaxime Sodium occurs as white to light yellow-white crystalline powder. It is freely soluble in water, sparingly soluble in methanol, and very slightly soluble in ethanol (95).
(1) Dissolve 2 mg of Cefotaxime Sodium in 0.01 mol/L hydrochloric acid TS to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both Amount [mg (potency)] of cefoperazone (C ＝ MS2 × QTb/QSb 25H27N9O8S2) spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of MS1: Amount [mg (potency)] of Sulbactam RS taken MS2: Amount [mg (potency)] of Cefoperazone RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate (7 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of 0.005 mol/L tetrabutylammo-nium hydroxide TS and acetonitrile for liquid chromatogra-phy (3:1). Flow rate: Adjust so that the retention time of sulbactam is about 7 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sulbactam, the internal standard, and cefoperazone are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak of sulbactam is not more than 1.0z.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefotaxime Sodium in 10 mL of water: the solution is clear, and its absorbance at 430 nm, determined as directed under Ultraviolet-visible Spectrophotometry <2.24>, is not more than 0.40. Sulfate <1.14>—Dissolve 2.0 g of Cefotaxime Sodium in 40 mL of water, add 2 mL of dilute hydrochloric acid and water to make 50 mL, shake well, and filter. Discard first 10 mL of the filtrate, and to the subsequent 25 mL of the fil-trate add water to make 50 mL. Perform the test with this JP XVIII Official Monographs / Cefotetan 685 ‌solution as the test solution. Prepare the control solution as follows: To 1.0 mL of 0.005 mol/L sulfuric acid VS add 1 mL of dilute hydrochloric acid and water to make 50 mL (not more than 0.048z). 600 mL of this solution add 400 mL of methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Heavy metals <1.07>—Proceed with 1.0 g of Cefotaxime Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefotaxime Sodium according to Method 3, and perform the test (not more than 2 ppm). Related substances—Perform the test with 10 mL of the sample solution obtained in the Assay as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area obtained from the chromatogram by the automatic integration method, and calculated the amounts of them by the area percentage method: the amount of the peak other than cefotaxime is not more than 1.0z and the total amount of these peaks is not more than 3.0z. Operating conditions— Detector, column, column temperature, mobile phase A, mobile phase B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3.5 times as long as the retention time of cefotaxime, beginning after the solvent peak. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the Assay. Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mobile phase A to make exactly 100 mL. Pipet 2 mL of this solution, and add the mobile phase A to make exactly 20 mL. Confirm that the peak area of cefotaxime obtained with 10 mL of this solution is equivalent to 0.15 to 0.25z of that with 10 mL of the standard solution. Loss on drying <2.41> Not more than 3.0z (1 g, 1059C, 3 hours).
Weigh accurately an amount of Cefotaxime Sodium and Cefotaxime RS, equivalent to about 40 mg (potency), dissolve each in the mobile phase A to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of cefotaxime in each solution. Amount [ mg (potency)] of cefotaxime (C16H17N5O7S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefotaxime RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: To 0.05 mol/L disodium hydrogen phos-phate TS add phosphoric acid to adjust the pH to 6.25. To 860 mL of this solution add 140 mL of methanol. Mobile phase B: To 0.05 mol/L disodium hydrogen phos-phate TS add phosphoric acid to adjust the pH to 6.25. To 0 – 7 100 0 7 – 9 100 → 80 0 → 20 9 – 16 80 20 16 – 45 80 → 0 20 → 100 45 – 50 0 100 Flow rate: 1.3 mL per minute (the retention time of cefotaxime is about 14 minutes). System suitability— System performance: To 1 mL of the standard solution add 7.0 mL of water and 2.0 mL of methanol, mix, then add 25 mg of sodium carbonate decahydrate, and shake. After allowing to stand for 10 minutes, add 3 drops of acetic acid (100) and 1 mL of the standard solution, and mix. When the procedure is run with 10 mL of this solution under the above operating conditions, desacetyl cefotaxime with the relative retention time being about 0.3 to cefotaxime and cefotaxime are eluted in this order with the resolution between these peaks being not less than 20, and the symmetry factor of the peak of cefotaxime is not more than 2. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cefotaxime is not more than 2.0z.
Cefpiramide Sodium contains not less than 900 mg (potency) and not more than 990 mg (potency) per mg, calculated on the anhydrous basis.
Cefpiramide Sodium occurs as white to yellow-ish white powder. It is very soluble in dimethylsulfoxide, freely soluble in water, sparingly soluble in methanol, and slightly soluble in JP XVIII Official Monographs / Cefpiramide Sodium 693 ethanol (95).
(1) Determine the absorption spectrum of a solution of Cefpiramide Sodium in 0.05 mol/L phosphate buffer solution (pH 7.0) (1 in 50,000) as directed under Ul- ASa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the standard solution ASb: Peak area of cefpiramide from the standard solution ATa: Peak area of 1-methyl-1H-tetrazole-5-thiol from the sample solution traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. ATc : Area of each peak other than 1-methyl-1H-tetrazole-5-thiol and cefpiramide from the sample solution Determine the 1H spectrum of a solution of Cef-piramide Sodium in deuterated dimethylsulfoxide for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits singlet signals, A, B and C, at around d 2.3 ppm, at around d 3.9 ppm and at around d 8.2 ppm, respectively. The ratio of the integrated intensity of each signal, A:B:C, is about 3:3:1. Cefpiramide Sodium responds to the Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －33 – －409(0.2 g calculated on the anhydrous basis, 0.05 mol/L phosphate buffer solu-tion (pH 7.0), 10 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 2.0 g of Cefpiramide Sodium in 20 mL of water is between 5.5 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Cefpiramide Sodium in 10 mL of 0.05 mol/L phosphate buffer solution (pH 7.0): the solution is clear, and colorless or light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Cef- piramide Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Weigh accurately about 25 mg of Cefpiramide Sodium, dissolve in 0.03 mol/L phosphate buffer solution (pH 7.5) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of 1-methyl-1H-tetrazole-5-thiol for liquid chromatography, previously dried in a desiccator (in vacu-um, silica gel) for 2 hours, and an amount of Cefpiramide RS, equivalent to about 75 mg (potency), dissolve them in 0.03 mol/L phosphate buffer solution (pH 7.5) to make ex-actly 100 mL. Pipet 2 mL of this solution, add 0.03 mol/L phosphate buffer solution (pH 7.5) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method. Calculate the amount of 1-methyl-1H-tetrazole-5-thiol, each of the other related substances and the total of the other related substances by the following equations: the amount of 1-methyl-1H-tetrazole-5-thiol, each of the other related sub-stances and the total of the other related substances are not more than 1.0z, not more than 1.5z and not more than 4.0z, respectively. Amount (z) of 1-methyl-1H-tetrazole-5-thiol (C2H4N4S) ＝ MSa/MT × ATa/ASa Amount (z) of each of other related substances ＝ MSb/MT × ATc/ASb MSa: Amount (mg) of 1-methyl-1H-tetrazole-5-thiol taken MSb: Amount [mg (potency)] of Cefpiramide RS taken MT: Amount (mg) of Cefpiramide Sodium taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.03 mol/L phosphate buffer solution (pH 7.5) and methanol (3:1). Flow rate: Adjust so that the retention time of cefpiramide is about 11 minutes. Time span of measurement: About 2 times as long as the retention time of cefpiramide. System suitability— Test for required detectability: Measure exactly 5 mL of the standard solution, and add 0.03 mol/L phosphate buffer solution (pH 7.5) to make exactly 50 mL. Confirm that the peak area of 1-methyl-1H-tetrazole-5-thiol obtained with 5 mL of this solution is equivalent to 8 to 12z of that with 5 mL of the standard solution. System performance: Dissolve 25 mg of Cefpiramide RS and 7 mg of cinnamic acid in the mobile phase to make 50 mL. When the procedure is run with 5 mL of this solution under the above operating conditions, cinnamic acid and cef-piramide are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of 1-methyl-1H-tetrazole-5-thiol is not more than 2.0z. Water <2.48> Not more than 7.0z (0.35 g, volumetric titration, direct titration).
Weigh accurately an amount of Cefpiramide So-dium and Cefpiramide RS, equivalent to about 50 mg (po-tency), add exactly 5 mL of the internal standard solution to dissolve, then add the mobile phase to make 100 mL, and use these solutions as the sample solution and standard solu-tion. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of cefpiramide to that of the internal standard. Amount [ mg (potency)] of cefpiramide (C25H24N8O7S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cefpiramide RS taken Internal standard solution—A solution of 4-dimethylamino-antipyrine (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 30 cm in length, packed with octylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.01 mol/L phosphate buffer ‌694 solution (pH 6.8), acetonitrile, methanol and tetrahydrofu-ran (22:1:1:1). Flow rate: Adjust so that the retention time of cefpiramide is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, cefpiramide and the internal standard are eluted in this order with the resolution between these peaks being not less than 7. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of cefpiramide to that of the internal standard is not more than 2.0z.
Cefsulodin Sodium contains not less than 900 mg (potency) and not more than 970 mg (potency) per mg, calculated on the anhydrous basis.
Cefsulodin Sodium occurs as white to light yel-low, crystals or crystalline powder. It is freely soluble in water and in formamide, slightly soluble in methanol, and very slightly soluble in ethanol (95). It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Cefsulodin Sodium (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cefsulodin Sodium RS prepared in the same manner as sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Cef-sulodin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cefsulodin Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Cef-sulodin Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropanesulfonate for nuclear magnetic reso-nance spectroscopy as an internal reference compound: it ex-hibits a multiplet signal A between d 7.3 ppm and d 7.7 ppm, and doublet signals, B and C, at around d 8.4 ppm and at around d 9.1 ppm, respectively. The ratio of integrated in-tensity of these signals, A:B:C, is about 5:2:2. Cefsulodin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋16.5 – ＋20.09(0.1 g calcu-lated on the anhydrous basis, water, 10 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cefsulodin Sodium in 10 mL of water: the pH of the solution is not less than 3.3 and not more than 4.8.
(1) Clarity of solution—Dissolve 1.0 g of Cef-sulodin Sodium in 10 mL of water: the solution is clear. Heavy metals <1.07>—To 1.0 g of Cefsulodin Sodium add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 5), mix, fire the ethanol to burn, then heat gradually to carbonize. After cooling, add 2 mL of nitric acid, heat carefully, then heat at 500 – 6009C to incin-erate. If a carbonized residue still retains, add a little amount of nitric acid, and heat again to incinerate. After cooling, add 6 mL of hydrochloric acid to the residue, heat to dryness on a water bath, then moisten the residue with 3 drops of hy-drochloric acid, add 10 mL of hot water, and heat on a water bath to dissolve. Add ammonia TS dropwise to adjust to pH 3 – 4, and add 2 mL of dilute acetic acid. If necessary, filter, wash the crucible and residue on the filter with 10 mL of water, transfer the filtrate and washings into a Nessler tube, add water to make 50 mL, and use this solution as the test solution. Prepare the control solution as follows: To 2.0 mL of Standard Lead Solution add 10 mL of a solution of mag-nesium nitrate hexahydrate in ethanol (95) (1 in 5), fire the ethanol to burn. After cooling, add 2 mL of nitric acid, heat carefully, then heat at 500 – 6009C. After cooling, add 6 mL of hydrochloric acid, then proceed in the same manner as for the preparation of the test solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cefsulodin Sodium according to Method 3, using a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in 5) and 15 mL of dilute hydrochloric acid instead of a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in 50) and 3 mL of hydrochloric acid, and perform the test (not more than 2 ppm). Related substances—Weigh accurately 0.10 g of Cef-sulodin Sodium, dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of isonicotinic acid amide and about 20 mg of Cefsulodin Sodium RS (separately determine the water <2.48> in the same manner as Cefsulodin Sodium), and dissolve in water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the areas of each peak by the automatic integration method. Calculate the amount of the related substances by the following for-mula: the amount of isonicotinic acid amide is not more than 1.0z, and the total amount of other related substances is not more than 1.2z. Amount (z) of isonicotinic acid amide ＝ A/BI × MI/MT × 5 Total amount (z) of the other related substances ＝ B/BS × MS/MT × 5 A: Peak area of isonicotinic acid amide from the sample solution B: Total peak area other than cefsulodin and other than isonicotinic acid amide from the sample solution BI: Peak area of isonicotinic acid amide from the standard solution BS: Peak area of cefsulodin from the standard solution MT: Amount (g) of Cefsulodin Sodium taken MS: Amount (g) of Cefsulodin Sodium RS taken MI: Amount (g) of isonicotinic acid amide taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of a solution of ammonium sulfate (1 in 100) and acetonitrile (97:3). Mobile phase B: A mixture of a solution of ammonium ‌702 sulfate (1 in 100) and acetonitrile (23:2). Flowing of mobile phase: Change the mobile phase A to B at 14 minutes after the injection of sample. Flow rate: Adjust so that the retention time of cefsulodin is about 9 minutes. Time span of measurement: About 4 times as long as the retention time of cefsulodin. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add water to make exactly 10 mL. Confirm that the peak areas of isonicotinic acid amide and cefsulodin ob-tained with 10 mL of this solution are equivalent to 7 to 13z of those with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, isonicotinic acid amide and cefsulodin are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefsulodin is not more than 1.0z. Water <2.48> Not more than 5.0z (1 g, volumetric titra-tion, direct titration, avoiding moisture absorption of the sample, using a mixture of formamide for water determina-tion and methanol for water determination (2:1) instead of methanol for water determination).
Weigh accurately an amount of Cefsulodin Sodium and Cefsulodin Sodium RS, equivalent to about 0.1 g (po-tency), dissolve each in water to make exactly 50 mL, and use these solutions as the sample solution and standard solu-tion. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine the peak areas, AT and AS, of cef-sulodin in each solution. Amount [ mg (potency)] of cefsulodin (C22H20N4O8S2) ＝ MS × AT/AS × 1000 MS: Amount [mg (potency)] of Cefsulodin Sodium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of ammonium sul-fate (1 in 100) and acetonitrile (97:3). Flow rate: Adjust so that the retention time of cefsulodin is about 9 minutes. System suitability— System performance: Dissolve 40 mg of isonicotinic acid amide in 25 mL of the standard solution. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, isonicotinic acid amide and cefsulodin are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of cefsulodin is not more than 1.0z.
Ceftizoxime Sodium contains not less than 925 mg (potency) and not more than 965 mg (potency) per mg, calculated on the anhydrous basis.
Ceftizoxime Sodium occurs as a white to light yellow, crystals or crystalline powder. It is very soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Ceftizoxime Sodium (1 in 63,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ceftizoxime Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Ceftizoxime Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethylsilylpropionate-d4 for nuclear magnetic resonance spectroscopy as an internal reference compound: it exhibits a singlet signal A at around d 4.0 ppm, a multiplet signal B around d 6.3 ppm, and a singlet signal C at around d 7.0 ppm. The ratio of integrated intensity of each signal, A:B:C, is about 3:1:1. Ceftizoxime Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋125 – ＋1459(0.25 g calcu-lated on the anhydrous bases, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Ceftizoxime Sodium in 10 mL of water: the pH of the solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Ceftizoxime Sodium in 10 mL of water: the solution is clear. Perform the test with this solution as directed under Method for Color Matching <2.65>: the color is not more colored than Matching Fluid M. Heavy metals <1.07>—Proceed with 2.0 g of Ceftizoxime Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Ceftizoxime Sodium according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.11 g of Ceftizoxime Sodium in 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and use this solution as the sample solution. Per-form the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the areas of each peak by the automatic integration method: each peak area other than ceftizoxime is not more than 0.5z of the peak area of ceftizoxime, and the total area of peaks other than ceftizoxime is not more than 1.0z of that of ceftizoxime. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Dissolve 2.31 g of disodium hydrogen-phosphate dodecahydrate and 1.42 g of citric acid monohy-drate in 1000 mL of water, adjust to pH 3.6 with diluted phosphoric acid (1 in 10) or dilute sodium hydroxide TS. To 200 mL of this solution add 10 mL of acetonitrile. Flow rate: Adjust so that the retention time of ceftizoxime is about 12 minutes. Time span of measurement: About 5 times as long as the retention time of ceftizoxime, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add 0.1 mol/L phosphate buffer solution (pH 7.0) to make exactly 100 mL, and use this solution as the solution for test for required detectability. Pipet 1 mL of the solu-tion, add 0.1 mol/L phosphate buffer solution (pH 7.0) to ‌710 make exactly 10 mL, and confirm that the peak area of ceftizoxime obtained with 5 mL of this solution is equivalent to 7 to 13z of that with 5 mL of the solution for test for re-quired detectability. System performance: Dissolve about 10 mg of Ceftizoxime RS in 100 mL of 0.1 mol/L phosphate buffer solution (pH 7.0) and use this solution as the solution for system suitability test. When the procedure is run with 5 mL of this solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of ceftizoxime are not less than 4000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak areas of ceftizoxime is not more than 2.0z. Water <2.48> Not more than 8.5z (0.4 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Ceftizoxime Sodium and Ceftizoxime RS, equivalent to about 25 mg (potency), dissolve each in 0.1 mol/L phosphate buffer solution (pH 7.0), add exactly 20 mL of the internal standard solution, then add 0.1 mol/L phosphate buffer solution (pH 7.0) to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of ceftizoxime to that of the internal standard. Storage—Light-resistant. Ceftriaxone Sodium Hydrate
Ceftriaxone Sodium Hydrate contains not less than 905 mg (potency) and not more than 935 mg (potency) per mg, calculated on the anhydrous basis.
Ceftriaxone Sodium Hydrate occurs as a white to yellow-white crystalline powder. It is freely soluble in water and in dimethylsulfoxide, spar- Amount [mg (potency)] of ceftizoxime (C H N O S ) ingly soluble in methanol, very slightly soluble in ethanol ＝ MS × QT/QS × 1000 13 13 5 5 2 (99.5), and practically insoluble in acetonitrile. MS: Amount [mg (potency)] of Ceftizoxime RS taken Internal standard solution—A solution of 3-hydroxybenzoic acid in 0.1 mol/L phosphate buffer solution (pH 7.0) (3 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Dissolve 2.31 g of disodium hydrogen phos-phate dodecahydrate and 1.42 g of citric acid monohydrate in 1000 mL of water, and adjust to pH 3.6 with diluted phos-phoric acid (1 in 10) or dilute sodium hydroxide TS. To 450 mL of this solution add 50 mL of acetonitrile. Flow rate: Adjust so that the retention time of ceftizoxime is about 4 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, ceftizoxime and the internal standard are eluted in this order with the resolution between these peaks being not less than 7.0 and the symmetry factor of each peak is not more than 1.5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ceftizoxime to that of the internal standard is not more than 1.0z.
(1) Determine the absorption spectrum of a solution of Ceftriaxone Sodium Hydrate (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ceftriaxone Sodium RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the 1H spectrum of a solution of Ceftriaxone Sodium Hydrate in deuterated dimethylsul-foxide for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using tetramethylsilane for nuclear magnetic reso- nance spectroscopy as an internal reference compound: it ex-hibits singlet signals, A, B, C and D, at around d 3.5 ppm, at around d 3.8 ppm, at around d 6.7 ppm and at around d 7.2 ppm, respectively. The ratio of integrated intensity of each signal, A: B: C: D, is about 3:3:1:2. When the signal at around d 3.5 ppm overlaps with the signal of water, perform the measurement in the probe kept at about 509C. Ceftriaxone Sodium Hydrate responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －153 – －1709(50 mg calcu-lated on the anhydrous basis, water, 2.5 mL, 20 mm). pH <2.54> Dissolve 0.6 g of Ceftriaxone Sodium Hydrate in 5 mL of water: the pH of the solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 0.6 g of Ceftriaxone Sodium Hydrate in 5 mL of water: the solu-tion is clear and light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Ceftriaxone Sodium Hydrate according to Method 2, and JP XVIII Official Monographs / Ceftriaxone Sodium Hydrate 711 perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Ceftriaxone Sodium Hydrate according to Method 3, and perform the test (not more than 2 ppm). Related substances 1—Dissolve 20 mg of Ceftriaxone Sodium Hydrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add a mixture of water and acetonitrile for liquid chromatography (11:9) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the peak areas of the impurity 1 having the relative retention time of about 0.5 and the impurity 2 having the relative retention time of about 1.3 to ceftriaxone obtained from the sample solution are not larger than the peak area of ceftriaxone from the standard solution. For the areas of the peaks, the impurity 1 and the impurity 2, multiply their correction fac-tors 0.9 and 1.2, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.796 g of anhydrous disodium hydrogen phosphate and 3.522 g of potassium dihydrogen phosphate in water to make exactly 1000 mL, and use this solution as the solution A. Separately, dissolve 20.256 g of citric acid monohydrate and 7.840 g of sodium hydroxide in water to make exactly 1000 mL, and use this solution as the solution B. Dissolve 4.00 g of tetra-n-heptylammonium bro-mide in 450 mL of acetonitrile for liquid chromatography, add 55 mL of the solution A, 5 mL of the solution B and 490 mL of water. Flow rate: Adjust so that the retention time of ceftriaxone is about 7 minutes. Time span of measurement: About 2 times as long as the retention time of ceftriaxone. System suitability— Test for required detectability: To 5 mL of the sample so-lution, add a mixture of water and acetonitrile for liquid chromatography (11:9) to make 200 mL, and use this solu-tion as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add a mixture of water and acetonitrile for liquid chromatography (11:9) to make exactly 100 mL. Confirm that the peak area of ceftriaxone obtained with 10 mL of this solution is equivalent to 0.9 to 1.1z of that with 10 mL of the solution for system suitability test. System performance: Dissolve 10 mg of Ceftriaxone So-dium Hydrate in a mixture of water and acetonitrile for liq-uid chromatography (11:9) to make 5 mL, add 5 mL of a so-lution of diethyl terephthalate in a mixture of water and acetonitrile for liquid chromatography (11:9) (9 in 5000), and add a mixture of water and acetonitrile for liquid chro-matography (11:9) to make 200 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ceftriaxone and diethyl terephthalate are eluted in this order, with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ceftriaxone is not more than 1.0z. Related substances 2—Dissolve 10 mg of Ceftriaxone Sodium Hydrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add a mixture of acetonitrile for liquid chromatogra-phy and water (23:11) to make exactly 100 mL, and use this solution as standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the each peak area of the impurities which appear after the peak of ceftriaxone ob-tained from the sample solution is not larger than the peak area of ceftriaxone from the standard solution, and the total peak area of these impurities is not larger than 2.5 times of the peak area from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.796 g of anhydrous disodium hydrogen phosphate and 3.522 g of potassium dihydrogen phosphate in water to make exactly 1000 mL, and use this solution as the solution A. Separately, dissolve 20.256 g of citric acid monohydrate and 7.840 g of sodium hydroxide in water to make exactly 1000 mL, and use this solution as the solution B. Dissolve 4.00 g of tetra-n-heptylammonium bro-mide in 450 mL of acetonitrile for liquid chromatography, and add 55 mL of the solution A, 5 mL of the solution B, 490 mL of water and 700 mL of acetonitrile for liquid chro-matography. Flow rate: Adjust so that the retention time of ceftriaxone is about 3 minutes. Time span of measurement: About 10 times as long as the retention time of ceftriaxone. System suitability— Test for required detectability: Measure 5 mL of the sam-ple solution, add a mixture of acetonitrile for liquid chroma-tography and water (23:11) to make 100 mL, and use this so-lution as the solution for system suitability test. Measure ex-actly 1 mL of the solution for system suitability test, and add a mixture of acetonitrile for liquid chromatography and water (23:11) to make exactly 100 mL. Confirm that the peak area of ceftriaxone obtained with 10 mL of this solution is equivalent to 0.9 to 1.1z of that with 10 mL of the solu-tion for system suitability test. System performance: Dissolve 10 mg of Ceftriaxone So-dium Hydrate in a mixture of acetonitrile for liquid chroma-tography and water (23:11) to make 5 mL, add 5 mL of a so-lution of diethyl terephthalate in a mixture of water and acetonitrile for liquid chromatography (11:9) (9 in 5000), and add a mixture of acetonitrile for liquid chromatography and water (23:11) to make 200 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, ceftriaxone and diethyl terephthalate are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of ceftriaxone is not more than 1.0z. ‌712 Water <2.48> Not less than 8.0z and not more than 11.0z (0.15 g, volumetric titration, direct titration).
Weigh accurately an amount of Ceftriaxone Sodium Hydrate and Ceftriaxone Sodium RS, equivalent to about 0.1 g (potency), dissolve each in a mixture of water and acetonitrile for liquid chromatography (11:9) to make ex-actly 50 mL. Pipet 5 mL of each solution, add exactly 5 mL of the internal standard solution and a mixture of water and acetonitrile for liquid chromatography (11:9) to make 200 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of ceftriaxone to that of the internal standard. Cefuroxime Axetil
Chloramphenicol Sodium Succinate contains not less than 711 mg (potency) and not more than 740 mg (potency) per mg, calculated on the anhydrous basis.
Chloramphenicol Sodium Succinate occurs as white to yellowish white, crystals or crystalline powder. It is very soluble in water, and freely soluble in methanol and in ethanol (99.5). It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Chloramphenicol Sodium Succinate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Chlo-ramphenicol Sodium Succinate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Chloramphenicol Sodium Succinate responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]25: ＋5 – ＋89(1.25 g calculated on the anhydrous basis, water, 25 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.4 g of Chloramphenicol Sodium Succinate in 5 mL of water is between 6.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Chloramphenicol Sodium Succinate in 10 mL of water: the solution is clear, and the absorbance at 420 nm of the so-lution determined as directed under Ultraviolet-visible Spec-trophotometry <2.24> is not more than 0.30. (2) Heavy metals <1.07>—Proceed with 1.0 g of Chlo- ramphenicol Sodium Succinate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Water <2.48> Not more than 2.0z (1.0 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Chloramphenicol Sodium Succinate, equivalent to about 20 mg (potency), dis-solve in water to make exactly 1000 mL, and use this solu-tion as the sample solution. Separately, weigh accurately an amount of Chloramphenicol Succinate RS, equivalent to about 20 mg (potency), add about 50 mL of water to make a ‌JP XVIII Official Monographs / Chloramphenicol and Colistin Sodium Methanesulfonate Ophthalmic Solution 731 suspension, and add gradually about 7 mL of 0.01 mol/L so-dium hydroxide TS while stirring to adjust the pH to 7.0. To this solution add water to make exactly 1000 mL, and use this solution as the standard solution. Determine the absor-bances, AT and AS, at 276 nm of the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Agar media for base layer and seed layer—Use the medium ii in 3) under (1) Agar media for seed and base layer. Agar medium for transferring test organisms—Use the medium i in 2) under (2) Agar media for transferring test organisms. Liquid medium for suspending test organisms—Use Amount [ mg (potency)] of chloramphenicol (C ＝ MS × AT/AS × 1000 11H12Cl2N2O5) the medium (2) Liquid media for suspending test organisms of 3.2. Culture media. Standard solutions—Weigh accurately an amount of MS: Amount [mg (potency)] of Chloramphenicol Suc-cinate RS taken
Cilastatin Sodium contains not less than 98.
Cilastatin Sodium occurs as a white to pale yel-lowish white powder. It is very soluble in water, freely soluble in methanol, and slightly soluble in ethanol (99.5). It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Cilastatin Sodium as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) A solution of Cilastatin Sodium (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. D Optical rotation <2.49> [a]20: ＋41.5 – ＋44.59(0.1 g calcu-lated on the anhydrous and residual solvent-free basis, a so-lution of hydrochloric acid in methanol (9 in 1000), 10 mL, 100 mL). pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Cilastatin Sodium in 100 mL of water is between 6.5 and 7.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Cilastatin Sodium in 100 mL of water: the solution is clear and the solution has no more color than the following control solution. Control solution: To a mixture of 2.4 mL of Iron (III) Chloride CS and 0.6 mL of Cobalt (II) Chloride CS add water to make 10 mL, pipet 5 mL of this solution, and add water to make exactly 100 mL. Heavy metals <1.07>—Proceed with 1.0 g of Cilasta- tin Sodium according to Method 2, and perform the test. After carbonization, add 0.5 mL of sulfuric acid instead of nitric acid. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 2.0 g of Cilastatin Sodium add 5 mL of nitric acid and 1 mL of sulfuric acid, and heat care-fully until white fumes are evolved. After cooling, heat with two 2-mL portions of nitric acid, then heat with several 2-mL portions of hydrogen peroxide (30) until a colorless or pale yellow solution is obtained. After cooling, heat again until white fumes are evolved. After cooling, add water to make 5 mL, and perform the test with this solution as the test solution: it shows no more color than the following color standard. Color standard: Prepare a solution according to the above procedure without using Cilastatin Sodium, add exactly 2 mL of Standard Arsenic Solution, and perform the test in the same manner as the test solution (not more than 1 ppm). Related substances—Dissolve about 40 mg of Cilasta-tin Sodium in 25 mL of water, and use this solution as the sample solution. Pipet 3 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed un-der Liquid Chromatography <2.01> according to the follow-ing conditions, and determine each peak area by the auto-matic integration method: the area of the peak other than cilastatin obtained from the sample solution is not larger than 1/6 times the peak area of cilastatin from the standard solution, and the total area of the peaks other than the peak of cilastatin from the sample solution is not larger than the peak area of cilastatin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.5 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 509C. Mobile phase A: A mixture of diluted phosphoric acid (1 in 1000) and acetonitrile (7:3). Mobile phase B: Diluted phosphoric acid (1 in 1000). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. ditions. Calculate the ratios of the peak areas of acetone, methanol and mesityl oxide and to the peak area of the inter-nal standard, QTa and QSa, QTb and QSb, QTc and QSc, and calculate the amounts of acetone, methanol and mesityl oxide by the following equation: they are not more than 1.0z, not more 0.5z and not more than 0.4z, respectively. Amount (z) of acetone (CH3COCH3) ＝ 1/MT × QTa/QSa × 400 × 0.79 Amount (z) of methanol (CH3OH) ＝ 1/MT × QTb/QSb × 100 × 0.79 Amount (z) of mesityl oxide (CH3COCH ＝ C(CH3)2) ＝ 1/MT × QTc/QSc × 100 × 0.86 MT: Amount (mg) of Cilastatin Sodium taken 0.79: Density (g/mL) of acetone and methanol 0.86: Density (g/mL) of mesityl oxide Internal standard solution—To 0.5 mL of 1-propanol add water to make 1000 mL. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3.2 mm in inside diameter and m in length, packed with tetrafluoroethylene polymer for gas chromatography (250 – 420 mm) coated with polyethy-lene glycol 20 M for gas chromatography at the ratio of 10z. Column temperature: A constant temperature of about 709C. Carrier gas: Helium. Flow rate: Adjust so that the retention time of the internal standard is about 5 minutes. Time after injection 0 – 30 15 → 100 85 → 0 30 – 40 of sample (min) Mobile phase A (volz) Mobile phase B (volz) Time span of measurement: About 3 times as long as the retention time of the internal standard. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, acetone, methanol, 1-propanol and mesityl oxide are Flow rate: 2.0 mL per minute. Time span of measurement: For 40 minutes. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add water to make exactly 30 mL. Confirm that the peak area of cilastatin obtained with 20 mL of this solution is equivalent to 2.3 to 4.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the retention time of cilastatin is about 20 minutes, and the number of theoretical plates and the symmetry fac-tor of the peak of cilastatin are not less than 10,000 and not more than 2.5, respectively. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cilastatin is not more than 2.0z. Residual solvents <2.46>—Weigh accurately about g of Cilastatin Sodium, add exactly 2 mL of the internal standard solution, dissolve in water to make 10 mL, and use this solution as the sample solution. Separately, measure ex-actly 2 mL of acetone, 0.5 mL of methanol and 0.5 mL of mesityl oxide, and add water to make exactly 1000 mL. Pipet 2 mL of this solution, add exactly 2 mL of the internal stand-ard solution, add water to make 10 mL, and use this solution as the standard solution. Perform the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following con- eluted in this order, and these peaks completely separate each other. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviations of the ratio of the peak area of acetone, methanol and mesityl oxide to that of the internal standard are not more than 4.0z, respec-tively. Water <2.48> Not more than 2.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately about 0.3 g of Cilastatin Sodium, dissolve in 30 mL of methanol, add 5 mL of water, and adjust to pH 3.0 with 0.1 mol/L hydrochloric acid TS. Titrate <2.50> with 0.1 mol/L sodium hydroxide VS from the first equivalence point to the third equivalence point (poten-tiometric titration), and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 19.02 mg of C16H25N2NaO5S
Clorazepate Dipotassium, when dried, contains not less than 98.
Clorazepate Dipotassium occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in water, and very slightly soluble in ethanol (99.5). It dissolves in acetic acid (100). The pH of a solution obtained by dissolving 1 g of Clorazepate Dipotassium in 100 mL of water is between 11.5 and 12.5. It gradually turns yellow on exposure to light.
(1) Carefully and gradually ignite to red-ness 30 mg of Clorazepate Dipotassium with 50 mg of so-dium. After cooling, add 3 drops of ethanol (99.5) and 5 mL of water, mix well, and filter: the filtrate responds to Qualitative Tests <1.09> for chloride. Determine the absorption spectrum of a solution of Clorazepate Dipotassium (1 in 200,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clorazepate Dipotassium as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Clorazepate Dipotassium responds to Qualitative Tests <1.09> (1) for potassium salt.
(1) Chloride <1.03>—Dissolve 1.0 g of Clorazepate Dipotassium in 20 mL of water, add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL. Perform the test with this solution as the test solution. Prepare the con-trol solution as follows: To 0.40 mL of 0.01 mol/L hydro-chloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Cloraze-pate Dipotassium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Clorazepate Dipotassium according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 15 mg of Clorazepate Dipotassium in 25 mL of a mixture of water, potassium car-bonate solution (97 in 1000) and acetonitrile (3:1:1), and use this solution as the sample solution. Pipet 1 mL of the sam-ple solution, add the mixture of water, potassium carbonate solution (97 in 1000) and acetonitrile (3:1:1) to make exactly 200 mL, and use this solution as the standard solution. Pre-pare these solutions quickly and perform the test within 3 minutes. Perform the test with exactly 5 mL each of the sam-ple solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the peak area of nordiazepam, having the relative retention time of about 3.0 to clorazepic acid, ob-tained from the sample solution is not larger than the peak area of clorazepic acid from the standard solution, the area of the peak other than clorazepic acid and nordiazepam is not larger than 1/5 times the peak area of clorazepic acid from the standard solution, and the total area of the peaks other than clorazepic acid is not larger than 2 times the peak area of clorazepic acid from the standard solution. For the area of the peak of nordiazepam, multiply the correction factor, 0.64. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 232 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 13.8 g of sodium dihydrogen phos-phate dihydrate in 500 mL of water, and adjust to pH 8.0 with sodium hydroxide TS. To 100 mL of this solution add 400 mL of acetonitrile and 300 mL of water. Flow rate: Adjust so that the retention time of clorazepic acid is about 1.3 minutes. Time span of measurement: About 10 times as long as the retention time of clorazepic acid, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add the mixture of water, potassium car-bonate solution (97 in 1000) and acetonitrile (3:1:1) to make exactly 25 mL. Confirm that the peak area of clorazepic acid obtained with 5 mL of this solution is equivalent to 15 to 25z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the number of theoretical plates and the symmetry fac-tor of the peak of clorazepic acid are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of clorazepic acid is not more than 1.5z. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, phosphorus (V) oxide, 609C, 5 hours). ‌JP XVIII Official Monographs / Clorazepate Dipotassium Capsules 797
Weigh accurately about 0.15 g of Clorazepate Dipotassium, previously dried, dissolve in 100 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of solution changes from violet to blue-green through blue (indicator: 3 drops of crystal violet TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 13.63 mg of C16H10ClKN2O3.KOH
Cloxacillin Sodium Hydrate contains not less than 900 mg (potency) and not more than 960 mg (potency) per mg, calculated on the anhydrous basis.
Cloxacillin Sodium Hydrate occurs as white to light yellow-white, crystals or crystalline powder. It is freely soluble in water, in N,N-dimethylformamide and in methanol, and sparingly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Cloxacillin Sodium Hydrate in methanol (1 in 2500) as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cloxacillin So-dium RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Clox-acillin Sodium Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Cloxacillin Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Cloxacillin Sodium Hydrate responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋163 – ＋1719(1 g calculated on the anhydrous basis, water, 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Cloxacillin Sodium Hydrate in 10 mL of water: the pH of the solution is between 5.0 and 7.5.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Cloxacillin Sodium Hydrate in 10 mL of water is clear, and its absorbance at 430 nm, deter-mined as directed under Ultraviolet-visible Spectrophotome-try <2.24>, is not more than 0.04. Heavy metals <1.07>—Proceed with 1.0 g of Cloxacil- lin Sodium Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Cloxacillin Sodium Hydrate according to Method 5, and perform the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Cloxacillin Sodium Hydrate in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test JP XVIII Official Monographs / Cloxazolam 801 ‌with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of the peak other than cloxacillin obtained from the sample so-lution is not larger than the peak area of cloxacillin from the standard solution, and the total area of the peaks other than cloxacillin from the sample solution is not larger than 3 times the peak area of cloxacillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of cloxacillin. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cloxacillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of cloxacillin are not less than 5000 and not more than 1.3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of cloxacillin is not more than 1.0z. Water <2.48> 3.0 – 4.5z (0.2 g, volumetric titration, direct titration).
Weigh accurately an amount of Cloxacillin Sodium Hydrate and Cloxacillin Sodium RS, equivalent to about 50 mg (potency), dissolve each in a suitable amount of the mo-bile phase, add exactly 5 mL of the internal standard solu-tion, then add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solu-tion, respectively. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of cloxacillin to that of the internal standard. Amount [ mg (potency)] of cloxacillin (C19H18ClN3O5S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Cloxacillin Sodium RS taken Internal standard solution—A solution of guaifenesin in the mobile phase (1 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 4.95 g of diammonium hydrogen phosphate in 700 mL of water, add 250 mL of acetonitrile, adjust to pH 4.0 with phosphoric acid, and add water to make 1000 mL. Flow rate: Adjust so that the retention time of cloxacillin is about 24 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, guaifenesin and cloxacillin are eluted in this order with the resolution between these peaks being not less than 25. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of cloxacillin to that of the internal standard is not more than 1.0z.
Colistin Sodium Methanesulfonate is the sodium salt of colistin derivatives. It is a mixture of colistin A sodium methanesul-fonate and colistin B sodium methanesulfonate. It, when dried, contains not less than 11,500 Units and not more than 15,500 Units per mg.
Colistin Sodium Methanesulfonate occurs as a white to light yellow-white powder. It is freely soluble in water, and practically insoluble in ethanol (95).
(1) Dissolve 20 mg of Colistin Sodium Methanesulfonate in 2 mL of water, add 0.5 mL of sodium hydroxide TS, and add 5 drops of copper (II) sulfate TS while shaking: a blue-purple color develops. Dissolve 40 mg of Colistin Sodium Methanesulfonate in 1 mL of 1 mol/L hydrochloric acid TS, and add 0.5 mL of dilute iodine TS: the color of iodine disappears. Determine the infrared absorption spectrum of Colistin Sodium Methanesulfonate, previously dried, as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with Sample solutions—Weigh accurately an amount of Colistin Sodium Methanesulfonate, previously dried, dis-solve in phosphate buffer solution (pH 6.0) to make a solu-tion containing about 100,000 Units per mL, and use this so-lution as the sample stock solution. Take exactly a suitable amount of the sample stock solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 10,000 Units and 2500 Units, and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
(1) Clarity and color of solution—Dissolve 0.16 g of Colistin Sodium Methanesulfonate in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Colistin Sodium Methanesulfonate according to Method 4, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Colistin Sodium Methanesulfonate according to Method 4, and perform the test (not more than 2 ppm). Free colistin—Dissolve 80 mg of Colistin Sodium Methanesulfonate in 3 mL of water, add 0.05 mL of a solu-tion of silicotungstic acid 26-water (1 in 10), and immedi-ately compare the solution with the reference suspension described under Test Methods for Plastic Containers <7.02>: the turbidity is not greater than that of the reference suspen-sion (not more than 0.25z). Loss on drying <2.41> Not more than 3.0z (0.1 g, reduced pressure, 609C, 3 hours).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Escherichia coli NIHJ Culture medium—To 10.0 g of peptone, 30.0 g of so-dium chloride, 3.0 g of meat extract and 20.0 g of agar add 1000 mL of water, then add a suitable amount of sodium hy-droxide TS so that the pH of the medium is being 6.5 to 6.6 after sterilization, sterile, and use this as the seeded agar me-dium and the agar medium for base layer. Standard solutions—Weigh accurately an amount of Colistin Sodium Methanesulfonate RS, previously dried, dis-solve in phosphate buffer solution (pH 6.0) to make a solu-tion containing 100,000 Units per mL, and use this solution as the standard stock solution. Keep the standard stock solu-tion at 109C or below and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, and add phosphate buffer solution (pH 6.0) to make solu-tions so that each mL contains 10,000 Units and 2500 Units, and use these solutions as the high concentration standard R? ＝ H; C 53H100 N16 O13 : 1169.46). solution and the low concentration standard solution, re-spectively. Description Colistin Sodium Methanesulfonate occurs as a white to light yellow-white powder. It is freely soluble in water, and practically insoluble in ethanol (95). Identification (1) Dissolve 20 mg of Colistin Sodium Methanesulfonate in 2 mL of water, add 0.5 mL of sodium hydroxide TS, and add 5 drops of copper (II) sulfate TS while shaking: a blue-purple color develops. Dissolve 40 mg of Colistin Sodium Methanesulfonate in 1 mL of 1 mol/L hydrochloric acid TS, and add 0.5 mL of dilute iodine TS: the color of iodine disappears. Determine the infrared absorption spectrum of Colistin Sodium Methanesulfonate, previously dried, as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with Sample solutions—Weigh accurately an amount of Colistin Sodium Methanesulfonate, previously dried, dis-solve in phosphate buffer solution (pH 6.0) to make a solu-tion containing about 100,000 Units per mL, and use this so-lution as the sample stock solution. Take exactly a suitable amount of the sample stock solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 10,000 Units and 2500 Units, and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
Compound Acrinol and Zinc Oxide Oil is light yellow to yellow in color.
(1) Shake well 1 g of Compound Acrinol and Zinc Oxide Oil with 10 mL of diethyl ether, 2 mL of acetic acid (100) and 10 mL of water, and separate the water layer. Shake the layer with 5 mL of hydrochloric acid and 2 to 3 drops of sodium nitrite TS, and allow to stand: a dark red color is produced (acrinol). Place 1 g of Compound Acrinol and Zinc Oxide Oil in a crucible, melt by warming, heat, gradually raising the temperature until the mass is thoroughly charred, and then ignite strongly: a yellow color is produced, and disappears on cooling. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the fil-trate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is produced (zinc oxide). Shake well 0.2 g of Compound Acrinol and Zinc Oxide Oil with 20 mL of ethanol (95) and 1 mL of acetic acid (100), centrifuge, filter, and use the filtrate as the sample solution. Separately, dissolve 5 mg of acrinol and 25 mg of ethyl aminobenzoate in 50 mL of ethanol (95) and in 2.5 mL of acetic acid (100), respectively, and use both solutions as the standard solutions (1) and (2). Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 5 mL each of the sample solution and stand-ard solutions on a plate of silica gel with fluorescent indica-tor for thin-layer chromatography. Develop the plate with a mixture of 2-propanol and acetic acid (100) (9:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 365 nm): the spots from the sample solution and standard solution (1) exhibit a blue fluorescence, and show the same Rf value. Also examine under ultraviolet light (main wavelength: 254 nm): the spots from the sample solution and standard solution (2) exhibit a purple color, and show the same Rf value.
Croscarmellose Sodium occurs as a white to yellowish white powder. It is practically insoluble in ethanol (99.5) and in diethyl ether. It swells with water and becomes a suspension. It is hygroscopic.◆
(1) To 1 g of Croscarmellose Sodium add 100 mL of a solution of methylene blue (1 in 250,000), stir well, and allow to stand: blue cotton-like precipitates ap-pear. To 1 g of Croscarmellose Sodium add 50 mL of water, and stir well to make a suspension. To 1 mL of this suspension add 1 mL of water and 5 drops of fleshly pre-pared solution of 1-naphtol in methanol (1 in 25), and gently add 2 mL of sulfuric acid along a wall of the vessel: a red-purple color appears at the zone of contact. The suspension obtained in (2) responds to Qualita-tive Tests <1.09> (1) for sodium salt. pH <2.54> To 1.0 g of Croscarmellose Sodium add 100 mL of water, and stir for 5 minutes: the pH of the supernatant liquid is between 5.0 and 7.0.
◆(1) Heavy metals <1.07>—Proceed with 2.0 g of Croscarmellose Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm).◆ (2) Sodium chloride and sodium glycolate—The total amount of sodium chloride and sodium glycolate is not more than 0.5z, calculated on the dried basis. Sodium chloride: Weigh accurately about 5 g of Croscarmellose Sodium, add 50 mL of water and 5 mL of hydrogen peroxide (30), and heat on a water bath for 20 minutes with occasional stirring. After cooling, add 100 mL of water and 10 mL of nitric acid, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration). Perform a blank determination in the same manner, and make any nec-essary correction. Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl Sodium glycolate: Weigh accurately about 0.5 g of Croscarmellose Sodium, add 2 mL of acetic acid (100) and 5 mL of water, and stir for 15 minutes. Add gradually 50 mL of acetone with stirring, then add 1 g of sodium chloride, stir for 3 minutes, and filter through a filter paper moistened with acetone. Wash the residue thoroughly with 30 mL of acetone, combine the filtrate and washings, add acetone to make exactly 100 mL, and use this solution as the sample stock solution. Separately, dissolve 0.100 g of glycolic acid in water to make 200 mL. Pipet 0.5 mL, 1 mL, 2 mL, 3 mL and 4 mL of this solution, add water to make them exactly 5 mL, then add 5 mL of acetic acid (100) and acetone to make exactly 100 mL, and designate them standard stock solution (1), standard stock solution (2), standard stock solution (3), standard stock solution (4) and standard stock solution (5), respectively. Pipet 2 mL each of the sample stock solution and the standard stock solutions (1), (2), (3), (4) and (5), and heat them in a water bath for 20 minutes to evaporate acetone. After cooling, add exactly 5 mL of 2,7-dihydrox-ynaphthalene TS, mix, then add 15 mL of 2,7-dihydrox-ynaphthalene TS, mix, cover the mouth of the vessels with aluminum foil, and heat in a water bath for 20 minutes. After cooling, add sulfuric acid to make exactly 25 mL, mix, and designate them sample solution, standard solution (1), standard solution (2), standard solution (3), standard solu-tion (4) and standard solution (5), respectively. Separately, to 10 mL of a mixture of water and acetic acid (100) (1:1) add acetone to make exactly 100 mL, and proceed with ex-actly 2 mL of this solution in the same manner for prepara-tion of the sample solution, and use the solution so obtained as the blank solution. Determine the absorbances, AT, AS1, AS2, AS3, AS4 and AS5, of the sample solution and the stand-ard solutions (1), (2), (3), (4) and (5), respectively, at 540 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the blank solution as the control. Determine the amount (g) of glycolic acid, X, in 100 mL of the sample stock solution from the calibration curve obtained with the standard solutions, and calculate the amount of sodium glycolate by the following formula. Amount (z) of sodium glycolate ＝ X/M × 100 × 1.289 M: Amount (g) of sample taken, calculated on the dried basis◆ (3) Water-soluble substance—Weigh accurately about 10 g of Croscarmellose Sodium, disperse in 800 mL of water by stirring for 1 minute every 10 minutes during 30 minutes, and allow to stand for at most 1 hour to precipitate. Filter by suction or centrifuge the clear upper portion, and weigh ac-curately the mass of about 150 mL of the filtrate or superna-tant liquid. Heat to concentrate this liquid avoiding to dry-ness, then dry at 1059C for 4 hours, and weigh the mass of the residue accurately. Calculate the amount of the water-soluble substance by the following formula: not less than 1.0z and not more than 10.0z. Amount (z) of water-soluble substance ＝ 100 M3 (800 ＋ M1)/M1M2 M1: Amount (g) of sample taken, calculated on the dried basis M2: Amount (g) of the filtrate or supernatant liquid of about 150 mL M3: Amount (g) of the residue◆ Precipitation test Put 75 mL of water in a 100-mL glass-stoppered graduated cylinder, and add portion by portion with 1.5 g of Croscarmellose Sodium divided into three por-tions while shaking vigorously at each time. Then, add water ‌to make 100 mL, shake until to get a homogenous disper-sion, and allow to stand for 4 hours: the volume of the set-tled layer is not less than 10.0 mL and not more than 30.0 mL. Degree of substitution Weigh accurately about 1 g of Croscarmellose Sodium, put in a 500-mL glass-stoppered conical flask, add 300 mL of sodium chloride TS, then add 25.0 mL of 0.1 mol/L sodium hydroxide VS, stopper, and allow to stand for 5 minutes with occasional shaking. Add 5 drops of m-cresol purple TS, then add exactly 15 mL of 0.1 mol/L hydrochloric acid VS using a buret, stopper the flask, and shake. If the color of the solution is purple, add exactly 1-mL portions of 0.1 mol/L hydrochloric acid VS using the buret, with shaking each time, until the color of the solution changes to yellow, then titrate <2.50> with 0.1 mol/L sodium hydroxide VS until the color changes from yellow to purple. Perform a blank determination in the same manner. Calcu-late the degrees of substitution of acid-carboxymethyl group and sodium-carboxymethyl group, A and S: A ＋ S is not less than 0.60 and not more than 0.85. A ＝ 1150M/(7102 － 412M － 80C ) S ＝ (162 ＋ 58A)C/(7102—80C ) M: Amount (mmol) of sodium hydroxide needed to neu-tralize 1 g of sample taken, calculated on the dried basis C: The value (z) obtained in Residue on ignition Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, 6 hours). Residue on ignition <2.44> 14.0 – 28.0z (after drying, 1 g).
Dantrolene Sodium Hydrate contains not less than 98.
Dantrolene Sodium Hydrate occurs as a yel-lowish orange to deep orange, crystalline powder. It is soluble in propylene glycol, sparingly soluble in meth- ‌JP XVIII Official Monographs / Daunorubicin Hydrochloride 829 anol, slightly soluble in ethanol (95), very slightly soluble in water and in acetic acid (100), and practically insoluble in acetone, in tetrahydrofuran and in diethyl ether.
(1) Determine the absorption spectrum of a solution of Dantrolene Sodium Hydrate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Dan-trolene Sodium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 0.1 g of Dantrolene Sodium Hydrate add 20 mL of water and 2 drops of acetic acid (100), shake well, and filter: the filtrate responds to Qualitative Tests <1.09> (1) for sodium salt.
(1) Alkalinity—To 0.7 g of Dantrolene Sodium Hydrate add 10 mL of water, shake well, and centrifuge or filter through a membrane filter. To 5 mL of the supernatant liquid or the filtrate add 45 mL of water, 3 drops of phenol-phthalein TS and 0.10 mL of 0.1 mol/L hydrochloric acid VS: a red color is not produced. Heavy metals <1.07>—Proceed with 1.0 g of Dantro- lene Sodium Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related Substances—Dissolve 50 mg of Dantrolene Time span of measurement: About twice as long as the retention time of dantrolene, beginning after the solvent peak. Water <2.48> 14.5 – 17.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately about 0.7 g of Dantrolene Sodium Hydrate, dissolve in 180 mL of a mixture of propylene glycol and acetone (1:1), and titrate <2.50> with 0.1 mol/L perchlo-ric acid VS (potentiometric titration). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.1 mol/L perchloric acid VS ＝ 33.62 mg of C14H9N4NaO5
The powder occurs as a fine, white powder, having a characteristic odor. The solution is a clear, yellow-brown to red-brown, viscous liquid, having the odor of cresol.
Dextran 40 is a product obtained by partial decom-position of polysaccharide, which is produced by fer-mentation of sucrose with Leuconostoc mesenteroides van Tieghem (Lactobacillaceae), and the average mo-lecular mass is about 40,000. When dried, it contains not less than 98.
Dextran 40 occurs as a white, amorphous pow-der. It is odorless and tasteless. It is practically insoluble in ethanol (95) and in diethyl ‌838 ether. It dissolves gradually in water. It is hygroscopic.
To 1 mL of a solution of Dextran 40 (1 in 3000) add 2 mL of anthrone TS: a blue-green color develops and turns gradually dark blue-green. Then to this solution add 1 mL of diluted sulfuric acid (1 in 2) or 1 mL of acetic acid (100): the solution does not change in color. pH <2.54> Dissolve 1.0 g of Dextran 40 in 10 mL of water: the pH of this solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Dextran 40 in 10 mL of water by warming: the solution is clear and colorless. Chloride <1.03>—Perform the test with 2.0 g of Dextran 40. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). the residue, and determine the intrinsic viscosity of the dried substance as directed in (1): the value is not more than 0.27. Low-molecular fraction—Weigh accurately about 6 g of Dextran 40, previously dried, dissolve in water to make exactly 100 mL, and transfer to a flask. Add slowly enough methanol to get 90z to 93z of the precipitate (usually 115 to 135 mL) at 25 ± 19C with stirring, centrifuge at 259C, and evaporate the supernatant liquid to dryness on a water bath. Dry the residue, and determined the intrinsic viscosity of the dried substance as directed in (1): the value is not less than 0.09.
Weigh accurately about 3 g of Dextran 40, previ-ously dried, dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Determine the opti-cal rotation aD with the sample solution as directed under Optical Rotation Determination <2.49> in a 100-mL cell at 20 ± 19C. Heavy metals <1.07>—Proceed with 1.0 g of Dextran 40 according to Method 1, and perform the test. Prepare the Amount (mg) of dextran 40 ＝ aD × 253.8 control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.5 g of Dextran 40 according to Method 1, and perform the test (not more than 1.3 ppm). Nitrogen—Weigh accurately about 2 g of Dextran 40, previously dried, and perform the test as directed under Nitrogen Determination <1.08>, where 10 mL of sulfuric acid is used for decomposition, and 45 mL of a solution of so-dium hydroxide (2 in 5) is added: the amount of nitrogen (N: 14.01) is not more than 0.010z. Reducing substances—Weigh exactly 3.00 g of Dex-tran 40, previously dried, dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh exactly 0.450 g of glucose, previously dried, dissolve in water to make exactly 500 mL, and use this solu-tion as the control solution. Pipet 5 mL each of the sample solution and the control solution, and add water to make exactly 50 mL, respectively. Pipet 5 mL each of these solu-tions, add 5 mL of alkaline copper TS, exactly measured, and heat for 15 minutes in a water bath. After cooling, add 1 mL of a solution of potassium iodine (1 in 40) and 1.5 mL of dilute sulfuric acid, and titrate <2.50> with 0.005 mol/L so-dium thiosulfate VS (indicator: 2 mL of starch TS). The titrant consumed for the sample solution is not less than that for the control solution. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Bacterial endotoxins <4.01> Less than 2.5 EU/g. Viscosity <2.53> (1) Dextran 40—Weigh accurately 0.2 to 0.5 g of Dextran 40, previously dried, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Perform the test with the sample solution and with water as directed in Method 1 at 259C : the intrinsic viscosity is between 0.16 and 0.19. (2) High-molecular fraction—Weigh accurately about 6 g of Dextran 40, previously dried, dissolve in water to make exactly 100 mL, and transfer to a flask. Add slowly enough methanol to get 7z to 10z of the precipitate (usually 80 to 90 mL) at 25 ± 19C with stirring. Dissolve the precipitate at 359C in a water bath with occasional shaking, and allow to stand for more than 15 hours at 25 ± 19C. Remove the supernatant liquid by decantation, and heat the precipitate of the lower layer to dryness on a water bath. Dry
Dextran 70 is a product obtained by partial decom-position of polysaccharide, which is produced by fer-mentation of sucrose with Leuconostoc mesenteroides van Tieghem (Lactobacillaceae), and the average mo-lecular mass is about 70,000. When dried, it contains not less than 98.
Dextran 70 occurs as a white, amorphous pow-der. It is odorless and tasteless. It is practically insoluble in ethanol (95) and in diethyl ether. It dissolves gradually in water. It is hygroscopic.
To 1 mL of a solution of Dextran 70 (1 in 3000) add 2 mL of anthrone TS: a blue-green color develops and turns gradually dark blue-green. Then to this solution add 1 mL of diluted sulfuric acid (1 in 2) or 1 mL of acetic acid (100): the solution does not change in color. pH <2.54> Dissolve 3.0 g of Dextran 70 in 50 mL of water: the pH of this solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Dextran 70 in 10 mL of water with warming: the solution is clear and colorless. Chloride <1.03>—With 2.0 g of Dextran 70, perform the test. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.018z). Heavy metals <1.07>—Proceed with 1.0 g of Dextran 70 according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.5 g of Dextran 70 according to Method 1, and perform the test (not more than 1.3 ppm). Nitrogen—Weigh accurately about 2 g of Dextran 70, previously dried, perform the test as directed under Nitrogen Determination <1.08>, where 10 mL of sulfuric acid is used for decomposition, and 45 mL of a solution of sodium hy-droxide (2 in 5) is added: the amount of nitrogen (N: 14.007) is not more than 0.010z. Reducing substances—Weigh exactly 3.00 g of Dex-tran 70, previously dried, dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh exactly 0.300 g of glucose, previously dried, dissolve in water to make exactly 500 mL, and use this solu-tion as the control solution. Pipet 5 mL each of the sample solution and the control solution, and add water to make exactly 50 mL, respectively. Pipet 5 mL of these diluted solu-tions, add exactly 5 mL of alkaline copper TS, and heat for 15 minutes in a water bath. After cooling, add 1 mL of a so-lution of potassium iodide (1 in 40) and 1.5 mL of dilute sul-furic acid, and titrate <2.50> with 0.005 mol/L sodium thio-sulfate VS (indicator: 2 mL of starch TS). The titrant consumed for the sample solution is not less than that for the control solution. Loss on drying <2.41> Not more than 5.0z (1 g, 1059C, 6 hours). Residue on ignition <2.44> Not more than 0.1z (1 g). Viscosity <2.53> (1) Dextran 70—Weigh accurately 0.2 to 0.5 g of Dextran 70, previously dried, dissolve in water to make exactly 100 mL, and use this solution as the sample so-lution. Perform the test with the sample solution and with water as directed in Method 1 at 259C : the intrinsic viscosity is between 0.21 and 0.26. High-molecular fraction—Weigh accurately about 6 g of Dextran 70, previously dried, dissolve in water to make exactly 100 mL, and transfer to a flask. Add slowly enough methanol to get 7z to 10z of the precipitate (usually, 75 to 85 mL) at 25 ± 19C with stirring. Dissolve the precipitate in a water bath at 359C with occasional shak-ing, and allow to stand for more than 15 hours at 25 ± 19C. Remove the supernatant liquid by decantation, and heat the precipitate of the lower layer on a water bath to dryness. Dry the residue, and determine the intrinsic viscosity of the dried residue as directed in (1): the value is not more than 0.35. Low-molecular fraction—Weigh accurately about 6 g of Dextran 70, previously dried, dissolve in water to make exactly 100 mL, and transfer to a flask. Add slowly enough methanol to get 90z to 93z of the precipitate (usually 110 to 130 mL) at 25 ± 19C with stirring, centrifuge at 259C, and evaporate the supernatant liquid to dryness on a water bath. Dry the residue, and determine the intrinsic viscosity of the dried residue as directed in (1): the value is not less than 0.10. Antigenicity Dissolve 6.0 g of Dextran 70 in isotonic so-dium chloride solution to make 100 mL, sterilize, and use this solution as the sample solution. Inject 1.0 mL of the sample solution on 3 occasions at intervals of 2 days intraperitoneally to each of 4 well-nourished, healthy guinea pigs weighing 250 to 300 g. Separately, inject 0.10 mL of horse serum intraperitoneally to each of 4 guinea pigs of another group as a control. Inject 0.20 mL of the sample so-lution intravenously to each of 2 guinea pigs of the first group 14 days after the first intraperitoneal injection and into each of the remaining 2 guinea pigs 21 days after the in-jection, and inject 0.20 mL of horse serum intravenously in the same manner into each guinea pigs of the second group. Observe the signs of respiratory distress, collapse or death of the animals for 30 minutes after each intravenous injection and 24 hours later: the animals of the first group exhibit no signs mentioned above. All the animals of the second group exhibit symptoms of respiratory distress or collapse and not less than 3 animals are killed. Pyrogen <4.04> Dissolve 6.0 g of Dextran 70 in isotonic so-dium chloride solution to make 100 mL, and perform the test: this solution meets the requirement.
Weigh accurately about 3 g of Dextran 70, previ-ously dried, dissolve in water to make exactly 50 mL, and use this solution as the sample solution. Determine the opti-cal rotation aD as directed under Optical Rotation Determi-nation <2.49> in a 100–mm cell at 20 ± 19C. ‌840 Amount (mg) of dextran 70 × aD ＝ 253.8 ‌Containers and storage Containers—Tight containers. Dextran Sulfate Sodium Sulfur 5
デキストラン硫酸エステルナトリウム イオウ 18
Dextran Sulfate Sodium Sulfur 18 occurs as a white to light yellow-white powder. It is odorless, and has a saline taste. It is freely soluble in water and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic.
(1) To 10 mL of a solution of toluidine blue (1 in 100,000) add 0.05 mL of a solution of Dextran Sul-fate Sodium Sulfur 18 (3 in 50) dropwise: a color of the solu-tion changes from blue to red-purple. To 1 mL of a solution of Dextran Sulfate Sodium Sulfur 18 (1 in 1500) add 2 mL of anthrone TS: a blue-green color develops, which turns dark blue-green gradually. Then, add 1 mL of diluted sulfuric acid (1 in 2) or 1 mL of acetic acid (100) to this solution: the solution remains dark blue-green. A solution of Dextran Sulfate Sodium Sulfur 18 (1 in 100) responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋90.0 – ＋110.09(1.5 g cal-culated on the dried basis, water, 25 mL, 100 mm). Viscosity <2.53> Weigh accurately about 1.5 g of Dextran Sulfate Sodium Sulfur 18, calculated on the dried basis, dis-solve in a solution of sodium chloride (29 in 500) to make exactly 100 mL, and use this solution as the sample solution. JP XVIII Official Monographs / Dextromethorphan Hydrobromide Hydrate 841 ‌Perform the test with the sample solution and a solution of sodium chloride (29 in 500) at 25 ± 0.029C as directed: the intrinsic viscosity is between 0.020 and 0.032. pH <2.54> Dissolve 1.0 g of Dextran Sulfate Sodium Sulfur 18 in 20 mL of water: the pH of this solution is between 5.5 and 7.5.
(1) Chloride <1.03>—Perform the test with 0.10 g of Dextran Sulfate Sodium Sulfer 18. Prepare the control so-lution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.106z). Sulfate <1.14>—Dissolve 0.10 g of Dextran Sulfate ‌Sodium Sulfur 18 in 6 mL of water, add 0.6 mL of barium chloride TS, and heat in a water bath for 4 minutes. After cooling, add 1 mL of dilute hydrochloric acid and water to make 50 mL, allow to stand for 10 minutes, and observe: the turbidity of the solution is not more intense than that of the control solution. Prepare the control solution as follows: to 1.0 mL of 0.005 mol/L sulfuric acid VS add 6 mL of water, and proceed in the same manner (not more than 0.480z). Heavy metals <1.07>—Proceed with 1.0 g of Dextran Sulfate Sodium Sulfur 18 according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dextran Sulfate Sodium Sulfur 18 according to Method 3, and perform the test (not more than 2 ppm). Sulfur content Weigh accurately about 0.5 g of Dextran Sulfate Sodium Sulfur 18, dissolve in 5 mL of water, add 1.5 mL of hydrochloric acid, and heat in a water bath for 1 hour. After cooling, add water to make exactly 100 mL, and use this solution as the sample solution. To exactly 10 mL of the sample solution add exactly 20 mL of 0.02 mol/L barium chloride VS, add 5 mL of methanol, and heat in a water bath for 30 minutes. After cooling, neutralize with sodium hy-droxide TS, and add 70 mL of water, 10 mL of a solution of zinc disodium ethylenediamine tetraacetate tetrahydrate (1 in 20), 3 mL of ammonium chloride TS and 7 mL of strong am-monium water, and titrate <2.50> with 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red to light blue (indicator: 5 drops of eriochrome black T TS). Perform a blank determi-nation in the same manner. Amount of sulfur (S: 32.07), cal-culated on the dried basis, is between 15.0 and 20.0z. Each mL of 0.02 mol/L barium chloride VS ＝ 0.6414 mg of S Loss on drying <2.41> Not more than 10.0z (0.5 g, in vacuum, phosphorus (V) oxide, 609C, 4 hours).
デキストラン硫酸エステルナトリウム イオウ 5
Dextran Sulfate Sodium Sulfur 5 occurs as a white to light yellow-white powder. It is odorless, and has a saline taste. It is freely soluble in water and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic.
(1) To 10 mL of a solution of toluidine blue (1 in 100,000) add 0.05 mL of a solution of Dextran Sul-fate Sodium Sulfur 5 (3 in 50) dropwise: a color of the solu-tion changes from blue to red-purple. To 1 mL of a solution of Dextran Sulfate Sodium Sulfur 5 (1 in 1500) add 2 mL of anthrone TS: a blue-green color develops, which turns dark blue-green gradually. Then, add 1 mL of diluted sulfuric acid (1 in 2) or 1 mL of acetic acid (100) to this solution: the solution remains dark blue-green. A solution of Dextran Sulfate Sodium Sulfur 5 (1 in 100) responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋135.0 – ＋155.09(1.5 g cal-culated on the dried basis, water, 25 mL, 100 mm). Viscosity <2.53> Weigh accurately about 1.5 g of Dextran Sulfate Sodium Sulfur 5, calculated on the dried basis, dis-solve in a solution of sodium chloride (29 in 500) to make exactly 100 mL, and use this solution as the sample solution. Perform the test with the sample solution and a solution of sodium chloride (29 in 500) at 25 ± 0.029C as directed: the intrinsic viscosity is between 0.030 and 0.040. pH <2.54> Dissolve 1.0 g of Dextran Sulfate Sodium Sulfur 5 in 20 mL of water: the pH of this solution is between 5.5 and 7.5.
(1) Clarity and color of solution—Dissolve 2.5 g of Dextran Sulfate Sodium Sulfur 5 in 50 mL of water: the solution is clear. And, determine the absorbance of the solu-tion at 420 nm as directed under Ultraviolet-visible Spectro-photometry <2.24>: not more than 0.090. Chloride <1.03>—Perform the test with 0.10 g of Dextran Sulfate Sodium Sulfur 5. Prepare the control solu-tion with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.106z). Sulfate <1.14>—Dissolve 0.10 g of Dextran Sulfate Sodium Sulfur 5 in 6 mL of water, add 0.6 mL of barium chloride TS, and heat in a water bath for 4 minutes. After cooling, add 1 mL of dilute hydrochloric acid and water to make 50 mL, allow to stand for 10 minutes, and observe: the turbidity of the solution is not more intense than that of the control solution. Prepare the control solution as follows: to 0.50 mL of 0.005 mol/L sulfuric acid VS add 6 mL of water, and proceed in the same manner (not more than 0.240z). Heavy metals <1.07>—Proceed with 1.0 g of Dextran Sulfate Sodium Sulfur 5 according to Method 2, and per- form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dextran Sulfate Sodium Sulfur 5 according to Method 3, and perform the test (not more than 2 ppm). Sulfur content Weigh accurately about 1.0 g of Dextran Sulfate Sodium Sulfur 5, dissolve in 5 mL of water, add 1.5 mL of hydrochloric acid, and heat in a water bath for 1 hour. After cooling, add water to make exactly 100 mL, and use this solution as the sample solution. To exactly 10 mL of the sample solution add exactly 20 mL of 0.02 mol/L barium chloride VS, add 5 mL of methanol, and heat in a water bath for 30 minutes. After cooling, neutralize with sodium hy-droxide TS, and add 70 mL of water, 10 mL of a solution of zinc disodium ethylenediamine tetraacetate tetrahydrate (1 in 20), 3 mL of ammonium chloride TS and 7 mL of strong am-monium water, and titrate <2.50> with 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red to light blue (indicator: 5 drops of eriochrome black T TS). Perform a blank determi-nation in the same manner. Amount of sulfur (S: 32.07), cal-culated on the dried basis, is between 3.0 and 6.0z. Each mL of 0.02 mol/L barium chloride VS ＝ 0.6414 mg of S Loss on drying <2.41> Not more than 10.0z (0.5 g, in vacuum, phosphorus (V) oxide, 609C, 4 hours).
Dibasic Calcium Phosphate Hydrate contains not less than 98.
Dibasic Calcium Phosphate Hydrate occurs as a white crystalline powder. It is practically insoluble in water and in ethanol (99.5). It dissolves in dilute hydrochloric acid and in dilute nitric acid.◆
(1) Dissolve 0.1 g of Dibasic Calcium Phosphate Hydrate in 10 mL of 2 mol/L hydrochloric acid TS by warming, add 2.5 mL of ammonia TS dropwise with shaking, and add 5 mL of ammonium oxalate TS: a white precipitate is produced. (2) Dissolve 0.1 g of Dibasic Calcium Phosphate Hy-drate in 5 mL of dilute nitric acid, and add 2 mL of hexaam-monium heptamolybdate TS after warming at 709C for 1 to 2 minutes: a yellow precipitate is produced.
(1) Acid-insoluble substance—Dissolve 5.0 g of Dibasic Calcium Phosphate Hydrate in 40 mL of water and 10 mL of hydrochloric acid, and boil gently for 5 minutes. After cooling, collect the insoluble substance using a filter paper for quantitative analysis. Wash with water until no more turbidity of the washing is produced when silver nitrate TS is added. Ignite to incinerate the residue and filter paper at 600 ± 509C: the mass is not more than 10 mg (not more than 0.2z). Chloride—To 0.20 g of Dibasic Calcium Phosphate Hydrate add 20 mL of water and 13 mL of dilute nitric acid, dissolve by warming, if necessary, add water to make 100 mL, and filter, if necessary. Put 50 mL of this solution in a Nessler tube, and use this as the test solution. Transfer 0.70 mL of 0.01 mol/L hydrochloric acid VS to another Nessler tube, add 6 mL of dilute nitric acid and water to make 50 mL, and use this solution as the control solution. Add 1 mL of silver nitrate TS to the test solution and the control solu-tion, mix well, and allow to stand for 5 minutes protecting from light. Compare the opalescence developed in both solu-tions against a black background by viewing downward or transversely. The opalescence developed in the test solution is not more than that of the control solution. (not more than 0.25z) Sulfate—Dissolve 0.50 g of Dibasic Calcium Phos-phate Hydrate in 5 mL of water and 5 mL of dilute hydro-chloric acid, add water to make 100 mL, and filter, if neces-sary. Put 20 mL of this solution in a Nessler tube, add 1 mL of dilute hydrochloric acid, and add water to make 50 mL, and use this as the test solution. Transfer 1.0 mL of 0.005 mol/L sulfuric acid VS to another Nessler tube, add 1 mL of dilute hydrochloride acid and water to make 50 mL, and use this solution as the control solution. Add 2 mL of barium chloride TS to the test solution and the control solution, mix well, and allow to stand for 10 minutes. Compare the white turbidity produced in both solutions against a black back-ground by viewing downward or transversely. The turbidity produced in the test solution is not thicker than that of the control solution. (not more than 0.48z) Carbonate—Mix 1.0 g of Dibasic Calcium Phosphate Hydrate with 5 mL of freshly boiled and cooled water, and add immediately 2 mL of hydrochloric acid: no efferves-cence occurs. (5) Heavy metals <1.07>—Dissolve 0.65 g of Dibasic ‌Calcium Phosphate Hydrate in a mixture of 5 mL of water and 5 mL of dilute hydrochloric acid by warming, cool, and add ammonia TS until precipitates begin to form in the solu-tion. Dissolve the precipitates by adding a small amount of dilute hydrochloric acid dropwise, add 10 mL of hydrochlo-ric acid-ammonium acetate buffer solution (pH 3.5) and water to make 50 mL, and perform the test using this solu-tion as the test solution. Prepare the control solution as follows: to 10 mL of hydrochloric acid-ammonium acetate buffer solution (pH 3.5) add 2.0 mL of Standard Lead Solu-tion and water to make 50 mL (not more than 31 ppm).◆ Barium—Heat 0.5 g of Dibasic Calcium Phosphate Hydrate with 10 mL of water, add 1 mL of hydrochloric acid dropwise with stirring, and filter after cooling, if necessary. Add 2 mL of potassium sulfate TS to this solution, and allow to stand for 10 minutes: no turbidity forms. (7) Arsenic <1.11>—Dissolve 1.0 g of Dibasic Calcium Phosphate Hydrate in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 2 ppm).◆ Loss on ignition <2.43> Not less than 24.5z and not more than 26.5z (1 g, 800 – 8259C, constant mass).
Weigh accurately about 0.4 g of Anhydrous Dibasic Calcium Phosphate, dissolve in 12 mL of dilute hydrochloric acid by heating on a water bath, if necessary, and add water to make exactly 200 mL. Pipet 20 mL of this solution, add exactly 25 mL of 0.02 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS, 50 mL of water and 5 mL of am-monia-ammonium chloride buŠer solution (pH 10.7), and titrate <2.50> the excess disodium dihydrogen ethylene-diamine tetraacetate with 0.02 mol/L zinc sulfate VS (indica-tor: 25 mg of eriochrome black T-sodium chloride indica-tor). Perform a blank determination in the same manner. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.721 mg of CaHPO4
Dibasic Sodium Phosphate Hydrate contains not less than 98.
Dibasic Sodium Phosphate Hydrate occurs as colorless or white crystals. It is odorless. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It effloresces in warm, dry air.
(1) A solution of Dibasic Sodium Phos-phate Hydrate (1 in 10) responds to Qualitative Tests <1.09> (1) and (2) for sodium salt. A solution of Dibasic Sodium Phosphate Hydrate (1 in 10) responds to Qualitative Tests <1.09> (1) and (3) for phosphate. Dissolve 0.1 g of Dibasic Sodium Phosphate Hydrate in 5 mL of dilute nitric acid, warm at 709C for 1 to 2 minutes, and add 2 mL of hexaammonium heptamolybdate TS: a yellow precipitate is formed. pH <2.54> Dissolve 1.0 g of Dibasic Sodium Phosphate Hydrate in 50 mL of water: the pH of this solution is be-tween 9.0 and 9.4.
(1) Clarity and color of solution—Dissolve 1.0 g of Dibasic Sodium Phosphate Hydrate in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Dibasic Sodium Phosphate Hydrate in 7 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Sulfate <1.14>—Dissolve 0.5 g of Dibasic Sodium Phosphate Hydrate in 2 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.005 mol/L sulfuric acid VS (not more than 0.038z). Carbonate—To 2.0 g of Dibasic Sodium Phosphate Hydrate add 5 mL of water, boil, and add 2 mL of hydro-chloric acid after cooling: the solution does not effervesce. Heavy metals <1.07>—Dissolve 2.0 g of Dibasic So- ‌dium Phosphate Hydrate in 4 mL of acetic acid (31) and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution by adding 2 mL of dilute acetic acid and water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Dibasic Sodium Phosphate Hydrate according to Method vacuum for 4 hours, as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Sodium Picosulfate Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. 1 cm 1, and perform the test (not more than 2 ppm). Absorbance <2.24> E1z (263 nm): 120 – 130 (4 mg calcu- Loss on drying <2.41> 57.0 – 61.0z (1 g, at 409C for 3 hours and then at 1059C for 5 hours, not exceeding 2 mm in sample layer).
Weigh accurately about 6 g of Dibasic Sodium Phos-phate Hydrate, dissolve in 50 mL of water, and then titrate <2.50> with 0.5 mol/L sulfuric acid VS at 159C until the green color of the solution changes to dark-greenish red-pur-ple (indicator: 3 to 4 drops of methyl orange-xylenecyanol FF TS). Each mL of 0.5 mol/L sulfuric acid VS ＝ 142.0 mg of Na HPO lated on the anhydrous basis, water, 100 mL). Purity (1) Clarity and color of solution—Dissolve 1.0 g of Sodium Picosulfate Hydrate in 10 mL of water: the solu-tion is clear and colorless to pale yellow. Chloride <1.03>—Perform the test with 0.5 g of So- dium Picosulfate Hydrate. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.028z). Sulfate <1.14>—Perform the test with 0.40 g of So- dium Picosulfate Hydrate. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 2 4 0.042z).
Diclofenac Sodium Suppositories contain not less than 93.0z and not more than 107.0z of the labeled as the sample solution. Pipet 2 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly amount of diclofenac sodium (C 318.13). 14H10Cl2NNaO2: 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of these solutions as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of these solutions by the automatic integration method: the area of each peak other than diclofenac obtained from the sample solution is not larger than the peak area of diclofenac from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of methanol and diluted acetic acid (100) (3 in 2500) (4:3). Flow rate: Adjust so that the retention time of diclofenac is about 20 minutes. Time span of measurement: About twice as long as the retention time of diclofenac, beginning after the solvent peak. System suitability— System performance: Dissolve 35 mg of ethyl parahy-droxybenzoate and 0.05 g of propyl parahydroxybenzoate in 100 mL of the mobile phase. To 1 mL of this solution add the mobile phase to make 50 mL. When the procedure is run Method of preparation Prepare as directed under Supposi-tories for Rectal Application, with Diclofenac Sodium. Identiˆcation To an amount of Diclofenac Sodium Sup-positories, equivalent to 25 mg of Diclofenac Sodium, add 200 mL of a mixture of methanol and 0.01 mol/L sodium hydroxide TS (99:1), and dissolve by warming. Cool while shaking, add a mixture of methanol and 0.01 mol/L sodium hydroxide TS (99:1) to make 250 mL, and ˆlter through a pledget of absorbent cotton if necessary. To 10 mL of this solution add a mixture of methanol and 0.01 mol/L sodium hydroxide TS (99:1) to make 100 mL. Determine the absorp-tion spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 280 nm and 284 nm. Uniformity of dosage unit <6.02> Perform the test accord-ing to the following method: it meets the requirement of the Content uniformity test. To 1 suppository of Diclofenac Sodium Suppositories add 5 mL of tetrahydrofuran, and sonicate to dissolve. Add a mixture of methanol and water (3:2) to make exactly 100 mL, shake, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.5 mm. Discard the ˆrst 5 mL of the ˆl-trate, pipet V mL of the subsequent ˆltrate, add a mixture of methanol and water (3:2) to make exactly V? mL so that each mL contains about 0.
Weigh accurately the mass of not less than 20 Diclofenac Sodium Suppositories, cut into small pieces care-fully, and mix uniformly. Weigh accurately a portion of the pieces, equivalent to about 25 mg of diclofenac sodium (C14H10Cl2NNaO2), add 5 mL of tetrahydrofuran, and sonicate to dissolve. Add a mixture of methanol and water (3:2) to make exactly 100 mL, shake, and ˆlter through a membrane ˆlter with a pore size not exceeding 0.5 mm. Discard the ˆrst 5 mL of the ˆltrate, pipet 10 mL of the sub-sequent ˆltrate, add a mixture of methanol and water (3:2) to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of diclofenac sodium for assay, previously dried, and dissolve in a mixture of methanol and water (3:2) to make exactly 100 mL. Pipet 5 mL of this solution, add a mixture of methanol and water (3:2) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of diclofenac in each solution. Amount (mg) of diclofenac sodium (C14H10Cl2NNaO2) ＝ MS × AT/AS × 1/2 MS: Amount (mg) of diclofenac sodium for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 12.5 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 13.6 g of sodium acetate trihydrate in water to make 1000 mL. To 200 mL of this solution add 300 mL of methanol. Flow rate: Adjust so that the retention time of diclofenac is about 3.5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of diclofenac are not less than 2000 and 0.7 to 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of diclofenac is not more than 1.0z.
Diclofenac Sodium, when dried, contains not less than 98.
Diclofenac Sodium occurs as white to pale yel- ‌JP XVIII Official Monographs / Diclofenac Sodium Suppositories 847 low-white, crystals or crystalline powder. It is freely soluble in methanol and in ethanol (95), spar-ingly soluble in water and in acetic acid (100), and practically insoluble in diethyl ether. It is hygroscopic.
(1) To 1 mL of a solution of Diclofenac Sodium in methanol (1 in 250) add 1 mL of nitric acid: a dark red color develops. Perform the test with 5 mg of Diclofenac Sodium as directed under Flame Coloration Test <1.04> (2): a light green color appears. Determine the infrared absorption spectrum of Diclofenac Sodium, previously dried, as directed in the po-tassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Diclofenac Sodium (1 in 100) responds to Qualitative Tests <1.09> for sodium salt.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Diclofenac Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Diclofenac Sodium according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.05 g of Diclofenac Sodium in 50 mL of the mobile phase, and use this solution areas of diclofenac is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 3 hours).
Weigh accurately about 0.5 g of Diclofenac Sodium, previously dried, dissolve in 40 mL of water in a separator, add 2 mL of dilute hydrochloric acid, and extract the pre-cipitate formed with 50 mL of chloroform. Extract again with two 20-mL portions of chloroform, and filter the ex-tract each time through a pledget of absorbent cotton moistened with chloroform. Wash the tip of the separator and the absorbent cotton with 15 mL of chloroform, com-bine the washing with the extracts, add 10 mL of a solution of 1 mol/L hydrochloric acid TS in ethanol (99.5) (1 in 100), and titrate <2.50> with 0.1 mol/L potassium hydroxide-ethanol VS from the first equivalent point to the second equivalent point (potentiometric titration). Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 31.81 mg of C14H10Cl2NNaO2
Dicloxacillin Sodium Hydrate contains not less than 910 mg (potency) and not more than 1020 mg (potency) per mg, calculated on the anhydrous basis.
Dicloxacillin Sodium Hydrate occurs as a white to light yellow-white crystalline powder. It is freely soluble in water and in methanol, and soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Dicloxacillin Sodium Hydrate (1 in 2500) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Dicloxacillin Sodium RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Dicloxacillin Sodium Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Dicloxacillin Sodium RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. Dicloxacillin Sodium Hydrate responds to Quali-tative Tests <1.09> (1) for sodium salt. Water <2.48> Not less than 3.0z and not more than 4.5z (0.1 g, volumetric titration, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 6.5 to 6.6 after sterilization. Standard solutions—Weigh accurately an amount of Dicloxacillin Sodium RS equivalent to about 50 mg (po-tency), dissolve in phosphate buffer solution (pH 6.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 24 hours. Take exactly a suitable amount of the standard stock solution before use, add phos-phate buffer solution (pH 6.0) to make solutions so that each mL contains 10 mg (potency) and 2.5 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. ‌JP XVIII Official Monographs / Diethylcarbamazine Citrate Tablets 849‌ Sample solutions—Weigh accurately an amount of Dicloxacillin Sodium Hydrate equivalent to about 50 mg (potency), dissolve in phosphate buffer solution (pH 6.0) to make exactly 50 mL. Take exactly a suitable amount of the solution, add phosphate buffer solution (pH 6.0) to make solutions so that each mL contains 10 mg (potency) and 2.5 mg (potency), and use these solutions as the high concentra-tion sample solution and the low concentration sample solu- Assay Weigh accurately about 0.75 g of Diethylcarbama-zine Citrate, previously dried, dissolve in 50 mL of acetic acid (100) by warming, cool, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.14 mg of C H N O.C H O tion, respectively. 10 21 3 6 8 7
Diphenhydramine, Phenol and Zinc Oxide Liniment is a white to whitish, pasty mass. It has a slight odor of phenol.
(1) To 3 g of Diphenhydramine, Phenol and Zinc Oxide Liniment add 20 mL of hexane, shake well, and separate the hexane layer. Shake thoroughly the hexane solution with 10 mL of 0.2 mol/L hydrochloric acid. Separate the aqueous layer, and adjust with sodium hydrox-ide TS to a pH of 4.6. Add 1 mL of bromophenol blue-potassium biphthalate TS and 10 mL of chloroform, and shake: a yellow color develops in the chloroform layer (diphenhydramine). Place 1 g of Diphenhydramine, Phenol and Zinc Oxide Liniment in a porcelain crucible, gradually raise the temperature by heating until the mass is charred, and ignite strongly: a yellow color is produced, and disappears on cool-ing. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. Add 2 to 3 drops of potassium hexacyanoferrate (II) TS to the filtrate: a white precipitate is produced (zinc oxide). Shake 0.5 g of Diphenhydramine, Phenol and Zinc Oxide Liniment with 1 mL of water and 5 mL of chlo-roform, filter, and use the filtrate as the sample solution. Dissolve 0.01 g each of diphenhydramine and phenol in 5 mL each of chloroform, and use these solutions as the stand-ard solution (1) and standard solution (2). Perform the test with these solutions as directed under Thin-layer Chroma-tography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia solution (28) (50:5:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: two spots obtained from the sample solution and each spot from the standard solution (1) and (2) show the same Rf value. Sublime iodine, and spray Dragendorff's TS evenly upon the plate: the spot from standard solution (1) and the corresponding spot from the sample solution reveal an orange color.
Disodium Edetate Hydrate contains not less than (3) A solution of Disodium Edetate Hydrate (1 in 20) responds to Qualitative Tests <1.
Disodium Edetate Hydrate occurs as white, crystals or crystalline powder. It is odorless and has a slight, acid taste. It is soluble in water, and practically insoluble in ethanol (95) and in diethyl ether.
(1) Dissolve 0.01 g of Disodium Edetate Hydrate in 5 mL of water, add 2 mL of a solution of potas-sium chromate (1 in 200) and 2 mL of arsenic trioxide TS, and heat in a water bath for 2 minutes: a purple color de-velops. (2) Dissolve 0.5 g of Disodium Edetate Hydrate in 20 mL of water, and add 1 mL of dilute hydrochloric acid: a white precipitate is produced. Collect the precipitate, wash with 50 mL of water, and dry at 1059C for 1 hour: the precipitate melts <2.60> between 2409C and 2449C (with decomposi-tion). ers. ‌Sodium Fusidate
(1) Clarity and color of solution—Dissolve 1.0 g of Disodium Edetate Hydrate in 50 mL of water: the solu-tion is clear and colorless. Cyanide—Transfer 1.0 g of Disodium Edetate Hy-drate to a round-bottomed flask, dissolve in 100 mL of water, add 10 mL of phosphoric acid, and distil. Place 15 mL of 0.5 mol/L sodium hydroxide VS in a 100-mL measur-ing cylinder, which is used as a receiver, and immerse the bottom end of the condenser into the solution. Distil the mixture until the distillate measures 100 mL, and use this so-lution as the sample solution. Transfer 20 mL of the sample solution to a glass-stoppered test tube, add 1 drop of phenol-phthalein TS, neutralize with dilute acetic acid, and add 5 mL of phosphate buffer solution (pH 6.8) and 1.0 mL of diluted sodium toluenesulfonchloramide TS (1 in 5). Imme-diately stopper the tube, mix gently, and allow to stand for a few minutes. Mix well with 5 mL of pyridine-pyrazolone TS, and allow to stand between 209C and 309C for 50 minutes: the solution has no more color than the following control so-lution. Control solution: Pipet 1.0 mL of Standard Cyanide Solu-tion, add 15 mL of 0.5 mol/L sodium hydroxide VS and water to make exactly 1000 mL, transfer 20 mL of this solu-tion to a glass-stoppered test tube, and proceed as directed for the sample solution. Heavy metals <1.07>—Proceed with 2.0 g of Disodi- um Edetate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Disodium Edetate Hydrate according to Method 1, and perform the test (not more than 2 ppm). Residue on ignition <2.44> 37.0 – 39.0z (1 g).
Weigh accurately about 1 g of Disodium Edetate Hydrate, dissolve in 50 mL of water, add 2 mL of ammonia-ammonium chloride buffer solution (pH 10.7) and 0.04 g of eriochrome black T-sodium chloride indicator, and titrate <2.50> with 0.1 mol/L zinc VS until the color of the solution changes from blue to red. Each mL of 0.1 mol/L zinc VS 99.0z of disodium edetate hydrate (C 2H2O). 10H14N2Na2O8. ＝ 37.22 mg of C10H14N2Na2O8.2H2O
Dried Aluminum Hydroxide Gel contains not less than 50.
Dried Aluminum Hydroxide Gel occurs as a white, amorphous powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. Most of it dissolves in dilute hydrochloric acid and in sodium hydroxide TS.
To 0.2 g of Dried Aluminum Hydroxide Gel add 20 mL of dilute hydrochloric acid, warm, and centri-fuge: the supernatant liquid responds to Qualitative Tests <1.09> for aluminum salt.
(1) Acidity or alkalinity—To 1.0 g of Dried Alu-minum Hydroxide Gel add 25 mL of water, shake well, and centrifuge: the supernatant liquid is neutral. Chloride <1.03>—To 1.0 g of Dried Aluminum Hy- droxide Gel add 30 mL of dilute nitric acid, heat gently to boil while shaking, cool, add water to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.284z). Sulfate <1.14>—To 1.0 g of Dried Aluminum Hy- droxide Gel add 15 mL of dilute hydrochloric acid, heat gently to boil while shaking, cool, add water to make 250 mL, and centrifuge. To 25 mL of the supernatant liquid add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 1.0 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.480z). Nitrate—To 0.10 g of Dried Aluminum Hydroxide Gel add 5 mL of water, then carefully add 5 mL of sulfuric acid, shake well to dissolve, and cool. Superimpose the solu-tion on 2 mL of iron (II) sulfate TS: no brown-colored ring is produced at the zone of contact. Heavy metals <1.07>—Dissolve 2.0 g of Dried Alumi- num Hydroxide Gel in 10 mL of dilute hydrochloric acid by heating, filter if necessary, and add water to make 50 mL. 0.05 mol/L zinc acetate VS until the color of the solution changes from light dark green to light red. (indicator: 2 mL of dithizone TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.549 mg of Al2O3
Weigh accurately about 2 g of Dried Aluminum Hy-droxide Gel, add 15 mL of hydrochloric acid, heat on a water bath with shaking for 30 minutes, cool, and add water to make exactly 500 mL. Pipet 20 mL of this solution, add exactly 30 mL of 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS and 20 mL of acetic acid (31)-am-monium acetate buffer solution (pH 4.8), boil for 5 minutes, 2 3 and cool. Add 55 mL of ethanol (95), and titrate <2.50> with Description Dried Aluminum Hydroxide Gel occurs as a white, amorphous powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. Most of it dissolves in dilute hydrochloric acid and in sodium hydroxide TS. Identification To 0.2 g of Dried Aluminum Hydroxide Gel add 20 mL of dilute hydrochloric acid, warm, and centri-fuge: the supernatant liquid responds to Qualitative Tests <1.09> for aluminum salt. Purity (1) Acidity or alkalinity—To 1.0 g of Dried Alu-minum Hydroxide Gel add 25 mL of water, shake well, and centrifuge: the supernatant liquid is neutral. Chloride <1.03>—To 1.0 g of Dried Aluminum Hy- droxide Gel add 30 mL of dilute nitric acid, heat gently to boil while shaking, cool, add water to make 100 mL, and centrifuge. To 5 mL of the supernatant liquid add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.284z). Sulfate <1.14>—To 1.0 g of Dried Aluminum Hy- droxide Gel add 15 mL of dilute hydrochloric acid, heat gently to boil while shaking, cool, add water to make 250 mL, and centrifuge. To 25 mL of the supernatant liquid add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Pre-pare the control solution with 1.0 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.480z). Nitrate—To 0.10 g of Dried Aluminum Hydroxide Gel add 5 mL of water, then carefully add 5 mL of sulfuric acid, shake well to dissolve, and cool. Superimpose the solu-tion on 2 mL of iron (II) sulfate TS: no brown-colored ring is produced at the zone of contact. Heavy metals <1.07>—Dissolve 2.0 g of Dried Alumi- num Hydroxide Gel in 10 mL of dilute hydrochloric acid by heating, filter if necessary, and add water to make 50 mL. 0.05 mol/L zinc acetate VS until the color of the solution changes from light dark green to light red. (indicator: 2 mL of dithizone TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.549 mg of Al2O3
Dried Sodium Carbonate occurs as white crys-tals or crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. A solution of Dried Sodium Carbonate (1 in 10) is alka-line. Dried Sodium Carbonate is hygroscopic.
A solution of Dried Sodium Carbonate (1 in 20) responds to Qualitative Tests <1.09> for sodium salt and for carbonate.
(1) Clarity and color of solution—Dissolve 1.0 g of Dried Sodium Carbonate in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of Dried Sodium Carbonate in 10 mL of water, add 12 mL of dilute nitric acid, dilute with water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.071z). Heavy metals <1.07>—Dissolve 1.0 g of Dried So- dium Carbonate in 10 mL of water, add 7.5 mL of dilute hy-drochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 35 mL of water and 2 mL of dilute acetic acid, dilute with water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 7.5 mL of dilute hy-drochloric acid on a water bath to dryness, add 2 mL of di-lute acetic acid and 2.0 mL of Standard Lead Solution, and dilute with water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.65 g of Dried Sodium Carbonate according to Method 1, and per-form the test (not more than 3.1 ppm). Loss on drying <2.41> Not more than 2.0z (2 g, 1069C, 4 hours).
Dissolve about 1.2 g of Dried Sodium Carbonate, weighed accurately, in 25 mL of water, and titrate with 0.5 mol/L sulfuric acid VS until the color of the solution changes from blue to yellow-green. Then boil cautiously, cool, and further titrate <2.50> until a greenish yellow color develops (indicator: 2 drops of bromocresol green TS). Each mL of 0.5 mol/L sulfuric acid VS ＝ 53.00 mg of Na CO Sodium Carbonate Hydrate
Dried Sodium Sulfite is white, crystals or pow-der. It is odorless. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. The pH of a solution of 1.0 g of Dried Sodium Sulfite in 10 mL of water is about 10. It gradually changes in moist air.
An aqueous solution of Dried Sodium Sul-fite (1 in 20) responds to Qualitative Tests <1.09> for sodium salt and sulfite.
(1) Thiosulfate—Dissolve 1.0 g of Dried Sodium Sulfite in 15 mL of water, add gradually 5 mL of hydrochlo-ric acid, shake, and allow to stand for 5 minutes: no tur-bidity is produced. Heavy metals <1.07>—Dissolve 1.0 g of Dried So- dium Sulfite in 5 mL of water, add 2 mL of hydrochloric acid gradually, and evaporate the mixture on a water bath to dryness. Add 3 mL of boiling water and 1 mL of hydrochlo-ric acid to the residue, and again evaporate to dryness on a water bath. Dissolve the residue in 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution as follows: evaporate 3 mL of hydrochloric acid to dryness, and add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Dissolve 0.5 g of Dried Sodium Sul- fite in 5 mL of water, add 1 mL of sulfuric acid, and evapo-rate on a sand bath until white fumes are evolved. Add water to make 5 mL, take this solution as the sample solution, and perform the test (not more than 4 ppm).
Weigh accurately about 0.2 g of Dried Sodium Sul-fite, transfer immediately to an iodine flask containing ex-actly 50 mL of 0.05 mol/L iodine VS, stopper, shake, and allow to stand for 5 minutes in a dark place. Add 1 mL of hydrochloric acid, and titrate <2.50> the excess iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 6.302 mg of Na2SO3
Ecabet Sodium Hydrate contains not less than 98.
Ecabet Sodium Hydrate is white crystals. It is freely soluble in methanol, and slightly soluble in water and in ethanol (99.5). It dissolves in sodium hydroxide TS. Dissolve 1.0 g of Ecabet Sodium Hydrate in 200 mL of water: the pH of the solution is about 3.5.
(1) Determine the absorption spectrum of a solution of Ecabet Sodium Hydrate in dilute sodium hydrox-ide TS (3 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensi-ties of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ecabet Sodium Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Place 1 g of Ecabet Sodium Hydrate in a porcelain crucible, and carbonize. After cooling, add 0.5 mL of nitric acid, heat gradually to incinerate, and dissolve the residue in 10 mL of water: the solution responds to Qualitative Tests <1.09> for sodium salt. D Optical rotation <2.49> [a]20: ＋69 – ＋769(0.25 g calcu-lated on the anhydrous basis, methanol, 25 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Ecabet Sodium Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 10 mg of Ecabet So-dium Hydrate in 10 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add the mobile phase to make exactly 20 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions. Deter-mine each peak area by the automatic integration method: the area of each peak other than ecabet obtained from the sample solution is not larger than the peak area of ecabet from the standard solution. ‌JP XVIII Official Monographs / Ecabet Sodium Granules 913 Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 225 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 0.1 mol/L potassium dihydrogen phos-phate TS add phosphoric acid to adjust the pH to 3.0. To 730 mL of this solution add 270 mL of acetonitrile. Flow rate: Adjust so that the retention time of ecabet is about 8 minutes. Time span of measurement: About 2 times as long as the retention time of ecabet, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of ecabet are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of ecabet is not more than 2.0z. Water <2.48> 17.3 – 19.2z (0.2 g, volumetric titration, direct titration).
Weigh accurately about 1.2 g of Ecabet Sodium Hy-drate, dissolve in 30 mL of methanol, add 30 mL of water, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (in-dicator: 4 drops of phenolphthalein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 40.25 mg of C20H27NaO5S
Etidronate Disodium, when dried, contains not less than 98.
Etidronate Disodium occurs as a white pow-der. It is freely soluble in water, and practically insoluble in ‌JP XVIII Official Monographs / Etidronate Disodium Tablets 981 ethanol (99.5). The pH of a solution prepared by dissolving 0.10 g of Etidronate Disodium in 10 mL of water is between 4.4 and 5.4. It is hygroscopic.
(1) To 5 mL of a solution of Etidronate Disodium (1 in 100) add 1 mL of copper (II) sulfate TS, and mix for 10 minutes: a blue precipitate is formed. Determine the infrared absorption spectrum of Etidronate Disodium, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. A solution of Etidronate Disodium (1 in 100) re-sponds to Qualitative Tests <1.09> for sodium salt.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Etidronate Disodium according to Method 4, and perform the test using the supernatant liquid obtained by centrifuging after addition of 2 mL of dilute acetic acid. Prepare the con-trol solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Etidronate Disodium according to Method 1, and per-form the test (not more than 2 ppm). Phosphite—Weigh accurately about 3.5 g of Etidronate Disodium, dissolve in 100 mL of 0.1 mol/L so-dium dihydrogen phosphate TS adjusted the pH to 8.0 with sodium hydroxide TS, add exactly 20 mL of 0.05 mol/L iodine VS, and immediately stopper tightly. Allow to stand in a dark place for 30 minutes, add 1 mL of acetic acid (100), and titrate <2.50> the excess of iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. The amount of phosphite (NaH2PO3) is not more than 1.0z. Each mL of 0.05 mol/L iodine VS ＝ 5.199 mg of NaH2PO3 Methanol—Weigh accurately about 0.5 g of Etidronate Disodium, dissolve in water to make exactly 5 mL, and use this solution as the sample solution. Separately, pipet 1 mL of methanol, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Per-form the test with exactly 1 mL each of the sample solution and standard solution as directed under Gas Chromatogra-phy <2.02> according to the following conditions, and deter-mine the peak areas, AT and AS, of methanol in each solu-tion and determine the amount of methanol (CH4O) by the following equation: not more than 0.1z. Amount (z) of methanol (CH4O) ＝ 1/M × AT/AS × 1/20 × 0.79 M: Amount (g) of Etidronate Disodium taken 0.79: Density (g/mL) of methanol Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 2 m in length, packed with porous copolymer beads for gas chro-matography (180 – 250 mm in particle diameter). Column temperature: A constant temperature of about 1309C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of methanol is about 2 minutes. System suitability— System performance: To 1 mL of methanol and 1 mL of ethanol (99.5) add water to make 100 mL. To 1 mL of this solution add water to make 100 mL. When the procedure is run with 1 mL of this solution under the above operating conditions, methanol and ethanol are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of methanol is not more than 5.0z. Loss on drying <2.41> Not more than 5.0z (0.5 g, 2109C, 2 hours).
Weigh accurately about 0.5 g of Etidronate Disodi-um, previously dried, and dissolve in water to make exactly 50 mL. Transfer exactly 15 mL of this solution to a chro-matographic column of 10 mm in internal diameter contain-ing 5 mL of strongly acidic ion exchange resin for column chromatography (H type), allow to flow at a flow rate of about 1.5 mL per minute, and wash the column with two 25-mL portions of water. Combine the eluate and the wash-ings, and titrate <2.50> with 0.1 mol/L sodium hydroxide VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 12.50 mg of C2H6Na2O7P2
Faropenem Sodium Hydrate contains not less than 870 mg (potency) and not more than 943 mg (potency) per mg, calculated on the anhydrous basis.
Faropenem Sodium Hydrate occurs as white to light yellow, crystals or crystalline powder. It is freely soluble in water and in methanol, slightly solu-ble in ethanol (95), and practically insoluble in diethyl ether.
(1) Dissolve 5 mg of Faropenem Sodium exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectra of Faro-penem Sodium Hydrate and Faropenem Sodium RS as di-rected in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectra: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋145 – ＋1509(0.5 g calcu-lated as the anhydrous basis, water, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Faropenem Sodium Hydrate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve a quantity of Faro-penem Sodium Hydrate equivalent to 0.10 g (potency) in 200 mL of water, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solu-tion. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and determine each peak area by the automatic in-tegration method: the peak area of the epimer, having the relative retention time of about 1.1 to faropenem, obtained from the sample solution is not larger than 3/10 times the peak area of faropenem from the standard solution, and the total area of the peaks other than faropenem from the sam-ple solution is not larger than 1/2 times the peak area of faropenem from the standard solution. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Time span of measurement: About 6 times as long as the retention time of faropenem, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add water to make exactly 20 mL. Con-firm that the peak area of faropenem obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution obtained in the Assay under the above operating conditions, the internal standard and faro-penem are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of faropenem is not more than 2.0z. Water <2.48> Not less than 12.6z and not more than 13.1z (20 mg, coulometric titration).
Weigh accurately an amount of Faropenem Sodium ‌Hydrate and Faropenem Sodium RS, equivalent to about 25 mg (potency), add exactly 10 mL each of the internal stand-ard solution, add water to make 50 mL, and use these solu-tions as the sample solution and the standard solution, re-spectively. Perform the test with 20 mL of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of faro-penem to that of the internal standard. Amount [ mg (potency)] of faropenem (C12H15NO5S) ＝ MS × QT/QS × 1000 MS: amount [mg (potency)] of Faropenem Sodium RS taken Internal standard solution—Dissolve 0.5 g of m-hydroxya-cetophenone in 20 mL of acetonitrile, and add water to make 200 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 305 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 4.8 g of potassium dihydrogen phosphate, 5.4 g of disodium hydrogen phosphate dodeca-hydrate and 1.0 g of tetra n-butyl ammonium bromide in water to make 1000 mL. To 870 mL of this solution add 130 mL of acetonitrile. Flow rate: Adjust so that the retention time of faropenem is about 11 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and faropenem are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- nm, and between 304 nm and 308 nm. Purity Related substances—Powder Faropenem Sodium for Syrup, if necessary. To a part of the powder, equivalent to about 25 mg (potency) of Faropenem Sodium Hydrate, add about 10 mL of water, shake well, then add water to make exactly 50 mL, and filter. Discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample so-lution. Pipet 2 mL of the sample solution, add water to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area of both solutions by the automatic integration method: the area of the peak of cleaved derivative, having the relative retention time of about 0.71 to faropenem, obtained from the sample solution is not larger than 1.5 times the peak area of faropenem from the standard solution, and the total area of the peaks other than faropenem from the sample solution is not larger than 2 times the peak area of faropenem from the standard solu-tion. For the area of the peak, having the relative retention time of about 0.71 to faropenem, multiply its correction fac-tor 0.37. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4 mm in inside diameter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Dissolve 6.12 g of potassium dihydrogen phosphate, 1.79 g of disodium hydrogen phosphate dodeca-hydrate and 1.61 g of tetra n-butylammonium bromide in water to make 1000 mL. Mobile phase B: A mixture of the mobile phase A and acetonitrile (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. ing conditions, the relative standard deviation of the ratios of the peak area of faropenem to that of the internal stand-ard is not more than 1.0z. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz)
Flavin Adenine Dinucleotide Sodium contains not less than 93.
Flavin Adenine Dinucleotide Sodium occurs as an orange-yellow to light yellow-brown powder. It is odor-less or has a slight, characteristic odor, and has a slightly bitter taste. It is freely soluble in water, and practically insoluble, in methanol, in ethanol (95), in ethyleneglycol and in diethyl ether. It is hygroscopic. It is decomposed by light.
(1) A solution of Flavin Adenine Dinucleo-tide Sodium (1 in 100,000) is light yellow-green in color, and shows a strong yellow-green fluorescence. To 5 mL of the so-lution add 0.02 g of hydrosulfite sodium: the color and the fluorescence of the solution disappear, and gradually reap-pear when the solution is shaken in air. Add dilute hydro-chloric acid or sodium hydroxide TS dropwise: the fluores-cence of the solution disappears. Determine the infrared absorption spectrum of Flavin Adenine Dinucleotide Sodium as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 0.1 g of Flavin Adenine Dinucleotide Sodium add 10 mL of nitric acid, evaporate on a water bath to dryness, and ignite. To the residue add 10 mL of diluted nitric acid (1 in 50), boil for 5 minutes, and after cooling, neutralize with ammonia TS, then filter the solution if necessary: the solution responds to Qualitative Tests <1.09> for sodium salt and Qualitative Tests <1.09> (1) and (3) for phosphate. D Optical rotation <2.49> [a]20: －21.0 – －25.59(0.3 g, calcu-lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Flavin Adenine Dinucleotide Sodium in 100 mL of water: the pH of this solution is be-tween 5.5 and 6.5.
(1) Clarity and color of solution—Dissolve 0.20 g of Flavin Adenine Dinucleotide Sodium in 10 mL of water: JP XVIII Official Monographs / Flavin Adenine Dinucleotide Sodium 1011 the solution is clear and orange-yellow in color. Free phosphoric acid—Weigh accurately about 0.02 g of Flavin Adenine Dinucleotide Sodium, dissolve in 10 mL of water, and use this solution as the sample solution. Sepa-rately, measure exactly 2 mL of Standard Phosphoric Acid Solution, add 10 mL of water, and use this solution as the standard solution. To each of the sample solution and stand-ard solution add 2 mL of diluted perchloric acid (100 in 117), then add 1 mL of hexaammonium heptamolybdate TS and 2 mL of 2,4-diaminophenol dihydrochloride TS, respec-tively, shake, add water to make exactly 25 mL, and allow to stand at 2019C for 30 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectropho-tometry <2.24>, using a solution prepared in the same man-ner with 2 mL of water, as the blank, and determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and the standard solution at 730 nm, respec-tively: the amount of free phosphoric acid is less than 0.25z. Amount (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 5.16 M: Amount (mg) of flavin adenine dinucleotide sodium taken, calculated on the anhydrous basis Heavy metals <1.07>—Proceed with 1.0 g of Flavin Adenine Dinucleotide Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Flavin Adenine Dinucleotide Sodium according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.10 g of Flavin Ade-nine Dinucleotide Sodium in 200 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 20 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Determine the peak area, A, of flavin adenine dinucleotide and the total area, S, of peaks other than flavin adenine dinucleotide by the automatic integration method: S/(A ＋ S ) is not more than 0.10. Operating conditions— Column, column temperature, mobile phase, flow rate, and time span of measurement: Proceed as directed in the operating conditions in the Procedure (ii) under the Assay (1). Detector: An ultraviolet absorption photometer (wave-length: 260 nm). System suitability— System performance: Proceed as directed in the system suitability in the Procedure (ii) under the Assay (1). Test for required detectability: To exactly 2 mL of the sample solution add the mobile phase to make exactly 20 mL, and use this solution as the solution for system suita-bility test. Confirm that the peak area of flavin adenine dinucleotide obtained with 20 mL of the solution for system suitability test is equivalent to 8 to 12z of that with 20 mL of the sample solution. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of flavin adenine dinucleotide is not more than 1.0z. Water <2.48> Take 50 mL of a mixture of methanol for water determination and ethyleneglycol for water determina-tion (1:1) into a dry titration flask, and titrate with Karl Fischer TS for water determination until end point. Weigh accurately about 0.1 g of Flavin Adenine Dinucleotide So-dium, transfer quickly to the titration flask, add an excess and constant volume of Karl Fischer TS for water determi-nation, dissolve by stirring for 10 minutes, and perform the test: the water content is not more than 10.0z.
(1) Procedure (i) Total flavin content—Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 0.1 g of Flavin Adenine Dinucleotide Sodium, and dissolve in water to make exactly 200 mL. Pipet 5 mL of this solution, add 5 mL of zinc chlo-ride TS, and heat in a water bath for 30 minutes. After cool-ing, add water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Riboflavin RS, previously dried at 1059C for 2 hours, dissolve in 200 mL of diluted acetic acid (100) (1 in 100) by warming, cool, add water to make exactly 500 mL. Pipet 10 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Deter-mine the absorbances, AT and AS, of the sample solution and standard solution at 450 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>, using water as the blank. Total amout (mg) of flavin ＝ MS × AT/AS × 4/5 MS: Amount (mg) of Riboflavin RS taken (ii) Peak area ratio of flavin adenine dinucleotide—Dis-solve 0.1 g of Flavin Adenine Dinucleotide Sodium in 200 mL of water, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under the Liquid Chromatography <2.01> according to the following conditions. Determine the peak area, A of flavin adenine dinucleotide, and the total area, S, of the peaks other than flavin adenine dinucleotide by the automatic integration method. Peak area ratio of flavin adenine dinucleotide ＝ 1.08A/(1.08A ＋ S ) Operating conditions— Detector: A visible spectrophotometer (wavelength: 450 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of a solution of potassium dihy-drogen phosphate (1 in 500) and methanol (4:1). Flow rate: Adjust so that the retention time of flavin ade-nine dinucleotide is about 10 minutes. Time span of measurement: About 4.5 times as long as the retention time of flavin adenine dinucleotide. System suitability— Test for required detectability: To exactly 2 mL of the sample solution add water to make exactly 20 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add water to make exactly 20 mL. Confirm that the peak area of flavin adenine dinucleotide obtained with 5 mL of this solu-tion is equivalent to 8 to 12z of that with 5 mL of the solu-tion for system suitability test. System performance: Dissolve 20 mg each of Flavin Ade-nine Dinucleotide Sodium and riboflavin sodium phosphate in 100 mL of water. When the procedure is run with 5 mL of this solution under the above operating conditions, flavin adenine dinucleotide and riboflavin phosphate are eluted in this order with the resolution between these peaks being not ‌1012‌ less than 2.0. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of flavin adenine dinucleotide is not more than 1.0z. (2) Calculation Amount (mg) of flavin adenine dinucleotide sodium (C27H31N9Na2O15P2) ＝ fT × fR × 2.2040 fT: Total amount (mg) of flavin in Flavin Adenine Dinucleotide Sodium obtained from the procedure (i) fR: Peak area ratio of flavin adenine dinucleotide in Flavin Adenine Dinucleotide Sodium obtained from the procedure (ii)
Flomoxef Sodium contains not less than 870 mg (potency) and not more than 985 mg (potency) per mg, calculated on the anhydrous basis.
Flomoxef Sodium occurs as white to light yel-low-white, powder or masses. It is very soluble in water, freely soluble in methanol, and sparingly soluble in ethanol (99.5).
(1) Decompose 0.01 g of Flomoxef Sodium as directed under Oxygen Flask Combustion Method <1.06>, using a mixture of 0.5 mL of 0.01 mol/L sodium hydroxide TS and 20 mL of water as the absorbing liquid. To 2 mL of the test solution so obtained add 1.5 mL of a mixture of alizarin complexone TS, acetic acid-potassium acetate buffer solution (pH 4.3) and cerium (III) nitrate TS (1:1:1): blue-purple color develops. Determine the absorption spectrum of a solution of Flomoxef Sodium (3 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Flomoxef Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Flomoxef Sodium in heavy water for nuclear magnetic resonance spec-troscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethyl- JP XVIII Official Monographs / Flomoxef Sodium 1015 silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits a single signal A at around d 3.5 ppm, a single signal or a sharp multiple signal B at around d 3.7 ppm, and a single signal C at around d 5.2 ppm. The ratio of the integrated in-tensity of these signals, A:B:C, is about 3:2:1. Flomoxef Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －8 – －139(1 g calculated on the anhydrous basis, a mixture of water and ethanol (99.5) (4:1), 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 0.5 g of Flomoxef Sodium in 5 mL of water is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Flomoxef Sodium in 10 mL of water: the solution is clear and has no more color than the following control solution. Control solution: To a mixture of 3.0 mL of Cobalt (II) Chloride CS and 12 mL of Iron (III) Chloride CS add 35 mL of diluted dilute hydrochloric acid (1 in 10). To 5.0 mL of this solution add 5.0 mL of diluted dilute hydrochloric acid (1:10). Heavy metals <1.07>—Proceed with 1.0 g of Flomoxef Sodium in a quartz crucible according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Flomoxef Sodium 5 mL of sulfuric acid and 5 mL of nitric acid, heat carefully until the solution changes to colorless to light yellow with occa-sional addition of 2 mL of nitric acid. After cooling, add 10 mL of ammonium oxalate TS, heat until white fumes evolve, and concentrate to 2 to 3 mL. After cooling, add water to make 10 mL, and perform the test using this solution as the test solution: the color is not darker than that of the control solution. Control solution: Proceed to prepare a solution in the same manner as the test solution without Flomoxef Sodium, and transfer 10 mL of the solution so obtained to the genera-tor bottle, add exactly 2 mL of Standard Arsenic Solution, and proceed in the same manner as the test solution (not more than 2 ppm). 1-(2-Hydroxyethyl)-1H-tetrazol-5-thiol—Use the sample solution obtained in the Assay as the sample solu-tions. Weigh accurately about 20 mg of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 25 mL of the internal standard solution and water to make 50 mL, and use this solution as the standard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internal standard: the amount of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol is not more than 1.0z of the amount of Flomoxef Sodium calculated on the anhy-drous basis. Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol (C3H6N4OS) ＝ MS × QT/QS × 1/10 MS: Amount (mg) of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol taken Internal standard solution—A solution of m-cresol (3 in 1000). Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add water to make exactly 20 mL. Confirm that the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol obtained with 5 mL of this solution is equivalent to 3.5 to 6.5z of that with 5 mL of the standard solution. System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol and the internal standard are eluted in this order with the resolution between these peaks being not less than 20. System repeatability: When the test is repeated 3 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of 1-(2-hydroxyethyl)-1H-tetrazol-5-thiol to that of the internal standard is not more than 1.0z. Water <2.48> Not more than 1.5z (0.5 g, volumetric titra-tion, back titration).
Weigh accurately an amount of Flomoxef Sodium and Flomoxef Triethylammonium RS, equivalent to about 50 mg (potency), and dissolve each in exactly 50 mL of the internal standard solution, add water to make 100 mL, and use these solutions as the sample solution and standard solu-tion. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatog-raphy <2.01> according to the following conditions, and cal-culate the ratios, QT and QS, of the peak area of flomoxef to that of the internal standard. Amount [ mg (potency)] of flomoxef (C15H18F2N6O7S2) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Flomoxef Triethylammo-nium RS taken Internal standard solution—A solution of m-cresol (3 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 246 nm). Column: A stainless steel column 4 mm in inside diameter and 20 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 – 10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.94 g of potassium dihydrogen phosphate, 3.22 g of disodium hydrogen phosphate dodeca-hydrate and 1.60 g of tetra-n-butylammonium bromide in water to make 1000 mL. To 750 mL of this solution add 250 mL of methanol. Flow rate: Adjust so that the retention time of flomoxef is about 9 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, flomoxef and the internal standard are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 3 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of flomoxef to that of the internal standard is not more than 1.0z.
Fluorescein Sodium contains not less than 98.
Fluorescein Sodium occurs as an orange pow-der. It is odorless, and tasteless. It is freely soluble in water, in methanol and in ethanol (95), and practically insoluble in diethyl ether. It is hygroscopic.
(1) To a solution of Fluorescein Sodium (1 in 100) having a strong green fluorescence, add a large quan-tity of water: the fluorescence remains. Acidify the solution with hydrochloric acid: the fluorescence disappears. Then render the solution alkaline with sodium hydroxide TS: the fluorescence reappears. Place 1 drop of a solution of Fluorescein Sodium (1 in 2000) on a piece of filter paper: a yellow spot develops. Expose the spot, while moist, to the vapor of bromine for 1 minute and then to ammonia vapor: the yellow color of the spot changes to red. Char 0.5 g of Fluorescein Sodium by ignition, cool, mix the residue with 20 mL of water, and filter: the filtrate responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity and color of solution—Dissolve 1 g of Fluorescein Sodium in 10 mL of water: the solution is clear, and shows a red color. Chloride <1.03>—Dissolve 0.15 g of Fluorescein So- dium in 20 mL of water, add 6 mL of dilute nitric acid and water to make 30 mL, and filter. To 20 mL of the filtrate add 2 mL of dilute nitric acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.355z). Sulfate <1.14>—Dissolve 0.20 g of Fluorescein So- dium in 30 mL of water, add 2.5 mL of dilute hydrochloric acid and water to make 40 mL, and filter. To 20 mL of the filtrate add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). Zinc—Dissolve 0.10 g of Fluorescein Sodium in 10 mL of water, add 2 mL of hydrochloric acid, and filter. To the filtrate add 0.1 mL of potassium hexacyanoferrate (II) TS: no turbidity is produced immediately. Related substances—Dissolve 0.20 g of Fluorescein Sodium in exactly 10 mL of methanol, and use this solution as the sample solution. Perform the test with this solution as directed under Thin-layer Chromatography <2.03>. Spot 5 mL of the sample solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and ammonia solution (28) (30:15:1) ‌1028 to a distance of about 10 cm, and air-dry the plate: any colored spot other than the principal spot does not appear. Loss on drying <2.41> Not more than 10.0z (1 g, 1059C, constant mass).
Transfer about 0.5 g of Fluorescein Sodium, accu-rately weighed, to a separator. Dissolve in 20 mL of water, add 5 mL of dilute hydrochloric acid, and extract with four 20-mL portions of a mixture of 2-methyl-1-propanol and chloroform (1:1). Wash each extract successively with the same 10 mL of water. Evaporate the combined extracts on a water bath with the aid of a current of air. Dissolve the residue in 10 mL of ethanol (99.5), evaporate the solution on a water bath to dryness, dry the residue at 1059C for 1 hour, and weigh as fluorescein (C20H12O5: 332.31). Amount (mg) of fluorescein sodium (C20H10Na2O5) ＝ amount (mg) of fluorescein (C20H12O5) × 1.132
Fosfomycin Calcium Hydrate is the calcium salt of a substance having antibacterial actively produced by the growth of Streptomyces fradiae or by the chemical synthesis. It contains not less than 725 mg (potency) and not more than 805 mg (potency) per mg, calculated on the anhydrous basis.
Fosfomycin Calcium Hydrate occurs as a white crystalline powder. It is slightly soluble in water, and practically insoluble in methanol and in ethanol (99.5).
(1) Determine the infrared absorption spec-trum of Fosfomycin Calcium Hydrate as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Fosfomy-cin Calcium Hydrate in heavy water for nuclear magnetic resonance spectroscopy (1 in 300), using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits a double signal at around d 1.5 ppm, a duple double signal at around d 2.9 ppm, a multiple signal at around d 3.3 ppm, and no signal at around d 1.4 ppm. A solution of Fosfomycin Calcium Hydrate (1 in 500) responds to Qualitative Tests <1.09> (3) for calcium salt. D Optical rotation <2.49> [a]20: －2.5 – －5.49(0.5 g calcu-lated on the anhydrous bases, 0.4 mol/L disodium dihydro-gen ethylenediamine tetraacetate TS (pH 8.5), 10 mL, 100 mm). JP XVIII Official Monographs / Fosfomycin Calcium Hydrate 1041 Phosphorus Content Weigh accurately about 0.1 g of Fos-fomycin Calcium Hydrate, add 40 mL of sodium periodate (107 in 10,000) and 2 mL of perchloric acid, and heat in a water bath for 1 hour. After cooling, add water to make exactly 200 mL. Pipet 10 mL of this solution, and add 1 mL of potassium iodide TS. To this solution add sodium thiosul-fate TS until the solution is colorless, add water to make exactly 100 mL, and use this solution as the sample stock so-lution. Separately, weigh accurately about 70 mg of potas-sium dihydrogen phosphate, proceed with this solution in the same manner as directed for the preparation of the sample stock solution, and use the solution so obtained as the stand-ard stock solution. Proceed and prepare a solution in the same manner for the preparation of the sample stock solu-tion without using Fosfomycin Calcium Hydrate, and use the solution so obtained as the blank stock solution. Pipet 5 mL each of the sample stock solution, the standard stock so-lution, and the blank stock solution, add 2.5 mL of ammo-nium molybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, mix, and add water to make exactly 25 mL, and use these solutions as the sample solu-tion, the standard solution, and the blank solution, respec-tively. After allowing these solutions to stand at 20 ± 19C for 30 minutes, perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as a blank, and determine the absorbances at 740 nm, AT, AS and AB, of the sample solution, the standard so-lution and the blank solution: the content of phosphorus is 15.2 – 16.7z. Amount (mg) of phosphorus (P) ＝ MS × (AT － AB)/(AS － AB) × 0.228 MS: Amount (mg) of potassium dihydrogen phosphate taken Calcium Content Weigh accurately about 0.2 g of Fosfo-mycin Calcium Hydrate, add 4 mL of 1 mol/L Hydrochloric acid TS, and shake well until the sample is completely dis-solved. To this solution add 100 mL of water, 9 mL of so-dium hydroxide TS and 0.1 g of methylthymol blue-sodium chloride indicator, and titrate <2.50> with 0.05 mol/L disodi-um dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from clear blue to gray or gray-purple: calcium content is 19.6 – 21.7z. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.004 mg of Ca
(1) Heavy metals <1.07>—To 1.0 g of Fosfomycin Calcium Hydrate add 40 mL of 0.25 mol/L acetic acid TS and water to make 50 mL. Proceed with this solution accord-ing to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Fosfomycin Calcium Hydrate according to Method 3, and perform the test (not more than 2 ppm). Glycol substance—Weigh accurately about 0.2 g of Fosfomycin Calcium Hydrate, transfer into a 250-mL iodine flask, add 100 mL of water, and dissolve by sonicating while cooling in ice. Add exactly 50 mL of phthalate buffer solu-tion (pH 5.8) and exactly 5 mL of sodium periodate solution (107 in 100,000), stopper, stir, and add 1 mL of water in the receiving part. Avoid exposure to light, allow to stand in a water bath at 309C for 60 minutes, add exactly 10 mL of a solution of potassium iodide (2 in 5) without haste, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS (indica-tor: 2 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction: amount of glycol substance (C3H7CaO5P) is not more than 1.5z. Each mL of 0.01 mol/L sodium thiosulfate VS ＝ 0.4854 mg of C3H7CaO5P Water <2.48> Not more than 12.0z (0.1 g, volumetric titration, direct titration. Use a mixture of formamide for water determination and methanol for water determination (2:1) instead of methanol for water determination).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Proteus sp. (MB838) Culture medium—Dissolve 5.0 g of peptone, 3.0 g of meat extract, 2.0 g of yeast extract, and 15 g of agar in 1000 mL of water, sterilize, and use as the agar media for base layer and seed layer with the pH of between 6.5 and 6.6 after sterilization. Seeded agar layer—Incubate the test organism on the slant of the agar medium for transferring test organisms at 379C for 40 – 48 hours. Subcultures at least 3 times. In-oculate the grown organisms onto the surface of 300 mL of the agar medium for transferring test organisms in a Roux bottle, incubate at 379C for 40 – 48 hours, and suspend the grown organisms in about 30 mL of water. To the suspen-sion add water, and use this as the stock suspension of test organism. The amount of the water to be added is adjust so that the percent transmission at 560 nm of the suspension diluted ten times with water is 17z. Keep the stock suspen-sion at 109C or below and use within 7 days. Add 1.0 – 2.0 mL of the stock suspension of test organism to 100 mL of the agar medium for seed layer previously kept at 489C, mix thoroughly, and use this as the deeded agar layer. Standard solutions—Weigh accurately an amount of Fosfomycin Phenethylammonium RS equivalent to about 20 mg (potency), dissolve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively. Sample solutions—Weigh accurately an amount of Fosfomycin Calcium Hydrate equivalent to about 20 mg (potency), and dissolve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 50 mL. To exactly a suitable amount of this solution add 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentration sample solution and the low concentra-tion sample solution, respectively.
Fosfomycin Sodium is the sodium salt of a sub-stance having antibacterial activity produced by the growth of Streptomyces fradiae or by the chemical synthesis. It contains not less than 725 mg (potency) and not more than 770 mg (potency) per mg, calculated on the anhydrous basis.
Fosfomycin Sodium occurs as a white crystal-line powder. It is very soluble in water, sparingly soluble in methanol, and practically insoluble in ethanol (99.5).
(1) Determine the infrared absorption spec-trum of Fosfomycin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Fosfo-mycin Sodium in heavy water for nuclear magnetic reso-nance spectroscopy (1 in 300), using sodium 3-trimethylsilyl-propanesulfonate for nuclear magnetic resonance spectros-copy as an internal reference compound, as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>: it exhibits a double signal at around d 1.5 ppm, a duple double signal at around d 2.8 ppm, a multiple signal at around d 3.3 ppm, and no signal at around d 1.3 ppm. A solution of Fosfomycin Sodium (1 in 500) responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －3.5 – －5.59(0.5 g calcu-lated on the anhydrous bases, water, 10 mL, 100 mm). pH <2.54> Dissolve 0.70 g of Fosfomycin Sodium in 10 mL of water: the pH of the solution is between 8.5 and 10.5. Phosphorus Content Weigh accurately about 0.1 g of Fos-fomycin Sodium, add 40 mL of a solution of sodium perio-date (107 in 10,000) and 2 mL of perchloric acid, and heat in a water bath for 1 hour. After cooling, add water to make exactly 200 mL. Pipet 10 mL of this solution, and add 1 mL of potassium iodide TS. To this solution add sodium thiosul-fate TS until the solution is colorless, add water to make exactly 100 mL, and use this solution as the sample stock so-lution. Separately, weigh accurately about 70 mg of potas-sium dihydrogen phosphate, proceed with this solution in the same manner as directed for the preparation of the sample stock solution, and use the solution so obtained as the stand-ard stock solution. Proceed and prepare a solution in the same manner for the preparation of the sample stock solu-tion without using Fosfomycin Sodium, and use the solution so obtained as the blank stock solution. Pipet 5 mL each of the sample stock solution, the standard stock solution, and the blank stock solution, add 2.5 mL of ammonium molyb- date-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sul-fonic acid TS, mix, and add water to make exactly 25 mL, and use these solutions as the sample solution, the standard solution, and the blank solution, respectively. After allowing these solutions to stand for 30 minutes at 20 ± 19C, perform the test with these solutions as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as a blank, and determine the absorbances at 740 nm, AT, AS and AB, of the sample solution, the standard solution and the blank solution: the content of phosphorus is 16.2 – 17.9z. Amount (mg) of phosphorus (P) ＝ M × (AT － AB)/(AS － AB) × 0.228 M: Amount (mg) of potassium dihydrogen phosphate taken
(1) Clarity and color of solution—Dissolve 1.0 g of Fosfomycin Sodium in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Fosfo- mycin Sodium according to Method 1, and perform the test. Prepare the control solution with 2.0 mL Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Fosfomycin Sodium according to Method 3, and perform the test (not more than 2 ppm). Glycol substance—Weigh accurately about 0.2 g of Fosfomycin Sodium, and dissolve in 100 mL of water in a 250-mL iodine flask. Add exactly 50 mL of phthalate buffer solution (pH 5.8) and exactly 5 mL of sodium periodate so-lution (107 in 100,000), stopper, stir, and add 1 mL of water in the receiving part. Allow to stand in a dark place for 90 minutes, add exactly 10 mL of a solution of potassium iodide (2 in 5) without haste, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS (indicator: 2 mL of starch TS). Perform a blank determination in the same manner, and make any necessary correction: amount of glycol substance (C3H7Na2O5P) is not more than 0.5z. Each mL of 0.01 mol/L sodium thiosulfate VS ＝ 0.5001 mg of C3H7Na2O5P Water <2.48> Not more than 3.0z (0.2 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Proteus sp. (MB838) Culture medium—Mix 5.0 g of peptone, 3.0 g of meat extract, 2.0 g of yeast extract, and 15 g of agar in 1000 mL of water, sterilize, and use as the agar media for base layer and seed layer with the pH of between 6.5 and 6.6 after sterilization. Seeded agar layer—Incubate the test organism on the slant of the agar medium for transferring test organisms at 379C for 40 – 48 hours. Subcultures at least 3 times. Inoculate the grown organisms onto the surface of 300 mL of the agar medium for transferring test organisms in a Roux bottle, incubate at 379C for 40 – 48 hours, and suspend the grown organisms in about 30 mL of water. To the suspen-sion add water, and use this as the stock suspension of test organism. The amount of the water to be added is adjust so that the percent transmission at 560 nm of the suspension diluted ten times with water is 17z. Keep the stock suspen-sion at 109C or below and use within 7 days. Add 1.0 – 2.0 mL of the stock suspension of test organism to 100 mL of the agar medium for seed layer previously kept at 489C, mix thoroughly, and use this as the deeded agar layer. ‌1044 Standard solutions—Weigh accurately an amount of Fosfomycin Phenethylammonium RS equivalent to about 20 mg (potency), dissolve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution at 59C or below and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentration standard solu-tion and the low concentration standard solution, respec-tively. Sample solutions—Weigh accurately an amount of Fosfomycin Sodium equivalent to about 20 mg (potency), and dissolve in 0.05 mol/L tris buffer solution (pH 7.0) to make exactly 50 mL. To exactly a suitable amount of this solution add 0.05 mol/L tris buffer solution (pH 7.0) to make solutions so that each mL contains 10 mg (potency) and 5 mg (potency), and use these solutions as the high concentra-tion sample solution and the low concentration sample solu-tion, respectively.
Hydrocortisone Sodium Phosphate contains not less than 96.
Hydrocortisone Sodium Phosphate occurs as a white to light yellow powder. It is freely soluble in water, sparingly soluble in methanol, and very slightly soluble in ethanol (95). It is hygroscopic. It shows crystal polymorphism.
(1) To 2 mg of Hydrocortisone Sodium Phosphate add 2 mL of sulfuric acid: a yellowish green fluorescence is exhibited initially, then gradually changes through orange-yellow to dark red. Examine the solution under ultraviolet light (main wavelength: 254 nm): an in-tense, light green fluorescence is exhibited. To this solution add carefully 10 mL of water: the color changes from yellow to orange-yellow with a light green fluorescence and a yel-low-brown, flocculent floating substance is formed. Determine the infrared absorption spectrum of Hy-drocortisone Sodium Phosphate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Hydrocortisone Sodium Phosphate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve Hydrocortisone Sodium Phosphate and Hydrocorti-sone Sodium Phosphate RS in methanol, respectively, then evaporate the methanol to dryness, and repeat the test on the residues. Moisten 1.0 g of Hydrocortisone Sodium Phosphate with a small quantity of sulfuric acid, and incinerate by gradual heating. After cooling, dissolve the residue in 10 mL of dilute nitric acid, and heat in a water bath for 30 minutes. After cooling, filter if necessary. This solution responds to Qualitative Tests <1.09> for sodium salt and for phosphate. D Optical rotation <2.49> [a]20: ＋123 – ＋1319(1 g calculated on the anhydrous basis, phosphate buffer solution (pH 7.0), 100 mL, 100 mm). JP XVIII Official Monographs / Hydrocortisone Sodium Succinate 1117 pH <2.54> Dissolve 1.0 g of Hydrocortisone Sodium Phos-phate in 100 mL of water: the pH of this solution is between 7.5 and 9.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Hydrocortisone Sodium Phosphate in 10 mL of water: the solution is clear and colorless to pale yellow. Chloride <1.03>—Dissolve 0.30 g of Hydrocortisone ‌Sodium Phosphate in 20 mL of water, and add 6 mL of dilute nitric acid and water to make 100 mL. To 5 mL of this solution add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.600z). Heavy metals <1.07>—Proceed with 0.5 g of Hydro- cortisone Sodium Phosphate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 40 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Hydrocortisone Sodium Phosphate according to Method 3, and perform the test (not more than 2 ppm). Free phosphoric acid—Weigh accurately about 0.25 g of Hydrocortisone Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample so-lution. Pipet 5 mL each of the sample solution and Standard Phosphoric Acid Solution into separate 25-mL volumetric flasks, add 2.5 mL of hexaammonium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, shake, add water to make exactly 25 mL, and allow to stand at 20 ± 19C for 30 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared with 5 mL of water in the same manner as the blank. Determine the absor-bances, AT and AS, at 740 nm of the sample solution and Standard Phosphoric Acid Solution: the amount of free phosphoric acid is not more than 1.0z. Content (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Hydrocortisone Sodium Phosphate taken, calculated on the anhydrous basis Free hydrocortisone—Dissolve 25 mg of Hydrocorti-sone Sodium Phosphate in the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Sepa-rately, weigh 25 mg of Hydrocortisone RS, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of hydrocortisone in each solution: AT is not larger than AS. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrocortisone is not more than 1.0z. Water <2.48> Not more than 5.0z (30 mg, coulometric titration).
Weigh accurately about 50 mg of Hydrocortisone Butyrate, previously dried, and dissolve in ethanol (99.5) to make exactly 100 mL. Pipet 2 mL of this solution, and add Description Hydrocortisone Sodium Phosphate occurs as a white to light yellow powder. It is freely soluble in water, sparingly soluble in methanol, and very slightly soluble in ethanol (95). It is hygroscopic. It shows crystal polymorphism. Identification (1) To 2 mg of Hydrocortisone Sodium Phosphate add 2 mL of sulfuric acid: a yellowish green fluorescence is exhibited initially, then gradually changes through orange-yellow to dark red. Examine the solution under ultraviolet light (main wavelength: 254 nm): an in-tense, light green fluorescence is exhibited. To this solution add carefully 10 mL of water: the color changes from yellow to orange-yellow with a light green fluorescence and a yel-low-brown, flocculent floating substance is formed. Determine the infrared absorption spectrum of Hy-drocortisone Sodium Phosphate as directed in the paste method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Hydrocortisone Sodium Phosphate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve Hydrocortisone Sodium Phosphate and Hydrocorti-sone Sodium Phosphate RS in methanol, respectively, then evaporate the methanol to dryness, and repeat the test on the residues. Moisten 1.0 g of Hydrocortisone Sodium Phosphate with a small quantity of sulfuric acid, and incinerate by gradual heating. After cooling, dissolve the residue in 10 mL of dilute nitric acid, and heat in a water bath for 30 minutes. After cooling, filter if necessary. This solution responds to Qualitative Tests <1.09> for sodium salt and for phosphate. D Optical rotation <2.49> [a]20: ＋123 – ＋1319(1 g calculated on the anhydrous basis, phosphate buffer solution (pH 7.0), 100 mL, 100 mm). JP XVIII Official Monographs / Hydrocortisone Sodium Succinate 1117 pH <2.54> Dissolve 1.0 g of Hydrocortisone Sodium Phos-phate in 100 mL of water: the pH of this solution is between 7.5 and 9.5. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Hydrocortisone Sodium Phosphate in 10 mL of water: the solution is clear and colorless to pale yellow. Chloride <1.03>—Dissolve 0.30 g of Hydrocortisone ‌Sodium Phosphate in 20 mL of water, and add 6 mL of dilute nitric acid and water to make 100 mL. To 5 mL of this solution add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.600z). Heavy metals <1.07>—Proceed with 0.5 g of Hydro- cortisone Sodium Phosphate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 40 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Hydrocortisone Sodium Phosphate according to Method 3, and perform the test (not more than 2 ppm). Free phosphoric acid—Weigh accurately about 0.25 g of Hydrocortisone Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample so-lution. Pipet 5 mL each of the sample solution and Standard Phosphoric Acid Solution into separate 25-mL volumetric flasks, add 2.5 mL of hexaammonium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, shake, add water to make exactly 25 mL, and allow to stand at 20 ± 19C for 30 minutes. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared with 5 mL of water in the same manner as the blank. Determine the absor-bances, AT and AS, at 740 nm of the sample solution and Standard Phosphoric Acid Solution: the amount of free phosphoric acid is not more than 1.0z. Content (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Hydrocortisone Sodium Phosphate taken, calculated on the anhydrous basis Free hydrocortisone—Dissolve 25 mg of Hydrocorti-sone Sodium Phosphate in the mobile phase to make exactly 20 mL, and use this solution as the sample solution. Sepa-rately, weigh 25 mg of Hydrocortisone RS, previously dried at 1059C for 3 hours, and dissolve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine the peak areas, AT and AS, of hydrocortisone in each solution: AT is not larger than AS. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of hydrocortisone is not more than 1.0z. Water <2.48> Not more than 5.0z (30 mg, coulometric titration). Assay Weigh accurately about 20 mg each of Hydrocorti- sone Sodium Phosphate and Hydrocortisone Sodium Phos-phate RS (previously determine the water <2.48> in the same manner as Hydrocortisone Sodium Phosphate), dissolve each in 50 mL of the mobile phase, add exactly 10 mL of the internal standard solution, then add the mobile phase to make 200 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of hydrocortisone phosphate to that of the internal standard, respectively. Amount (mg) of hydrocortisone sodium phosphate (C21H29Na2O8P) ＝ MS × QT/QS MS: Amount (mg) of Hydrocortisone Sodium Phosphate RS taken, calculated on the anhydrous basis Internal standard solution—A solution of isopropyl parahy-droxybenzoate in the mobile phase (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of 0.05 mol/L sodium dihydro-gen phosphate TS (pH 2.6) and methanol (1:1). Flow rate: Adjust so that the retention time of hydrocorti-sone phosphate is about 10 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, hydrocortisone phosphate and the internal standard are eluted in this order with the resolution between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of hydrocortisone phosphate to that of the internal standard is not more than 1.0z.
Hydrocortisone Sodium Succinate, calculated on the dried basis, contains not less than 97.
Hydrocortisone Sodium Succinate occurs as a white, powder or masses. It is freely soluble in water, in methanol and in ethanol (95). It is hygroscopic. It is gradually colored by light. It shows crystal polymorphism.
(1) Dissolve 0.2 g of Hydrocortisone So-dium Succinate in 20 mL of water, and add 0.5 mL of dilute hydrochloric acid with stirring: a white precipitate is formed. Collect the precipitate, wash it with two 10-mL portions of water, and dry at 1059C for 3 hours. To 3 mg of this dried matter add 2 mL of sulfuric acid: the solution shows a yel-lowish green fluorescence immediately, and the color of the solution gradually changes through orange-yellow to dark red. This solution shows a strong light green fluorescence under ultraviolet light. Add carefully 10 mL of water to this solution: the color changes from yellow to orange-yellow with a light green fluorescence, and a yellow-brown floccu-lent precipitate is formed. Dissolve 10 mg of the dried matter obtained in (1) in 1 mL of methanol, add 1 mL of Fehling's TS, and heat: an orange to red precipitate is formed. To 0.1 g of the dried matter obtained in (1) add 2 mL of sodium hydroxide TS, and allow to stand for 10 minutes. Filter the solution to remove the precipitate formed, mix the filtrate with 1 mL of dilute hydrochloric acid, filter if neces-sary, then adjust the solution to a pH of about 6 with diluted ammonia TS (1 in 10), and add 2 to 3 drops of iron (III) chloride TS: a brown precipitate is formed. Determine the infrared absorption spectrum of the dried matter obtained in (1) as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of previously dried Hydrocortisone Succinate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve Hydrocortisone Sodium Succinate and Hydrocortisone Succinate RS in methanol, respectively, then evaporate the methanol to dryness, and repeat the test on the residues. Hydrocortisone Sodium Succinate responds to Quali-tative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋135 – ＋1459(0.1 g calcu-lated on the dried basis, ethanol (95), 10 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 0.5 g of Hydrocortisone Sodium Succinate in 5 mL of water: the solution is clear and colorless. (2) Related substances—Dissolve 25 mg of Hydrocorti-sone Sodium Succinate in methanol to make exactly 10 mL, and use this solution as the sample solution. Separately, dis-solve 25 mg of hydrocortisone in methanol to make exactly 10 mL. Pipet 1 mL of this solution, add methanol to make exactly 20 mL, and use this solution as the standard solution (1). Pipet 6 mL of the standard solution (1), add methanol to make exactly 10 mL, and use this solution as the standard solution (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 3 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chlo-roform, ethanol (99.5) and formic acid (150:10:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spot ob-tained from the sample solution corresponding to the spot from the standard solution (1) is not more intense than the spot from the standard solution (1). Any spot other than the principal spot and the above spot from the sample solution is not more than one, and is not more intense than the spot from the standard solution (2). Loss on drying <2.41> Not more than 2.0z (0.5 g, 1059C, 3 hours).
Weigh accurately about 10 mg of Hydrocortisone Sodium Succinate, and dissolve in methanol to make exactly 100 mL. Pipet 5 mL of this solution, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 10 mg of Hydrocortisone Succinate RS, previously dried at 1059C for 3 hours, proceed in the same manner as directed for the sample solution, and use this solution as the standard solution. Determine the ab-sorbances, AT and AS, of the sample solution and standard solution at 240 nm as directed under Ultraviolet-visible Spec-trophotometry <2.24>. Amount (mg) of hydrocortisone sodium succinate (C25H33NaO8) ＝ MS × AT/AS × 1.048 MS: Amount (mg) of Hydrocortisone Succinate RS taken
Kaolin occurs as white or nearly white, frag-mentary masses or powder. It has a slightly clay-like odor. It is practically insoluble in water, in ethanol (99.5) and in diethyl ether. It is insoluble in dilute hydrochloric acid and in sodium hydroxide TS. When moistened with water, it darkens and becomes plas-tic.
(1) Heat 1 g of Kaolin with 10 mL of water and 5 mL of sulfuric acid in a porcelain dish, and evaporate the mixture nearly to dryness. Cool, add 20 mL of water, boil for 2 to 3 minutes, and filter: the color of the residue is gray. (2) The filtrate obtained in (1) responds to Qualitative Tests <1.09> (1), (2) and (4) for aluminum salt.
(1) Acid or alkali—Add 25 mL of water to 1.0 g of Kaolin, agitate thoroughly, and filter: the pH <2.54> of the filtrate is between 4.0 and 7.5. Acid-soluble substances—Add 20 mL of dilute hy-drochloric acid to 1.0 g of Kaolin, agitate for 15 minutes, and filter. Evaporate 10 mL of the filtrate to dryness, and heat strongly between 4509C and 5509C to constant mass: the mass of the ignited residue is not more than 10 mg. Carbonate—Stir 1.0 g of Kaolin with 5 mL of water, then add 10 mL of diluted sulfuric acid (1 in 2): no efferves-cence occurs. Heavy metals <1.07>—Boil 1.5 g of Kaolin gently with 50 mL of water and 5 mL of hydrochloric acid for 20 minutes with frequent agitation, cool, centrifuge, and sepa-rate the supernatant liquid. Wash the precipitate twice with 10 mL of water, centrifuge each time, and combine the su-pernatant liquid and the washings. Add dropwise ammonia solution (28) to this solution until a slight precipitate occurs, then add dilute hydrochloric acid dropwise while agitating strongly to complete solution. Add 0.45 g of hydroxylammo-nium chloride, and heat. Cool, add 0.45 g of sodium acetate trihydrate and 6 mL of dilute acetic acid, filter if necessary, and wash with 10 mL of water. Combine the filtrate and the washings, and add water to make 150 mL. Perform the test using 50 mL of this solution as the test solution. To 2.5 mL of Standard Lead Solution add 0.15 g of hydroxylammo-nium chloride, 0.15 g of sodium acetate trihydrate, 2 mL of dilute acetic acid and water to make 50 mL, and use this so-lution as the control solution (not more than 50 ppm). Iron <1.10>—Add 10 mL of dilute hydrochloric acid to 40 mg of Kaolin, and heat for 10 minutes with shaking in a water bath. After cooling, add 0.5 g of L-tartaric acid, dis-solve with shaking, prepare the test solution with this solu-tion according to Method 2, and perform the test according to Method B. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 500 ppm). Arsenic <1.11>—Add 5 mL of water and 1 mL of sul- furic acid to 1.0 g of Kaolin, and heat on a sand bath until white fumes begin to evolve. Cool, and add water to make 5 mL. Perform the test with this solution as the test solution (not more than 2 ppm). Foreign matter—Place 5 g of Kaolin in a beaker, add 100 mL of water, stir, and decant to leave sand. Repeat this procedure several times with 100-mL portions of water: no ‌JP XVIII Official Monographs / Ketoconazole 1221‌ sandy residue remains. Loss on ignition <2.43> Not more than 15.0z (1 g, 6009C, 5 hours). Plasticity Add 7.5 mL of water to 5.0 g of Kaolin, and agitate thoroughly: the resultant mass has no remarkable fluidity.
Latamoxef Sodium contains not less than 830 mg (potency) and not more than 940 mg (potency) per mg, calculated on the anhydrous basis.
Latamoxef Sodium occurs as white to light yel-low-white, powder or masses. It is very soluble in water, freely soluble in methanol, and slightly soluble in ethanol (95).
(1) Determine the absorption spectrum of a solution of Latamoxef Sodium (3 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Latamoxef Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Determine the 1H spectrum of a solution of Latamox-ef Sodium in heavy water for nuclear magnetic resonance spectroscopy (1 in 10) as directed under Nuclear Magnetic Resonance Spectroscopy <2.21>, using sodium 3-trimethyl-silylpropanesulfonate for nuclear magnetic resonance spec-troscopy as an internal reference compound: it exhibits single signals, A and B, at around d 3.5 ppm and at around d 4.0 ppm. The ratio of the integrated intensity of these sig-nals, A:B, is about 1:1. Latamoxef Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: －32 – －409(0.5 g calculated on the anhydrous basis, phosphate buffer solution (pH 7.0), 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Latamoxef Sodium in 10 mL of water is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Latamoxef Sodium in 10 mL of water: the solution is clear and has no more color than the following control solu-tion. Control solution: To a mixture of 3.0 mL of Cobalt (II) Chloride CS and 36 mL of Iron (III) Chloride CS add 11 mL of diluted dilute hydrochloric acid (1 in 10). To 2.5 mL of this solution add 7.5 mL of diluted dilute hydrochloric acid (1:10). Heavy metals <1.07>—Carbonize 1.0 g of Latamoxef Sodium by heating gently, previously powdered if it is masses. After cooling, add 10 mL of a solution of magne-sium nitrate hexahydrate in ethanol (1 in 10), and burn the ethanol. After cooling, add 1 mL of sulfuric acid. Proceed according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution by dissolv- ing 1.0 g of Latamoxef Sodium in 20 mL of water, and per-form the test (not more than 2 ppm). Related substances—Dissolve 25 mg of Latamoxef Sodium in water to make 50 mL, and use this solution as the sample solution. Pipet 2 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the peak area of 1-methyl-1H-tetrazole-5-thiol, having the relative retention time of about 0.5 to the first eluted peak of the two peaks of latamoxef, obtained from the sample solution is not larger than the peak area of latamoxef from the standard solution, and the peak area of decarboxylatamoxef, having the relative retention time of about 1.7 to the first peak of the two peaks of latamoxef, is not larger than 2 times that of latamoxef from the standard solution. For the peak area of 1-methyl-1H-tetrazole-5-thiol, multiply its correction factor, 0.52. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: Proceed as directed in the system suitability in the Assay. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of latamoxef is not more than 2.0z. Water <2.48> Not more than 5.0z (0.5 g, volumetric titra-tion, back titration). Isomer ratio Dissolve 25 mg of Latamoxef Sodium in water to make 50 mL, and use this solution as the sample solution. Perform the test with 5 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the areas, Aa and Ab, of the two peaks in order of elution, which appear close to each other at the retention time of about 10 minutes: Aa/Ab is be-tween 0.8 and 1.4. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 7.7 g of ammonium acetate in water to make 1000 mL. To 950 mL of this solution add 50 mL of methanol. Flow rate: Adjust so that the retention time of the first eluted peak of latamoxef is about 8 minutes. System suitability— System performance: When the procedure is run with 5 mL of the sample solution under the above operating conditions, ‌1246‌ the resolution between the two peaks of latamoxef is not less than 3. System repeatability: When the test is repeated 3 times with 5 mL of the sample solution under the above operating conditions, the relative standard deviation of the area of the first eluted peak of latamoxef is not more than 2.0z.
Weigh accurately an amount of Latamoxef Sodium and Latamoxef Ammonium RS, equivalent to about 25 mg (potency) each, dissolve in exactly 5 mL of the internal standard solution, add water to make 50 mL, and use these solutions as the sample solution and standard solution. Per-form the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of latamoxef to that of the internal standard. Amount [ mg (potency)] of latamoxef (C20H20N6O9S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Latamoxef Ammonium RS taken Internal standard solution—A solution of m-cresol (3 in 200). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.94 g of potassium dihydrogen phosphate, 3.22 g of disodium hydrogen phosphate dodeca-hydrate and 1.60 g of tetra-n-butylammonium bromide in water to make exactly 1000 mL. To 750 mL of this solution add 250 mL of methanol. Flow rate: Adjust so that the retention time of latamoxef is about 7 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, latamoxef and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of latamoxef to that of the internal standard is not more than 1.0z.
Containers—Tight containers. Storage—Not exceeding 59C.
Levothyroxine Sodium Hydrate contains not less well.
Levothyroxine Sodium Hydrate occurs as a pale yellow-white to light yellow-brown powder. It is odor-less. It is slightly soluble in ethanol (95), and practically insolu-ble in water and in diethyl ether. It dissolves in sodium hydroxide TS. It is gradually colored by light.
(1) Heat 0.1 g of Levothyroxine Sodium Hydrate over a flame: a purple gas evolves. To 0.5 mg of Levothyroxine Sodium Hydrate add 8 mL of a mixture of water, ethanol (95), hydrochloric acid and sodium hydroxide TS (6:5:2:2), warm in a water bath for 2 minutes, cool, and add 0.1 mL of sodium nitrite TS. Allow to stand in a dark place for 20 minutes, and add 1.5 mL of ammonia solution (28): a yellowish red color is pro-duced. Determine the absorption spectrum of a solution of Levothyroxine Sodium Hydrate in dilute sodium hydroxide TS (1 in 10,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Moisten Levothyroxine Sodium Hydrate with sulfu-ric acid, and ignite: the residue responds to Qualitative Tests <1.09> (1) and (2) for sodium salt. D Optical rotation <2.49> [a]20: －5 – －69(0.3 g calculated on the dried basis, a mixture of ethanol (95) and sodium hy-droxide TS (2:1), 10 mL, 100 mm). ‌1262
(1) Clarity and color of solution—To 0.5 g of Levomepromazine Maleate add 10 mL of methanol, and dis-solve by warming: the solution is clear, and colorless or pale yellow. Chloride <1.03>—Dissolve 0.5 g of Levomepromazine Maleate in 40 mL of methanol, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS, 40 mL of methanol, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.028z). Heavy metals <1.07>—Proceed with 2.0 g of Levomepromazine Maleate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Description Levothyroxine Sodium Hydrate occurs as a pale yellow-white to light yellow-brown powder. It is odor-less. It is slightly soluble in ethanol (95), and practically insolu-ble in water and in diethyl ether. It dissolves in sodium hydroxide TS. It is gradually colored by light. Identification (1) Heat 0.1 g of Levothyroxine Sodium Hydrate over a flame: a purple gas evolves. To 0.5 mg of Levothyroxine Sodium Hydrate add 8 mL of a mixture of water, ethanol (95), hydrochloric acid and sodium hydroxide TS (6:5:2:2), warm in a water bath for 2 minutes, cool, and add 0.1 mL of sodium nitrite TS. Allow to stand in a dark place for 20 minutes, and add 1.5 mL of ammonia solution (28): a yellowish red color is pro-duced. Determine the absorption spectrum of a solution of Levothyroxine Sodium Hydrate in dilute sodium hydroxide TS (1 in 10,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Moisten Levothyroxine Sodium Hydrate with sulfu-ric acid, and ignite: the residue responds to Qualitative Tests <1.09> (1) and (2) for sodium salt. D Optical rotation <2.49> [a]20: －5 – －69(0.3 g calculated on the dried basis, a mixture of ethanol (95) and sodium hy-droxide TS (2:1), 10 mL, 100 mm). ‌1262 Purity (1) Clarity and color of solution—Dissolve 0.3 g of Levothyroxine Sodium Hydrate in 10 mL of a mixture of ethanol (95) and sodium hydroxide TS (2:1) by warming: the solution is clear and pale yellow to pale yellow-brown in color. Soluble halides—Dissolve 0.01 g of Levothyroxine Sodium Hydrate in 10 mL of water and 1 drop of dilute nitric acid, shake for 5 minutes, and filter. To the filtrate Levothyroxine Sodium Tablets
Liothyronine Sodium contains not less than 95.
Liothyronine Sodium occurs as a white to light brown powder. It is odorless. It is slightly soluble in ethanol (95), and practically insolu-ble in water and in diethyl ether. It dissolves in sodium hydroxide TS and in ammonia TS.
(1) To 5 mL of a solution of Liothyronine Sodium in ethanol (95) (1 in 1000) add 1 mL of ninhydrin TS, and warm in a water bath for 5 minutes: a purple color develops. Heat 0.02 g of Liothyronine Sodium with a few drops of sulfuric acid over a flame: a purple gas is evolved. Determine the absorption spectrum of a solution of Liothyronine Sodium in ethanol (95) (1 in 10,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Ignite 0.02 g of Liothyronine Sodium until thor-oughly charred. After cooling, add 5 mL of water to the residue, shake, and filter: the filtrate responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋18 – ＋229(0.2 g calculated on the dried basis, a mixture of ethanol (95) and 1 mol/L hy-drochloric acid TS (4:1), 10 mL, 100 mm).
(1) Soluble halide—To 10 mg of Liothyronine So-dium add 10 mL of water and 1 drop of dilute nitric acid, shake for 5 minutes, and filter. Add water to the filtrate to make 10 mL, and mix with 3 drops of silver nitrate TS: the solution shows no more turbidity than the following control solution. Control solution: To 0.35 mL of 0.01 mol/L hydrochloric acid VS add 1 drop of dilute nitric acid and water to make 10 mL, and add 3 drops of silver nitrate TS. Iodine and iodide—Dissolve 0.10 g of Liothyronine Sodium in 10 mL of dilute sodium hydroxide TS and 15 mL of water, add 5 mL of dilute sulfuric acid, and allow to stand for 10 minutes with occasional shaking. Filter the mixture into a Nessler tube, add 10 mL of chloroform and 3 drops of a solution of potassium iodate (1 in 100) to the filtrate, mix for 30 seconds, and allow to stand: the chloroform layer has Amount [mg (potency)] of lincomycin (C ＝ MS × AT/AS × 15 18H34N2O6S) no more color than the following control solution. Control solution: Weigh exactly 0.111 g of potassium iodide, and dissolve in water to make 1000 mL. Pipet 1 mL MS: Amount [mg (potency)] of Lincomycin Hydrochlo-ride RS taken
Weigh accurately about 25 mg of Liothyronine So-dium, and proceed as directed under Oxygen Flask Combus-tion Method <1.06>, using a mixture of 10 mL of a solution of sodium hydroxide (1 in 100) and 1 mL of a freshly pre-pared solution of sodium bisulfate (1 in 100) as the absorb-ing liquid, and prepare the test solution. Apply a small amount of water to the upper part of apparatus A, pull out C carefully, and wash C, B and the inner wall of A with 40 mL of water. To the test solution add 1 mL of bromine-acetic acid TS, insert the stopper C, and shake vigorously for 1 minute. Remove the stopper, rinse the stopper, the sample holder and the inner wall of the flask with 40 mL of water, and add 0.5 mL of formic acid. Stopper the flask with C, and shake vigorously for 1 minute again. Remove the stop-per, and rinse the stopper, the sample holder and the inner wall of the flask with 40 mL of water again. Bubble the solu-tion with enough nitrogen gas in the flask to remove the oxygen and excess bromine, add 0.5 g of potassium iodide to the solution, and dissolve. Add immediately 3 mL of dilute sulfuric acid, mix, and allow to stand for 2 minutes. Titrate <2.50> the solution with 0.02 mol/L sodium thiosulfate VS (indicator: 3 mL of starch TS). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.02 mol/L sodium thiosulfate VS ＝ 0.7477 mg of C15H11I3NNaO4
Lobenzarit Sodium, when dried, contains not less than 98.
Lobenzarit Sodium occurs as a white to pale yellow-white crystalline powder. It is soluble in water, and practically insoluble in ethanol (99.5).
(1) A solution of Lobenzarit Sodium (1 in 50) responds to Qualitative Tests <1.09> (1) for chloride. Determine the absorption spectrum of a solution of Lobenzarit Sodium (1 in 100,000) as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum: both spectra exhibit simi-lar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Lobenzarit Sodium, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Lobenzarit Sodium (1 in 50) responds to Qualitative Tests <1.09> (2) for sodium salt.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Lobenzarit Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Lobenzarit Sodium according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 50 mg of Lobenzarit Sodium in 2.5 mL of water, and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, and add water to make exactly 100 mL. Pipet 4 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of tetra-hydrofuran, water and triethylamine (50:15:8) to a distance of about 10 cm, and air-dry the plate. Examine under ultra-violet light (main wavelength: 254 nm): the spot other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 2 hours).
Weigh accurately about 0.1 g of Lobenzarit Sodium, previously dried, dissolve in exactly 40 mL of water, add ex-actly 60 mL of a mixture of diethyl ether and tetrahydrofu-ran (1:1), and titrate <2.50> with 0.1 mol/L hydrochloric acid VS while well shaking (indicator: 10 drops of bromophenol blue TS) until the blue color of the water layer changes to a persistent light blue-green. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L hydrochloric acid VS ＝ 16.78 mg of C14H8ClNNa2O4
Losartan Potassium contains not less than 98.
Losartan Potassium occurs as a white crystal-line powder. It is very soluble in water, and freely soluble in methanol and in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Losartan Potassium in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Losartan Potassium RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Losartan Potassium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Losartan Potassium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Losartan Potassium responds to Qualitative Tests <1.09> (1) for potassium salt. Perform the test with Losartan Potassium as directed under Flame Coloration Test <1.04> (2): a green color ap-pears.
(1) Chloride <1.03>—To 1.0 g of Lorazepam add 50 mL of water, allow to stand for 1 hour with occasional shaking, and filter. To 25 mL of the filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.20 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 1.0 g of Lora- zepam according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Lorazepam according to Method 3, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Lorazepam in 20 mL of ethanol (95), and use this solution as the sample so-lution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chroma-tography. Develop the plate with a mixture of chloroform, 1,4-dioxane and acetic acid (100) (91:5:4) to a distance of about 15 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, 1059C, 3 hours). Residue on ignition <2.44> Not more than 0.3z (1 g).
Weigh accurately about 0.4 g of Lorazepam, previ-ously dried, dissolve in 50 mL of acetone, and titrate <2.50> with 0.1 mol/L tetrabutylammonium hydroxide VS (poten-tiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L tetrabutylammonium hydroxide VS C22H22ClKN6O: 461.00 Monopotassium 5-{[4?-(2-butyl-4-chloro-5-hydroxymethyl-1H-imidazol-1-yl)methyl]biphenyl-2-yl}-1H-tetrazol-1-ide [124750-99-8] Losartan Potassium contains not less than 98.5z and not more than 101.0z of losartan potassium (C22H22ClKN6O), calculated on the anhydrous basis. Description Losartan Potassium occurs as a white crystal-line powder. It is very soluble in water, and freely soluble in methanol and in ethanol (99.5). Identification (1) Determine the absorption spectrum of a solution of Losartan Potassium in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Losartan Potassium RS pre-pared in the same manner as the sample solution: both spec-tra exhibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of Losartan Potassium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Losartan Potassium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Losartan Potassium responds to Qualitative Tests <1.09> (1) for potassium salt. Perform the test with Losartan Potassium as directed under Flame Coloration Test <1.04> (2): a green color ap-pears. Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Losartan Potassium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard ＝ 32.12 mg of C15H10 Cl2N2O2 Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 30 mg of Losartan
Loxoprofen Sodium Hydrate contains not less than 98.
Loxoprofen Sodium Hydrate occurs as white to yellowish white, crystals or crystalline powder. It is very soluble in water and in methanol, freely soluble in ethanol (95), and practically insoluble in diethyl ether. A solution of Loxoprofen Sodium Hydrate (1 in 20) does not show optical rotation. The pH of a solution of 1.0 g of Loxoprofen Sodium Hy-drate in 20 mL of freshly boiled and cooled water is between 6.5 and 8.5.
(1) Determine the absorption spectrum of a solution of Loxoprofen Sodium Hydrate (1 in 55,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Lox-oprofen Sodium Hydrate as directed in the potassium bro-mide disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Loxoprofen Sodium Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Loxoprofen Sodium Hydrate in 10 mL of water: the solu-tion is clear and colorless or pale yellow. The color is not darker than that of diluted Matching Fluid A (1 in 2). Heavy metals <1.07>—Proceed with 2.0 g of Lox- oprofen Sodium Hydrate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 1.0 g of Loxoprofen Sodium Hydrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add methanol to make exactly 200 mL, and use this solution ‌1280 as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1,2-dichloroethane and acetic acid (100) (9:1) to a distance of about 15 cm, and air-dry the plate. Examine under ultra- Loxoprofen Sodium Tablets
メタケイ酸アルミン酸マグネシウム
Magnesium Aluminometasilicate contains not less than 29.
Magnesium Aluminometasilicate occurs as a white, powder or grain. It is practically insoluble in water and in ethanol (99.5). When heat 1 g of Magnesium Aluminometasilicate with 10 mL of dilute hydrochloric acid, most of it dissolves.
(1) To 0.5 g of Magnesium Alumino-metasilicate add 5 mL of diluted sulfuric acid (1 in 3), heat until white fumes are evolved, cool, add 20 mL of water, and filtrate. Neutralize the filtrate with ammonia TS, and filter the precipitate produced. Dissolve the residue in dilute hy-drochloric acid: the solution responds to Qualitative Tests thiocyanate (3 in 10), and shake: the solution is not more colored than the following control solution (not more than 0.03z). Control solution: Pipet 1 mL of Standard Iron Solution, add water to make 45 mL, add 2 mL of hydrochloric acid, shake, and proceed in the same manner. Arsenic <1.11>—To 1.0 g of Magnesium Alumino- metasilicate add 10 mL of water and 1 mL of sulfuric acid, and shake thoroughly. After cooling, perform the test using this solution as the test solution (not more than 2 ppm). Loss on drying <2.41> Not more than 20.0z (1 g, 1109C, 7 hours). Acid-consuming capacity <6.04> Weigh accurately about 0.2 g of Magnesium Aluminometasilicate, transfer to a glass- ‌stoppered flask, add exactly 100 mL of 0.1 mol/L hydro-chloric acid VS, stopper the flask tightly, shake at 37 ± 29C for 1 hour, and filter. Pipet 50 mL of the filtrate, and titrate <2.50> the excess hydrochloric acid, while stirring thor-oughly, with 0.1 mol/L sodium hydroxide VS until the pH of the solution becomes 3.5. Perform a blank determination in the same manner. The consumed volume of 0.1 mol/L hy-drochloric acid VS is not less than 210 mL per g of Magnesi-um Aluminometasilicate calculated on the dried basis.
(1) Soluble salts—To 10.0 g of Magnesium Alumi-nometasilicate add 150 mL of water, boil gently for 15 minutes while shaking thoroughly. After cooling, add water to make 150 mL, and centrifuge. To 75 mL of the superna-tant liquid add water to make 100 mL, and use this solution as the sample solution. Evaporate 25 mL of the sample solu-tion on a water bath to dryness, then ignite the residue at 7009C for 2 hours: the mass of the residue is not more than 20 mg. Alkalinity—To 20 mL of the sample solution ob-tained in (1), add 2 drops of phenolphthalein TS, and add 0.1 mol/L hydrochloric acid VS until the solution becomes colorless: the consumed volume is not more than 0.50 mL. Chloride <1.03>—To 10 mL of the sample solution obtained in (1), add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.75 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.053z). Sulfate <1.14>—To 2 mL of the sample solution ob- tained in (1), add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). Heavy metals <1.07>—To 2.67 g of Magnesium Alu-minometasilicate add 20 mL of water and 8 mL of hydro-chloric acid, evaporate to dryness on a water bath. To the residue add 5 mL of dilute acetic acid and 20 mL of water, boil for 2 minutes, add 0.4 g of hydroxylammonium chlo-ride, and heat to boiling. After cooling, add water to make exactly 100 mL, and filter. Pipet 25 mL of the filtrate, adjust to pH 3.0 with dilute acetic acid or ammonia TS, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 2 mL of hydrochloric acid to dryness on a water bath, add 2.0 mL of Standard Lead Solution, 0.1 g of hy-droxylammonium chloride and water to make 25 mL, adjust to pH 3.0 with dilute acetic acid or ammonia TS, and add water to make 50 mL (not more than 30 ppm). Iron—To 0.11 g of Magnesium Aluminometasilicate add 8 mL of 2 mol/L nitric acid TS, boil for 1 minute, cool, add water to make exactly 100 mL, and centrifuge. Pipet 30 mL of the supernatant liquid, add water to make 45 mL, add 2 mL of hydrochloric acid, and shake. Add 50 mg of ammo- 2 3 nium peroxodisulfate and 3 mL of a solution of ammonium more than 14.0z of magnesium oxide (MgO: 40.30), and not less than 29.2z and not more than 35.6z of silicon dioxide (SiO2: 60.08), calculated on the dried basis. Description Magnesium Aluminometasilicate occurs as a white, powder or grain. It is practically insoluble in water and in ethanol (99.5). When heat 1 g of Magnesium Aluminometasilicate with 10 mL of dilute hydrochloric acid, most of it dissolves. Identification (1) To 0.5 g of Magnesium Alumino-metasilicate add 5 mL of diluted sulfuric acid (1 in 3), heat until white fumes are evolved, cool, add 20 mL of water, and filtrate. Neutralize the filtrate with ammonia TS, and filter the precipitate produced. Dissolve the residue in dilute hy-drochloric acid: the solution responds to Qualitative Tests thiocyanate (3 in 10), and shake: the solution is not more colored than the following control solution (not more than 0.03z). Control solution: Pipet 1 mL of Standard Iron Solution, add water to make 45 mL, add 2 mL of hydrochloric acid, shake, and proceed in the same manner. Arsenic <1.11>—To 1.0 g of Magnesium Alumino- metasilicate add 10 mL of water and 1 mL of sulfuric acid, and shake thoroughly. After cooling, perform the test using this solution as the test solution (not more than 2 ppm). Loss on drying <2.41> Not more than 20.0z (1 g, 1109C, 7 hours). Acid-consuming capacity <6.04> Weigh accurately about 0.2 g of Magnesium Aluminometasilicate, transfer to a glass- ‌stoppered flask, add exactly 100 mL of 0.1 mol/L hydro-chloric acid VS, stopper the flask tightly, shake at 37 ± 29C for 1 hour, and filter. Pipet 50 mL of the filtrate, and titrate <2.50> the excess hydrochloric acid, while stirring thor-oughly, with 0.1 mol/L sodium hydroxide VS until the pH of the solution becomes 3.5. Perform a blank determination in the same manner. The consumed volume of 0.1 mol/L hy-drochloric acid VS is not less than 210 mL per g of Magnesi-um Aluminometasilicate calculated on the dried basis.
(1) Aluminum oxide—Weigh accurately about 1.25 g of Magnesium Aluminometasilicate, transfer to a con-ical flask, add 10 mL of 3 mol/L hydrochloric acid TS and 50 mL of water, and heat on a water bath for 15 minutes. To the solution add 8 mL of hydrochloric acid, heat on a water bath for 10 minutes. After cooling, transfer to a 250-mL volumetric flask, rinse the flask with water, and add water to make 250 mL. Centrifuge the solution, and use the superna-tant liquid as the sample solution. Pipet 20 mL of the sample solution, add exactly 20 mL of 0.05 mol/L disodium dihy-drogen ethylenediamine tetraacetate VS. To this solution add 15 mL of acetic acid–ammonium acetate buffer solution (pH 4.8) and 20 mL of water, and boil for 5 minutes. After cooling, add 50 mL of ethanol (95), and titrate <2.50> with 0.05 mol/L zinc sulfate VS until the color of the solution changes from light dark green to light red (indicator: 2 mL of dithizone TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.549 mg of Al O
Magnesium Aluminosilicate contains not less than 27.
Magnesium Aluminosilicate occurs as a white, powder or grain. It is practically insoluble in water and in ethanol (99.5). When heat 1 g of Magnesium Aluminosilicate with 10 mL of dilute hydrochloric acid, most of it dissolves.
(1) To 0.5 g of Magnesium Aluminosilicate add 5 mL of diluted sulfuric acid (1 in 3), heat until white fumes are evolved, cool, add 20 mL of water, and filtrate. Neutralize the filtrate with ammonia TS, and filter the pre-cipitate produced. Dissolve the residue in dilute hydrochloric acid: the solution responds to Qualitative Tests <1.09> for aluminum salt. The filtrate obtained in (1) responds to Qualitative Tests <1.09> (2) for magnesium salt. Wash the residue obtained in (1) with 30 mL of water, add 2 mL of a solution of methylene blue trihydrate (1 in 10,000), and wash with 30 mL of water: the precipitate has a blue color.
(1) Soluble salts—To 10.0 g of Magnesium Aluminosilicate add 150 mL of water, boil gently for 15 minutes while shaking thoroughly. After cooling, add water to make 150 mL, and centrifuge. To 75 mL of the superna-tant liquid add water to make 100 mL, and use this solution as the sample solution. Evaporate 25 mL of the sample solu-tion on a water bath to dryness, then ignite the residue at 7009C for 2 hours: the mass of the residue is not more than 20 mg. Alkalinity—To 20 mL of the sample solution ob-tained in (1), add 2 drops of phenolphthalein TS, and add 0.1 mol/L hydrochloric acid VS until the solution becomes colorless: the consumed volume is not more than 0.50 mL. Chloride <1.03>—To 10 mL of the sample solution obtained in (1), add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.75 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.053z). Sulfate <1.14>—To 2 mL of the sample solution ob- tained in (1), add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). Heavy metals <1.07>—To 2.67 g of Magnesium Aluminosilicate add 20 mL of water and 8 mL of hydrochlo-ric acid, evaporate to dryness on a water bath. To the residue add 5 mL of dilute acetic acid and 20 mL of water, boil for 2 minutes, add 0.4 g of hydroxylammonium chloride, and heat to boiling. After cooling, add water to make exactly 100 mL, and filter. Pipet 25 mL of the filtrate, adjust to pH 3.0 with dilute acetic acid or ammonia TS, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 2 mL of hydrochloric acid to dryness on a water bath, add 2.0 mL of Standard Lead Solution, 0.1 g of hydroxylammonium chlo-ride and water to make 25 mL, adjust to pH 3.0 with dilute acetic acid or ammonia TS, and add water to make 50 mL (not more than 30 ppm). Iron—To 0.11 g of Magnesium Aluminosilicate add 8 mL of 2 mol/L nitric acid TS, boil for 1 minute, cool, add water to make exactly 100 mL, and centrifuge. Pipet 30 mL of the supernatant liquid, add water to make 45 mL, add 2 mL of hydrochloric acid, and shake. Add 50 mg of ammo-nium peroxodisulfate and 3 mL of a solution of ammonium thiocyanate (3 in 10), and shake: the solution is not more colored than the following control solution (not more than 0.03z). Control solution: Pipet 1 mL of Standard Iron Solution, add water to make 45 mL, add 2 mL of hydrochloric acid, shake, and proceed in the same manner. Arsenic <1.11>—To 1.0 g of Magnesium Aluminosili- cate add 10 mL of water and 1 mL of sulfuric acid, and shake thoroughly. After cooling, perform the test using this solution as the test solution (not more than 2 ppm). Loss on drying <2.41> Not more than 20.0z (1 g, 1109C, 7 hours). Acid-consuming capacity <6.04> Weigh accurately about 0.2 g of Magnesium Aluminosilicate, transfer to a glass-stoppered flask, add exactly 100 mL of 0.1 mol/L hydro-chloric acid VS, stopper the flask tightly, shake at 37 ± 29C for 1 hour, and filter. Pipet 50 mL of the filtrate, and titrate <2.50> the excess hydrochloric acid, while stirring thor- oughly, with 0.1 mol/L sodium hydroxide VS until the pH of the solution becomes 3.5. Perform a blank determination in the same manner. The consumed volume of 0.1 mol/L hy-drochloric acid VS is not less than 250 mL per g of Magnesi-um Aluminosilicate calculated on the dried basis.
(1) Aluminum oxide—Weigh accurately about 1.25 g of Magnesium Aluminosilicate, transfer to a conical flask, add 10 mL of 3 mol/L hydrochloric acid TS and 50 mL of water, and heat on a water bath for 15 minutes. To the solu-tion add 8 mL of hydrochloric acid, heat on a water bath for 10 minutes. After cooling, transfer to a 250-mL volumetric flask, rinse the flask with water, and add water to make 250 mL. Centrifuge the solution, and use the supernatant liquid as the sample solution. Pipet 20 mL of the sample solution, add exactly 20 mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS. To this solution add 15 mL of acetic acid–ammonium acetate buffer solution (pH 4.8) and 20 mL of water, and boil for 5 minutes. After cooling, add 50 mL of ethanol (95), and titrate <2.50> with 0.05 mol/L zinc sulfate VS until the color of the solution changes from light dark green to light red (indicator: 2 mL of dithi-zone TS). Perform a blank determination in the same man-ner. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.549 mg of Al2O3 ‌Magnesium oxide—Pipet 50 mL of the sample solu-tion obtained in (1), add 50 mL of water and 25 mL of a so-lution of 2,2?,2!-nitrilotriethanol (1 in 2), shake thoroughly, then add 25 mL of ammonia-ammonium chloride buffer so-lution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodi-um dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue lasting for 30 seconds (indicator: 40 mg of eriochrome black T-sodi-um chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.015 mg of MgO Silicon dioxide—Weigh accurately about 1 g of Mag-nesium Aluminosilicate, add 30 mL of dilute hydrochloric acid, and evaporate to dryness on a water bath. Moisten the residue with hydrochloric acid, evaporate to dryness on a water bath. To the residue add 8 mL of hydrochloric acid, stir, then add 25 mL of hot water, and stir again. After allowing to stand, filter the supernatant liquid through a filter paper for quantitative analysis, add 10 mL of hot water to the residue, stir, and decant the supernatant liquid on a filter paper to filter. Then wash the residue with three 10-mL portions of hot water, add 50 mL of water to the residue, and heat on a water bath for 15 minutes. Transfer the residue onto the filter paper, wash the residue with hot water until the last 5 mL of washing yields no precipitate on addi-tion of 1 mL of silver nitrate TS, place the residue and the filter paper in a platinum crucible, ignite to ash, and then ignite at 800 ± 259C for 1 hour. After cooling, weigh the crucible, and designate the mass as a (g). Then add 6 mL of hydrofluoric acid, evaporate to dryness, ignite for 5 minutes, weigh the crucible after cooling, and designate the mass as b (g). Amount (g) of silicon dioxide (SiO2) ＝ a – b
Magnesium Carbonate is a basic hydrated magne-sium carbonate or a normal hydrated magnesium car-bonate. Magnesium Carbonate contains not less than 40.
Magnesium Carbonate occurs as white, friable masses or powder. It is odorless. It is practically insoluble in water, in ethanol (95), in 1-propanol and in diethyl ether. It dissolves in dilute hydrochloric acid with effervescence. Its saturated solution is alkaline.
(1) Dissolve 1 g of Magnesium Carbonate in 10 mL of dilute hydrochloric acid, boil, then cool, neu-tralize with sodium hydroxide TS, and filter, if necessary: the solution responds to Qualitative Tests <1.09> for magne-sium salt. 2 3 (2) Magnesium Carbonate responds to Qualitative Tests Magnesium oxide—Pipet 50 mL of the sample solu-tion obtained in (1), add 50 mL of water and 25 mL of a so-lution of 2,2?,2!-nitrilotriethanol (1 in 2), shake thoroughly, then add 25 mL of ammonia-ammonium chloride buffer so-lution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodi-um dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue lasting for 30 seconds (indicator: 40 mg of eriochrome black T-sodi-um chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.015 mg of MgO Silicon dioxide—Weigh accurately about 1 g of Mag-nesium Aluminometasilicate, add 30 mL of dilute hydro-chloric acid, and evaporate to dryness on a water bath. Moisten the residue with hydrochloric acid, evaporate to dryness on a water bath. To the residue add 8 mL of hydro-chloric acid, stir, then add 25 mL of hot water, and stir again. After allowing to stand, filter the supernatant liquid through a filter paper for quantitative analysis, add 10 mL of hot water to the residue, stir, and decant the supernatant liquid on a filter paper to filter. Then wash the residue with three 10-mL portions of hot water, add 50 mL of water to the residue, and heat on a water bath for 15 minutes. Trans-fer the residue onto the filter paper, wash the residue with hot water until the last 5 mL of washing yields no precipitate on addition of 1 mL of silver nitrate TS, place the residue and the filter paper in a platinum crucible, ignite to ash, and then ignite at 800 ± 259C for 1 hour. After cooling, weigh the crucible, and designate the mass as a (g). Then add 6 mL of hydrofluoric acid, evaporate to dryness, ignite for 5 minutes, weigh the crucible after cooling, and designate the mass as b (g). Amount (g) of silicon dioxide (SiO2) ＝ a – b <1.09> (1) for carbonate.
(1) Soluble salts—To 2.0 g of Magnesium Car-bonate add 40 mL of 1-propanol and 40 mL of water, heat to boil with constant stirring, cool, and filter. Wash the residue with water, combine the washings with the filtrate, and add water to make exactly 100 mL. Evaporate 50 mL of the solution on a water bath to dryness, and dry at 1059C for 1 hour: the mass of the residue does not exceed 10.0 mg. Heavy metals <1.07>—Moisten 1.0 g of Magnesium Carbonate with 4 mL of water, dissolve by addition of 10 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 35 mL of water, 2 mL of dilute acetic acid, 1 drop of ammonia TS, filter, if necessary, wash the filter paper with water, combine the washings with the filtrate, and add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 10 mL of dilute hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solu-tion, and dilute with water to make 50 mL (not more than 30 ppm). Iron <1.10>—Prepare the test solution with 0.10 g of Magnesium Carbonate according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 200 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Magnesium Carbonate, previously moistened with 1.5 mL of water, add 3.5 mL of dilute hydrochloric acid, and per-form the test (not more than 5 ppm). Calcium oxide—Weigh accurately about 0.6 g of Magnesium Carbonate, and dissolve in 35 mL of water and 6 mL of dilute hydrochloric acid. Add 250 mL of water and 5 mL of a solution of L-tartaric acid (1 in 5), then add 10 mL of a solution of 2,2?,2!-nitrilotriethanol (3 in 10) and 10 mL of 8 mol/L potassium hydroxide TS, allow to stand for 5 minutes, and titrate <2.50> with 0.01 mol/L disodium dihy-drogen ethylenediamine tetraacetate VS until the color of the solution changes form red-purple to blue (indicator: 0.1 g of NN indicator). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 0.5608 mg of CaO The content of calcium oxide (CaO: 56.08) is not more than 0.6z. ‌Acid-insoluble substances—Mix 5.0 g of Magnesium Carbonate and 75 mL of water, add 10 mL of hydrochloric acid dropwise while stirring, boil for 5 minutes, and cool. Collect the insoluble residue using filter paper for quantita-tive analysis, wash well with water until the last washing shows no turbidity with silver nitrate TS, and ignite the residue together with the filter paper: the mass of the residue is not more than 2.5 mg. Precipitation test Transfer 1.0 g of Magnesium Carbonate, previously sifted through a No. 100 (150 mm) sieve to a glass-stoppered measuring cylinder with a 50-mL graduation line at 150 mm from the bottom, and add water to make 50 mL. Shake vigorously for exactly 1 minute, allow to stand for 15 minutes, and measure the height of the precipitate (in gradu-ation in ml).
Weigh accurately about 0.4 g of Magnesium Car-bonate, dissolve in 10 mL of water and 3.5 mL of dilute hy-drochloric acid, and add water to make exactly 100 mL. Pipet 25 mL of the solution, add 50 mL of water and 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7) and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS (indicator: 40 mg of erio-chrome black T-sodium chloride indicator). Perform a blank determination in the same manner, and make any necessary correction. From the volume of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS consumed deduct the volume of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS corresponding to the content of calcium oxide (CaO) obtained in the Purity (5). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.015 mg of MgO Each mg of calcium oxide (CaO) ＝ 0.36 mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS
Magnesium Oxide, when ignited, contains not less than 96.
Magnesium Oxide occurs as a white, powder or granules. It is odorless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. It dissolves in dilute hydrochloric acid. It absorbs moisture and carbon dioxide in air.
A solution of Magnesium Oxide in dilute hy-drochloric acid (1 in 50) responds to Qualitative Tests <1.09> for magnesium salt.
(1) Alkali and soluble salts—Transfer 2.0 g of Magnesium Oxide to a beaker, add 100 mL of water, cover the beaker with a watch-glass, heat on a water bath for 5 minutes, and filter immediately. After cooling, to 50 mL of the filtrate add 2 drops of methyl red TS and 2.0 mL of 0.05 mol/L sulfuric acid VS: a red color develops. Evaporate 25 mL of the remaining filtrate to dryness, and dry the residue at 1059C for 1 hour: the mass of the residue is not more than 10 mg. Carbonate—Boil 0.10 g of Magnesium Oxide with 5 mL of water, cool, and add 5 mL of acetic acid (31): almost no effervescence occurs. Heavy metals <1.07>—Dissolve 1.0 g of Magnesium Oxide in 20 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 35 mL of water, add 1 drop of phenolphthalein TS, neutralize with ammonia TS, add 2 mL of dilute acetic acid, and filter, if necessary. Wash the filter paper with water, add water to the combined washing and the filtrate to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: to 20 mL of dilute hydro-chloric acid add 1 drop of phenolphthalein TS, neutralize with ammonia TS, and add 2 mL of dilute acetic acid, 4.0 mL of Standard Lead Solution and water to make 50 mL (not more than 40 ppm). Iron <1.10>—Prepare the test solution with 40 mg of ‌Magnesium Oxide according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 500 ppm). Calcium oxide—Weigh accurately about 0.25 g of Magnesium Oxide, previously ignited, dissolve in 6 mL of dilute hydrochloric acid by heating. Cool, add 300 mL of water and 3 mL of a solution of L-tartaric acid (1 in 5), then add 10 mL of a solution of 2,2?,2!-nitrilotriethanol (3 in 10) and 10 mL of 8 mol/L potassium hydroxide TS, allow to stand for 5 minutes, and titrate <2.50> with 0.01 mol/L diso-dium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue (indi-cator: 0.1 g of NN indicator). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 0.5608 mg of CaO The mass of calcium oxide (CaO: 56.08) is not more than 1.5z. Arsenic <1.11>—Dissolve 0.20 g of Magnesium Oxide in 5 mL of dilute hydrochloric acid, and perform the test with this solution as the test solution (not more than 10 ppm). Acid-insoluble substances—Mix 2.0 g of Magnesium Oxide with 75 mL of water, add 12 mL of hydrochloric acid dropwise, while shaking, and boil for 5 minutes. Collect the insoluble residue using filter paper for quantitative analysis, wash well with water until the last washing shows no tur-bidity with silver nitrate TS, and ignite the residue together with the filter paper: the mass of the ignited residue does not more than 2.0 mg. Fluoride—(i) Apparatus: Use a hard glass appa-ratus as illustrated in the figure. Ground-glass joints may be used. (ii) Procedure: Transfer 5.0 g of Magnesium Oxide to the distilling flask A with the aid of 20 mL of water, add about 1 g of glass wool and 50 mL of diluted purified sulfu-ric acid (1 in 2), and connect A to the distillation apparatus, previously washed with steam streamed through the steam introducing tube E. Connect the condenser C with the receiver D containing 10 mL of 0.01 mol/L sodium hydrox-ide VS and 10 mL of water so that the lower end of C is immersed in the solution. Heat A gradually until the temper-ature of the solution in A reaches 1309C, then open the rub-ber tube F, close the rubber tube G, boil water in the steam generator B vigorously, and introduce the generated steam into F. Simultaneously, heat A, and maintain the tempera-ture of the solution in A between 1359C and 1459C. Adjust the distilling rate to about 10 mL per minute. Collect about 170 mL of the distillate, then stop the distillation, wash C with a small quantity of water, combine the washings with the distillate, add water to make exactly 200 mL, and use this solution as the test solution. Perform the test with the test solution as directed in the procedure of determination for fluoride under Oxygen Flask Combustion Method <1.06>. No corrective solution is used in this procedure. The content of fluoride (F) is not more than 0.08z. Amount (mg) of fluoride (F: 19.00) in the test solution ＝ amount (mg) of fluoride in 5 mL of the standard solution × AT/AS × 200/V Loss on ignition <2.43> Not more than 10z (0.25 g, 9009C, constant mass).
Ignite Magnesium Oxide to constant mass at 9009C, weigh accurately about 0.2 g of the residue, dissolve in 10 mL of water and 4.0 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Pipet 25 mL of this solu-tion, add 50 mL of water and 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7) and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetra-acetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Perform a blank determination in the same manner, and make any necessary correction. From the volume of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS consumed, deduct the volume of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS corresponding to the content of calcium oxide (CaO) obtained in the Purity (5). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.015 mg of MgO Each mg of calcium oxide (CaO) ＝ 0.36 mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS
Magnesium Silicate contains not less than 45.
Magnesium Silicate occurs as a white, fine powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether.
(1) Mix 0.5 g of Magnesium Silicate with 10 mL of dilute hydrochloric acid, filter, and neutralize the filtrate with ammonia TS: the solution responds to Qualita-tive Tests <1.09> for magnesium salt. (2) Prepare a bead by fusing ammonium sodium hydro-genphosphate tetrahydrate on a platinum loop. Place the bead in contact with Magnesium Silicate, and fuse again: an infusible matter appears in the bead, which changes to an opaque bead with a web-like structure upon cooling.
(1) Soluble salts—Add 150 mL of water to 10.0 g of Magnesium Silicate, heat on a water bath for 60 minutes with occasional shaking, then cool, dilute with water to 150 mL, and centrifuge. Dilute 75 mL of the resultant transpar-ent liquid with water to 100 mL, and use this solution as the sample solution. Evaporate 25 mL of the sample solution on a water bath to dryness, and ignite the residue at 7009C for 2 hours: the mass of the ignited residue is not more than 0.02 g. Alkalinity—To 20 mL of the sample solution ob-tained in (1) add 2 drops of phenolphthalein TS and 1.0 mL of 0.1 mol/L hydrochloric acid VS: no color develops. Chloride <1.03>—Take 10 mL of the sample solution obtained in (1), add 6 mL of dilute nitric acid, dilute with water to 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.75 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.053z). Sulfate <1.14>—To the residue obtained in (1) add about 3 mL of dilute hydrochloric acid, and heat on a water bath for 10 minutes. Add 30 mL of water, filter, wash the residue on the filter with water, combine the washings with the filtrate, and dilute to 50 mL with water. To 4 mL of the solution add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). ‌Heavy metals <1.07>—To 1.0 g of Magnesium Silicate add 20 mL of water and 3 mL of hydrochloric acid, and boil for 2 minutes. Filter, and wash the residue on the filter with two 5-mL portions of water. Evaporate the combined filtrate and washings on a water bath to dryness, add 2 mL of dilute acetic acid to the residue, warm until solution is complete, filter, if necessary, add water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 3.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 30 ppm). Arsenic <1.11>—To 0.40 g of Magnesium Silicate add 5 mL of dilute hydrochloric acid, heat gently to boiling while shaking well, cool rapidly, and centrifuge. Mix the residue with 5 mL of dilute hydrochloric acid with shaking, centri-fuge, then add 10 mL of water to the residue, and repeat the extraction in the same manner. Concentrate the combined extracts on a water bath to 5 mL. Use this solution as the test solution, and perform the test (not more than 5 ppm). Loss on ignition <2.43> Not more than 34z (0.5 g, 8509C, 3 hours). Acid-consuming capacity <6.04> Place about 0.2 g of Mag-nesium Silicate, accurately weighed, in a glass-stoppered flask, add exactly 30 mL of 0.1 mol/L hydrochloric acid VS and 20 mL of water, shake at 37 ± 29C for 1 hour, and cool. Pipet 25 mL of the supernatant liquid, and titrate <2.50> the excess hydrochloric acid, while stirring well, with 0.1 mol/L sodium hydroxide VS until the pH becomes 3.5. 1 g of Magnesium Silicate, calculated on the anhydrous basis by making allowance for the observed loss on ignition determined as directed in the preceding Loss on ignition, consumes not less than 140 mL and not more than 160 mL of 0.1 mol/L hydrochloric acid VS.
(1) Silicon dioxide—Weigh accurately about 0.7 g of Magnesium Silicate, add 10 mL of 0.5 mol/L sulfuric acid TS, evaporate on a water bath to dryness, add 25 mL of water to the residue, and heat on a water bath for 15 minutes with occasional stirring. Filter the supernatant liquid through filter paper for quantitative analysis, add 25 mL of hot water to the residue, stir, and decant the supernatant liq-uid on the filter paper to filter. Wash the residue in the same manner with two 25-mL portions of hot water, transfer the residue onto the filter paper, and wash with hot water until the last washing does not respond to Qualitative Tests <1.09> (1) for sulfate. Place the residue and the filter paper in a platinum crucible, incinerate with strong heating, and ignite between 7759C and 8259C for 30 minutes, then cool, and weigh the residue as a (g). Moisten the residue with water, and add 6 mL of hydrofluoric acid and 3 drops of sulfuric acid. Evaporate to dryness, ignite for 5 minutes, cool, and weigh the residue as b (g). Content (z) of silicon dioxide (SiO2) ＝ (a － b)/M × 100 M: Mass (g) of the Magnesium Silicate taken Magnesium oxide—Weigh accurately about 0.3 g of Magnesium Silicate, transfer to a 50-mL conical flask, add 10 mL of 0.5 mol/L sulfuric acid VS, and heat on a water bath for 15 minutes. Cool, transfer to a 100-mL volumetric flask, wash the conical flask with water, add the washings to the volumetric flask, dilute with water to 100 mL, and filter. Pipet 50 mL of the filtrate, shake with 50 mL of water and 5 mL of diluted 2,2?,2!-nitrilotriethanol (1 in 2), add 2.0 mL of ammonia TS and 10 mL of ammonia-ammonium chloride buffer solution (pH 10.7) and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS (indi-cator: 40 mg of eriochrome black T-sodium chloride indi-cator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.015 mg of MgO Ratio of percentage (z) of magnesium oxide (MgO) to silicon dioxide (SiO2)—Calculate the quotient from the percentages obtained in (1) and (2).
Magnesium Stearate is a compound of magnesium with a mixture of solid fatty acids, and consists chiefly of variable proportions of magnesium stearate and magnesium palmitate obtained from sources of vegeta-ble or animal origin. It contains not less than 4.
Magnesium Stearate occurs as a white, light, bulky powder. It is smooth to the touch and sticky to the skin. It has no odor or a faint, characteristic odor. It is practically insoluble in water and in ethanol (99.5).◆
Mix 5.0 g of Magnesium Stearate with 50 mL of peroxide-free diethyl ether, 20 mL of dilute nitric acid, and 20 mL of water in a round-bottom flask, and heat to dissolve completely under a reflux condenser. After cool-ing, transfer the contents of the flask to a separator, shake, allow the layers to separate, and transfer the aqueous layer to a flask. Extract the diethyl ether layer with two 4-mL por-tions of water, and combine these extracts to the main aque-ous extract. After washing the combined aqueous extract with 15 mL of peroxide-free diethyl ether, transfer to a 50-mL volumetric flask, add water to make 50 mL, and use this solution as the sample solution. To 1 mL of the sample solution add 1 mL of ammonia TS: A white precipitate is formed that dissolves on addition of 1 mL of ammonium chloride TS. By further addition of 1 mL of a solution of disodium hydrogen phosphate dodecahydrate (4 in 25) a white crystalline precipitate is formed.
(1) Acidity or alkalinity—Heat 1.0 g of Magnesi-um Stearate in 20 mL of freshly boiled and cooled water on a water bath for 1 minute while shaking, cool, and filter. Add 0.05 mL of bromothymol blue TS to 10 mL of the filtrate: not more than 0.05 mL of 0.1 mol/L hydrochloric acid VS or 0.1 mol/L sodium hydroxide VS is required to change the color of the solution. Chloride <1.03>—Perform the test with 10.0 mL of the sample solution obtained in Identification. Prepare the control solution with 1.4 mL of 0.02 mol/L hydrochloric acid VS (not more than 0.1z). Sulfate <1.14>—Perform the test with 6.0 mL of the sample solution obtained in Identification. Prepare the con-trol solution with 3.0 mL of 0.02 mol/L sulfuric acid VS (not more than 1.0z). (4) Heavy metals <1.07>—Heat 1.0 g of Magnesium Stearate weakly first, then incinerate at about 500 ± 259C. After cooling, add 2 mL of hydrochloric acid, evaporate on a water bath to dryness, add 20 mL of water and 2 mL of dilute acetic acid to the residue, and heat for 2 minutes. After cooling, filter this solution through a filter paper, wash the filter paper with 15 mL of water, and combine the washing with the filtrate. To the filtrate add water to make 50 mL, and perform the test with this solution as the test so-lution. Prepare the control solution as follows: evaporate 2 mL of hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm).◆ Loss on drying <2.41> Not more than 6.0z (2 g, 1059C, constant mass). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 103 CFU/g and 5 × 102 CFU/g, respectively. Salmonella and Escherichia coli are not observed.◆ Relative content of stearic acid and palmitic acid Transfer 0.10 g of Magnesium Stearate to a small conical flask fitted with a reflux condenser. Add 5.0 mL of boron trifluoride-methanol TS, mix, and reflux for 10 minutes to dissolve the solids. Add 4 mL of heptane through the condenser, and reflux for 10 minutes. After cooling, add 20 mL of saturated sodium chloride solution, shake, and allow the layers to separate. Pass the heptane layer through about 0.1 g of an-hydrous sodium sulfate, previously washed with heptane, into another flask. Transfer 1.0 mL of this solution to a 10-mL volumetric flask, dilute with heptane to volume, and use this solution as the sample solution. Perform the test with 1 mL of the sample solution as directed under Gas chro-matography <2.02> according to the following conditions, and determine the area, A, of the methyl stearate peak and the sum of the areas, B, of all of the fatty acid ester peaks. Calculate the percentage of stearic acid in the fatty acid frac-tion of Magnesium Stearate by the following formula. Content (z) of stearic acid ＝ A/B × 100 Similarly, calculate the percentage of palmitic acid in the portion of Magnesium Stearate taken. The methyl stearate peak, and the sum of the stearate and palmitate peaks are not less than 40z and not less than 90z of the total area of all fatty acid ester peaks, respectively. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A fused silica capillary column 0.32 mm in inside diameter and 30 m in length, the inside coated with a 0.5-mm layer of polyethylene glycol 15000-diepoxide for gas chroma-tography. Column temperature: Maintain at 709C for 2 minutes after injection, then program to increase the temperature at the rate of 59C per minute to 2409C and to maintain 2409C for 5 minutes. Injection port temperature: A constant temperature of about 2209C. Detector temperature: A constant temperature of about 2609C. Carrier gas: Helium. Flow rate: 2.4 mL per minute. Splitless. Time span of measurement: For 41 minutes after the sol-vent peak.◆ System suitability— Test for required detectability:◆ Place about 50 mg each of stearic acid for gas chromatography and palmitic acid for gas chromatography in a small conical flask fitted with a reflux condenser. Add 5.0 mL of boron trifluoride-methanol TS, mix, and proceed in the same manner as directed for the preparation of the sample solution, and use the solution so obtained as the solution for system suitability test. ◆To exactly 1 mL of the solution for system suitability test add heptane to make exactly 10 mL. To exactly 1 mL of this so-lution add heptane to make exactly 10 mL. Further, to ex-actly 1 mL of this solution add heptane to make exactly 10 mL. Confirm that the peak area of methyl stearate obtained with 1 mL of this solution is equivalent to 0.05 to 0.15z of that with 1 mL of the solution for system suitability test.◆ System performance: When the procedure is run with 1 mL of the solution for system suitability test under the above op-erating conditions, the relative retention time of methyl palmitate to methyl stearate is about 0.9, and the resolution between these peaks is not less than 5.0. System repeatability: When the test is repeated 6 times with the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak areas of methyl palmitate and methyl stearate are not more than 3.0z, respectively, and the relative standard de-viation of the ratios of the peak area of methyl palmitate to methyl stearate is not more than 1.0z.
Transfer about 0.5 g of Magnesium Stearate, accu-rately weighed, to a 250-mL flask, add 50 mL of a mixture of ethanol (99.5) and 1-butanol (1:1), 5 mL of ammonia so-lution (28), 3 mL of ammonium chloride buffer solution (pH 10), 30.0 mL of 0.1 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS and 1 to 2 drops of eriochrome black T TS, and mix. Heat at 45 – 509C to make the solution clear, and after cooling, titrate <2.50> the excess disodium di-hydrogen ethylenediamine tetraacetate with 0.1 mol/L zinc sulfate VS until the solution changes from blue to violet in color. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 2.431 mg of Mg ◆Containers and storage Containers—Tight containers.◆ ‌Magnesium Sulfate Hydrate‌
Magnesium Sulfate Hydrate occurs as color-less or white crystals. It has a cooling, saline, bitter taste. It is very soluble in water, and practically insoluble in ethanol (95). It dissolves in dilute hydrochloric acid.
A solution of Magnesium Sulfate Hydrate (1 in 40) responds to Qualitative Tests <1.09> for magnesium salt and for sulfate. pH <2.54> Dissolve 1.0 g of Magnesium Sulfate Hydrate in 20 mL of water: the pH of this solution is between 5.0 and 8.2.
(1) Clarity and color of solution—Dissolve 1.0 g of Magnesium Sulfate Hydrate in 20 mL of water: the solu-tion is clear and colorless. Chloride <1.03>—Perform the test with 1.0 g of Mag- nesium Sulfate Hydrate. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.014z). Heavy metals <1.07>—Proceed with 2.0 g of Magnesi- um Sulfate Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Zinc—Dissolve 2.0 g of Magnesium Sulfate Hydrate in 20 mL of water, and add 1 mL of acetic acid (31) and 5 drops of potassium hexacyanoferrate (II) TS: no turbidity is produced. Calcium—Dissolve 1.0 g of Magnesium Sulfate Hy-drate in 5.0 mL of dilute hydrochloric acid, add water to make 100 mL, and use this solution as the sample solution. Separately, dissolve 1.0 g of Magnesium Sulfate Hydrate in 2.0 mL of Standard Calcium Solution and 5.0 mL of dilute hydrochloric acid, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions, and determine the absor-bances, AT and AS, of both solutions: AT is smaller than AS － AT (not more than 0.02z). Gas: Combustible gas—Acetylene or hydrogen. Supporting gas—Air. Lamp: Calcium hollow-cathod lamp. Wavelength: 422.7 nm. Arsenic <1.11>—Prepare the test solution with 1.0 g <2.50> with 0.05 mol/L disodium dihydrogen ethylene-diamine tetraacetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Perform a blank deter-mination in the same manner, and make any necessary cor-rection. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 6.018 mg of MgSO4
Magnesium Sulfate Mixture is a light yellow clear liquid. It has a bitter and acid taste.
(1) Magnesium Sulfate Mixture responds to Qualitative Tests <1.09> for magnesium salt. (2) Magnesium Sulfate Mixture responds to Qualitative Tests <1.09> (2) for chloride.
Pipet 10 mL of Magnesium Sulfate Mixture, and add water to make exactly 100 mL. Pipet 10 mL of this solu-tion, add 50 mL of water and 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylenediamine tetraa-cetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 12.32 mg of MgSO4.7H2O
Mitiglinide Calcium Hydrate contains not less than 98.
Mitiglinide Calcium Hydrate occurs as a white powder. It is freely soluble in methanol and in ethanol (99.5), and slightly soluble in water. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Mitiglinide Calcium Hydrate in methanol (1 in 1000) as directed under Ultraviolet-visible Spectrophotome-try <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mitiglinide Cal-cium RS prepared in the same manner as the sample solu-tion: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mitiglinide Calcium Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Mitiglinide Calcium RS: both spectra exhibit similar intensi-ties of absorption at the same wave numbers. To 0.5 g of Mitiglinide Calcium Hydrate add 3 mL of 1 mol/L hydrochloric acid TS and 5 mL of diethyl ether, shake, then separate the aqueous layer, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> (2) for calcium salt. D Optical rotation <2.49> [a]20: ＋ 8.4 – ＋ 9.09(0.38 g calcu-lated on the anhydrous basis, methanol, 20 mL, 100 mm).
(1) Heavy metals <1.07>—Place 1.0 g of Mitiglinide Calcium Hydrate in a crucible, cover the crucible loosely, and ignite at a low temperature until charred. After cooling, add 2 mL of nitric acid and 5 drops of sulfuric acid to the content of the crucible, heat carefully until white fumes are no longer evolved, and ignite between 500 and 6009C. After cooling, moisten the residue with a little amount of sulfuric acid, and incinerate again by ignition. After cooling, add 2 mL of hydrochloric acid, evaporate to peaks other than mitiglinide from sample solution is not larger than 3/10 times the peak area of mitiglinide from the standard solution. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase: Adjust to pH 2.0 of a mixture of water, acetonitrile for liquid chromatography and n-amyl alcohol (66:33:1) with phosphoric acid. Flow rate: Adjust so that the retention time of mitiglinide is about 12 minutes. Time span of measurement: About 2 times as long as the retention time of mitiglinide, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add a mixture of water and acetonitrile (2:1) to make exactly 50 mL. Confirm that the peak area of mitiglinide obtained with 15 mL of this solution is equivalent to 7 to 13 z of that with 15 mL of the standard solution. System performance: When the procedure is run with 15 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of mitiglinide are not less than 4000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 15 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitiglinide is not more than 2.0z. Water <2.48> 4.5 – 6.0z (50 mg, coulometric titration).
Weigh accurately about 50 mg each of Mitiglinide Calcium Hydrate and Mitiglinide Calcium RS (separately de-termine the water <2.48> in the same manner as Mitiglinide Calcium Hydrate), add a mixture of water and acetonitrile (2:1) to them, dissolve by sonicating while occasional shak-ing, and add the mixture of water and acetonitrile (2:1) to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 10 mL of the internal standard solution, add the mixture of water and acetonitrile (2:1) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of mitiglinide to that of the internal standard. Amount (mg) of mitiglinide calcium hydrate (C H CaN O .2H O) ＝ M × Q /Q × 1.054 conditions: the principal peaks in the chromatograms ob-tained from the sample solution and standard solution show the same retention time, and both spectra of these peaks ex-hibit similar intensities of absorption at the same wave-lengths. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Purity. Detector: Photodiode array detector (wavelength: 210 nm, spectrum range of measurement: 200 – 360 nm). 38 48 2 6 2 S T S System suitability— MS: Amount (mg) of Mitiglinide Calcium RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of 2-nitrophenol in acetonitrile (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). ‌Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with palmitamide propyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Adjust to pH 2.0 of a mixture of water, acetonitrile for liquid chromatography and n-amyl alcohol (62:37:1) with phosphoric acid. Flow rate: Adjust so that the retention time of mitiglinide is about 7.5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and mitiglinide are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of mitiglinide is not more than 1.0z.
Weigh accurately about 0.4 g of Monobasic Calcium Phosphate Hydrate, previously dried, dissolve in 3 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add exactly 25 mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS, 50 mL of water and 5 mL of ammonia-ammonium chlo-ride buffer solution (pH 10.7), and titrate <2.50> the excess ‌JP XVIII Official Monographs / Calcium Polystyrene Sulfonate 607 disodium dihydrogen ethylenediamine tetraacetate with 0.02 mol/L zinc acetate VS (indicator: 25 mg of eriochrome black T-sodium chloride indicator). Perform a blank determina-tion in the same manner. Each mL of 0.02 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 5.041 mg of Ca(H2PO4)2.H2O
Montelukast Sodium contains not less than 98.
Montelukast Sodium occurs as a white to pale yellow-white powder. It is very soluble in methanol and in ethanol (99.5), and freely soluble in water. It is hygroscopic. It turns yellow on exposure to light. It shows crystal polymorphism.
(1) Place 0.1 g of Montelukast Sodium in a crucible, and ignite until a white residue is formed. To the residue add 2 mL of water, and then filter. To the filtrate add 2 mL of potassium carbonate solution (3 in 20), and heat to boiling: no precipitate is observed. To this solution add 4 mL of potassium hexahydroxoantimonate (V) TS, heat to boiling, and cool immediately in ice water: a white pre-cipitate is formed. Rub the inside wall of the test tube with a glass rod, if necessary. Determine the absorption spectrum of a solution of Montelukast Sodium in a mixture of methanol and water (3:1) (1 in 100,000) as directed under Ultraviolet-visible Spec-trophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Mon-telukast Sodium for Identification RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Mon-telukast Sodium as directed in the paste method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Montelukast Sodium for Identification RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Or, per-form the test by the potassium bromide disk method or the ATR method, and compare the spectrum with the spectrum of Montelukast Sodium for Identification RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dis-solve Montelukast Sodium and Montelukast Sodium for Identification RS in toluene, add heptane, shake, then allow to stand, and remove the supernatant liquid by decantation. Dry the residue at 759C for 16 hours under reduced pressure, and perform the test by paste method, potassium bromide disk method or the ATR method.
(1) Heavy metals—Dissolve 0.5 g of Montelukast Sodium in 20 mL of a mixture of acetone and water (4:1), and use this solution as the sample solution. Separately, take 0.5 mL of Standard Lead Solution, add 20 mL of the mix-ture of acetone and water (4:1), and use this solution as the standard solution. To the sample solution and the standard solution add 2 mL of acetate buffer solution (pH 3.5), and shake. To these solutions add 1.2 mL of thioacetamide-alka-line glycerin TS, shake immediately, then allow to stand for 2 minutes, and filter through a membrane filter with a pore size 0.45 mm (about 13 mm in diameter). Compare the color on the membrane filters through which each solution is filtered: the color obtained from the sample solution is not darker than that obtained from the standard solution (not more than 10 ppm). Related substances—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Montelukast So-dium in 50 mL of a mixture of methanol and water (9:1), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi- mentioned above is not more than 0.10z. Furthermore, the total amount of the peaks other than montelukast is not more than 0.6z. Operating conditions— Detector, column, column temperature, mobile phases A and B, flowing of mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: For 16 minutes after injec-tion, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 1 mL of the sample solution, add the mixture of methanol and water (9:1) to make exactly 100 mL. Pipet 1 mL of this solution, add the mixture of methanol and water (9:1) to make exactly 20 mL, and use this solution as the solution for system suitability test. When the procedure is run with 10 mL of the solution for system suitability test under the above operating condi-tions, the SN ratio of the peak of montelukast is not less than 10. For the calculations mentioned above, the peak areas smaller than that of montelukast, founded in the chromato-gram obtained with 10 mL of the solution for system suitabil-ity test, are excluded. Enantiomer—Conduct this procedure using light-resistant vessels. Dissolve 50 mg of Montelukast Sodium in 50 mL of a mixture of water and acetonitrile (1:1), and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method, and calculate the amounts of them by the area percentage method: the amount of the peak of the enantiomer having the relative retention time of about 0.7 to montelukast is not more than 0.2z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 280 nm). Column: A stainless steel column 4.0 mm in inside diame-ter and 15 cm in length, packed with a1-acid glycoprotein binding silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: Dissolve 2.3 g of ammonium acetate in 1000 mL of water, and adjust to pH 5.7 with acetic acid (100). Mobile phase B: A mixture of methanol and acetonitrile (3:2). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. tions. Determine each peak area by the automatic integration method, and calculate the amount of them by the area per-centage method: the amount of the peak of related substance A, having the relative retention time of about 0.4 to mon-telukast is not more than 0.2z, the amounts of the peaks, related substance B and related substance E, having respec- Time after injection 0 – 30 70 → 60 30 → 40 30 – 35 of sample (min) Mobile phase A (volz) Mobile phase B (volz) tively the relative retention times of about 0.8 and about 1.2 are not more than 0.15z, the total amount of the two peaks, related substances C and D, both having the relative reten-tion time about 0.9 is not more than 0.15z, the amount of the peak of related substance F having the relative retention time of about 1.9 is not more than 0.3z, and the amount of the peak other than montelukast and other than the peaks Flow rate: 0.9 mL per minute (the retention time of mon- telukast is about 25 minutes). System suitability— Test for required detectability: Pipet 1 mL of the sample solution, add the mixture of water and acetonitrile (1:1) to make exactly 100 mL. Pipet 1 mL of this solution, add the mixture of water and acetonitrile (1:1) to make exactly 10 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, the SN ratio of the peak of montelukast is not less than 10. System performance: When the procedure is run with 10 mL of a solution of Montelukast Racemate for System Suitabillity RS in the mixture of water and acetonitrile (1:1) (1 in 10,000) under the above operating conditions, the reso-lution between the peak of montelukast and the enantiomer is not less than 2.9. Water <2.48> Not more than 4.0z (0.3 g, volumetric titra-tion, direct titration).
Conduct this procedure using light-resistant vessels. Weigh accurately about 50 mg of Montelukast Sodium, and dissolve in a mixture of methanol and water (9:1) to make exactly 50 mL. Pipet 10 mL of this solution, add the mixture of methanol and water (9:1) to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 26 mg of Montelukast Dicyclohexylamine RS, dissolve in the mixture of methanol and water (9:1) to make exactly 50 mL. Pipet 5 mL of this solution, add the mixture of methanol and water (9:1) to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of montelukast in each solution. Amount (mg) of montelukast sodium (C35H35ClNNaO3S) ＝ MS × AT/AS × 5/2 × 0.792 MS: Amount (mg) of Montelukast Dicyclohexylamine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 5 cm in length, packed with phenylsilanized silica gel for liquid chromatography (1.8 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: A mixture of water and trifluoroacetic acid (2000:3). Mobile phase B: A mixture of acetonitrile and trifluoroa-cetic acid (2000:3). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 3 3 – 16 60 → 49 40 → 51 Flow rate: 1.2 mL per minute (the retention time of mon-telukast is about 7 minutes). System suitability— System performance: Use a solution of Montelukast for System Suitability RS in the mixture of methanol and water (9:1) (1 in 1000) as the solution A for peak identification. Perform the test with 10 mL of the solution A for peak iden-tification under the above operating conditions, and identify the peaks having the relative retention times to montelukast of about 0.4 (related substance A), about 0.9 (related sub-stances C and D), about 1.2 (related substance E), and about 1.9 (related substance F). Place 1 mL of the solution A for peak identification in a clear glass container, allow to stand for about 20 minutes, and use this solution as the solution B for peak identification. When the procedure is run with 10 mL of the solution B for peak identification under the above operating conditions, and identify the peak having the rela-tive retention time of about 0.8 to montelukast (related sub-stance B), the resolution between the peaks of related sub-stance B and montelukast is not less than 2.5, and between the peaks of montelukast and related substance E is not less than 1.5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of montelukast is not more than 0.73z.
Containers—Tight containers Storage—Light-resistant.
Mupirocin Calcium Hydrate is the calcium salt of a substance having antibacterial activity produced by the growth of Pseudomonas fluorescens. It contains not less than 895 mg (potency) and not more than 970 mg (potency) per mg, calculated on the anhydrous basis.
Mupirocin Calcium Hydrate occurs as a white powder and has a bitter taste. It is freely soluble in methanol, slightly soluble in water and in ethanol (95), and practically insoluble in diethyl ether.
(1) To 1 mL of a solution of Mupirocin Calcium Hydrate in methanol (1 in 200) add 4 mL of hydroxylamine perchlorate-ethanol TS and 1 mL of N,N?-dicyclohexylcarbodiimide-ethanol TS, shake well, and allow to stand in lukewarm water for 20 minutes. After cooling, add 1 mL of iron (III) perchorate-ethanol TS to the solution, and shake: a dark purple color develops. Determine the absorption spectrum of a solution of Mupirocin Calcium Hydrate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 219 nm and 224 nm. Determine the infrared absorption spectrum of Mupirocin Calcium Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 1708 cm－1, 1648 cm－1, 1558 cm－1, 1231 cm－1, 1151cm－1 and 894 cm－1. A solution of Mupirocin Calcium Hydrate (3 in 1000) responds to Qualitative Tests <1.09> (3) for calcium salt. D Optical rotation <2.49> [a]20: －16 – －209(1 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm).
(1) Related substances—Dissolve 50 mg of Mupirocin Calcium Hydrate in a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make 10 mL, and use this solution as the sample solution (1). Pipet 2 mL of the sample solution (1), add a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetra-hydrofuran (3 in 4) (1:1) to make exactly 100 mL, and use this solution as the sample solution (2). Preserve these sam-ple solutions at a temperature between 49C and 89C. Per-form the test with exactly 20 mL of the sample solution (1) and the sample solution (2) as directed under Liquid Chro- solution (2), and add a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make exactly 20 mL. Con-firm that the peak area of mupirocin obtained with 20 mL of this solution is equivalent to 4 to 6z of that with 20 mL of the sample solution (2). System repeatability: When the test is repeated 6 times with 20 mL of the sample solution (2) under the above oper-ating conditions, the relative standard deviation of the peak areas of mupirocin is not more than 2.0z. (2) Inorganic salt from manufacturing process—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not less than 3.0z and not more than 4.5z (0.5 g, volumetric titration, direct titration).
Weigh accurately the mass of not less than 20 Mosapride Citrate Tablets, and powder. Weigh accurately a portion of the powder, equivalent to about 10 mg of mosapride citrate (C21H25ClFN3O3.C6H8O7), and moisten with 2 mL of water. Add 70 mL of methanol, shake for 20 minutes, add methanol to make exactly 100 mL, and centri-fuge. Pipet 10 mL of the supernatant liquid, add methanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 53 mg of Description Mupirocin Calcium Hydrate occurs as a white powder and has a bitter taste. It is freely soluble in methanol, slightly soluble in water and in ethanol (95), and practically insoluble in diethyl ether. Identification (1) To 1 mL of a solution of Mupirocin Calcium Hydrate in methanol (1 in 200) add 4 mL of hydroxylamine perchlorate-ethanol TS and 1 mL of N,N?-dicyclohexylcarbodiimide-ethanol TS, shake well, and allow to stand in lukewarm water for 20 minutes. After cooling, add 1 mL of iron (III) perchorate-ethanol TS to the solution, and shake: a dark purple color develops. Determine the absorption spectrum of a solution of Mupirocin Calcium Hydrate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 219 nm and 224 nm. Determine the infrared absorption spectrum of Mupirocin Calcium Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 1708 cm－1, 1648 cm－1, 1558 cm－1, 1231 cm－1, 1151cm－1 and 894 cm－1. A solution of Mupirocin Calcium Hydrate (3 in 1000) responds to Qualitative Tests <1.09> (3) for calcium salt. D Optical rotation <2.49> [a]20: －16 – －209(1 g calculated on the anhydrous basis, methanol, 20 mL, 100 mm). Purity (1) Related substances—Dissolve 50 mg of Mupirocin Calcium Hydrate in a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make 10 mL, and use this solution as the sample solution (1). Pipet 2 mL of the sample solution (1), add a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetra-hydrofuran (3 in 4) (1:1) to make exactly 100 mL, and use this solution as the sample solution (2). Preserve these sam-ple solutions at a temperature between 49C and 89C. Per-form the test with exactly 20 mL of the sample solution (1) and the sample solution (2) as directed under Liquid Chro- solution (2), and add a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make exactly 20 mL. Con-firm that the peak area of mupirocin obtained with 20 mL of this solution is equivalent to 4 to 6z of that with 20 mL of the sample solution (2). System repeatability: When the test is repeated 6 times with 20 mL of the sample solution (2) under the above oper-ating conditions, the relative standard deviation of the peak areas of mupirocin is not more than 2.0z. (2) Inorganic salt from manufacturing process—Being specified separately when the drug is granted approval based on the Law. Water <2.48> Not less than 3.0z and not more than 4.5z (0.5 g, volumetric titration, direct titration). Assay Weigh accurately an amount of Mupirocin Calcium Hydrate and Mupirocin Lithium RS, equivalent to about 20 mg (potency), dissolve in a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make exactly 200 mL, and use these solutions as the sample solution and the standard solution. Preserve these solutions at a temperature between 49C and 89C. Perform the test with exactly 20 mL of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of mupirocin in each solution. Amount [ mg (potency)] of mupirocin (C H O ) matography <2.01> according to the following conditions, and determine the areas of each peak of the sample solution ＝ MS × AT/AS × 1000 26 44 9 and the sample solution (2) by the automatic integration method. Calculate the amount of the related substances by the following formula: the amount of principal related sub-stance (appeared at about 0.7 of the relative retention time to mupirocin) is not more than 4.0z, and the total amount of related substances (the total area of the peaks other than of the solvent and mupirocin) is not more than 6.0z. Amount (z) of principal related substance MS: Amount [mg (potency)] of Mupirocin Lithium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). ＝ Ai A ＋ Am × 100 × P × 100 100 － A × 100 A ＋ Am Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 7.71 g of ammonium acetate in 750 mL of water, adjust the pH to 5.7 with acetic acid (100), Total amount (z) of related substances and add water to make 1000 mL. To 300 mL of this solution ＝ A A ＋ Am × 100 × P × 100 100 － A × 100 A ＋ Am add 100 mL of tetrahydrofuran. Flow rate: Adjust so that the retention time of mupirocin is about 12.5 minutes. System suitability— A: Total peak areas other than the solvent and mupirocin from the sample solution (1) Ai: Peak area of the relative retention time of about 0.7 to mupirocin from the sample solution (1) Am: A value of 50 times of peak area of mupirocin from the sample solution (2) P: Potency per mg obtained from the assay Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of mupirocin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 1 mL of the sample System performance: Dissolve about 20 mg of Mupirocin Lithium RS and about 5 mg of ethyl parahydroxybenzoate in a mixture of 0.1 mol/L acetic acid-sodium acetate buffer solution (pH 4.0) and a solution of tetrahydrofuran (3 in 4) (1:1) to make 200 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, mupirocin and ethyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of mupirocin is not more than 1.0z.
Natural Aluminum Silicate occurs as a white or slightly colored powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. Natural Aluminum Silicate (1 g) dissolves when heated in 20 mL of a solution of sodium hydroxide (1 in 5), with some decomposition, leaving a large amount of insoluble sub-stance.
(1) To 0.5 g of Natural Aluminum Silicate add 3 mL of diluted sulfuric acid (1 in 3), heat until white fumes evolve, cool, add 20 mL of water, and filter. Render the filtrate slightly acid with ammonia TS: the solution re-sponds to Qualitative Tests <1.09> for aluminum salt. (2) Prepare a bead by fusing ammonium sodium hydrogenphosphate tetrahydrate on a platinum loop. Place the bead in contact with Natural Aluminum Silicate, and fuse again: an infusible material appears in the bead, producing, upon cooling, an opaque bead with a web-like structure.
(1) Acidity or alkalinity—Shake 5.0 g of Natural Aluminum Silicate with 100 mL of water, and centrifuge: the supernatant liquid so obtained is neutral. Chloride <1.03>—To 5.0 g of Natural Aluminum Sili- cate add 100 mL of water, boil gently for 15 minutes while shaking, then cool, add water to restore the original volume, and centrifuge. To 10 mL of the supernatant liquid add 6 mL of dilute nitric acid, dilute to 50 mL with water, and per-form the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochlo-ric acid VS (not more than 0.021z). Sulfate <1.14>—To the residue obtained in (6) add 3 mL of dilute hydrochloric acid, heat on a water bath for 10 minutes, dilute to 50 mL with water, and filter. To 2.0 mL of the filtrate add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). Heavy metals <1.07>—To 1.5 g of Natural Aluminum Silicate add 50 mL of water and 5 mL of hydrochloric acid, boil gently for 20 minutes while shaking, then cool, centri-fuge, remove the supernatant liquid, wash the residue with two 10-mL portions of water, centrifuging each time, com-bine these washings with the filtrate, and add ammonia solu-tion (28) dropwise, until a precipitate just appears. Add dropwise dilute hydrochloric acid with vigorous shaking and redissolve the precipitate. Heat the mixture with 0.45 g of hydroxylammonium chloride, cool, and add 0.45 g of so-dium acetate trihydrate, 6 mL of dilute acetic acid and water to make 150 mL. Perform the test, using 50 mL of this solu-tion as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution, 0.15 g of hydroxylam-monium chloride, 0.15 g of sodium acetate trihydrate, 2 mL of dilute acetic acid and water to make 50 mL (not more than 40 ppm). Arsenic <1.11>—To 1.0 g of Natural Aluminum Sili- cate, add 5 mL of dilute hydrochloric acid, heat gently to boil while shaking well, cool rapidly, and centrifuge. Mix the residue with 5 mL of dilute hydrochloric acid with shaking, centrifuge, then add 10 mL of water to the residue, and repeat the extraction in the same manner. Concentrate the ‌JP XVIII Official Monographs / Synthetic Aluminum Silicate 443 combined extracts on a water bath to 5 mL. Use this solution as the test solution, and perform the test (not more than 2 ppm). Soluble salts—Evaporate 50 mL of the supernatant liquid obtained in (1) on a water bath to dryness, and ignite the residue at 7009C for 2 hours: the mass of the ignited residue is not more than 40 mg. Fluoride—(i) Apparatus: Use a hard glass appa-ratus as illustrated in the figure. Ground-glass joints may be used. (ii) Procedure: Transfer 5.0 g of Natural Aluminum Sili-cate to the distilling flask A with the aid of 20 mL of water, add about 1 g of glass wool and 50 mL of diluted purified sulfuric acid (1 in 2), and connect A to the distillation appa-ratus, previously washed with steam streamed through the steam introducing tube E. Connect the condenser C with the receiver D containing 10 mL of 0.01 mol/L sodium hydroxide VS and 10 mL of water so that the lower end of C is immersed in the solution. Heat A gradually until the tem-perature of the solution in A reaches 1309C, then open the rubber tube F, close the rubber tube G, boil water in the steam generator B vigorously, and introduce the generated steam into F. Simultaneously, heat A, and maintain the tem-perature of the solution in A between 1359C and 1459C. Adjust the distilling rate to about 10 mL per minute. Collect about 170 mL of the distillate, then stop the distillation, wash C with a small quantity of water, combine the washings with the distillate, add water to make exactly 200 mL, and use this solution as the test solution. Perform the test with the test solution as directed in the procedure of determina-tion for fluoride under Oxygen Flask Combustion Method <1.06>. No corrective solution is used in this procedure. The content of fluoride (F) is not more than 0.01z. Amount (mg) of fluoride (F: 19.00) in the test solution ＝ amount (mg) of fluoride in 5 mL of the standard solution × AT/AS × 200/V Loss on drying <2.41> Not more than 20.0z (1 g, 1059C, 3 hours). Adsorptive power To 0.10 g of Natural Aluminum Silicate add 20 mL of a solution of methylene blue trihydrate (3 in 2000), shake for 15 minutes, allow to stand for 5 hours at 37 ± 29C, and centrifuge. Dilute 1.0 mL of the supernatant liquid with water to 200 mL. Place 50 mL of the solution in a Nessler tube and observe horizontally or vertically against a white background: the color of the solution is not deeper than that of the following control solution. Control solution: Dilute 1.0 mL of a solution of methy-lene blue trihydrate (3 in 2000) with water to 400 mL, and use 50 mL of this solution.
Ozagrel Sodium, when dried, contains not less than 98.
Ozagrel Sodium occurs as white, crystals or crystalline powder. It is freely soluble in water, soluble in methanol, and prac- tically insoluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Ozagrel Sodium (1 in 200,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Ozagrel Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ozagrel Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ozagrel Sodium RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. A solution of Ozagrel Sodium (1 in 20) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> The pH of a solution prepared by dissolving 0.5 g of Ozagrel Sodium in 10 mL of water is between 9.5 and 10.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Ozagrel Sodium in 10 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 2.0 g of Ozagrel Sodium in 30 mL of water, add 1 mL of acetic acid (100) and water to make 50 mL, shake, and allow to stand for 30 minutes. Filter the solution, discard the first 5 mL of the filtrate, and to 25 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test with this so-lution as the test solution. Prepare the control solution as follows: To 0.35 mL of 0.01 mol/L hydrochloric acid VS add 0.5 mL of acetic acid (100), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.012z). Heavy metals <1.07>—Proceed with 2.0 g of Ozagrel Sodium according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 10 ppm). Related substances—Dissolve 50 mg of Ozagrel So-dium in 100 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 5 mL of the sam-ple solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method, and calcu-late the amount of them by the area percentage method: each of the amount other than ozagrel is not more than 0.2z, and the total amount other than ozagrel is not more than 0.5z. Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave- length: 220 nm). Time span of measurement: About 2 times as long as the retention time of ozagrel, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the sample solution, and add the mobile phase to make exactly 200 mL, and use this solution as the solution for system suitability test. Pipet 2 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of ozagrel obtained with 5 mL of this solu-tion is equivalent to 15 to 25z of that with 5 mL of the solu-tion for system suitability test. System performance: When the procedure is run with 5 mL of the solution for system suitability test under the above op-erating conditions, the number of theoretical plates and the symmetry factor of the peak of ozagrel are not less than 6000 and not more than 2.0, respectively. System repeatability: When the test is repeated 6 times with 5 mL of the solution for system suitability test under the above operating conditions, the relative standard deviation of the peak area of ozagrel is not more than 2.0z. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours). ‌Assay Weigh accurately about 25 mg each of Ozagrel So-dium and Ozagrel Sodium RS, both previously dried, and dissolve each in methanol to make exactly 25 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, and use these solutions as the sample so-lution and the standard solution, respectively. Perform the test with 1 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of ozagrel to that of the internal standard. Amount (mg) of ozagrel sodium (C13H11N2NaO2) ＝ MS × QT/QS MS: Amount (mg) of Ozagrel Sodium RS taken Internal standard solution—A solution of benzoic acid in methanol (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 272 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of a solution of ammonium ace-tate (3 in 1000) and methanol (4:1). Flow rate: Adjust so that the retention time of ozagrel is about 10 minutes. System suitability— System performance: When the procedure is run with 1 mL Description Ozagrel Sodium Injection occurs as a clear and colorless liquid. Identification To a suitable volume of Ozagrel Sodium In-jection add water so that each mL contains 5 mg of Ozagrel Sodium. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum between 269 nm and 273 nm. pH Being specified separately when the drug is granted ap-proval based on the Law. Purity Related substance—To a suitable volume of Ozagrel Sodium Injection add the mobile phase so that each mL con-tains 0.4 mg of Ozagrel Sodium, and use this solution as the sample solution. Then, proceed as directed in the Purity (4) under Ozagrel Sodium. Bacterial endotoxins <4.01> Less than 3.7 EU/mg. Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
Perform the test following the test 1). The test 2) may be performed instead of 1), if possible. To exactly a volume of Ozagrel Sodium Injection, equivalent to about 4 mg of ozagrel sodium (C13H11N2NaO2), add exactly 5 mL of the internal standard solution and methanol to make 100 mL, and use this solution as the sam-ple solution. Separately, weigh accurately about 40 mg of Ozagrel Sodium RS, previously dried at 1059C for 4 hours, and dissolve in methanol to make exactly 50 mL. Pipet 5 mL of this solution, add exactly 5 mL of the internal standard solution, add 10 mL of water, then add methanol to make 100 mL, and use this solution as the standard solution. Then, proceed as directed in the Assay under Ozagrel Sodi-um. of the standard solution under the above operating condi-tions, the internal standard and ozagrel are eluted in this order with the resolution between these peaks being not less Amount (mg) of ozagrel sodium (C ＝ MS × QT／QS × 1／10 13H11N2NaO2) than 2.0, and the symmetry factor of the peak of ozagrel is not more than 2.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ozagrel to that of the internal standard is not more than 1.0z.
Parnaparin Sodium occurs as a white or light yellow powder. It is freely soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic.
(1) Mix 0.1 mL of a solution of Parnaparin Sodium (1 in 20) and 10 mL of a solution of tritoluidine blue O (1 in 100,000), and shake the mixture: the blue color of so-lution immediately changes to purple. (2) A solution of Parnaparin Sodium (1 in 20) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> Dissolve 0.1 g of Parnaparin Sodium in 10 mL of water: the pH of this solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Parnaparin Sodium in 10 mL of water: the solution is clear and colorless or pale yellow. Heavy metals <1.07>—Proceed with 1.0 g of Par- naparin Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 8.0z (0.2 g, in vacu-um, phosphorus (V) oxide, 609C, 3 hours). Molecular mass Calculate the molecular mass of Parnapa-rin Sodium by the following methods: The mass-average mo-lecular mass ranges between 4500 and 6500. Creation of calibration curve—Weigh 20 mg of low-molecular mass heparin for calibration of molecular mass, and dissolve it in 2.0 mL of the mobile phase as the standard solution. Perform the test with 50 mL of the standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak height, HUV, in chromatogram obtained by the ultraviolet absorption photometer, and determine the peak height, HRI, in chromatogram obtained by the differential refractometer. Calculate the ratio of HUV to HRI, HRI/HUV, at each peak. Assume the molecular mass in the 4th peak from the low molecular mass in chromatogram obtained by the ultraviolet absorption photometer as 2400, and make the calculation of the standard coefficient from dividing 2400 by the HRI/HUV at the corresponding peak. Make the calculation to multiply the HRI/HUV at each peak by the standard coefficient, and determine the molecular mass of each peak by the calcula-tion. Prepare the calculation curve by plotting the logarithm of molecular masses at each peak on the vertical axis and the retention time on the chromatogram obtained by the differential refractometer on the horizontal axis. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 234 nm) and a differential refractometer. Column: Connect 2 stainless steel columns which are 7.5 mm in inside diameter and 30 cm in length, and are packed with porous silica gel for liquid chromatography; one column, the molecular mass of limited size exclusion is about 500,000; the other, the molecular mass of limited size exclu-sion is about 100,000. Connect a pump, the about 500,000-molecular mass of limited size exclusion column, the about 100,000-molecular mass of limited size exclusion column, the ultraviolet absorption photometer and the differential refractometer in this order. Column temperature; A constant temperature of about 409C. Mobile phase: Dissolve 28.4 g of sodium sulfate anhydride in 1000 mL of water, and 5.0 with 0.05 mol/L sulfuric acid TS. Flow rate: 0.5 mL per minute. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, confirm that more than 10 peaks in chromatogram obtained as directed under either the Ultraviolet-visible Spec-trophotometry, or the Differential Refractometry are ob-served. System repeatability: When the tests repeated 6 times with 50 mL of the standard solution under the above operating conditions, relative standard deviation of the 4th peak height in chromatogram (HUV and HRI) is not more than 3.0z. Determination of molecular mass—Dissolve the 20 mg of Parnaparin Sodium with 2.0 mL of mobile phase, and use this solution as the sample solution. Perform the test with 50 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following condi-tions. Divide the main peak observed between 30 minutes and 45 minutes to 30 sec-interval fractions, and determine the strength of differential refractometer of each 30 sec-interval fraction. Determine the molecular mass of each frac-tion using the calibration curve and the retention time of each fraction. Determine the mean of molecular mass in the entire peak using the strength of differential refractometer and the molecular mass in every fractions. Mean molecular mass of parnaparin sodium ＝ S(ni･Mi)/Sni ni: The differential refractometer strength of fraction i in the main peak of chromatogram Mi: Molecular mass of fraction i in main peak Operating conditions— Detector: A differential refractometer. Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in (i) Creation of calibration curve. System suitability— Proceed as directed in (i) Creation of calibration curve. Distribution of molecular mass The molecular mass of Parnaparin Sodium is calculated as directed in the determi-nation of molecular mass and the distribution of molecular mass is calculated by the following equation: the molecular mass of not less than 80z parnaparin sodium is between 1500 and 10,000. Distribution of molecular mass (z) ＝ (Snj/Sni) × 100 ni: The differential refractometer strength of fraction i in the main peak of chromatogram Snj: Sum of differential refractometer strength in the each fraction between 1500 and 10,000 molecular mass in the main peak The degree of sulfate ester Dissolve 0.5 g of Parnaparin Sodium with 10 mL water. Treat the solution with 5 mL of a strongly basic ion exchange resin, and subsequently with 10 mL of a strongly acidic ion exchange resin. Dilute the solu-tion with water to 50 mL, and titrate <2.50> with 0.1 mol/L Sodium hydroxide VS (potentiometric titration). Calculate the degree of sulfate ester of Parnaparin Sodium from the equivalence point by the following equation; it is between 2.0 and 2.4. The degree of sulfate ester ＝ the first equivalence point (mL)/[the second equivalence point (mL) – first equivalence point (mL)] Total nitrogen Weigh accurately about 0.10 g of Parnapa-rin Sodium which is dried, and perform the test as directed under Nitrogen Determination <1.08>: it contains not less than 1.9z and not more than 2.3z of nitrogen (N:14.01). Anti-factor IIa activity Determine the potency of anti-factor IIa activity of Parnaparin Sodium according to the following method, it contains not less than 35 and not more than 60 low-molecular-mass-heparin unit per mg, calculated on the dried basis. Standard solution—Dissolve Low-molecular Mass Heparin RS with isotonic sodium chloride solution to make solutions which contain 0.1, 0.2 and 0.3 low-molecular-mass-heparin unit (anti-factor IIa activity) in 1 mL, respec-tively. Sample solution—Weigh accurately about 50 mg of Parnaparin Sodium, and dissolve it with isotonic sodium chloride solution to adjust the solution which contains 4 mg parnaparin sodium in 1 mL. Procedure—To each plastic tube add 0.10 mL of the sample solution and the standard solution, separately. To each tube add 0.10 mL of human normal plasma and mix, and incubate at 37 ± 19C accurately for 1 minute. Next, to each test tube add 0.10 mL of activated thromboplastin-time assay solution, which is pre-warmed at 37 ± 19C, and after the mixing incubate accurately for 5 minutes at 37 ± 19C. Then, to each tube add 0.10 mL of calcium chloride solution (277 in 100,000) which is pre-warmed at 37 ± 19C, mix, start a stop watch simultaneously, and permit to stand at the same temperature. Determine the time for the first appear-ance of fibrin clot. Calculation—Determine the low-molecular-mass-heparin unit (anti-factor IIa activity) of the sample solution from calibration curve obtained plots of clotting times for each standard solution; calculate the low-molecular-mass- heparin unit (anti-factor IIa activity) for 1 mg of parnaparin sodium as following equation. The low-molecular-mass-heparin unit (anti-factor IIa activity) for 1 mg of parnaparin sodium ＝ the low-molecular-mass-heparin unit (anti-factor IIa activity) in 1 mL of sample solution × b/a a: Amount (mg) of Parnaparin Sodium b: The total volume (mL) in which Parnaparin Sodium has been dissolved with isotonic sodium chloride solu-tion for the preparation of sample solution The ratio of anti-factor Xa activity to anti-factor IIa activity Divide the anti-factor Xa activity, obtained in the Assay, by the anti-factor IIa activity which has been obtained from the test according to the method of anti-factor IIa activity; the ratio of anti-factor Xa activity to anti-factor IIa activity is between 1.5 and 2.5.
Standard solution—Dissolve Low-molecular Mass-Heparin RS in isotonic sodium chloride solution to make so-lutions which contain 0.4, 0.6 and 0.8 low-molecular-mass-heparin units (anti-factor Xa activity) in 1 mL, respectively. Sample solution—Weigh accurately about 50 mg of Parnaparin Sodium, and dissolve it in isotonic sodium chlo-ride solution to make a solution which contains 7 mg par-naparin sodium in 1 mL. Procedure—To each plastic tube add 0.10 mL of either the sample solution or the standard solution, sepa-rately. Subsequently to the every tubes add 0.70 mL of Tris-buffered solution (pH 8.4), 0.10 mL of anti-thrombin III TS, and 0.10 mL of normal human plasma, and mix them. To another plastic tube transfer 0.20 mL of these solutions, separately, and incubate for accurate 3 minutes at 37 ± 19C. Next, to each tube add 0.10 mL of facter Xa TS and mix it, permit to stand 37 ± 19C accurately for 30 seconds, and im-mediately add 0.20 mL of chromogenic synthetic substrate solution (3 in 4000) and mix it, and subsequently incubate accurately for 3 min at 37 ± 19C. To each test tube add 0.30 mL of diluted acetic acid (100) solution (1 in 2) to stop the reaction. Separately, to plastic tube add 0.10 mL of isotonic sodium chloride solution, 0.70 mL of Tris-buffered solution (pH 8.4), 0.10 mL of anti-thrombin III TS, and 0.10 mL of normal human plasma to every tubes, and mix well. To another plastic tube transfer 0.20 mL of the solution, sepa-rately, and add both 0.30 mL of water and 0.30 mL of di-luted acetic acid (100) (1 in 2). Determine the absorbance of both the sample solution and the standard solution at 405 nm as directed under Ultraviolet-visible Spectrophotometry <2.24> using a solution obtained from this solution as the blank. Calculation method—Determine the low-molecular-mass unit (anti-factor Xa activity) of the sample solution using the calibration curve prepared from the absorbance of the standard solutions and their logarithmic concentrations, and calculate the low-molecular-mass unit (anti-factor Xa activity) in 1 mg of Parnaparin Sodium. Low-molecular-mass-heparin unit (anti-factor Xa activity) in 1 mg of Parnaparin Sodium ＝ the low-molecular-mass-heparin unit (anti-factor Xa activity) in 1 mL of the sample solution × b/a a: Amount (mg) of Parnaparin Sodium taken b: The total volume (mL) in which Parnaparin Sodium has been dissolved with isotonic sodium chloride solu-tion for the preparation of sample solution ‌Containers and storage Containers—Well-closed contain-ers. Paroxetine Hydrochloride Hydrate
Calcium Paraaminosalicylate Hydrate occurs as a white to slightly colored powder. It has a slightly bitter taste. It is very slightly soluble in water, and practically insoluble in methanol and in ethanol (99.5). It is gradually colored to brown by light.
(1) To 50 mg of Calcium Paraaminosalicy-late Hydrate add 100 mL of water, shake well, and filter. To 10 mL of the filtrate add 1 mL of 1 mol/L hydrochloric acid TS, shake, and add 1 drop of iron (III) chloride TS: a red-purple color develops. Determine the infrared absorption spectrum of Cal-cium Paraaminosalicylate Hydrate as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. To 3 g of Calcium Paraaminosalicylate Hydrate add 15 mL of ammonium chloride TS and 15 mL of water, heat on a water bath until almost dissolved, and filter after cool-ing: the filtrate responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt.
(1) Chloride <1.03>—Dissolve 1.0 g of Calcium Paraaminosalicylate Hydrate in 15 mL of dilute nitric acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.70 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.025z). Heavy metals <1.07>—Proceed with 1.0 g of Calcium Paraaminosalicylate Hydrate according to method 3, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 0.40 g of Calcium Para- aminosalicylate Hydrate in 20 mL of 0.1 mol/L hydrochloric acid TS by warming on a water bath, use this solution as the test solution, and perform the test (not more than 5 ppm). 3-Aminophenol—To 0.10 g of Calcium Para-aminosalicylate Hydrate add 5 mL of 0.1 mol/L disodium dihydrogen ethylenediamine tetraacetate TS, previously cooled in ice-water, and dissolve by shaking vigorously. Add immediately 3 mL of ammonia-ammonium chloride buffer solution (pH 11.0) previously cooled in ice water, and shake. Add 2 mL of 4-amino-N,N-diethylaniline sulfate TS, shake, add 10.0 mL of cyclohexane and 4 mL of diluted potassium hexacyanoferrate (III) TS (1 in 10), and shake immediately for 20 seconds. Centrifuge this solution, wash the separated cyclohexane layer with two 5-mL portions of diluted ammo-nia TS (1 in 14), add 1 g of anhydrous sodium sulfate, shake, and allow to stand for 5 minutes: the clear cyclohexane layer is not more colored than the following control solution. Control solution: Dissolve 50 mg of 3-aminophenol in water, and dilute with water to exactly 500 mL. Measure ex-actly 20 mL of this solution, and add water to make exactly 100 mL. Take 5.0 mL of this solution, add 3 mL of ammo-nia-ammonium chloride buffer solution (pH 11.0) previously cooled in ice-water, and treat this solution in the same man-ner as the sample. Water <2.48> 23.3 – 26.3z (0.1 g, volumetric titration, direct titration).
Weigh accurately about 0.2 g of Calcium Para-aminosalicylate Hydrate, dissolve in 60 mL of water and 0.75 mL of dilute hydrochloric acid by warming on a water bath. After cooling, add water to make exactly 100 mL, and use this solution as the sample solution. Measure exactly 30 mL of the sample solution, transfer to an iodine flask, and add exactly 25 mL of 0.05 mol/L bromine VS and 20 mL of a solution of potassium bromide (1 in 4). Add immediately 14 mL of a mixture of acetic acid (100) and hydrochloric acid (5:2), stopper the flask immediately, and allow to stand for 10 minutes with occasional shaking. Add cautiously 6 mL of potassium iodide TS, and shake gently. After 5 minutes, titrate <2.50> the produced iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Per-form a blank determination in the same manner. Each mL of 0.05 mol/L bromine VS ＝ 3.187 mg of C7H5CaNO3
Pemirolast Potassium contains not less than 98.
Pemirolast Potassium occurs as a light yellow crystalline powder. It is freely soluble in water, slightly soluble in methanol, and very slightly soluble in ethanol (99.5). It dissolves in potassium hydroxide TS. Melting point: about 3229C (with decomposition).
(1) Determine the absorption spectrum of a solution of Pemirolast Potassium in diluted potassium hy-droxide TS (1 in 10,000) (1 in 100,000) as directed under Ul-traviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pemirolast Potassium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pemirolast Potassium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Pemirolast Potassium RS: both spectra exhibit mL each of the sample solution and standard solution as di-rected under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pemirolast to that of the internal standard. similar intensities of absorption at the same wave numbers. Pemirolast Potassium responds to Qualitative Tests <1.09> (1) for potassium salt. Amount (mg) of pemirolast potassium (C ＝ MS × QT/QS 10H7KN6O)
(1) Clarity of solution—A solution obtained by dissolving 0.5 g of Pemirolast Potassium in 10 mL of water is clear. Heavy metals <1.07>—Proceed with 0.5 g of Pemirolast Potassium according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Stand-ard Lead Solution (not more than 20 ppm). ‌Related substances—Dissolve 50 mg of Pemirolast Potassium in 50 mL of a mixture of phosphate buffer solu-tion (pH 8.0) and methanol (3:2), and use this solution as the sample solution. Pipet 2 mL of the sample solution, and add a mixture of phosphate buffer solution (pH 8.0) and metha-nol (3:2) to make exactly 100 mL. To exactly 2.5 mL of this solution add a mixture of phosphate buffer solution (pH 8.0) and methanol (3:2) to make exactly 50 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the area of the peak other than pemirolast obtained from the sample solution is not larger than the peak area of pemirolast from the standard so-lution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 9 times as long as the retention time of pemirolast. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add a mixture of phosphate buffer solu-tion (pH 8.0) and methanol (3:2) to make exactly 25 mL. Confirm that the peak area of pemirolast obtained with 10 mL of this solution is equivalent to 15 to 25z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pemirolast are not less than 3000 and not more than 1.7, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pemirolast is not more than 2.0z. Water <2.48> Not more than 0.5z (0.1 g, coulometric titration).
Weigh accurately about 50 mg each of Pemirolast Potassium and Pemirolast Potassium RS (separately deter-mine the water <2.48> in the same manner as Pemirolast Potassium), dissolve in a mixture of phosphate buffer solu-tion (pH 8.0) and methanol (3:2) to make them exactly 50 mL. Pipet 5 mL each of these solutions, add exactly 5 mL of the internal standard solution, then add a mixture of phos-phate buffer solution (pH 8.0) and methanol (3:2) to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 MS: Amount (mg) of Pemirolast Potassium RS taken, cal-culated on the anhydrous basis Internal standard solution—A solution of ethyl aminobenzo-ate in methanol (1 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, methanol and acetic acid (100) (30:20:1). Flow rate: Adjust so that the retention time of pemirolast is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, pemirolast and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of pemirolast to that of the internal standard is not more than 1.0z.
Benzylpenicillin Potassium occurs as white, crystals or crystalline powder. It is very soluble in water, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Benzylpenicillin Potassium (1 in 1000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Benzylpenicillin Potassium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Ben-zylpenicillin Potassium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Benzylpenicillin Potassium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Benzylpenicillin Potassium responds to Qualitative Tests <1.09> (1) for potassium salt. D Optical rotation <2.49> [a]20: ＋270 – ＋3009(1.0 g calcu-lated on the dried basis, water, 50 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Benzylpenicillin Potassium in 100 mL of water is be-tween 5.0 and 7.5.
(1) Clarity and color of solution—A solution ob-tained by dissolving 1.0 g of Benzylpenicillin Potassium in 10 mL of water is clear, and its absorbance at 400 nm deter-mined as directed under Ultraviolet-visible Spectrophotome-try <2.24> is not more than 0.10. Heavy metals <1.07>—Proceed with 2.0 g of Benzyl- penicillin Potassium according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution by inciner- ating 1.0 g of Benzylpenicillin Potassium according to Method 4, and perform the test. In the incineration, use a crucible of porcelain, and after addition of 10 mL of a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) add 1 mL of hydrogen peroxide (30), then burn the ethanol (not more than 2 ppm). Related substances—Dissolve 40 mg of Benzylpeni-cillin Potassium in 20 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than benzylpenicillin obtained from the sample solution is not larger than the peak area of benzylpenicillin from the stand-ard solution, and the total area of the peaks other than ben-zylpenicillin from the sample solution is not larger than 3 times the peak area of benzylpenicillin from the standard so-lution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 5 times as long as the retention time of benzylpenicillin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 10 mL of the stand-ard solution, and add water to make exactly 100 mL. Con-firm that the peak area of benzylpenicillin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of benzylpenicillin are not less than 4000 and 0.7 to 1.2, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of benzylpenicillin is not more than 1.0z. Loss on drying <2.41> Not more than 1.0z (3 g, reduced pressure not exceeding 0.67 kPa, 609C, 3 hours).
Weigh accurately amounts of Benzylpenicillin Potassium and Benzylpenicillin Potassium RS, equivalent to about 6 × 104 Units, dissolve each in water to make exactly 20 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine the peak areas, AT and AS, of benzylpenicillin in each solution. Amount (unit) of benzylpenicillin potassium (C16H17KN2O4S) ＝ MS × AT/AS ‌528 MS: Amount (unit) of benzylpenicillin potassium RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of diammonium hydrogen phos-phate solution (33 in 5000) and acetonitrile (19:6), adjusted to pH 8.0 with phosphoric acid. Flow rate: Adjust so that the retention time of benzyl-penicillin is about 7.5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of benzylpenicillin are not less than 2000 and not more than 3.0, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of benzylpenicillin is not more than 1.0z.
Pentobarbital Calcium contains not less than 98.
Pentobarbital Calcium occurs as a white pow-der. It is sparingly soluble in water, slightly soluble in ethanol 1 cm Absorbance <2.24> E 1z (278 nm): 67.5 – 71.5 (after dry- (95), and practically insoluble in acetonitrile. ing, 0.1 g, 0.01 mol/L hydrochloric acid TS, 1000 mL).
(1) Determine the infrared absorption spec-trum of Pentobarbital Calcium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 1 g of Pentobarbital Calcium add 5 mL of ethanol (95) and 5 mL of dilute hydrochloric acid, dissolve by warm-ing with shaking, shake with 5 mL of dilute hydrochloric acid and 10 mL of water, allow to cool, and filter. To the fil-trate add 1 drop of methyl red TS, and add ammonia TS until a slight yellow color develops: the solution responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt.
(1) Clarity and color of solution—Dissolve 0.10 g of Pentazocine in 20 mL of 0.1 mol/L hydrochloric acid TS: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Pentazo- cine according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Pentazocine according to Method 3, and perform the test with a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) (not more than 2 ppm). Related substances—Dissolve 0.20 g of Pentazocine in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add chloroform to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and isopro-pylamine (94:3:3) to a distance of about 13 cm, and air-dry A solution of Pentobarbital Calcium (1 in 100) shows no optical rotation. Identification (1) Determine the infrared absorption spec-trum of Pentobarbital Calcium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 1 g of Pentobarbital Calcium add 5 mL of ethanol (95) and 5 mL of dilute hydrochloric acid, dissolve by warm-ing with shaking, shake with 5 mL of dilute hydrochloric acid and 10 mL of water, allow to cool, and filter. To the fil-trate add 1 drop of methyl red TS, and add ammonia TS until a slight yellow color develops: the solution responds to Qualitative Tests <1.09> (1), (2) and (3) for calcium salt. Purity (1) Chloride <1.03>—To 1.0 g of Pentobarbital Calcium add 5 mL of ethanol (95) and 2.5 mL of dilute nitric acid, dissolve by warming with shaking, cool, add water to make 50 mL, shake well, and filter. Discard the first 10 mL of the filtrate, and to 15 mL of the subsequent filtrate add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 0.30 mL of 0.01 mol/L hy- ‌drochloric acid VS add 1.5 mL of ethanol (95), 6 mL of di-lute nitric acid and water to make 50 mL (not more than 0.035z). Heavy metals <1.07>—To 2.0 g of Pentobarbital Cal- lution and standard solution as directed under Liquid Chro-matography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pen-tobarbital to that of the internal standard. cium add 5 mL of ethanol (95) and 5 mL of dilute hydro-chloric acid, dissolve by warming with shaking, cool, add water to make 80 mL, shake well, and filter. Discard the first Amount (mg) of pentobarbital calcium (C ＝ MS × QT/QS × 1.084 22H34CaN4O6) 10 mL of the filtrate, to 40 mL of the subsequent filtrate add 1 drop of phenolphthalein TS, add dropwise ammonia TS until a pale red color develops, and add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: To 2.5 mL of ethanol (95) add 2.5 mL of dilute hy-drochloric acid and water to make 30 mL. Add 1 drop of phenolphthalein TS, add dropwise ammonia TS until a pale red color develops, then add 2.0 mL of Standard Lead Solu-tion, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Related substances—Dissolve 10 mg of Pentobarbital Calcium in 100 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the areas of each peak by the automatic integration method: the area of any peak other than the peak of pentobarbital obtained from the sam-ple solution is not larger than 3/10 times the peak area of pentobarbital from the standard solution, and the total of these peak area is not larger than the peak area of pentobar-bital from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of pentobarbital, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system performance in the Assay. Test for required detectability: Pipet 2 mL of the standard solution, add water to make exactly 20 mL, and confirm that the peak area of pentobarbital obtained with 20 mL of this solution is equivalent to 5 to 15z of that with 20 mL of the standard solution. System repeatability: When the test is repeated 6 times MS: Amount (mg) of Pentobarbital RS taken Internal standard solution—Dissolve 0.2 g of isopropyl par-ahydroxybenzoate in 20 mL of acetonitrile for liquid chro-matography, and add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1.36 g of potassium dihydrogen phosphate in 1000 mL of water, and adjust to pH 4.0 with diluted phosphoric acid (1 in 10). To 650 mL of this solution add 350 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of pentobar-bital is about 7 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, pentobarbital and the internal standard are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of pentobarbital to that of the internal standard is not more than 1.0z.
Phenethicillin Potassium contains not less than 1400 units and not more than 1480 units per mg, calculated on the dried basis.
Phenethicillin Potassium occurs as a white to light yellow-white crystalline powder. It is freely soluble in water, and slightly soluble in ethanol (99.5).
(1) Determine the absorption spectrum of a solution of Phenethicillin Potassium (1 in 5000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Phenethicillin Potassium as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Phenethicillin Potassium responds to Qualitative Tests <1.09> (1) for potassium salt. D Optical rotation <2.49> [a]20: ＋217 – ＋2449(1 g calculated on the dried basis, phosphate TS, 100 mL, 100 mm). Phenethicillin potassium Dissolve about 50 mg of Phenethicillin Potassium in the mobile phase to make 50 mL, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AD and AL, of D-a-phenethicillin and L-a-phenethicillin by the automatic inte-gration method: AL/(AD ＋ AL) is between 0.50 and 0.70. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 6 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Adjust the pH of a mixture of a solution of diammonium hydrogen phosphate (1 in 150) and acetonitrile (41:10) to 7.0 with phosphoric acid. Flow rate: Adjust so that the retention time of L-a-phenethicillin is about 25 minutes. System suitability— System performance: When the procedure is run with 10 mL of the sample solution under the above operating condi-tions, D-a-phenethicillin and L-a-phenethicillin are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the sample solution under the above operating conditions, the relative standard deviation of the peak area of L-a-phenethicillin is not more than 2.0z.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Phenethicillin Potassium according to Method 2, and per-form the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Phenethicillin Potassium according to Method 4 and, per-form the test (not more than 2 ppm). Related substances—Dissolve 50 mg of Phenethicillin Potassium in 50 mL of the mobile, and use this solution as ‌the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solu-tion as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the total of the peak areas other than D-a-phenethicillin and L-a-phenethicillin obtained from the sample solution is not larger than 5 times the total of the peak areas of D-a-phenethicillin and L-a-phenethicillin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the L-a-Phenethicillin potassium. Time span of measurement: About 1.5 times as long as the retention time of L-a-phenethicillin. System suitability— System performance, and system repeatability: Proceed as directed in the system suitability in the L-a-Phenethicillin po-tassium. Test for required detectability: Measure exactly 2 mL of the standard solution, and add the mobile phase to make ex-actly 10 mL. Confirm that the peak area of L-a-phenethicil-lin obtained with 10 mL of this solution is equivalent to 14 to 26z of that with 10 mL of the standard solution. Loss on drying <2.41> Not more than 1.0z (0.1 g, in vacu-um, 609C, 3 hours).
Weigh accurately an amount of Phenethicillin Po-tassium and dried Phenethicillin Potassium RS, equivalent to about 40,000 units, dissolve each in phosphate buffer solution (pH 6.0) to make exactly 20 mL, and use these solu-tions as the sample solution and the standard solution, respectively. Pipet 2 mL each of these solutions in 100-mL glass-stoppered flasks, add 2.0 mL of sodium hydroxide TS to them, and allow to stand for exactly 15 minutes. To them add 2.0 mL of diluted hydrochloric acid (1 in 10) and exactly 10 mL of 0.005 mol/L iodine VS, and allow them to stand for exactly 15 minutes. Add 0.2 – 0.5 mL of starch TS, and titrate <2.50> with 0.01 mol/L sodium thiosulfate VS until the color of the solution disappears. Separately, to exactly 2 mL each of the sample solution and standard solution add exactly 10 mL of 0.005 mol/L iodine VS, then proceed in the same manner as above without allowing to stand for 15 minutes as a blank determination, and make any necessary correction. Determine the volumes, VT and VS, of 0.005 mol/L iodine VS consumed in the sample solution and standard solution. Phenobarbital
Phenol and Zinc Oxide Liniment is a white pasty mass. It has a slight odor of phenol.
(1) Shake well 1 g of Phenol and Zinc Oxide Liniment with 10 mL of diethyl ether, and filter. To the filtrate add 10 mL of dilute sodium hydroxide TS, shake well, and separate the water layer. To 1 mL of the water layer add 1 mL of sodium nitrite TS and 1 mL of dilute hy-drochloric acid, shake, and add 3 mL of sodium hydroxide TS: a yellow color develops (phenol). Place 1 g of Phenol and Zinc Oxide Liniment in a porcelain crucible, heat gradually raising the temperature until the content is charred, and then ignite it strongly: a yellow color develops, and disappears on cooling. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the filtrate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is produced (zinc oxide). Shake 0.5 g of Phenol and Zinc Oxide Liniment with 1 mL of water and 5 mL of chloroform, separate the chlo-roform layer, and use this solution as the sample solution. Separately, dissolve 0.01 g of phenol in 5 mL of chloroform, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chro-matography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and ammonia solution (28) (50:5:1) to a distance of about 10 cm, and air-dry the plate. Allow the plate to stand in iodine vapor: the spots obtained from the sample solution and the standard solution show the same Rf value.
Piperacillin Sodium contains not less than 863 mg (potency) and not more than 978 mg (potency) per mg, calculated on the anhydrous basis.
Piperacillin Sodium occurs as a white, powder or mass. It is very soluble in water, freely soluble in methanol and in ethanol (95), and practically insoluble in acetonitrile.
(1) Determine the infrared absorption spec-trum of Piperacillin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Piperacillin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋175 – ＋1909(0.8 g calcu-lated on the anhydrous basis, water, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Piperacillin Sodium in 4 mL of water: the pH of the solution is between 5.0 and 7.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Piperacillin Sodium in 10 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Piper- acillin Sodium according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Piperacillin Sodium according to Method 4, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.10 g of Piperacillin Sodium in 50 mL of the mobile phase A, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the areas of each peak by the automatic integration method: the area of the peak of ampicillin appeared at the retention time of about 7 minutes obtained from the sample solution is not larger than 1/2 times that of piperacillin from the standard solution, the total area of related compounds 1 appeared at the retention times of about 17 minutes and about 21 minutes is not larger than 2 times of the peak area of piper-acillin from the standard solution, the peak area of related compound 2 appeared at the retention time of about 56 minutes is not larger than that of piperacillin from the stand-ard solution, and the total area of the peaks other than factor of the peak of piperacillin are not less than 15,000 and not more than 1.5, respectively. System repeatability:When the test is repeated 3 times with 20 mL of the standard solution under the above operating conditions, the relative standard deviation of the peak area of piperacillin is not more than 2.0z. Water <2.48> Not more than 1.0z (3 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Piperacillin Sodium, equivalent to about 0.1 g (potency), and dissolve in water to make exactly 100 mL. To exactly 5 mL of this solution add exactly 5 mL of the internal standard solution, and use this solution as the sample solution. Separately, weigh accurately an amount of Piperacillin RS, equivalent to about 0.1 g (po-tency), and dissolve in the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add exactly 5 mL of the in-ternal standard solution, and use this solution as the stand-ard solution. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak height of piperacillin to that of the internal standard. piperacillin is not larger than 5 times of the peak area of Amount [ mg (potency)] of piperacillin (C H N O S) piperacillin from the standard solution. For the peak areas of ampicillin, related compound 1 and related compound 2, ＝ MS × QT/QS × 1000 23 27 5 7 multiply their correction factors, 1.39, 1.32 and 1.11, respec-tively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase A: A mixture of water, acetonitrile and 0.2 mol/L potassium dihydrogen phosphate (45:4:1). Mobile phase B: A mixture of acetonitrile, water and 0.2 mol/L potassium dihydrogen phosphate (25:24:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 7 7 – 13 100 → 83 0 → 17 13 – 41 41 – 56 83 → 20 17 → 80 56 – 60 Flow rate: 1.0 mL per minute (the retention time of piper-acillin is about 33 minutes). Time span of measurement: About 1.8 times as long as the retention time of piperacillin, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase A to make exactly 20 mL. Confirm that the peak area of piperacillin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry MS: Amount [mg (potency)] of Piperacillin RS taken Internal standard solution—A solution of acetanilide in the mobile phase (1 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: To 60.1 g of acetic acid (100) and 101.0 g of triethylamine add water to make exactly 1000 mL. To 25 mL of this solution add 25 mL of dilute acetic acid and 210 mL of acetonitrile, and add water to make exactly 1000 mL. Flow rate: Adjust so that the retention time of piperacillin is about 5 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, the internal standard and piperacillin are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak height of piperacillin to that of the internal stand-ard is not more than 1.0z.
Pitavastatin Calcium Hydrate contains not less than 98.
Pitavastatin Calcium Hydrate occurs as a white to pale yellow powder. It is slightly soluble in methanol, very slightly soluble in water and in ethanol (99.5). It dissolves in dilute hydrochloric acid. It shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Pitavastatin Calcium Hydrate in methanol (1 in 125,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Pitavastatin Calcium Hydrate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorption at the wave numbers of 3400 – 3300 cm－1, about 1560 cm－1, 1490 cm－1, 1219 cm－1, 1066 cm－1 and 766 cm－1. Dissolve 0.25 g of Pitavastatin Calcium Hydrate in 5 mL of dilute hydrochloric acid, neutralize with ammonia TS, and filter: the filtrate responds to Qualitative Tests <1.09> (1), (2), and (3) for calcium. D Optical rotation <2.49> [a]20: ＋22.0 – ＋24.59(0.1 g calcu-lated on the anhydrous basis, a mixture of water and aceto-nitrile (1:1), 10 mL, 100 mm).
(1) Heavy metals <1.07>—To 1.0 g of Pitavastatin Calcium Hydrate in a quartz crucible add 10 mL of a solu-tion of magnesium nitrate hexahydrate in ethanol (95) (1 in 10) and mix well, then fire the ethanol to burn, and heat gradually to carbonize. After cooling, moisten the residue with 1.5 mL of sulfuric acid, heat carefully, then ignite at 5509C until the residue is incinerated. After cooling, moisten the residue with 1.5 mL of nitric acid, heat carefully, then ignite at 5509C until the residue is completely incinerated. After cooling, dissolve the residue in 3 mL of hydrochloric acid, and evaporate the solvent to dryness on a water bath. Moisten the residue with 3 drops of hydrochloric acid, dis-solve in 10 mL of hot water with the aid of gentle heat, and filter. Wash the residue with 20 mL of water, and pour the filtrates and washings into a Nessler tube. Add 1 drop of phenolphthalein TS, add ammonia TS dropwise until the solution develops a pale red color, then add 2 mL of dilute acetic acid, add water to make 50 mL, and use this solution as the test solution. The control solution is prepared as follows: Take 10 mL of a solution of magnesium nitrate hex-ahydrate in ethanol (95) (1 in 10), and fire the ethanol to burn. Hereafter, proceed as for the test solution, then add 2.0 mL of Standard Lead Solution, 2 mL of acetic acid and water to make 50 mL (not more than 20 ppm). (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 0.10 g of Pitavastatin Cal-cium Hydrate in 100 mL of a mixture of acetonitrile and water (3:2), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of aceto-nitrile and water (3:2) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak of related substance A, having the relative retention time of about 1.1 to pitavastatin, obtained from the sample solution is not more than 1/2 times the peak area of pitavastatin from the standard solution, and the area of the peak other than pitavastatin and the peak mentioned above from the sample solution is not more than 1/10 times the peak area of pitavastatin from the standard solution. Fur-thermore, the total area of the peaks other than pitavastatin from the sample solution is not larger than the peak area of pitavastatin from the standard solution. For the area of the peak of related substance B, having the relative retention time of about 1.4, multiply the correction factor, 1.8. Operating conditions— Detector, column, and column temperature: Proceed as directed in the operating conditions in the Assay. Mobile phase A: To 10 mL of dilute acetic acid add water to make 1000 mL. To 800 mL of this solution add diluted so-dium acetate TS (1 in 100) to adjust to pH 3.8. Mobile phase B: Acetonitrile for liquid chromatography. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 20 20 – 40 60 → 10 40 → 90 40 – 60 Flow rate: Adjust so that the retention time of pitavastatin is about 23 minutes. Time span of measurement: About 2.5 times as long as the retention time of pitavastatin, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, add a mixture of acetonitrile and water (3:2) to make exactly 20 mL. Confirm that the peak area of pitavastatin obtained with 10 mL of this solution is equiva-lent to 4 to 6z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of pitavastatin are not less than 17,000 and not more than 1.3, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pitavastatin is not more than 2.0z. ‌Water <2.48> 9.0 – 13.0z (0.2 g, volumetric titration, direct titration. Use a mixture of pyridine for water determi-nation and ethylene glycol for water determination (83:17) instead of methanol for water determination).
Conduct this procedure using light-resistant vessels. Weigh accurately about 0.1 g of Pitavastatin Calcium Hy-drate, dissolve in a mixture of acetonitrile and water (3:2) to make exactly 100 mL. Pipet 5 mL of this solution, add ex-actly 5 mL of the internal standard solution, then add a mix-ture of acetonitrile and water (3:2) to make 50 mL, and use this solution as the sample solution. Separately, weigh accu-rately about 30 mg of Pitavastatin Methylbenzylamine RS (separately determine the water <2.48> by coulometric titra-tion using 0.1 g), dissolve in a mixture of acetonitrile and water (3:2) to make exactly 25 mL. Pipet 5 mL of this solu-tion, add exactly 5 mL of the internal standard solution, then add a mixture of acetonitrile and water (3:2) to make 50 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of pitavastatin to that of the internal standard. Amount (mg) of pitavastatin calcium (C50H46CaF2N2O8) ＝ MS × QT/QS × 4 × 0.812 MS: Amount (mg) of Pitavastatin Methylbenzylamine RS taken, calculated on the anhydrous basis Internal standard solution—Butyl parahydroxybenzoate in a mixture of acetonitrile and water (3:2) (3 in 2000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: To 10 mL of dilute acetic acid add water to make 1000 mL. To 350 mL of this solution add 650 mL of methanol, and dissolve 0.29 g of sodium chloride in this so-lution. Flow rate: Adjust so that the retention time of pitavastatin is about 17 minutes. System suitability— System performance: When the procedure is run with 10 Others Related substance A: (3RS,5RS )-7-[2-Cyclopropyl-4-(4-fluorophenyl)quinolin-3-yl]-3,5-dihydroxyhept-6-enoic acid Related substance B: 7-[2-Cyclopropyl-4-(4-fluorophenyl)quinolin-3-yl]-3-hydroxy-5-oxohept-6-enoic acid Pitavastatin Calcium Orally Disintegrating Tablets
Polaprezinc occurs as a white to pale yellow-white crystalline powder. It is practically insoluble in water, in methanol and in ethanol (99.5). It dissolves in dilute hydrochloric acid. Identiˆcation (1) To 2 mL of a solution of Polaprezinc in 0.2 mol/L hydrochloric acid TS (1 in 1000) add 0.5 mL of a solution of sulfanilic acid in 1 mol/L hydrochloric acid TS (1 in 200), 0.5 mL of a solution of sodium nitrite (1 in 20) and 3 mL of sodium carbonate TS: a red color is produced. Amount [mg (potency)] of mecillinam (C ＝ MS × QT/QS × 25/V 15H23N3O3S) (2) A solution of Polaprezinc in 0.2 mol/L hydrochloric acid TS (1 in 1000) responds to Qualitative Tests <1.09> for zinc salt. MS: Amount [mg (potency)] of Pivmecillinam Hydrochlo-ride RS taken Internal standard solution—A solution of diphenyl in the mobile phase (1 in 12,500). Determine the infrared absorption spectrum of Polaprezinc as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋8 – ＋99(1 g calculated on the anhydrous basis, 3 mol/L hydrochloric acid TS, 50 mL, 100 mm).
(1) Lead—Weigh accurately about 0.5 g of Polaprezinc, dissolve in 3 mL of dilute nitric acid, add water to make exactly 10 mL, and use this solution as the sample solution. Separately, pipet 0.5 mL, 1.0 mL, 1.5 mL and 2.0 mL of Standard Lead Solution, to each solution add 3 mL of dilute nitric acid and water to make exactly 10 mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions, and calculate the amount of lead in the sample solution using a calibration curve ob-tained from the absorbances of the standard solutions: not more than 10 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Lead hollow-cathode lamp. Wavelength: 283.3 nm. (2) Related substances—Dissolve 50 mg of Polaprezinc in 10 mL of 0.1 mol/L hydrochloric acid TS, add the mobile phase to make 100 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area by the auto-matic integration method: the peak area of L-histidine, hav-ing the relative retention time of about 0.38 to L-carnosine, obtained from the sample solution is not larger than 1/5 times of the peak area of L-carnosine from the standard so-lution, the area of the peak other than L-carnosine and the peak mentioned above from the sample solution is not larger than 1/10 times of the peak area of L-carnosine from the standard solution. Furthermore, the total area of the peaks other than L-carnosine from the sample solution is not larger than the peak area of L-carnosine from the standard solu-tion. Operating conditions— Detector, column, column temperature, mobile phase, and ‰ow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 4 times as long as the retention time of L-carnosine, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 2 mL of the standard solution, add the mobile phase to make exactly 20 mL. Con-ˆrm that the peak area of L-carnosine obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the standard solution. System performance: Dissolve 50 mg each of Polaprezinc and L-histidine in 10 mL of 0.1 mol/L hydrochloric acid TS, and add the mobile phase to make 100 mL. When the proce-dure is run with 10 mL of this solution under the above oper-ating conditions, L-histidine and L-carnosine are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of L-carnosine is not more than 2.0z. Water <2.48> Not more than 5.0z (0.2 g, volumetric titra-tion, direct titration, stir for 30 minutes).
(1) Polaprezinc—Weigh accurately about 25 mg of Polaprezinc, dissolve in 5 mL of 0.1 mol/L hydrochloric acid TS, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of L-Carnosine RS, previously dried at 1059C for 3 hours, dissolve in 5 mL of 0.1 mol/L hydro-chloric acid TS, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine the peak areas, AT and AS, of L-carnosine in each solution. Amount (mg) of polaprezinc (C9H12N4O3Zn) ＝ MS × AT/AS × 1.292 MS: Amount (mg) of L-Carnosine RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 459C. Mobile phase: Dissolve 1.4 g of potassium dihydrogen phosphate in 1000 mL of water, adjust to pH 3.5 with diluted phosphoric acid (1 in 100). Dissolve 2 g of sodium 1-octane sulfonate in 900 mL of this solution, and add 100 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of L-carnosine is about 15 minutes. System suitability— System performance: Dissolve 5 mg of L-histidine in 20 mL of the standard solution. When the procedure is run with 10 mL of this solution under the above operating conditions, L-histidine and L-carnosine are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of L-carnosine is not more than 1.0z. (2) Zinc—Weigh accurately about 0.2 g of Polaprezinc, dissolve in 3 mL of dilute hydrochloric acid, and add water to make exactly 100 mL. Pipet 25 mL of this solution, add 10 mL of ammonia-ammonium chloride buŠer solution (pH 10.7), and titrate <2.50> with 0.01 mol/L disodium dihydro-gen ethylenediamine tetraacetate VS (indicator: 40 mg of eriochrome black T-sodium chloride indicator). Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 0.6538 mg Zn
Potassium Bromide, when dried, contains not less than 99.
Potassium Bromide occurs as colorless or white crystals, granules or crystalline powder. It is odorless. It is freely soluble in water and in glycerin, soluble in hot ethanol (95), and slightly soluble in ethanol (95).
A solution of Potassium Bromide (1 in 10) responds to Qualitative Tests <1.09> for potassium salt and for bromide.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Bromide in 3 mL of water: the solution is clear and colorless. Alkalinity—Dissolve 1.0 g of Potassium Bromide in 10 mL of water, add 0.10 mL of 0.05 mol/L sulfuric acid VS and 1 drop of phenolphthalein TS, heat to boiling, and cool: no color develops. Chloride—Make a calculation from the result ob-tained in the Assay: not more than 84.5 mL of 0.1 mol/L silver nitrate VS is consumed for 1 g of Potassium Bromide. Sulfate <1.14>—Proceed with 2.0 g of Potassium Bromide, and perform the test. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.024z). Iodide—Dissolve 0.5 g of Potassium Bromide in 10 mL of water, add 2 to 3 drops of iron (III) chloride TS and 1 mL of chloroform, and shake: no red-purple to purple color develops in the chloroform layer. Bromate—Dissolve 1.0 g of Potassium Bromide in 10 mL of freshly boiled and cooled water, and add 0.1 mL of potassium iodide TS, 1 mL of starch TS and 3 drops of dilute sulfuric acid. Shake the mixture gently, and allow to stand for 5 minutes: no blue color develops. Heavy metals <1.07>—Proceed with 2.0 g of Potas- sium Bromide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Barium—Dissolve 0.5 g of Potassium Bromide in 10 mL of water, add 0.5 mL of dilute hydrochloric acid and 1 mL of potassium sulfate TS, and allow to stand for 10 minutes: no turbidity is produced. Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Bromide according to Method 1, and perform the test (not more than 2 ppm). ‌Loss on drying <2.41> Not more than 1.0z (1 g, 1109C, 4 hours).‌
Weigh accurately about 0.4 g of Potassium Bromide, previously dried, and dissolve in 50 mL of water. Add 10 mL of dilute nitric acid and exactly measured 50 mL of 0.1 mol/L silver nitrate VS, and titrate <2.50> the excess silver nitrate with 0.1 mol/L ammonium thiocyanate VS (in-dicator: 2 mL of ammonium iron (III) sulfate TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L silver nitrate VS ＝ 11.90 mg of KBr
Potassium Canrenoate, when dried, contains not less than 98.
Potassium Canrenoate occurs as a pale yellow-white to pale yellow-brown, crystalline powder. It is freely soluble in water, soluble in methanol, sparingly soluble in ethanol (95), and practically insoluble in chlo-roform and in diethyl ether.
(1) Dissolve 2 mg of Potassium Canrenoate in 2 drops of sulfuric acid: an orange color develops. Ob-serve under ultraviolet light (main wavelength: 365 nm): the solution shows a yellow-green fluorescence. Add 1 drop of acetic anhydride to this solution: the color of the solution changes to red. Determine the absorption spectrum of a solution of Potassium Canrenoate in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Po-tassium Canrenoate, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. The solution of Potassium Canrenoate (1 in 10) re-sponds to Qualitative Tests <1.09> (1) for potassium salt. D Optical rotation <2.49> [a]20: －71 – －769(after drying, 0.2 g, methanol, 20 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Potassium Canrenoate in 20 mL of water: the pH of this solution is between 8.4 and 9.4.
(1) Clarity and color of solution—Dissolve 0.5 g of Potassium Canrenoate in 5 mL of water: the solution is clear, and shows a pale yellow to light yellow color. Heavy metals <1.07>—Proceed with 2.0 g of Potas-sium Canrenoate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Canrenoate according to Method 3, and per-form the test (not more than 2 ppm). Canrenone—Place 0.40 g of Potassium Canrenoate in a glass-stoppered centrifuge tube, cool in ice-water to a temperature not higher than 59C, add 6 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 10.0) being cooled to a temperature not higher than 59C to dissolve, and add 8 mL of water being cooled to a tempera-ture not higher than 59C. Add exactly 10 mL of chloroform, allow to stand for 3 minutes at a temperature not higher than 59C, shake vigorously for 2 minutes, and centrifuge. Drain off the water layer, collect 5 mL of the chloroform layer, transfer to a glass-stoppered centrifuge tube containing 3 mL of boric acid-potassium chloride-sodium hydroxide buffer solution (pH 10.0) cooled to a temperature not higher than 59C, and 4 mL of water cooled to a temperature not higher than 59C, shake for 1 minute, and centrifuge. Drain off the water layer, pipet 2 mL of the chloroform layer, and add chloroform to make exactly 10 mL. Determine the absor-bance of this solution at 283 nm as directed under Ultravio-let-visible Spectrophotometry <2.24>: it is not more than 0.67. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 4 hours).
Weigh accurately about 0.2 g of Potassium Can-renoate, previously dried, dissolve in 75 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Use a solution of saturated potas-sium chloride-acetic acid (100) as the internal liquid.). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 39.66 mg of C22H29KO4
Potassium Carbonate, when dried, contains not less than 99.
Potassium Carbonate occurs as white granules or powder. It is odorless. It is very soluble in water, and practically insoluble in ethanol (95). A solution of Potassium Carbonate (1 in 10) is alkaline. It is hygroscopic.
A solution of Potassium Carbonate (1 in 10) responds to Qualitative Tests <1.09> for potassium salt and for carbonate.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Carbonate in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Dissolve 1.0 g of Potassium ‌Carbonate in 2 mL of water and 6 mL of dilute hydrochloric acid, and evaporate to dryness on a water bath. Dissolve the residue in 35 mL of water and 2 mL of dilute acetic acid, di-lute with water to 50 mL, and perform the test using this so-lution as the test solution. Prepare the control solution as follows: evaporate 6 mL of dilute hydrochloric acid on a water bath to dryness, add 2 mL of dilute acetic acid and 2.0 mL of Standard Lead Solution, and dilute with water to 50 mL (not more than 20 ppm).‌ Sodium—Dissolve 1.0 g of Potassium Carbonate in 20 mL of water, and perform the test as directed under Flame Coloration Test <1.04> (1): no persisting yellow color is produced. Arsenic <1.11>—Prepare the test solution with 0.5 g of Potassium Carbonate, according to Method 1, and per-form the test (not more than 4 ppm). Loss on drying <2.41> Not more than 1.0z (3 g, 1809C, 4 hours).
Dissolve about 1.5 g of Potassium Carbonate, previ-ously dried and accurately weighed, in 25 mL of water, titrate with 0.5 mol/L sulfuric acid VS until the blue color of the solution changes to yellow-green, boil cautiously, then cool, and titrate <2.50> until a greenish yellow color develops (indicator: 2 drops of bromocresol green TS). Each mL of 0.5 mol/L sulfuric acid VS ＝ 69.11 mg of K2CO3
Potassium Chloride, when dried, contains not less than 99.
Potassium Chloride occurs as colorless or white crystals or crystalline powder. It is odorless, and has a saline taste. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. A solution of Potassium Chloride (1 in 10) is neutral.
A solution of Potassium Chloride (1 in 50) responds to Qualitative Tests <1.09> for potassium salt and for chloride.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Chloride in 5 mL of water: the solution is clear and colorless. Acidity and alkalinity—Dissolve 5.0 g of Potassium Chloride in 50 mL of freshly boiled and cooled water, and add 3 drops of phenolphthalein TS: no red color develops. Then add 0.50 mL of 0.01 mol/L sodium hydroxide VS: a red color develops. Bromide—Dissolve 1.0 g of Potassium Chloride in water to make 100 mL. To 5 mL of the solution add 3 drops of dilute hydrochloric acid and 1 mL of chloroform, and add 3 drops of sodium toluensulfonchloramide TS dropwise while shaking: no yellow to yellow-red color develops in the chloroform layer. Iodide—Dissolve 0.5 g of Potassium Chloride in 10 mL of water, add 3 drops of iron (III) chloride TS and 1 mL of chloroform, shake, allow to stand for 30 minutes, and shake again: no red-purple to purple color develops in the chloroform layer. Heavy metals <1.07>—Proceed with 4.0 g of Potas- sium Chloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 5 ppm). Calcium and magnesium—Dissolve 0.20 g of Potas-sium Chloride in 20 mL of water, add 2 mL of ammonia TS, 2 mL of ammonium oxalate TS and 2 mL of disodium hydrogenphosphate TS, and then allow to stand for 5 minutes: no turbidity is produced. Sodium—Dissolve 1.0 g of Potassium Chloride in 20 mL of water, and perform Flame Coloration Test <1.04> (1): no persistent, yellow color develops. Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Chloride according to Method 1, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1309C, 2 hours).
Weigh accurately about 0.2 g of Potassium Chlo-ride, previously dried, dissolve in 50 mL of water, and titrate <2.50> with 0.1 mol/L silver nitrate VS while shaking vigor- ously (indicator: 3 drops of fluorescein sodium TS). Each mL of 0.1 mol/L silver nitrate VS ＝ 7.455 mg of KCl
Potassium Clavulanate is the potassium salt of a substance having b-lactamase inhibiting activity pro-duced by the growth of Streptomyces clavuligerus. It contains not less than 810 mg (potency) and not more than 860 mg (potency) per mg, calculated on the anhydrous basis.
Potassium Clavulanate occurs as a white to light yellow-white, crystalline powder. It is very soluble in water, soluble in methanol, and slightly soluble in ethanol (95). It is hygroscopic.
(1) To 1 mL of a solution of Potassium Clavulanate (1 in 50,000) add 5 mL of imidazole TS, and warm in a water bath at 309C for 12 minutes. After cooling, determine the absorption spectrum of this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Po-tassium Clavulanate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Potassium Clavulanate responds to Qualitative Tests <1.09> (1) for potassium salt. D Optical rotation <2.49> [a]20: ＋53 – ＋639(0.5 g calculated on the anhydrous basis, water, 50 mL, 100 mm).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Potassium Clavulanate according to Method 2, and perform the test. Prepare the control solution with 4.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Clavulanate according to Method 3, and per-form the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Potassium Clavulanate in 10 mL of the mobile phase A, and use this so-lution as the sample solution. Pipet 1 mL of the sample solu-tion, add the mobile phase A to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> acid and amoxycillin are eluted in this order with the resolu-tion between these peaks being not less than 8 and the num-ber of theoretical plates of the peak of clavulanic acid is not less than 2500. System repeatability: When the test is repeated 3 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of clavulanic acid is not more than 2.0z. Water <2.48> Not more than 1.5z (5 g, volumetric titra-tion, direct titration).
Weigh accurately an amount of Potassium Clavu-lanate and Lithium Clavulanate RS, equivalent to about 12.5 mg (potency), dissolve each in 30 mL of water, add exactly 5 mL of the internal standard solution and water to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of clavularic acid to that of the internal stand-ard. Amount [ mg (potency)] of clavularic acid (C H NO ) according to the following conditions, and determine each peak area by the automatic integration method: the area of ＝ MS × QT/QS × 1000 8 9 5 each peak other than clavulanic acid obtained from the sam-ple solution is not larger than the peak area of clavulanic acid from the standard solution, and the total area of the peaks other than clavulanic acid from the sample solution is not larger than 2 times of the peak area of clavulanic acid from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: Adjust the pH of 0.05 mol/L sodium dihydrogen phosphate TS to 4.0 with phosphoric acid. Mobile phase B: A mixture of the mobile phase A and methanol (1:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 4 4 – 15 100 → 0 0 → 100 15 – 25 Flow rate: 1.0 mL per minute. Time span of measurement: About 6 times as long as the retention time of clavulanic acid. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add the mobile phase A to make exactly 10 mL. Confirm that the peak area of clavulanic acid obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: Dissolve 10 mg each of Potassium Clavulanate and amoxycillin hydrate in 100 mL of the mo-bile phase A. When the procedure is run with 20 mL of this solution under the above operating conditions, clavulanic MS: Amount [mg (potency)] of Lithium Clavulanate RS taken Internal standard solution—Dissolve 0.3 g of sulfanilamide in 30 mL of methanol, and add water to make 100 mL. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.36 g of sodium acetate trihydrate in 900 mL of water, adjust to pH 4.5 with diluted acetic acid (31) (2 in 5), and add 30 mL of methanol and water to make 1000 mL. Flow rate: Adjust so that the retention time of clavularic acid is about 6 minutes. System suitability— System performance: When the procedure is run with 5 mL of the standard solution under the above operating condi-tions, clavularic acid and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratios of the peak area of clavularic acid to that of the internal stand-ard is not more than 1.0z.
Potassium Guaiacolsulfonate occurs as white crystals or crystalline powder. It is odorless or has a slight, characteristic odor and a slightly bitter taste. It is freely soluble in water and in formic acid, soluble in methanol, and practically insoluble in ethanol (95), in acetic anhydride and in diethyl ether.
(1) To 10 mL of a solution of Potassium Guaiacolsulfonate (1 in 100) add 2 drops of iron (III) chlo-ride TS: a blue-purple color develops. Dissolve 0.25 g of Potassium Guaiacolsulfonate in water to make 500 mL, and to 10 mL of this solution add phosphate buffer solution (pH 7.0) to make 100 mL. Deter-mine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. A solution of Potassium Guaiacolsulfonate (1 in 10) responds to Qualitative Tests <1.09> for potassium salt. pH <2.54> Dissolve 1.0 g of Potassium Guaiacolsulfonate in 20 mL of water: the pH of the solution is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Guaiacolsulfonate in 20 mL of water: the solu-tion is clear and colorless. Sulfate <1.14>—Perform the test with 0.8 g of Potas- sium Guaiacolsulfonate. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.030z). Heavy metals <1.07>—Proceed with 1.0 g of Potas- sium Guaiacolsulfonate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Potassium Guaiacolsulfonate according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.20 g of Potassium Guaiacolsulfonate in 200 mL of mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area by the automatic integration method: the total area of peaks other than potassium guaiacolsulfonate obtained from the sample solution is not larger than the peak area of potas-sium guaiacolsulfonate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 279 nm). Column: A stainless steel column 4 mm in inside diameter and 20 to 25 cm in length, packed with dimethylamino-propylsilanized silica gel for liquid chromatography (5 to 10 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of 0.05 mol/L potassium dihy-drogenphosphate TS and methanol (20:1). Flow rate: Adjust so that the retention time of potassium guaiacolsulfonate is about 10 minutes. Selection of column: Weigh 50 mg each of potassium guaiacolsulfonate and guaiacol, and dissolve in 50 mL of the mobile phase. Proceed with 5 mL of this solution under the above operating conditions, and calculate the resolution. Use a column giving elution of guaiacol and potassium guaiacolsulfonate in this order with the resolution of these peaks being not less than 4. Detection sensitivity: Adjust the sensitivity so that the peak height of potassium guaiacolsulfonate from 5 mL of the standard solution is not less than 10 mm. Time span of measurement: About twice as long as the retention time of potassium guaiacolsulfonate. Water <2.48> 3.0 – 4.5z (0.3 g, volumetric titration, direct titration).
Weigh accurately about 0.3 g of Potassium Guaia-colsulfonate, dissolve in 2.0 mL of formic acid, add 50 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L per-chloric acid VS (potentiometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 24.23 mg of C7H7KO5S
Potassium Hydroxide contains not less than 85.
Potassium Hydroxide occurs as white fused masses, in small pellets, in flakes, in sticks and in other forms. It is hard and brittle, and shows a crystalline fracture. It is freely soluble in water and in ethanol (95), and practi- cally insoluble in diethyl ether. It rapidly absorbs carbon dioxide in air. It deliquesces in the presence of moisture.
(1) A solution of Potassium Hydroxide (1 in 500) is alkaline. (2) A solution of Potassium Hydroxide (1 in 25) re-sponds to Qualitative Tests <1.09> for potassium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Hydroxide in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 2.0 g of Potassium Hy- ‌droxide in water to make 100 mL. To 25 mL of the solution add 8 mL of dilute nitric acid and water to make 50 mL. Per-form the test using this solution as the test solution. Prepare the control solution with 0.7 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.050z). Heavy metals <1.07>—Dissolve 1.0 g of Potassium Hydroxide in 5 mL of water, add 7 mL of dilute hydrochlo-ric acid, and evaporate on a water bath to dryness. Dissolve the residue in 35 mL of water, 2 mL of dilute acetic acid and 1 drop of ammonia TS, add water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: evaporate 7 mL of dilute hy-drochloric acid on a water bath to dryness, dissolve the residue in 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solution, and add water to make 50 mL (not more than 30 ppm). Sodium—Dissolve 0.10 g of Potassium Hydroxide in 10 mL of dilute hydrochloric acid, and perform the test as directed under Flame Coloration Test <1.04> (1): no persis-tent yellow color develops. Potassium carbonate—The amount of potassium car-bonate (K2CO3: 138.21) is not more than 2.0z when calcu-lated by the following equation using B (mL) obtained in the Assay. Amount of potassium carbonate (mg) ＝ 138.21 × B
Weigh accurately about 1.5 g of Potassium Hydrox-ide, and dissolve in 40 mL of freshly boiled and cooled water. Cool the solution to 159C, add 2 drops of phenol-phthalein TS, and titrate <2.50> with 0.5 mol/L sulfuric acid VS until the red color of the solution disappears. Record the amount A (mL) of 0.5 mol/L sulfuric acid VS consumed, then add 2 drops of methyl orange TS, and titrate <2.50> again with 0.5 mol/L sulfuric acid VS until the solution changes to a persistent light red color. Record the amount B (mL) of 0.5 mol/L sulfuric acid VS consumed. Calculate the amount potassium hydroxide (KOH) from the amount, A (mL) － B (mL). Each mL of 0.5 mol/L sulfuric acid VS ＝ 56.11 mg of KOH
Potassium Iodide, when dried, contains not less than 99.
Potassium Iodide occurs as colorless or white crystals, or a white crystalline powder. It is very soluble in water, soluble in ethanol (95), and practically insoluble in diethyl ether. It is slightly deliquescent in moist air.
A solution of Potassium Iodide (1 in 20) responds to Qualitative Tests <1.09> for potassium salt and for iodide.
(1) Clarity and color of solution—Dissolve 1.0 g of Potassium Iodide in 2 mL of water: the solution is clear and colorless. Alkalinity—Dissolve 1.0 g of Potassium Iodide in 10 mL of freshly boiled and cooled water, and add 0.50 mL of 0.005 mol/L sulfuric acid VS and 1 drop of phenolphthalein TS: no color develops. Chloride, bromide and thiosulfate—Dissolve 0.20 g of Potassium Iodide in 5 mL of ammonia TS, add 15.0 mL of 0.1 mol/L silver nitrate VS, shake for 2 to 3 minutes, and filter. To 10 mL of the filtrate, add 15 mL of dilute nitric acid: no brown color develops. The solution has no more turbidity than that of the following control solution. Control solution: To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 2.5 mL of ammonia TS, and 7.5 mL of 0.1 mol/L silver nitrate VS and 15 mL of dilute nitric acid. Nitrate, nitrite and ammonium—Place 1.0 g of Po-tassium Iodide in a 40-mL test tube, and add 5 mL of water, 5 mL of sodium hydroxide TS and 0.2 g of aluminum wire. Insert the absorbent cotton in the mouth of the test tube, and place a piece of moistened red litmus paper on it. Heat the test tube carefully on a water bath for 15 minutes: the gas evolved does not turn red litmus paper to blue. Cyanide—Dissolve 0.5 g of Potassium Iodide in 10 mL of water. To 5 mL of this solution add 1 drop of iron (II) sulfate TS and 2 mL of sodium hydroxide TS, warm, then add 4 mL of hydrochloric acid: no green color develops. Iodate—Dissolve 0.5 g of Potassium Iodide in 10 mL of freshly boiled and cooled water, and add 2 drops of dilute sulfuric acid and 1 drop of starch TS: no blue color develops immediately. Heavy metals <1.07>—Proceed with 2.0 g of Potas- sium Iodide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Barium—Dissolve 0.5 g of Potassium Iodide in 10 mL of water, add 1 mL of dilute sulfuric acid, and allow to stand for 5 minutes: no turbidity is produced. Sodium—Dissolve 1.0 g of Potassium Iodide in 10 mL of water, and perform Flame Coloration Test <1.04> (1): a yellow color develops, but does not persist. Arsenic <1.11>—Prepare the test solution with 0.40 g of Potassium Iodide according to Method 1, and perform the test (not more than 5 ppm). Loss on drying <2.41> Not more than 1.0z (2 g, 1059C, 4 hours).
Weigh accurately about 0.5 g of Potassium Iodide, previously dried, in an iodine flask, dissolve in 10 mL of water, add 35 mL of hydrochloric acid and 5 mL of chlo-roform, and titrate <2.50> with 0.05 mol/L potassium iodate VS with shaking until the red-purple color of the chloroform layer disappears. The end point is reached when the red-pur-ple color does not reappear in the chloroform layer within 5 minutes after the layer has been decolorized. Each mL of 0.05 mol/L potassium iodate VS ＝ 16.60 mg of KI
Potassium Permanganate occurs as dark pur-ple crystals and has a metallic luster. It is soluble in water. A solution of Potassium Permanganate (1 in 1000) has a slightly sweet, astringent taste.
A solution of Potassium Permanganate (1 in 100) responds to Qualitative Tests <1.09> for permanganate.
(1) Water-insoluble substances—Dissolve 2.0 g of Potassium Permanganate, previously powdered, in 200 mL of water. Filter the insoluble substances through a tared glass filter (G4), wash with water until the last washing shows no color, and dry at 1059C for 2 hours: the mass of the residue is not more than 4 mg. (2) Arsenic <1.11>—Dissolve 0.40 g of Potassium Per- manganate in 10 mL of water, add 1 mL of sulfuric acid, add hydrogen peroxide (30) dropwise until the solution remains colorless, and evaporate on a sand bath nearly to dryness. Dissolve the residue in 5 mL of water, and perform the test with this solution as the test solution: the color pro-duced is not more intense than the following color standard. Color standard: To 10 mL of water add 1 mL of sulfuric acid and the same volume of hydrogen peroxide (30) as used for the preparation of the test solution. Evaporate the solu-tion on a sand bath nearly to dryness, add 2.0 mL of Stand-ard Arsenic Solution and water to make 5 mL, and carry out the test with this solution in the same manner as the test solu- tion (not more than 5 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, silica gel, 18 hours).
Weigh accurately about 0.6 g of Potassium Perman-ganate, previously dried, dissolve in water to make exactly 200 mL, and use this solution as the sample solution. Pipet 25 mL of 0.05 mol/L oxalic acid VS into a 500-mL conical flask, add 200 mL of diluted sulfuric acid (1 in 20), and keep at a temperature between 309C and 359C. Transfer the sam-ple solution to a buret. Add quickly 23 mL of the sample so-lution from the buret to the flask while shaking gently, and then allow the flask to stand until the red color disappears. Potassium Sulfate
Potassium Sulfate, when dried, contains not less than 99.
Potassium Sulfate occurs as colorless crystals or a white, crystalline powder. It has a slightly saline, some-what bitter taste. It is soluble in water and practically insoluble in ethanol (95).
A solution of Potassium Sulfate (1 in 20) re-sponds to Qualitative Tests <1.09> for potassium salt and for sulfate.
(1) Clarity and color of solution, and acid or alkali—Dissolve 1.0 g of Potassium Sulfate in 20 mL of water: the solution is clear, colorless and neutral. Chloride <1.03>—Perform the test with 0.5 g of Potassium Sulfate. Prepare the control solution with 0.40 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.028z). Heavy metals <1.07>—Proceed with 2.0 g of Potas- sium Sulfate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Sodium—Dissolve 1.0 g of Potassium Sulfate in 20 mL of water, and perform the test as directed under Flame Coloration Test <1.04> (1): no persistent yellow color devel-ops. Arsenic <1.11>—Prepare the test solution with 0.40 g of Potassium Sulfate according to Method 1, and perform the test (not more than 5 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, 1109C, 4 hours).
Weigh accurately about 0.5 g of Potassium Sulfate, previously dried, boil with 200 mL of water and 1.0 mL of hydrochloric acid, and add gradually 8 mL of boiling barium chloride TS. Heat the mixture on a water bath for 1 hour, collect the precipitate, and wash the precipitate with water until the last washing shows no opalescence on the addition of silver nitrate TS. Dry, heat strongly to constant mass be-tween 5009C and 6009C by raising the temperature gradu-ally, and weigh as barium sulfate (BaSO4: 233.39). Amount (mg) of potassium sulfate (K SO ) Warm the mixture to a temperature between 559C and 609C, 2 4 and continue the titration <2.50> slowly until the red color persists for 30 seconds. Each mL of 0.05 mol/L oxalic acid VS ＝ 3.161 mg of KMnO4
Prasterone Sodium Sulfate Hydrate occurs as white crystals or crystalline powder. It is odorless. It is soluble in methanol, sparingly soluble in water and in ethanol (95), and practically insoluble in acetone and in diethyl ether. The pH of a solution of 1.0 g of Prasterone Sodium Sul-fate Hydrate in 200 mL of water is between 4.5 and 6.5. Melting point: about 1609C (with decomposition, after drying).
(1) Dissolve 0.01 g of Prasterone Sodium Sulfate Hydrate in 4 mL of ethanol (95), add 2 mL of 1,3-dinitrobenzene TS and 2 mL of a solution of sodium hydrox-ide (1 in 8): a red-purple color develops, and gradually changes to brown. To 10 mL of a solution of Prasterone Sodium Sulfate Heavy metals <1.07>—Proceed with 2.0 g of Praster-one Sodium Sulfate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 0.10 g of Prasterone Sodium Sulfate Hydrate in 10 mL of methanol, and use this solution as the sample solution. Pipet 1 mL of the sample so-lution, add methanol to make exactly 200 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of chloroform, methanol and water (75:22:3) to a distance of about 10 cm, and air-dry the plate. Spray evenly a mixture of sulfuric acid and ethanol (95) (1:1) on the plate, and heat the plate at 809C for 5 minutes: the spots other than the princi-pal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> 8.0 – 9.0z (0.5 g, in vacuum, phos-phorus (V) oxide, 609C, 3 hours).
(1) Clarity and color of solution—Dissolve 0.25 g of Prasterone Sodium Sulfate Hydrate in 50 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Prasterone So- dium Sulfate Hydrate in 20 mL of acetone and 20 mL of water, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.011z). Sulfate <1.14>—To 1.2 g of Prasterone Sodium Sul-
Weigh accurately about 0.25 g of Prasterone So-dium Sulfate Hydrate, dissolve in 30 mL of water. Apply this solution to a chromatographic column 10 mm in inside diameter, previously prepared by pouring 5 mL of strongly acidic ion-exchange resin (H type) for column chromatogra-phy, and elute at the rate of 4 mL per minute. Wash the chromatographic column with 100 mL of water, combine the washings with above effluent solution, and titrate <2.50> with 0.05 mol/L sodium hydroxide VS (potentiometric titra-tion). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L sodium hydroxide VS Hydrate (1 in 200) add 0.5 mL of bromine TS: the color of bromine TS immediately disappears. ＝ 19.52 mg of C19 H27 NaO5S Determine the infrared absorption spectrum of Prasterone Sodium Sulfate Hydrate as directed in the potas-sium bromide disk method under the Infrared Spectro-photometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Prasterone Sodium Sulfate Hydrate (1 in 200) responds to Qualitative Tests <1.09> for sodium salt. D Optical rotation <2.49> [a]20: ＋10.7 – ＋12.19(0.73 g cal-culated on the dried basis, methanol, 20 mL, 100 mm). Purity (1) Clarity and color of solution—Dissolve 0.25 g of Prasterone Sodium Sulfate Hydrate in 50 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 1.0 g of Prasterone So- dium Sulfate Hydrate in 20 mL of acetone and 20 mL of water, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution as follows: to 0.30 mL of 0.01 mol/L hydrochloric acid VS add 20 mL of acetone, 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.011z). Sulfate <1.14>—To 1.2 g of Prasterone Sodium Sul-
Pravastatin Sodium contains not less than 98.
Pravastatin Sodium occurs as a white to yel-lowish white, powder or crystalline powder. It is freely soluble in water and in methanol, and soluble in ethanol (99.5). It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Pravastatin Sodium (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pravastatin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it ex-hibits absorption at the wave numbers of about 2970 cm－1, 2880 cm－1, 1727 cm－1 and 1578 cm－1. Dissolve 50 mg of Pravastatin Sodium in 5 mL of methanol, and use this solution as the sample solution. Separately, dissolve 24 mg of Pravastatin 1,1,3,3-Tetra-methylbutylammonium RS in 2 mL of methanol, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 2 mL each of the sample solution and stand-ard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate, ethanol (99.5) and acetic acid (100) (80:16:1) to a distance of about 8 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the color tone and the Rf value of the principal spot ob-tained from the sample solution are not different with them of the spot from the standard solution. A solution of Pravastatin Sodium (1 in 10) responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋153 – ＋1599(0.1 g calcu-lated on the anhydrous and residual solvent-free basis, water, 20 mL, 100 mm). pH <2.54> The pH of a solution obtained by dissolving 1.0 g of Pravastatin Sodium in 20 mL of freshly boiled and cooled water is between 7.2 and 8.2.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Pravastatin Sodium according to Method 2, and perform the test. Prepare the control solution with 1.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 0.10 g of Pravastatin Sodium in 100 mL of a mixture of water and methanol (11:9), and use this solution as the sample solution. Pipet 10 mL of the sample solution, add the mixture of water and methanol (11:9) to make exactly 100 mL. Pipet 5 mL of this solution, add the mixture of water and methanol (11:9) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic inte-gration method: the area of the peak other than pravastatin from the sample solution is not larger than 1/5 times the peak area of pravastatin from the standard solution, and the total area of the peaks other than pravastatin from the sam-ple solution is not larger than the peak area of pravastatin from the standard solution. Keep the sample solution and standard solution at not over than159C. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.5 times as long as the retention time of pravastatin, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 5 mL of the standard solution add a mixture of water and methanol (11:9) to make exactly 50 mL. Confirm that the peak area of pravastatin obtained with 10 mL of this solution is equivalent to 7 to 13z of that obtained with 10 mL of the standard so-lution. System performance: Dissolve 5 mg of pravastatin sodium in 50 mL of the mixture of water and methanol (11:9). When the procedure is run with 10 mL of this solution under the above operating conditions, the number of theoretical plates and the symmetry factor of the peak of pravastatin are not less than 3500 and not more than 1.6, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of pravastatin is not more than 2.0z. Water <2.48> Not more than 4.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately about 0.1 g of Pravastatin So-dium, and dissolve in a mixture of water and methanol (11:9) to make exactly 100 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution and the mixture of water and methanol (11:9) to make 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 30 mg of Pravastatin 1,1,3,3-Tetramethyl-butylammonium RS (previously determine the water <2.48> with 0.5 g by direct titration in volumetric titration) dissolve in the mixture of water and methanol (11:9) to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 10 mL of the internal standard solution, add a mixture of water and methanol (11:9) to make 100 mL, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of pravastatin to that of the internal standard. Amount (mg) of pravastatin sodium (C23H35NaO7) ＝ MS × QT/QS × 4 × 1.052 MS: Amount (mg) of pravastatin in taken Pravastatin 1,1,3,3-Tetramethylbutylammonium RS, calculated on the anhydrous basis Internal standard solution—A solution of ethyl parahydrox-ybenzoate in the mixture of water and methanol (11:9) (3 in 4000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 238 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water, methanol, acetic acid (100) and triethylamine (550:450:1:1). Flow rate: Adjust so that the retention time of pravastatin is about 21 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and pravastatin are eluted in this order with the resolution between these peaks being not less than 10. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 50 50 – 75 50 → 0 50 → 100 ‌the peak area of pravastatin to that of the internal standard is not more than 1.0z.
Precipitated Calcium Carbonate occurs as a white, fine crystalline powder. It is odorless and tasteless. It is practically insoluble in water, but its solubility in water is increased in the presence of carbon dioxide. It is practically insoluble in ethanol (95) and in diethyl ether. It dissolves with effervescence in dilute acetic acid, in dilute hydrochloric acid and in dilute nitric acid.
(1) Dissolve 0.5 g of Precipitated Calcium Carbonate in 10 mL of dilute hydrochloric acid, boil, then cool, and neutralize with ammonia TS: the solution responds to Qualitative Tests <1.09> for calcium salt. (2) Precipitated Calcium Carbonate responds to Qualita-tive Tests <1.09> (1) for carbonate.
(1) Acid-insoluble substances—To 5.0 g of Precipitated Calcium Carbonate add 50 mL of water, then add 20 mL of hydrochloric acid dropwise with stirring, boil for 5 minutes, cool, add water to make 200 mL, and filter through filter paper for quantitative analysis. Wash the residue until the last washing shows no turbidity with silver nitrate TS, and ignite the residue together with the filter paper: the mass of the residue is not more than 10.0 mg. Heavy metals <1.07>—Mix 2.0 g of Precipitated Cal- cium Carbonate with 5 mL of water, add slowly 6 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 50 mL of water, and filter. To 25 mL of the filtrate add 2 mL of dilute acetic acid, 1 drop of ammonia TS and water to make 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: evaporate 3 mL of hydro-chloric acid on a water bath to dryness, and add 2.0 mL of Standard Lead Solution, 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Barium—Mix 1.0 g of Precipitated Calcium Car-bonate with 10 mL of water, add dropwise 4 mL of hydro-chloric acid with stirring, boil for 5 minutes, cool, add water to make 40 mL, and filter. With the filtrate, perform the test tion, and perform the test (not more than 5 ppm). Loss on drying <2.41> Not more than 1.0z (1 g, 1809C, 4 hours).
Weigh accurately about 0.12 g of Precipitated Cal-cium Carbonate, previously dried, and dissolve in 20 mL of water and 3 mL of dilute hydrochloric acid. Add 80 mL of water, 15 mL of a solution of potassium hydroxide (1 in 10) and 0.05 g of NN indicator, and titrate <2.50> immediately with 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS until the color of the solution changes from red-purple to blue. Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 5.005 mg of CaCO3
Prednisolone Sodium Phosphate occurs as a white to pale yellow powder. It is freely soluble in water, soluble in methanol, and prac-tically insoluble in ethanol (99.5). It is hygroscopic.
(1) Moisten 1.0 g of Prednisolone Sodium Phosphate with a small amount of sulfuric acid, and gradu-ally heat to incinerate. After cooling, dissolve the residue in 10 mL of dilute nitric acid, and heat in a water bath for 30 minutes. After cooling, filter if necessary. This solution re-sponds to Qualitative Tests <1.09> for phosphate. Dissolve 2 mg of Prednisolone Sodium Phosphate in 2 mL of sulfuric acid, and allow to stand for 2 minutes: a deep red color, without fluorescence, develops. Determine the absorption spectrum of a solution of Prednisolone Sodium Phosphate (1 in 50,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pred-nisolone Sodium Phosphate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. The solution obtained in (1) responds to Qualitative Tests <1.09> for sodium salt. D Optical rotation <2.49> [a]20: ＋96 – ＋1039(1 g calculated on the anhydrous basis, phosphate buffer solution (pH 7.0), 100 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Prednisolone Sodium Phos-phate in 100 mL of water: the pH of the solution is between 7.5 and 9.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Prednisolone Sodium Phosphate in 10 mL of water: the solution is clear and not more colored than the following control solution. Control solution: To a mixture of 3.0 mL of Cobalt (II) Chloride CS, 3.0 mL of Iron (III) Chloride CS and 2.4 mL of Copper (II) Sulfate CS add diluted hydrochloric acid (1 in 40) to make 10 mL. To 2.5 mL of this solution add diluted hydrochloric acid (1 in 40) to make 100 mL. Heavy metals <1.07>—Proceed with 0.5 g of Pred- nisolone Sodium Phosphate according to Method 3, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 40 ppm). Free phosphoric acid—Weigh accurately about 0.25 g of Prednisolone Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Pipet 5 mL each of the sample solution and Phos-phoric Acid Standard Solution, add 2.5 mL of hexaammo-nium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphtol-4-sulfonic acid TS, shake, add water to make ex-actly 25 mL, and allow to stand at 20 ± 19C for 30 minutes. Perform the test with these solutions as directed under Ultra-violet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of water in the same manner as the blank. Determine the absorbances, AT and AS, of each solu-tion from the sample solution and standard solution at 740 nm: the content of free phosphoric acid is not more than 1.0z. Content (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Prednisolone Sodium Phosphate taken, calculated on the anhydrous basis Related substances—Dissolve 10 mg of Prednisolone Sodium Phosphate in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 2 mL of the sam-ple solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions. Determine each peak area of both solutions by the automatic integration method: the area of each peak other than prednisolone phos-phate obtained from the sample solution is not larger than 1.5 times the peak area of prednisolone phosphate from the standard solution, and the total area of the peaks other than prednisolone phosphate from the sample solution is not larger than 2.5 times the peak area of prednisolone phos-phate from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 245 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 10 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 6.80 g of potassium dihydrogen phosphate in water to make 1000 mL, and adjust the pH to 2.5 with phosphoric acid. To 1000 mL of this solution add 250 mL of acetonitrile. Flow rate: Adjust so that the retention time of predniso-lone phosphate is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of prednisolone phosphate. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add the mobile phase to make exactly 50 mL. Confirm that the peak area of prednisolone phosphate ob-tained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of prednisolone phosphate are not less than 3000 and not more than 2.0, respectively. ‌System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of prednisolone phosphate is not more than 2.0z.‌ Water <2.48> Not more than 8.0z (0.1 g, volumetric titra-tion, direct titration).
Weigh accurately about 0.1 g of Prednisolone So-dium Phosphate, and dissolve in water to make exactly 100 mL. Pipet 2 mL of this solution, add 1 mL of alkaline phos-phatase TS, and allow to stand for 2 hours with occasional shaking. To this solution add exactly 20 mL of 1-octanol, and shake vigorously. Centrifuge this solution, pipet 10 mL of the 1-octanol layer, add 1-octanol to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 25 mg of Prednisolone RS, previ-ously dried at 1059C for 3 hours, and dissolve in 1-octanol to make exactly 100 mL. Pipet 6 mL of this solution, add a so-lution prepared by adding 1 mL of alkaline phosphatase TS to 2 mL water and being allowed to stand for 2 hours with occasional gentle shaking, add exactly 14 mL of 1-octanol, and shake vigorously. Proceed in the same manner as the sample solution to make the standard solution. Perform the test with the sample solution and standard solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using 1-octanol as the blank, and determine the absorbances, AT and AS, at 245 nm. Amount (mg) of prednisolone sodium phosphate (C21H27Na2O8P) ＝ MS × AT/AS × 3 × 1.344 MS: Amount (mg) of Prednisolone RS taken
Purified Sodium Hyaluronate is the sodium salt of glycosaminoglycans composed of disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine ob-tained from cockscomb or microorganisms. It contains not less than 90.
Purified Sodium Hyaluronate occurs as white, powder, granules or fibrous masses. It is sparingly soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Purified Sodium Hyaluronate, previously dried, as directed in the potassium bromide disk method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. (2) A solution of Purified Sodium Hyaluronate (1 in 1000) responds to Qualitative Tests <1.09> (1) for sodium salt. Viscosity <2.53> Weigh accurately an amount of Purified Sodium Hyaluronate so that the downflowing time of its so-lution in 100 mL of 0.2 mol/L sodium chloride TS is 2.0 to 2.4 times longer than that of 0.2 mol/L sodium chloride TS, dissolve in 0.2 mol/L sodium chloride TS to make exactly 100 mL, and use this solution as the sample solution (1). Pipet 16 mL, 12 mL and 8 mL of the sample solution (1), to each add 0.2 mol/L sodium chloride TS to make exactly 20 mL, and use these solutions as the sample solutions (2), (3) and (4), respectively. Perform the test with the sample solu-tions (1), (2), (3) and (4) as directed under Method 1 at 30 ± 0.19C using an Ubbelohde-type viscometer in which the downflowing time for 0.2 mol/L sodium chloride TS is 200 to 300 seconds: the intrinsic viscosity calculated on the dried basis is between 10.0 dL/g and 24.9 dL/g or between 25.0 dL/g and 55.0 dL/g.
(1) Clarity and color of solution—Dissolve 0.10 g of Purified Sodium Hyaluronate in 10 mL of water: the solu-tion is clear and colorless. Chloride <1.03>—Dissolve 0.20 g of Purified Sodium Hyaluronate in 15 mL of water, add 6 mL of dilute nitric acid, and heat on a water bath for 30 minutes. After cooling, add water to make 50 mL. Perform the test using this solu-tion as the test solution. Prepare the control solution with 0.70 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.124z). Heavy metals <1.07>—Proceed with 1.0 g of Purified Sodium Hyaluronate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Protein—Weigh accurately about 20 mg of Purified Sodium Hyaluronate, calculated on the dried basis, dissolve in 1.0 mL of dilute sodium hydroxide TS, and use this solu-tion as the sample solution. Separately, weigh accurately about 10 mg of bovine serum albumin, dissolve in dilute so- (C14 H20NNaO 11)n dium hydroxide TS to make exactly 1000 mL, and use this solution as the standard solution. To 1.0 mL each of the [9067-32-7] Purified Sodium Hyaluronate is the sodium salt of glycosaminoglycans composed of disaccharide units of D-glucuronic acid and N-acetyl-D-glucosamine ob-tained from cockscomb or microorganisms. It contains not less than 90.0z and not more than 105.5z of sodium hyaluronate [(C H NNaO ) ], sample solution and standard solution add 5.0 mL of alka-line copper TS (2), immediately stir, allow to stand at room temperature for 10 minutes, add 0.5 mL of diluted Folin's TS (1 in 2), immediately stir, and allow to stand at room temperature for 30 minutes. Perform the test with these solu-tions as directed under Ultraviolet-visible Spectrophotome-try <2.24>, using a solution, prepared with 1.0 mL of dilute calculated on the dried basis. 14 20 n sodium hydrochloride in the same manner, as the blank: the absorbance of the sample solution at 750 nm does not exceed It is composed of an average molecular mass of the sodium salt of hyaluronic acid between 500,000 and 1,490,000 or between 1,500,000 and 3,900,000. The average molecular mass of Purified Sodium Hyaluronate should be labeled. the absorbance of the standard solution (not more than 0.05z). Nucleic acid—Determine the absorbance of a solu-tion of 0.10 g Purified Sodium Hyaluronate in 50 mL of water as directed under Ultraviolet-visible Spectrophotome- ‌try <2.24>, using water as the blank: the absorbance at 260 nm is not more than 0.02. Other acidic mucopolysaccharides—(In the case of chicken-derived samples) Dissolve 0.25 g of Purified Sodium Hyaluronate in 100 mL of water, and use this solution as the sample solution. Immerse a cellulose acetate membrane 6 cm in length in 0.2 mol/L pyridine-formic acid buffer solution (pH 3.0). Take out the membrane and remove excessive buffer solution using a filter paper. Place the membrane in an electrophoresis vessel saturated with 0.2 mol/L pyridine-formic acid buffer solution (pH 3.0) and run at 0.5 mA/cm for 1 minute. Apply 2 mL of the sample solution to the mem-brane in an area 1 cm in width at 1.5 cm from the anode. Carry out electrophoresis at 0.5 mA/cm for 1 hour. After the electrophoresis, stain the membrane by immersing it in alcian blue staining solution for 10 to 20 minutes. After staining, decolorize sufficiently with diluted acetic acid (100) (3 in 100): no bands other than the principal band appears. Hemolytic streptococci—(In the case of microor-ganism-derived samples) Dissolve 0.5 g of Purified Sodium
Weigh accurately about 50 mg of Purified Sodium Hyaluronate, and dissolve in water to make exactly 50 mL. Pipet 1 mL of this solution, add water to make exactly 20 mL, and use this solution as the sample solution. Separately, weigh accurately about 20 mg of D-Glucuronolactone RS, previously dried (under reduced pressure not exceeding 0.67 kPa, silica gel, 24 hours), and dissolve in water to make ex-actly 100 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this solution as the standard solution. Pipet 1 mL each of the sample solution and standard solu-tion, gently add into the 5.0 mL of sodium tetraborate-sulfuric acid TS, previously cooled in ice water, stir while cooling, heat in a water bath for 10 minutes, and cool in ice water. To each solution add exactly 0.2 mL of carbazole TS, stir well, heat in a water bath for 15 minutes, and cool in ice water to room temperature. Determine the absorbances, AT and AS, of the sample solution and standard solution at 530 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using a solution, prepared with 1 mL of water in the same manner, as the blank. Hyaluronate in sterile isotonic sodium chloride solution to make exactly 100 mL. Take 0.5 mL of this solution, apply to 2 blood agar plates, respectively, using a Conradi stick, and Amount (mg) of sodium hyaluronate [(C ＝ MS × AT/AS × 2.279 14H 20NNaO 11)n] incubate at 379C for 48 hours: no hemolytic colonies appear, or if any, no streptococci are observed in the colony under a microscope. Hemolysis—(In the case of microorganism-derived samples) Dissolve 0.40 g of Purified Sodium Hyaluronate in sterile isotonic sodium chloride solution to make exactly 100 mL. To 0.5 mL of this solution add 0.5 mL of 1z blood sus-pension, mix, allow to stand at 379C for 2 hours, and, if nec-essary, centrifuge at 3000 revolutions per minute for 10 minutes: the erythrocytes precipitate and the supernatant liq-uid is clear as in a blank determination performed in the same manner using 0.5 mL of sterile isotonic sodium chlo-ride solution as the blank and 0.5 mL of sterile purified water as the positive control. Loss on drying <2.41> Not more than 15.0z (0.1 g, reduced pressure not exceeding 0.67 kPa, phosphorus (V) oxide, 609C, 5 hours). Microbial limit <4.05> The acceptance criteria of TAMC and TYMC are 102 CFU/g and 101 CFU/g, respectively. In the case of the sample of the labeled average molecular mass between 500,000 and 1,490,000, perform the test with 1 g, and of the labeled average molecular mass between 1,500,000 and 3,900,000, perform the test with 0.3 g. Average molecular mass In the case of the labeled average molecular mass of between 500,000 and 1,490,000. Calculate the average molecular mass of Purified Sodium Hyaluronate according to the following equation: it is be-tween 500,000 and 1,490,000. For [h], use the maximum vis-cosity under Viscosity. Ø[h] × 105 » 1 MS: Amount (mg) of D-Glucuronolactone RS taken
Containers—Tight containers. Storage—Light-resistant, at not exceeding 159C.
Rabeprazole Sodium occurs as a white to pale yellow-white powder. It is very soluble in water, and freely soluble in ethanol (99.5). It dissolves in 0.01 mol/L sodium hydroxide TS. It is hygroscopic. A solution of Rabeprazole Sodium (1 in 20) shows no opti-cal rotation. Rabeprazole Sodium shows crystal polymorphism.
(1) Determine the absorption spectrum of a solution of Rabeprazole Sodium in 0.01 mol/L sodium hy-droxide TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Rabeprazole Sodium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Rabeprazole Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Rabeprazole Sodium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. If any difference appears between the spectra, dissolve the sample, or the sample and the RS separately in ethanol (99.5), evaporate the ethanol at 409C, dry the residues in vacuum at 559C for 24 hours, and perform the test with the residues. A solution of Rabeprazole Sodium (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. sample solution is not larger than 4/5 times the peak area of rabeprazole from the standard solution, the area of the peak other than rabeprazole and the peak mentioned above from the sample solution is not larger than 1/10 times the peak area of rabeprazole from the standard solution, and the total area of the peaks other than rabeprazole from the sample so-lution is not larger than the peak area of rabeprazole from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of rabeprazole, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, and add a mixture of methanol and 0.01 mol/L so-dium hydroxide TS (3:2) to make exactly 100 mL. Confirm that the peak area of rabeprazole obtained with 10 mL of this solution is equivalent to 3.5 to 6.5z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of rabeprazole are not less than 3000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rabeprazole is not more than 2.0z. Loss on drying <2.41> Not more than 1.0z (1 g, in vacu-um, phosphorus (V) oxide, 24 hours. Take the sample to be tested while avoiding moisture absorption.).
(1) Heavy metals <1.07>—Proceed with 2.0 g of Rabeprazole Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand- internal standard. Amount (mg) of sodium rabeprazole (C ＝ MS × QT/QS 18H20N3NaO3S) ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Rabeprazole Sodium in 50 mL of a mixture of methanol and 0.01 mol/L sodium hydroxide TS (3:2), and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add a mix-ture of methanol and 0.01 mol/L sodium hydroxide TS (3:2) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak having the relative retention time of about 0.7 to rabeprazole obtained from the MS: Amount (mg) of Rabeprazole Sodium RS taken, cal-culated on the dried basis Internal standard solution—A solution of 1-amino-2-methyl-naphthalene in a mixture of methanol and 0.01 mol/L so-dium hydroxide TS (3:2) (1 in 250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 290 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ‌309C.‌ Mobile phase: A mixture of methanol and 0.05 mol/L phosphate buffer solution (pH 7.0) (3:2). Flow rate: Adjust so that the retention time of rabeprazole is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, rabeprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 4, and the symmetry factor of the peak of rabepra-zole is not more than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of rabeprazole to that of the internal standard is not more than 1.0z.
Take the sample to be tested while avoiding moisture absorption. Weigh accurately about 0.1 g each of Rabepra-zole Sodium and Rabeprazole Sodium RS (separately deter-mine the loss on drying <2.41> under the same conditions as Rabeprazole Sodium), dissolve each in a mixture of metha-nol and 0.01 mol/L sodium hydroxide TS (3:2) to make ex-actly 25 mL. Pipet 5 mL each of these solutions, add exactly 10 mL of the internal standard solution to each, then add a mixture of methanol and 0.01 mol/L sodium hydroxide TS (3:2) to make 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of rabeprazole to that of the Purity (1) Heavy metals <1.07>—Proceed with 2.0 g of Rabeprazole Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand- internal standard. Amount (mg) of sodium rabeprazole (C ＝ MS × QT/QS 18H20N3NaO3S) ard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 50 mg of Rabeprazole Sodium in 50 mL of a mixture of methanol and 0.01 mol/L sodium hydroxide TS (3:2), and use this solution as the sam-ple solution. Pipet 1 mL of the sample solution, add a mix-ture of methanol and 0.01 mol/L sodium hydroxide TS (3:2) to make exactly 100 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak having the relative retention time of about 0.7 to rabeprazole obtained from the MS: Amount (mg) of Rabeprazole Sodium RS taken, cal-culated on the dried basis Internal standard solution—A solution of 1-amino-2-methyl-naphthalene in a mixture of methanol and 0.01 mol/L so-dium hydroxide TS (3:2) (1 in 250). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 290 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about ‌309C.‌ Mobile phase: A mixture of methanol and 0.05 mol/L phosphate buffer solution (pH 7.0) (3:2). Flow rate: Adjust so that the retention time of rabeprazole is about 5 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, rabeprazole and the internal standard are eluted in this order with the resolution between these peaks being not less than 4, and the symmetry factor of the peak of rabepra-zole is not more than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of rabeprazole to that of the internal standard is not more than 1.0z.
Rosuvastatin Calcium contains not less than 97.
Rosuvastatin Calcium occurs as a white pow-der. It is freely soluble in acetonitrile, soluble in methanol, and slightly soluble in water and in ethanol (99.5). It is hygroscopic.
(1) Determine the absorption spectrum of a solution of Rosuvastatin Calcium in methanol (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry Amount (mg) of ritodrine hydrochloride (C ＝ MS × QT/QS × 4 17H21NO3.HCl) <2.24>, and compare the spectrum with the Reference Spec- trum or the spectrum of a solution of Rosuvastatin Calcium RS prepared in the same manner as the sample solution: MS: Amount (mg) of Ritodrine Hydrochloride RS taken Internal standard solution—A solution of methyl parahy-droxybenzoate in methanol (3 in 5000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 274 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 6.6 g of diammonium hydrogen phosphate and 1.1 g of sodium 1-heptanesulfonate in 700 mL of water, and add 300 mL of methanol. Adjust to pH 3.0 with phosphoric acid. Flow rate: Adjust so that the retention time of ritodrine is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, ritodrine and the internal standard are eluted in this order with the resolution between these peaks being not less than 3. System repeatability: When the test is repeated 6 times both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Rosuvastatin Calcium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Rosuvastatin Calcium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Rosuvastatin Calcium in a mixture of water and methanol (1:1) (1 in 125) responds to Qualitative Tests <1.09> (3) for calcium salt.
(1) Inorganic impurities (chloride)—Being speci-fied separately when the drug is granted approval based on the Law. Heavy metals <1.07>—Proceed with 1.0 g of Rosuvastatin Calcium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 20 ppm). Related substances—Conduct this procedure using light-resistant vessels. Use the sample solution obtained in the Assay as the sample solution. Separately, pipet 1 mL of the standard solution obtained in the Assay, add a mixture of water and acetonitrile (3:1) to make exactly 10 mL. Pipet 1 mL of this solution, add a mixture of water and aceto- nitrile (3:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions. Determine each peak area of related sub-stances, AT, in the sample solution and the peak area of rosuvastatin, AS, in the standard solution by the automatic integration method, and calculate the amount of the related substances by the following equation: the amount of related substance A having the relative retention time of about 0.90 to rosuvastatin is not more than 0.2z, the amount of related substance B (diastereomer) having the relative retention time of about 1.1 is not more than 0.5z, the amount of related substance C having the relative retention time of about 1.5 is not more than 0.7z, the amount of related substance D hav-ing the relative retention time of about 1.7 is not more than 0.15z, and each amount of other related substance is not more than 0.1z. Furthermore the total amount of the related substances is not more than 1.1z. For the area of the peak of related substance C, multiply the correction factor 1.4. Amount (z) of related substance ＝ MS/MT × AT/AS × 1/5 MS: Amount (mg) of Rosuvastatin Calcium RS taken, cal-culated on the anhydrous basis MT: Amount (mg) of Rosuvastatin Calcium taken, calcu-lated on the anhydrous basis Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2.8 times as long as the retention time of rosuvastatin, beginning after the solvent peak. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: Pipet 5 mL of the standard solution obtained in the Assay, add 24 mL of acetonitrile, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add 24 mL of acetonitrile, and add water to make exactly 100 mL. Confirm that the peak area of rosuvastatin obtained with 10 mL of this solution is equivalent to 0.035 to 0.065z of that with 10 mL of the standard solution in the Assay. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution in the Assay under the above operating conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 2.0z. Enantiomer—Dissolve 25 mg of Rosuvastatin Cal-cium in 6 mL of acetonitrile, add water to make exactly 25 mL, and use this solution as the sample solution. Pipet 1 mL of this solution, add a mixture of water and acetonitrile (3:1) to make exactly 200 mL, and use this solution as the stand-ard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine each peak area by the automatic integration method: the area of related substance E (enan-tiomer) having the relative retention time of about 0.92 to rosuvastatin obtained from the sample solution is not larger than 1/5 times the peak area of rosuvastatin from the stand-ard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave- length: 242 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with silica gel coated with cellulose tris(4-methylbenzoate) for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of diluted trifluoroacetic acid (1 in 1000) and acetonitrile (3:1). Flow rate: Adjust so that the retention time of rosuvasta-tin is about 26.5 minutes. System suitability— Test for required detectability: Pipet 5 mL of the standard solution, add a mixture of water and acetonitrile (3:1) to make exactly 50 mL. Confirm that the peak area of rosuvastatin obtained with 10 mL of this solution is equiva-lent to 7 to 13z of that with the standard solution. System performance: To 5 mg of rosuvastatin calcium enantiomer add 12 mL of acetonitrile and 10 mL of water, sonicate to dissolve, and add water to make 50 mL. To 1 mL of this solution and 6 mL of acetonitrile add 25 mg of Rosuvastatin Calcium, sonicate to dissolve, and add water to make 25 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, rosuvastatin enantiomer and rosuvastatin are eluted in this order with the resolution between these peaks being not less than 1.5, and the symmetry factor of the peak of rosuvastatin is 1.0 – 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 2.0z. Water <2.48> Not more than 6.1z (20 mg, coulometric titration).
Conduct this procedure using light-resistant vessels. Weigh accurately about 35 mg each of Rosuvastatin Calcium and Rosuvastatin Calcium RS (separately determine the water <2.48> in the same manner as Rosuvastatin Calcium), dissolve each in 12 mL of acetonitrile, add water to make ex-actly 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of rosuvastatin in each solution. Amount (mg) of rosuvastatin calcium [(C22H27FN3O6S)2Ca] ＝ MS × AT/AS MS: Amount (mg) of Rosuvastatin Calcium RS taken, cal-culated on the anhydrous basis Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 242 nm). Column: A stainless steel column 3 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase A: A mixture of water, acetonitrile and diluted trifluoroacetic acid (1 in 100) (70:29:1). Mobile phase B: A mixture of acetonitrile, water, and diluted trifluoroacetic acid (1 in 100) (75:24:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. 0 – 30 30 – 50 100 → 60 0 → 40 50 – 60 60 → 0 40 → 100 60 – 70 ‌Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) Related substance B (diastereomer): (3RS,5RS,6E )-7-{4-(4-Fluorophenyl)-6-(1-methylethyl)-2-[methyl(methylsulfonyl)amino]pyrimidin-5-yl}-3,5-dihydroxyhept-6-enoic acid Flow rate: 0.75 mL per minute. System suitability— System performance: Dissolve 10 mg of Rosuvastatin Cal-cium in 10 mL of a solution of trifluoroacetic acid in aceto-nitrile (1 in 100), and allow to stand at 409C for 1 hour. After cooling, add 20 mL of water, adjust to pH 6 – 8 with sodium hydroxide TS, and add water to make 50 mL. To 3 mL of this solution, add water to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, rosuvastatin and the related substance B (diastereomer) are eluted in this order with the resolution between these peaks being not less than 2.5, and the symme-try factor of the peak of rosuvastatin is not more than 1.5. System repeatability: When the test is repeated 5 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of rosuvastatin is not more than 2.0z.
Saccharin Sodium Hydrate contains not less than ＝ MS × AT/AS × 2 7 5 3 98.
Saccharin Sodium Hydrate occurs as colorless crystals or a white crystalline powder. It is freely soluble in water and in methanol, and sparingly soluble in ethanol (95). It effloresces slowly and loses about half the amount of water of crystallization in air.
(1) Determine the infrared absorption spec-trum of Saccharin Sodium Hydrate, previously dried at 1059C to constant mass, as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the spectrum of Saccharin Sodium RS dried in the same manner as Saccharin Sodium Hydrate: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. A solution of Saccharin Sodium Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz)
(1) Clarity and color of solution—Dissolve 2.0 g of Saccharin Sodium Hydrate in water to make 10 mL, and 0 – 7.0 90 10 7.0 – 8.0 90 → 5 10 → 95 8.0 – 10.0 5 95 10.0 – 10.1 5 → 90 95 → 10 10.1 – 10.5 90 10 Flow rate: 1.0 mL per minute (the retention time of sac-charin is about 7.3 minutes). use this solution as the test solution. Perform the test with the test solution as directed under Turbidity Measurement <2.61>: the solution is clear. Perform the test with the test so- lution according to Method 2 under Methods for Color Matching <2.65>: the solution is colorless. Acidity or alkalinity—Dissolve 1.0 g of Saccharin Sodium Hydrate in 10 mL of water, and add 1 drop of phe-nolphthalein TS: the solution is colorless. Add 1 drop of 0.1 mol/L sodium hydroxide VS to the solution: the color changes to red. Heavy metals <1.07>—Dissolve 2.0 g of Saccharin So-dium Hydrate in 40 mL of water, add 0.7 mL of dilute hy-drochloric acid, dilute with water to make 50 mL, and rub the inner wall of the vessel with a glass rod until crystalliza-tion begins. Allow the solution to stand for 1 hour after the beginning of crystallization, and then filter through dry filter paper. Reject the first 10 mL of the filtrate, and take 25 mL of the subsequent filtrate. Add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test, using this solu-tion as the test solution. To 1.0 mL of Standard Lead Solu-tion add 2 mL of dilute acetic acid and water to make 50 mL, and use this solution as the control solution (not more than 10 ppm). Benzoate and salicylate—Dissolve 0.5 g of Saccharin Sodium Hydrate in 10 mL of water, add 5 drops of acetic acid (31) and 3 drops of iron (III) chloride TS: no turbidity is produced, and no red-purple to purple color develops. o-Toluene sulfonamide—Dissolve 10 g of Saccharin Sodium Hydrate in 50 mL of water, and extract with three 30-mL portions of ethyl acetate. Combine all the ethyl ace-tate extracts, wash with 30 mL of a solution of sodium chlo-ride (1 in 4), dehydrate with 5 g of anhydrous sodium sul- lution to stand between 489C and 509C for 10 minutes: the solution has no more color than Matching Fluid A. Water <2.48> Not more than 15.0z (0.1 g, volumetric titration, direct titration).
Weigh accurately about 50 mg of Saccharin Sodium Hydrate, and dissolve in a mixture of water and methanol (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the sample solution. Sepa-rately, weigh accurately about 25 mg of Saccharin Sodium RS (separately determine the water <2.48> in the same man-ner as Saccharin Sodium Hydrate), dissolve in a mixture of water and methanol (1:1) to make exactly 25 mL, and use this solution as the standard stock solution. Pipet 5 mL of the standard stock solution, add a mixture of water and methanol (1:1) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of saccharin in each solution. Amount (mg) of saccharin sodium (C H NNaO S) fate, and evaporate ethyl acetate. To the residue add exactly 5 mL of the internal standard solution to dissolve, and use ＝ MS × AT/AS × 2 7 4 3 this solution as the sample solution. Separately, dissolve 0.10 g of o-toluene sulfonamide in ethyl acetate to make exactly 100 mL. Pipet 1 mL of this solution, evaporate on a water bath to dryness, dissolve the residue in exactly 5 mL of the internal standard solution, and use this solution as the stand-ard solution. Perform the test with 1 mL each of the sample solution and standard solution as directed under Gas Chro-matography <2.02> according to the following conditions, and calculate the ratios, QT and QS, of the peak height of o-toluene sulfonamide to that of the internal standard: QT is not more than QS. Internal standard solution—A solution of caffeine in ethyl acetate (1 in 500). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 1 m in length, packed with siliceous earth for gas chromatogra-phy (180 to 250 mm in diameter), coated with diethylene glycol succinate polyester for gas chromatography at the ratio of 3z. Column temperature: A constant temperature of about 2009C. Injection port temperature: A constant temperature of about 2259C. Detector temperature: A constant temperature of about 2509C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of caffeine is about 6 minutes. System suitability— System performance: When the procedure is run with 1 mL of the standard solution under the above operating condi-tions, the internal standard and o-toluene sulfonamide are eluted in this order with the resolution between these peaks being not less than 2.0. System repeatability: When the test is repeated 6 times with 1 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak height of o-toluene sulfonamide to that of the internal standard is not more than 2.0z. Readily carbonizable substances <1.15>—Perform the test with 0.20 g of Saccharin Sodium Hydrate. Allow the so- MS: Amount (mg) of Saccharin Sodium RS taken, calcu-lated on the anhydrous basis Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 230 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase A: Dissolve 8.7 g of dipotassium hydrogen phosphate in diluted phosphoric acid (1 in 1000) to make 1000 mL. Mobile phase B: Methanol. Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 7.0 7.0 – 8.0 90 → 5 10 → 95 8.0 – 10.0 10.0 – 10.1 5 → 90 95 → 10 10.1 – 10.5 Flow rate: 1.0 mL per minute (the retention time of sac-charin is about 7.3 minutes). System suitability— System performance: Dissolve 25 mg of phthalic anhy-dride in a mixture of water and methanol (1:1) to make 25 mL. To 5 mL of this solution add 5 mL of the standard stock solution and a mixture of water and methanol (1:1) to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating conditions, phthalic an-hydride and saccharin are eluted in this order with the reso-lution between these peaks being not less than 1.5. When the procedure is run with 10 mL of the standard solution under the above operating conditions, the symmetry factor of the peak of saccharin is not more than 1.5. System repeatability: When the test is repeated 5 times ‌with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of saccharin is not more than 0.73z.
Sivelestat Sodium Hydrate contains not less than 98.
Sivelestat Sodium Hydrate occurs as a white crystalline powder. It is freely soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in water. It dissolves in sodium hydroxide TS. Melting point: about 1909C (with decomposition, after drying in vacuum, 609C, 2 hours).
(1) Determine the absorption spectrum of a solution of Sivelestat Sodium Hydrate in boric acid-potassi-um chloride-sodium hydroxide buffer solution (pH 9.0) (1 in 40,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wavelengths. Determine the infrared absorption spectrum of Sivelestat Sodium Hydrate as directed in the paste method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 50 mg of Sivelestat Sodium Hydrate in 5 mL of water with one drop of ammonia TS: the solution re-sponds to Qualitative Tests <1.09> for sodium salt.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Sivelestat Sodium Hydrate according to Method 4, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). (2) Related substances—Dissolve 10 mg of Sivelestat So-dium Hydrate in 10 mL of a mixture of water and aceto-nitrile (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of water and acetonitrile (1:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 10 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and determine each peak area by the automatic integration method: the area of the peak, having the relative retention time of about 1.2 to sivelestat, obtained from the sample solution is not larger than 1/2 times the peak area of sivelestat from the standard solution, the areas of the peaks, having the relative retention time of about 0.25, about 0.60, and about 2.7, from the sample solution is not larger than 3/10 times the peak area of sivelestat from the standard solution, the area of the peaks other than sivelestat and peaks mentioned above from the sample solution is not larger than 1/10 times the peak area of sivelestat from the standard solution, and the total area of the peaks other than sivelestat from the sample solu-tion is not larger than the peak area of sivelestat from the standard solution. Operating conditions— Column, column temperature, mobile phase and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Time span of measurement: About 4 times as long as the retention time of sivelestat, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of water and acetonitrile (1:1) to make exactly 20 mL. Confirm that the peak area of sivelestat obtained with 10 mL of this solution is equivalent to 4 to 6z of that with 10 mL of the standard solution. System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of sivelestat are not less than 5000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak ‌area of sivelestat is not more than 2.0z. Water <2.48> 12.0 – 14.0z (0.2 g, volumetric titration, direct titration).
Weigh accurately about 50 mg of Sivelestat Sodium Hydrate, dissolve in a mixture of water and acetonitrile (1:1) to make exactly 50 mL. Pipet 5 mL of this solution, and add exactly 5 mL of the internal standard solution. To 4 mL of this solution, add 7 mL of acetonitrile and 9 mL of water, and use this solution as the sample solution. Separately, weigh accurately about 40 mg of Sivelestat RS, previously dried (in vacuum, 609C, 2 hours), and dissolve in acetonitrile to make exactly 50 mL. Pipet 5 mL of this solution, and add exactly 5 mL of the internal standard solution. To 2 mL of this solution, add 3 mL of acetonitrile and 5 mL of water, and use this solution as the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of sivelestat to that of the inter-nal standard. tions, with Sivelestat Sodium Hydrate. Description Sivelestat Sodium for Injection occurs as white, masses or powder. Identification (1) Dissolve an amount of Sivelestat So-dium for Injection, equivalent to 0.1 g of Sivelestat Sodium Hydrate, in 10 mL of water. To 1 mL of this solution add boric acid-potassium chloride-sodium hydroxide buffer solu-tion (pH 9.0) to make 100 mL. Determine the absorption spectrum of this solution as directed under Ultraviolet-visible Spectrophotometry <2.24>: it exhibits a maximum be-tween 311 nm and 315 nm. (2) Take an amount of Sivelestat Sodium for Injection, equivalent to 0.1 g of Sivelestat Sodium Hydrate, add 10 mL of methanol, and shake. Take 1 mL of the supernatant liq-uid, add methanol to make 10 mL, and use this solution as the sample solution. Separately, dissolve 10 mg of sivelestat sodium hydrate in 10 mL of methanol, and use this solution as the standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution Amount (mg) of sivelestat sodium (C ＝ MS × QT/QS × 1.051 20H21N2NaO7S) on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of ethyl acetate and acetic acid (100) (20:1) to a distance of MS: Amount (mg) of Sivelestat RS taken Internal standard solution—A solution of propyl para-hydroxybenzoate in acetonitrile (1 in 2500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 5.44 g of potassium dihydrogen phosphate in water to make 1000 mL, then adjust to pH 3.5 with phosphoric acid. To 5 volumes of this solution, add 4 volumes of acetonitrile. Flow rate: Adjust so that the retention time of sivelestat is about 10 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, the internal standard and sivelestat are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of sivelestat to that of the internal standard is not more than 1.0z.
Sodium Acetate Hydrate, when dried, contains not less than 99.
Sodium Acetate Hydrate occurs as colorless crystals or a white crystalline powder. It is odorless or has a slight, acetous odor. It has a cool, saline and slightly bitter taste. It is very soluble in water, freely soluble in acetic acid (100), soluble in ethanol (95), and practically insoluble in diethyl ether. It is efflorescent in warm, dry air.
A solution of Sodium Acetate Hydrate (1 in 10) responds to Qualitative Tests <1.09> for acetate and for sodium salt.
(1) Clarity and color of solution—Dissolve 2.0 g of Sodium Acetate Hydrate in 20 mL of water: the solution is clear and colorless. Acidity or alkalinity—Dissolve 1.0 g of Sodium Ace-tate Hydrate in 20 mL of freshly boiled and cooled water, and add 3 drops of phenolphthalein TS: a red color devel-ops. When cooled to 109C, or 1.0 mL of 0.01 mol/L hydro-chloric acid VS is added after cooling to 109C, the red color disappears. Chloride <1.03>—Perform the test with 1.0 g of Sodium Acetate Hydrate. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.011z). Sulfate <1.14>—Perform the test with 1.0 g of So- dium Acetate Hydrate. Prepare the control solution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.017z). Heavy metals <1.07>—Proceed with 2.0 g of Sodium Acetate Hydrate according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Calcium and magnesium—Dissolve 4.0 g of Sodium Acetate Hydrate in 25 mL of water, add 6 g of ammonium chloride, 20 mL of ammonia solution (28) and 0.25 mL of a solution of sodium sulfite heptahydrate (1 in 10), and titrate <2.50> with 0.01 mol/L disodium dihydrogen ethylene- diamine tetraacetate VS until the blue color changes to grayish blue (indicator: 0.1 g of methylthymol blue-potas-sium nitrate indicator): the amount of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS consumed is not more than 0.5 mL. Arsenic <1.11>—Prepare the test solution with 1.0 g of Sodium Acetate Hydrate, according to Method 1, and perform the test (not more than 2 ppm). Potassium permanganate-reducing substance—Dis-solve 1.0 g of Sodium Acetate Hydrate in 100 mL of water, add 5 mL of dilute sulfuric acid, boil, add 0.50 mL of 0.002 mol/L potassium permanganate VS, and further boil for 5 minutes: the red color of the solution does not disappear. Loss on drying <2.41> 39.0 – 40.5z (1 g, first at 809C for 2 hours, and then at 1309C for 2 hours). ‌Assay Weigh accurately about 0.2 g of Sodium Acetate Hydrate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS until the color of the solution changes from yellow to green (indicator: 1 mL of p-naphtholbenzein TS). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 8.203 mg of C2H3NaO2
Sodium Aurothiomalate contains not less than 49.
Sodium Aurothiomalate occurs as a white to light yellow, powder or granules. It is very soluble in water, and practically insoluble in ethanol (99.5). It is hygroscopic. It changes in color by light to greenish pale yellow.
(1) To 2 mL of a solution of Sodium Aurothiomalate (1 in 10) add 1 mL of a solution of calcium nitrate tetrahydrate (1 in 10): a white precipitate is produced, and it dissolves in dilute nitric acid and reappears on the ad-dition of ammonium acetate TS. To 2 mL of a solution of Sodium Aurothiomalate (1 in 10) add 3 mL of silver nitrate TS: a yellow precipitate is produced, and it dissolves in an excess of ammonia TS. Place 2 mL of a solution of Sodium Aurothiomalate (1 in 10) in a porcelain crucible, add 1 mL of ammonia TS and 1 mL of hydrogen peroxide (30), evaporate to dryness, and ignite. Add 20 mL of water to the residue, and filter: the residue on the filter paper occurs as a yellow or dark yellow, powder or granules. The filtrate obtained in (3) responds to Qualitative Tests <1.09> for sodium salt. The filtrate obtained in (3) responds to Qualitative Tests <1.09> for sulfate. pH <2.54> Dissolve 1.0 g of Sodium Aurothiomalate in 10 mL of water: the pH of this solution is between 5.8 and 6.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Aurothiomalate in 10 mL of water: the solution is clear and light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Sodium Aurothiomalate according to Method 2, and perform the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sodium Aurothiomalate according to Method 3, and per-form the test (not more than 2 ppm). Ethanol—Weigh accurately about 0.2 g of Sodium Aurothiomalate, add exactly 3 mL of the internal standard solution and 2 mL of water to dissolve, and use this solution as the sample solution. Separately, pipet 3 mL of ethanol (99.5), and add water to make exactly 1000 mL. Pipet 2 mL of this solution, add exactly 3 mL of the internal standard solution, and use this solution as the standard solution. Per-form the test with 2 mL each of the sample solution and standard solution as directed under Gas Chromatography <2.02> according to the following conditions, and calculate the ratios of the peak area of ethanol to that of the internal standard, QT and QS: the amount of ethanol is not more than 3.0z. Amount (mg) of ethanol ＝ QT/QS × 6 × 0.793 0.793: Density (g/mL) of ethanol (99.5) at 209C Internal standard solution—A solution of 2-propanol (1 in 500). Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A column 3 mm in inside diameter and 3 m in length, packed with porous styrene-divinylbenzene copoly-mer for gas chromatography (particle diameter: 150 – 180 mm) (average pore size: 0.0085 mm; 300 – 400 m2/g). Column temperature: A constant temperature of about 1809C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of the internal standard is about 7 minutes. System suitability— System performance: When the procedure is run with 2 mL of the standard solution under the above operating condi-tions, ethanol and the internal standard are eluted in this order with the resolution between these peaks being not less than 4. System repeatability: When the test is repeated 6 times with 2 mL of the standard solution under the above operating conditions, the relative standard deviation of the ratio of the peak area of ethanol to that of the internal standard is not more than 2.0z. Water <2.48> Not more than 5.0z (0.1 g, coulometric titration). Use a water vaporizer (heating temperature: 1059C; heating time: 30 minutes).
Weigh accurately about 25 mg of Sodium Aurothiomalate, and dissolve in 2 mL of aqua regia by heat-ing. After cooling, add water to make exactly 100 mL. Pipet 2 mL of the solution, add water to make exactly 25 mL, and use this solution as the sample solution. Separately, pipet 5 mL, 10 mL and 15 mL of Standard Gold Solution for atomic absorption spectrophotometry, add water to make exactly 25 mL, and use these solutions as the standard solutions (1), (2) and (3), respectively. Perform the test with the sample solu-tion and standard solutions (1), (2) and (3) as directed under Atomic Absorption Spectrophotometry <2.23> under the fol-lowing conditions. Determine the amount of gold in the sam-ple solution using the calibration curve obtained from the absorbances of the standard solutions. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Gold hollow-cathode lamp. Wavelength: 242.8 nm.
Sodium Benzoate, when dried, contains not less than 99.
Sodium Benzoate occurs as white, granules, crystals or crystalline powder. It is odorless, and has a sweet and saline taste. It is freely soluble in water, slightly soluble in ethanol (95), and practically insoluble in diethyl ether.
A solution of Sodium Benzoate (1 in 100) re-sponds to Qualitative Tests <1.09> for benzoate and Qualita-tive Tests <1.09> (1) and (2) for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Benzoate in 5 mL of water: the solution is clear and colorless. Acidity or alkalinity—Dissolve 2.0 g of Sodium Ben-zoate in 20 mL of freshly boiled and cooled water, and add 2 drops of phenolphthalein TS and 0.20 mL of 0.05 mol/L sul-furic acid VS: the solution remains colorless. To this solu-tion add 0.40 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. Sulfate <1.14>—Dissolve 0.40 g of Sodium Benzoate in 40 mL of water, add slowly 3.5 mL of dilute hydrochloric acid with thorough stirring, allow to stand for 5 minutes, and filter. Discard the first 5 mL of the filtrate, take the sub-sequent 20 mL of the filtrate, add water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 0.40 mL of 0.005 mol/L sulfu-ric acid VS (not more than 0.120z). Heavy metals <1.07>—Dissolve 2.0 g of Sodium Ben- zoate in 44 mL of water, add gradually 6 mL of dilute hydro-chloric acid with thorough stirring, and filter. Discard the first 5 mL of the filtrate, take the subsequent 25 mL of the filtrate, neutralize with ammonia TS, add 2 mL of dilute ace-tic acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Mix well 1.0 g of Sodium Benzoate with 0.40 g of calcium hydroxide, ignite, dissolve the residue in 10 mL of dilute hydrochloric acid, and perform the test using this solution as the test solution (not more than 2 ppm). Chlorinated compounds—Dissolve 1.0 g of Sodium Benzoate in 10 mL of water, add 10 mL of dilute sulfuric acid, and extract with two 20-mL portions of diethyl ether. Combine the diethyl ether extracts, and evaporate the diethyl ether on a water bath. Place 0.5 g of the residue and 0.7 g of calcium carbonate in a crucible, mix with a small amount of water, and dry. Ignite it at about 6009C, dissolve in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washing, add water to make 50 mL, and add 0.5 mL of silver nitrate TS: this solu-tion has no more turbidity than the following control solu-tion. Control solution: Dissolve 0.7 g of calcium carbonate in 20 mL of dilute nitric acid, and filter. Wash the residue with 15 mL of water, combine the filtrate and the washings, add 1.2 mL of 0.01 mol/L Hydrochloric acid VS and water to make 50 mL, and add 0.5 mL of silver nitrate TS. Phthalic acid—To 0.10 g of Sodium Benzoate add 1 mL of water and 1 mL of resorcinol-sulfuric acid TS, and heat the mixture in an oil bath heated at a temperature be-tween 1209C and 1259C to evaporate the water, then heat the residue for further 90 minutes, cool, and dissolve in 5 mL of water. To 1 mL of the solution add 10 mL of a solu-tion of sodium hydroxide (43 in 500), shake, then examine under light at a wavelength between 470 nm and 490 nm: the green fluorescence of the solution is not more intense than that of the following control solution. Control solution: Dissolve 61 mg of potassium hydrogen phthalate in water to make exactly 1000 mL. Pipet exactly 1 mL of the solution, add 1 mL of resorcinol-sulfuric acid TS, and proceed as directed above. Loss on drying <2.41> Not more than 1.5z (2 g, 1109C, 4 hours).
Weigh accurately about 1.5 g of Sodium Benzoate, previously dried, and transfer to a 300-mL glass-stoppered flask. Dissolve in 25 mL of water, add 75 mL of diethyl ether and 10 drops of bromophenol blue TS, and titrate <2.50> with 0.5 mol/L hydrochloric acid VS, while mixing the aqueous and diethyl ether layers by vigorous shaking, until a persistent, light green color is produced in the aqueous layer. Each mL of 0.5 mol/L hydrochloric acid VS ＝ 72.05 mg of C7H5NaO2
Sodium Bicarbonate and Bitter Tincture Mix-ture is a clear, yellowish liquid, having a bitter taste.
Sodium Bisulfite is a mixture of sodium hydrogen-sulfite and sodium pyrosulfite. It contains not less than 64.
Sodium Bisulfite occurs as white, granules or powder, having the odor of sulfur dioxide. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. A solution of Sodium Bisulfite (1 in 20) is acid. Sodium Bisulfite is slowly affected by air or by light.
A solution of Sodium Bisulfite (1 in 20) re-sponds to Qualitative Tests <1.09> for sodium salt and for bi-sulfite.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Bisulfite in 10 mL of water: the solution is clear and colorless. Thiosulfate—Dissolve 1.0 g of Sodium Bisulfite in 15 mL of water, add slowly 5 mL of dilute hydrochloric acid, shake, and allow to stand for 5 minutes: no turbidity is pro-duced. Heavy metals <1.07>—Dissolve 1.0 g of Sodium Bi- sulfite in 10 mL of water, add 5 mL of hydrochloric acid, and evaporate on a water bath to dryness. To the residue add 2 mL of dilute acetic acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution as follows: evaporate 5 mL of hy-drochloric acid on a water bath to dryness, and add 2 mL of dilute acetic acid and 2.0 mL of Standard Lead Solution, and dilute with water to make 50 mL (not more than 20 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Sodium Bisulfite according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 0.5 g of Sodium Bisulfite in mL of water. Add 1 mL of sulfuric acid, heat on a sand bath until white fumes are evolved, add water to make 5 mL, and perform the test with this solution as the test solution (not more than 4 ppm).
Weigh accurately about 0.15 g of Sodium Bisulfite, and transfer immediately into an iodine flask containing ‌exactly 50 mL of 0.05 mol/L iodine VS, stopper, shake, and allow to stand for 5 minutes in a dark place. Add 1 mL of hydrochloric acid, and titrate <2.50> the excess iodine with‌ 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 3.203 mg of SO2
Sodium Borate contains not less than 99.
Sodium Bromide, when dried, contains not less than 99.
Sodium Bromide occurs as colorless or white, crystals or crystalline powder. It is odorless. It is freely soluble in water, and soluble in ethanol (95). It is hygroscopic, but not deliquescent.
A solution of Sodium Bromide (1 in 10) re-sponds to Qualitative Tests <1.09> for sodium salt and for bromide.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Bromide in 3 mL of water: the solution is clear 10H2O). 2 4 7 and colorless. Alkalinity—Dissolve 1.0 g of Sodium Bromide in 10 Description Sodium Borate occurs as colorless or white crystals or a white crystalline powder. It is odorless, and has a slightly characteristic, saline taste. It is freely soluble in glycerin, soluble in water, and practi-cally insoluble in ethanol (95), in ethanol (99.5) and in diethyl ether. When placed in dry air, Sodium Borate effloresces and is coated with a white powder. Identification A solution of Sodium Borate (1 in 20) re-sponds to Qualitative Tests <1.09> for sodium salt and for borate. pH <2.54> Dissolve 1.0 g of Sodium Borate in 20 mL of water: the pH of this solution is between 9.1 and 9.6. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Sodium Borate in 20 mL of water by warming slightly: the solution is clear and colorless. Carbonate or bicarbonate—Dissolve 1.0 g of pow-dered Sodium Borate in 20 mL of freshly boiled and cooled water, and add 3 mL of dilute hydrochloric acid: the solu-tion does not effervesce. Heavy metals <1.07>—Dissolve 1.5 g of Sodium Borate in 25 mL of water and 7 mL of 1 mol/L hydrochloric acid TS, add 1 drop of phenolphthalein TS, and add ammo-nia TS until a pale red color develops. Then add dilute acetic acid until the solution becomes colorless again, add 2 mL of dilute acetic acid, and add water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 3.0 mL of Standard Lead So-lution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 0.40 g of Sodium Borate according to Method 1, and perform the test (not more than 5 ppm).
Weigh accurately about 2 g of Sodium Borate, dis-solve in 50 mL of water, and titrate <2.50> with 0.5 mol/L hydrochloric acid VS (indicator: 3 drops of methyl red TS). Each mL of 0.5 mol/L hydrochloric acid VS ＝ 95.34 mg of Na2B4O7.10H2O
Sodium Carbonate Hydrate contains not less than 99.
Sodium Carbonate Hydrate occurs as colorless or white crystals. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. A solution of Sodium Carbonate Hydrate (1 in 10) is alkaline. Sodium Carbonate Hydrate is efflorescent in air. It liquefies in its water of crystallization at 349C, and becomes anhydrous at above 1009C.
A solution of Sodium Carbonate Hydrate (1 in 20) responds to Qualitative Tests <1.09> for sodium salt and for carbonate.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Carbonate Hydrate in 5 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 0.5 g of Sodium Car- bonate Hydrate in 10 mL of water, add 7 mL of dilute nitric acid, dilute with water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.071z). Heavy metals <1.07>—Dissolve 2.0 g of Sodium Car- bonate Hydrate in 10 mL of water, add 8 mL of dilute hy-drochloric acid, and evaporate to dryness on a water bath. Dissolve the residue in 35 mL of water and 2 mL of dilute acetic acid, dilute with water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 8 mL of dilute hydro-chloric acid on a water bath to dryness, add 2 mL of dilute acetic acid and 2.0 mL of Standard Lead Solution, and di-lute with water to make 50 mL (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 0.65 of Sodium Carbonate Hydrate according to Method 1, and per-form the test (not more than 3.1 ppm). Loss on drying <2.41> 61.0 – 63.0z (1 g, 1059C, 4 hours).
Dissolve about 3 g of Sodium Carbonate Hydrate, weighed accurately, in 25 mL of water, and titrate with 0.5 mol/L sulfuric acid VS until the color of the solution changes from blue to yellow-green. Boil cautiously, cool, and further titrate <2.50> until a greenish yellow color ap-pears (indicator: 2 drops of bromocresol green TS). 2 3
Sodium Chloride occurs as colorless or white, crystals or crystalline powder. It is freely soluble in water, and practically insoluble in ethanol (99.5).◆
(1) A solution of Sodium Chloride (1 in 20) responds to Qualitative Tests <1.09> for sodium salt. (2) A solution of Sodium Chloride (1 in 20) responds to Qualitative Tests <1.09> for chloride.
◆(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Chloride in 5 mL of water: the solution is clear and colorless.◆ Acidity or alkalinity—Dissolve 20.0 g of Sodium Chloride in freshly boiled and cooled water to make exactly 100 mL, and use this solution as the sample solution. To 20 mL of the sample solution add 0.1 mL of bromothymol blue-sodium hydroxide-ethanol TS and 0.5 mL of 0.01 mol/L hydrochloric acid VS: the color of the solution is yel-low. Separately, to 20 mL of the sample solution add 0.1 mL of bromothymol blue-sodium hydroxide-ethanol TS and 0.5 mL of 0.01 mol/L sodium hydroxide VS: the color of the so-lution is blue. Sulfates—To 7.5 mL of the sample solution obtained in (2) add water to make 30 mL, and use this solution as the sample solution. Separately, dissolve 0.181 g of potassium sulfate in diluted ethanol (3 in 10) to make exactly 500 mL. Pipet 5 mL of this solution, and add diluted ethanol (3 in 10) to make exactly 100 mL. To 4.5 mL of this solution add 3 mL of a solution of barium chloride dihydrate (1 in 4), shake, and allow to stand for 1 minutes. To 2.5 mL of this solution add 15 mL of the sample solution and 0.5 mL of acetic acid (31), and allow to stand for 5 minutes: any tur-bidity produced does not more than that produced in the fol-lowing control solution. Control solution: Dissolve 0.181 g of potassium sulfate in water to make exactly 500 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and proceed in the same manner as directed above using this solution instead of the sample solution. Phosphates—To 2.0 mL of the sample solution ob-tained in (2) add water to make exactly 100 mL, then add 4 mL of molybdenium-sulfuric acid TS, mix, add 0.1 mL of tin (II) chloride-hydrochloric acid TS, and allow to stand for 10 minutes: the color of the solution is not darker than the following control solution. Control solution: To 1.0 mL of Standard Phosphoric Acid Solution add 12.5 mL of 2 mol/L sulfuric acid TS and water to make exactly 250 mL. Then, proceed in the same manner as above with 100 mL of this solution. Bromides—To 0.50 mL of the sample solution ob-tained in (2) add 4.0 mL of water, 2.0 mL of dilute phenol red TS and 1.0 mL of a freshly prepared solution of sodium toluenesulfonchloramide trihydrate (1 in 10,000), and mix immediately. After allowing to stand for 2 minutes, add 0.15 mL of 0.1 mol/L sodium thiosulfate VS, mix, add water to make exactly 10 mL, and use this solution as the sample so-lution. Separately, to 5.0 mL of a solution of potassium bro-mide (3 in 1,000,000) add 2.0 mL of dilute phenol red TS and 1.0 mL of a solution of sodium toluenesulfonchloramide trihydrate (1 in 10,000), and mix immediately. Proceed in the same manner as the preparation of the sample solution, and use the solution so obtained as the standard solution. Per-form the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24> using water as the control: the absorbance at 590 nm of the sample solution is not more than that of the standard solution. Iodides—Wet 5 g of Sodium Chloride by adding dropwise a freshly prepared mixture of soluble starch TS, 0.5 mol/L sulfuric acid TS and sodium nitrite TS (1000:40:3), allow to stand for 5 minutes, and examine: a blue color does not appear. Ferrocyanides—Dissolve 2.0 g of Sodium Chloride in 6 mL of water, and add 0.5 mL of a mixture of a solution of iron (II) sulfate heptahydrate (1 in 100) and a solution of ammonium iron (III) sulfate dodecahydrate in diluted sulfu-ric acid (1 in 400) (1 in 100) (19:1): a blue color does not de-velop within 10 minutes. (8) Heavy metals <1.07>—Proceed with 5.0 g of So- dium Chloride according to Method 1, and perform the test. Prepare the control solution with 1.5 mL of Standard Lead Solution (not more than 3 ppm).◆ Iron—To 10 mL of the sample solution obtained in (2) add 2 mL of a solution of citric acid monohydrate (1 in 5) and 0.1 mL of mercapto acetic acid, alkalize with ammonia TS, add water to make exactly 20 mL, and allow to stand for 5 minutes: the solution has not more color than the follow-ing control solution. Control solution: Pipet 1 mL of Standard Iron Solution, and add water to make exactly 25 mL. To 10 mL of this so-lution add 2 mL of a solution of citric acid monohydrate (1 in 5) and 0.1 mL of mercapto acetic acid, and proceed in the same manner as directed for the sample solution. Barium—To 5.0 mL of the sample solution ob-tained in (2) add 5.0 mL of water and 2.0 mL of dilute sulfu-ric acid, and allow to stand for 2 hours: the solution has not more turbidity than the following control solution. Control solution: To 5.0 mL of the sample solution ob-tained in (2) add 7.0 mL of water, and allow to stand for 2 hours. Magnesium and alkaline-earth materials—To 200 mL of water add 0.1 g of hydroxylammonium chloride, 10 mL of ammonium chloride buffer solution (pH 10), 1 mL of 0.1 mol/L zinc sulfate VS and 0.15 g of eriochrome black T-sodium chloride indicator, and warm to 409C. Add 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS dropwise until the purple color of the solution changes to blue. To this solution add a solution prepared by dissolving 10.0 g of Sodium Chloride in 100 mL of water, and add 2.5 mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS: the color of the solution is a blue. (12) Arsenic <1.11>—Prepare the test solution with 1.0 g of Sodium Chloride according to Method 1, and per-form the test (not more than 2 ppm).◆ ‌Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours).‌
Weigh accurately about 50 mg of Sodium Chloride, dissolve in 50 mL of water, and titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration). Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl ◆Containers and storage Containers—Tight containers.◆ 10 Sodium Chloride Injection
Sodium Citrate Hydrate, when dried, contains not less than 99.
Sodium Citrate Hydrate occurs as colorless crystals, or a white crystalline powder. It is odorless, and has a cooling, saline taste. It is freely soluble in water, and practically insoluble in ethanol (95) and in diethyl ether.
A solution of Sodium Citrate Hydrate (1 in 20) responds to Qualitative Tests <1.09> for citrate and for sodium salt. pH <2.54> Dissolve 1.0 g of Sodium Citrate Hydrate in 20 mL of water: the pH of this solution is between 7.5 and 8.5.
(1) Clarity and color of solution—A solution of 1.0 g of Sodium Citrate Hydrate in 10 mL of water is clear and colorless. Chloride <1.03>—Take 0.6 g of Sodium Citrate Hy- drate, and perform the test. Prepare the control solution with 0.25 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.015z). Sulfate <1.14>—To 0.5 g of Sodium Citrate Hydrate add water to make 40 mL, then add 3.0 mL of dilute hydro-chloric acid and water to make 50 mL, and perform the test. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.048z). Heavy metals <1.07>—Proceed with 2.5 g of Sodium Citrate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.5 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sodium Citrate Hydrate according to Method 1, and per-form the test (not more than 2 ppm). Tartrate—To a solution of 1.0 g of Sodium Citrate Hydrate in 2 mL of water add 1 mL of potassium acetate TS and 1 mL of acetic acid (31): no crystalline precipitate is formed after the sides of the tube have been rubbed with a glass rod. Oxalate—Dissolve 1.0 g of Sodium Citrate Hydrate in a mixture of 1 mL of water and 3 mL of dilute hydrochlo-ric acid, add 4 mL of ethanol (95) and 0.2 mL of calcium chloride TS, and allow to stand for 1 hour: the solution is clear. Readily carbonizable substances <1.15>—Take 0.5 g of Sodium Citrate Hydrate, and perform the test by heating at 909C for 1 hour: the solution has no more color than Matching Fluid K. Loss on drying <2.41> 10.0 – 13.0z (1 g, 1809C, 2 hours).
Weigh accurately about 0.2 g of Sodium Citrate Hy-drate, previously dried, add 30 mL of acetic acid for nona-queous titration, warm to dissolve, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 8.602 mg of C6H5Na3O7
Sodium Cromoglicate contains not less than 98.
Sodium Cromoglicate occurs as a white crys-talline powder. It is odorless and tasteless at first, and later develops a slightly bitter taste. It is freely soluble in water, sparingly soluble in propylene glycol, very slightly soluble in ethanol (95), and practically insoluble in 2-propanol and in diethyl ether. It is hygroscopic. It gradually acquires a yellow color by light.
(1) Dissolve 0.1 g of Sodium Cromoglicate in 2 mL of water, add 2 mL of sodium hydroxide TS, and boil for 1 minute: a yellow color is produced. After cooling, add 0.5 mL of concentrated diazobenzene sulfonic acid TS: a dark red color is produced. Determine the absorption spectrum of a solution of Sodium Cromoglicate in phosphate buffer solution (pH 7.4) (1 in 100,000) as directed under Ultraviolet-visible Spectro-photometry <2.24>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wavelengths. Sodium Cromoglicate responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity and color of solution—Dissolve 0.50 g of Sodium Cromoglicate in 10 mL of water: the solution is clear and colorless to pale yellow. Acidity or alkalinity—Dissolve 2.0 g of Sodium Cromoglicate in 40 mL of freshly boiled and cooled water, add 6 drops of bromothymol blue TS, and use this solution as the sample solution. To 20 mL of the sample solution add 0.25 mL of 0.1 mol/L sodium hydroxide VS: a blue color is produced. To another 20 mL of the sample solution add 0.25 mL of 0.1 mol/L hydrochloric acid VS: a yellow color is produced. Heavy metals <1.07>—Proceed with 1.0 g of Sodium Cromoglicate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Oxalate—Dissolve 0.25 g of Sodium Cromoglicate in water to make exactly 50 mL, and use this solution as the sample solution. Separately, dissolve 49 mg of oxalic acid di-hydrate, exactly weighed, in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Pipet 20 mL each of the sample solution and standard solution, add exactly 5 mL of iron salicylate TS to each solution, and add water to make 50 mL. Determine the absorbances of these solutions as directed under Ultraviolet-visible Spectropho-tometry <2.24> using water as the blank: the absorbance of ‌the sample solution at 480 nm is not smaller than that of the standard solution. Related substances—Dissolve 0.20 g of Sodium Cromoglicate in 10 mL of water, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make exactly 10 mL, pipet 1 mL of this solution, add water to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of metha-nol, chloroform and acetic acid (100) (9:9:2) to a distance of about 10 cm, and air-dry the plate. Examine under ultravio-let light (main wavelength: 254 nm): spots other than the principal spot obtained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 10.0z (1 g, in vacu-um, 1059C, 4 hours).
Weigh accurately about 0.18 g of Sodium Cromogli-cate, and dissolve in a mixture of 25 mL of propylene glycol and 5 mL of 2-propanol by warming. After cooling, add 30 mL of 1,4-dioxane, and titrate <2.50> with 0.1 mol/L per-chloric acid-1,4-ioxane VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid-1,4-dioxane VS ＝ 25.62 mg of C23H14Na2O11
Sodium Fusidate occurs as white, crystals or crystalline powder. It is freely soluble in water, in methanol and in ethanol (99.5).
(1) Determine the infrared absorption spec-tra of Sodium Fusidate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Sodium Fusidate responds to Qualitative Tests <1.09> for sodium salt.
(1) Heavy metals <1.07>—Proceed with 2.0 g of Sodium Fusidate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Related substances—Dissolve 25 mg of Sodium Fusi-date in a mixture of acetonitrile for liquid chromatography, diluted phosphoric acid (3 in 1000) and methanol (5:4:1) to make 10 mL, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add water to make ex-actly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solu-tion and standard solution as directed under Liquid Chroma-tography <2.01> according to the following conditions. De-termine each peak area by the automatic integration method: the peak area of the related substance A, having the relative retention time of about 0.4 to fusidic acid, obtained from the sample solution is not larger than 3/10 times the peak area of fusidic acid from the standard solution, the peak area of the related substance B, having the relative retention time of about 0.5, from the sample solution is not larger than 2/5 times the peak area of fusidic acid from the standard solu-tion, the peak areas of the related substance C having the relative retention time of about 0.6, the related substance D having the relative retention time of about 0.63, the unknown substance having the relative retention time of about 0.65, the related substance E having the relative reten-tion time of about 0.7, the related substance G having the relative retention time of about 0.96 and the related sub-stance H having the relative retention time of about 1.18, from the sample solution are not larger than 1/5 times the peak area of fusidic acid from the standard solution, the peak area of the related substance F, having the relative retention time of about 0.82, from the sample solution is not larger than 7/10 times the peak area of fusidic acid from the standard solution, the peak area of the related substance I, having the relative retention time of about 1.23, from the sample solution is not larger than 1/2 times the peak area of fusidic acid from the standard solution, the peak area of the related substance J, having the relative retention time of about 1.4, from the sample solution is not larger than the peak area of fusidic acid from the standard solution, the area of the peak other than fusidic acid and the peaks men-tioned above from the sample solution is not larger than 1/10 times the peak area of fusidic acid from the standard solution. Furthermore, the total area of the peaks other than fusidic acid from the sample solution is not larger than 2 times the peak area of fusidic acid from the standard solu-tion. For the areas of the peaks, the related substances C, D, E, G and H, multiply their correction factors, 0.7, 0.7, 0.3, 0.6 and 0.6, respectively. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 235 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (3.5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase A: A mixture of diluted phosphoric acid (3 in 1000), acetonitrile for liquid chromatography and metha-nol (2:2:1). Mobile phase B: A mixture of acetonitrile for liquid chro-matography, methanol and diluted phosphoric acid (3 in 1000) (7:2:1). Flowing of mobile phase: Control the gradient by mixing the mobile phases A and B as directed in the following table. Time after injection of sample (min) Mobile phase A (volz) Mobile phase B (volz) 0 – 3 3 – 28 100 → 0 0 → 100 28 – 33 Flow rate: 1.0 mL per minute. Time span of measurement: For 33 minutes after injec-tion, beginning after the solvent peak. System suitability— Test for required detectability: Pipet 1 mL of the standard solution, and add a mixture of acetonitrile for liquid chro-matography, diluted phosphoric acid (3 in 1000) and metha-nol (5:4:1) to make exactly 20 mL. Conˆrm that the peak area of fusidic acid obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of fusidic acid are not less than 43,000 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of fusidic acid is not more than 2.0z. Water <2.48> Not more than 2.0z (1 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Staphylococcus aureus ATCC 6538 P Culture medium—Use the medium ii in 3) under (1) Agar media for seed and base layer. Standard solutions—Weigh accurately an amount of Diethanolamine Fusidate RS, equivalent to about 20 mg (po-tency), dissolve in 2 mL of ethanol (95), add water to make exactly 20 mL, and use this solution as the standard stock so-lution. Keep the standard stock solution at a temperature not exceeding 59C and use within 7 days. Take exactly a suitable amount of the standard stock solution before use, add phos-phate buffer solution (pH 6.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration standard solution and the low concentration standard solution, respectively. Sample solutions—Weigh accurately an amount of Sodium Fusidate, equivalent to about 20 mg (potency), and dissolve in water to make exactly 20 mL. Take exactly a suitable amount of this solution, add phosphate buffer solu-tion (pH 6.0) to make solutions so that each mL contains 4 mg (potency) and 1 mg (potency), and use these solutions as the high concentration sample solution and the low concen-tration sample solution, respectively.
Containers—Tight containers. Storage—Light-resistant, and at a temperature 2 to 89C.
Sodium Bicarbonate occurs as white, crystals or crystalline powder. It is odorless, and has a characteristic, saline taste. It is soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It slowly decomposes in moist air.
A solution of Sodium Bicarbonate (1 in 30) responds to Qualitative Tests <1.09> for sodium salt and for bicarbonate. pH <2.54> Dissolve 1.0 g of Sodium Bicarbonate in 20 mL of water: the pH of this solution is between 7.9 and 8.4.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Bicarbonate in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—To 0.40 g of Sodium Bicarbonate ‌add 4 mL of dilute nitric acid, heat to boil, cool, and add 6 mL of dilute nitric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 0.45 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.040z).‌ Carbonate—Dissolve 1.0 g of Sodium Bicarbonate in 20 mL of freshly boiled and cooled water with very gentle swirling at a temperature not exceeding 159C. Add 2.0 mL of 0.1 mol/L hydrochloric acid VS and 2 drops of phenol-phthalein TS: no red color develops immediately. Ammonium—Heat 1.0 g of Sodium Bicarbonate: the gas evolved does not change moistened red litmus paper to blue. Heavy metals <1.07>—Dissolve 4.0 g of Sodium Bi- carbonate in 5 mL of water and 4.5 mL of hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the residue in 2 mL of dilute acetic acid, 35 mL of water and 1 drop of ammonia TS, dilute with water to 50 mL, and per-form the test using this solution as the test solution. Prepare the control solution as follows: evaporate 4.5 mL of hydro-chloric acid to dryness, and add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 5 ppm). Arsenic <1.11>—Dissolve 1.0 g of Sodium Bicar- bonate in 3 mL of water and 2 mL of hydrochloric acid, and perform the test using this solution as the test solution (not more than 2 ppm).
Weigh accurately about 2 g of Sodium Bicarbonate, dissolve in 25 mL of water, and titrate with 0.5 mol/L sulfu-ric acid VS. When the color of the solution changes from blue to yellow-green, boil with caution, cool, and continue the titration <2.50> until a greenish yellow color develops (in-dicator: 2 drops of bromocresol green TS). Each mL of 0.5 mol/L sulfuric acid VS ＝ 84.01 mg of NaHCO3
Sodium Hydroxide contains not less than 95.
Sodium Hydroxide occurs as white, fused masses, in small pellets, in flakes, in sticks, and in other forms. It is hard and brittle, and shows a crystalline fracture. It is freely soluble in water and in ethanol (95), and practi- Average molecular mass ＝ 0.78 cally insoluble in diethyl ether. It rapidly absorbs carbon dioxide in air.
(1) A solution of Sodium Hydroxide (1 in 500) is alkaline. (2) A solution of Sodium Hydroxide (1 in 25) responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Hydroxide in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 2.0 g of Sodium Hydrox- ide in water, and add water to make 100 mL. To 25 mL of the solution add 10 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.7 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.050z). Heavy metals <1.07>—Dissolve 1.0 g of Sodium Hy-droxide in 5 mL of water, add 11 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the 14 20 11 n residue in 35 mL of water, add 2 mL of dilute acetic acid and ＝ MS × AT/AS × 1/V × 3/100 MS: Amount (mg) of sodium hyaluronate for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with porous polymethacry-late for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 32.2 g of sodium sulfate decahy-drate in water to make 1000 mL. drop of ammonia TS, add water to make 50 mL, and per-form the test using this solution as the test solution. Evaporate 11 mL of dilute hydrochloric acid on a water bath to dryness, dissolve the residue in 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solution, add water to make 50 mL, and use this solution as the control solution (not more than 30 ppm). Potassium—Dissolve 0.10 g of Sodium Hydroxide in water and dilute with water to make 40 mL. Add 1.0 mL of dilute acetic acid to 4.0 mL of this solution, and shake. Add 5.0 mL of a solution of sodium tetraphenylboron (1 in 30), shake immediately, and allow to stand for 10 minutes: the ‌solution has no more turbidity than the following control so-lution. Control solution: Dissolve 9.5 mg of potassium chloride in water, and dilute with water to make 1000 mL. Add 1.0 mL of dilute acetic acid to 4.0 mL of this solution, shake, and proceed as directed above. Sodium carbonate—The amount of sodium carbon-ate (Na2CO3: 105.99) is not more than 2.0z, when calcu-lated by the following equation using B (mL) which is ob-tained in the Assay. Amount (mg) of sodium carbonate ＝ 105.99 × B Mercury—Dissolve 2.0 g of Sodium Hydroxide in 1 mL of a solution of potassium permanganate (3 in 50) and 30 mL of water, neutralize gradually with purified hydro-chloric acid, and add 5 mL of diluted sulfuric acid (1 in 2). To this solution add a solution of hydroxylammonium chlo-ride (1 in 5) until the precipitate of manganese dioxide disap-pears, add water to make exactly 100 mL, and use this solu-tion as the sample solution. Perform the tests according to Atomic Absorption Spectrophotometry <2.23> (Cold vapor type) with the sample solution. Place the sample solution in the sample bottle of an atomic absorption spectrophotome-ter, add 10 mL of tin (II) chloride-sulfuric acid TS, connect the bottle immediately to the atomic absorption spectropho-tometer, and circulate air. Read the absorbance AT of the sample solution when the indication of the recorder rises rapidly and becomes constant at the wavelength of 253.7 nm. On the other hand, to 2.0 mL of Standard Mercury So-lution add 1 mL of a solution of potassium permanganate (3 in 50), 30 mL of water and a volume of purified hydrochlo-ric acid equal to that used in the preparation of the sample solution, and read the absorbance AS of the solution ob-tained by the same procedure as used for the sample solu-tion: AT is smaller than AS.
To exactly V mL of Purified Sodium Hyaluronate Ophthalmic Solution, equivalent to about 1.5 mg of purified sodium hyaluronate [(C14H20NNaO11)n], add the mobile phase to make exactly 30 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of sodium hyaluronate for assay, previously dried under reduced pressure (not exceeding 0.67 kPa) at 609C for 5 hours using phosphorus (V) oxide as a desiccant, and dis-solve in a solution of sodium chloride (9 in 1000) to make ex-actly 50 mL. Pipet 1 mL of this solution, add the mobile phase to make exactly 20 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of hyaluronic acid in each solution. Amount (mg) of purified sodium hyaluronate [(C H NNaO ) ] It deliquesces in moist air. Identification (1) A solution of Sodium Hydroxide (1 in 500) is alkaline. (2) A solution of Sodium Hydroxide (1 in 25) responds to Qualitative Tests <1.09> for sodium salt. Purity (1) Clarity and color of solution—Dissolve 1.0 g of Sodium Hydroxide in 20 mL of water: the solution is clear and colorless. Chloride <1.03>—Dissolve 2.0 g of Sodium Hydrox- ide in water, and add water to make 100 mL. To 25 mL of the solution add 10 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.7 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.050z). Heavy metals <1.07>—Dissolve 1.0 g of Sodium Hy-droxide in 5 mL of water, add 11 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Dissolve the 14 20 11 n residue in 35 mL of water, add 2 mL of dilute acetic acid and ＝ MS × AT/AS × 1/V × 3/100 MS: Amount (mg) of sodium hyaluronate for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 210 nm). Column: A stainless steel column 7.8 mm in inside diame-ter and 30 cm in length, packed with porous polymethacry-late for liquid chromatography (7 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 32.2 g of sodium sulfate decahy-drate in water to make 1000 mL. drop of ammonia TS, add water to make 50 mL, and per-form the test using this solution as the test solution. Evaporate 11 mL of dilute hydrochloric acid on a water bath to dryness, dissolve the residue in 2 mL of dilute acetic acid and 3.0 mL of Standard Lead Solution, add water to make 50 mL, and use this solution as the control solution (not more than 30 ppm). Potassium—Dissolve 0.10 g of Sodium Hydroxide in water and dilute with water to make 40 mL. Add 1.0 mL of dilute acetic acid to 4.0 mL of this solution, and shake. Add 5.0 mL of a solution of sodium tetraphenylboron (1 in 30), shake immediately, and allow to stand for 10 minutes: the ‌solution has no more turbidity than the following control so-lution. Control solution: Dissolve 9.5 mg of potassium chloride in water, and dilute with water to make 1000 mL. Add 1.0 mL of dilute acetic acid to 4.0 mL of this solution, shake, and proceed as directed above. Sodium carbonate—The amount of sodium carbon-ate (Na2CO3: 105.99) is not more than 2.0z, when calcu-lated by the following equation using B (mL) which is ob-tained in the Assay. Amount (mg) of sodium carbonate ＝ 105.99 × B Mercury—Dissolve 2.0 g of Sodium Hydroxide in 1 mL of a solution of potassium permanganate (3 in 50) and 30 mL of water, neutralize gradually with purified hydro-chloric acid, and add 5 mL of diluted sulfuric acid (1 in 2). To this solution add a solution of hydroxylammonium chlo-ride (1 in 5) until the precipitate of manganese dioxide disap-pears, add water to make exactly 100 mL, and use this solu-tion as the sample solution. Perform the tests according to Atomic Absorption Spectrophotometry <2.23> (Cold vapor type) with the sample solution. Place the sample solution in the sample bottle of an atomic absorption spectrophotome-ter, add 10 mL of tin (II) chloride-sulfuric acid TS, connect the bottle immediately to the atomic absorption spectropho-tometer, and circulate air. Read the absorbance AT of the sample solution when the indication of the recorder rises rapidly and becomes constant at the wavelength of 253.7 nm. On the other hand, to 2.0 mL of Standard Mercury So-lution add 1 mL of a solution of potassium permanganate (3 in 50), 30 mL of water and a volume of purified hydrochlo-ric acid equal to that used in the preparation of the sample solution, and read the absorbance AS of the solution ob-tained by the same procedure as used for the sample solu-tion: AT is smaller than AS. Assay Weigh accurately about 1.5 g of Sodium Hydroxide, and dissolve in 40 mL of freshly boiled and cooled water. Cool the solution to 159C, add 2 drops of phenolphthalein TS, and titrate <2.50> with 0.5 mol/L sulfuric acid VS until the red color of the solution disappears. Record the amount, A (mL), of 0.5 mol/L sulfuric acid VS consumed. Then add 2 drops of methyl orange TS to the solution, and further titrate <2.50> with 0.5 mol/L sulfuric acid VS until the solu-tion shows a persistent light red color. Record the amount, B (mL), of 0.5 mol/L sulfuric acid VS consumed. Calculate the amount of NaOH from the difference, A (mL) － B (mL). Each mL of 0.5 mol/L sulfuric acid VS ＝ 40.00 mg of NaOH
Sodium Iodide, when dried, contains not less than 99.
Sodium Iodide occurs as colorless crystals or a white crystalline powder. It is odorless. It is very soluble in water, and freely soluble in glycerin and in ethanol (95). It deliquesces in moist air.
A solution of Sodium Iodide (1 in 20) re-sponds to Qualitative Tests <1.09> for sodium salt and for iodide.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Iodide in 2 mL of water: the solution is clear and colorless. Alkalinity—Dissolve 1.0 g of Sodium Iodide in 10 mL of freshly boiled and cooled water, and add 1.0 mL of 0.005 mol/L sulfuric acid VS and 1 drop of phenolphthalein TS: no color is produced. Chloride, bromide and thiosulfate—Dissolve 0.20 g of Sodium Iodide in 5 mL of ammonia TS, add 15.0 mL of 0.1 mol/L silver nitrate VS, shake for a few minutes, and filter. To 10 mL of the filtrate add 15 mL of dilute nitric acid: no brown color appears. The solution has no more tur-bidity than the following control solution. Control solution: To 0.30 mL of 0.01 mol/L hydrochloric acid VS add 2.5 mL of ammonia TS, 7.5 mL of 0.1 mol/L silver nitrate VS and 15 mL of dilute nitric acid. Nitrate, nitrite and ammonium—Place 1.0 g of So-dium Iodide in a 40-mL test tube, and add 5 mL of water, 5 mL of sodium hydroxide TS and 0.2 g of aluminum wire. In-sert a pledget of absorbent cotton in the mouth of the test tube, and place a piece of moistened red litmus paper on the cotton. Heat the test tube on a water bath for 15 minutes: the evolved gas does not turn moistened red litmus paper to blue. Cyanide—Dissolve 0.5 g of Sodium Iodide in 10 mL of water. To 5 mL of this solution add 1 drop of iron (II) sulfate TS and 2 mL of sodium hydroxide TS, warm, and add 4 mL of hydrochloric acid: no green color develops. Iodate—Dissolve 0.5 g of Sodium Iodide in 10 mL of freshly boiled and cooled water, and add 2 drops of dilute sulfuric acid and 1 drop of starch TS: no blue color develops immediately. Heavy metals <1.07>—Proceed with 2.0 g of Sodium Iodide according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Barium—Dissolve 0.5 g of Sodium Iodide in 10 mL of water, add 1 mL of dilute sulfuric acid, and allow to stand for 5 minutes: no turbidity is produced. Potassium—Dissolve 1.0 g of Sodium Iodide in water, and add water to make 100 mL. To 4.0 mL of this so-lution add 1.0 mL of dilute acetic acid, shake, add 5.0 mL of a solution of sodium tetraphenylboron (1 in 30), immediately shake, and allow to stand for 10 minutes: the solution has no more turbidity than the following control solution. Control solution: Dissolve 9.5 mg of potassium chloride in water, and add water to make 1000 mL. To 4.0 mL of this solution add 1.0 mL of dilute acetic acid, shake, and then proceed as directed above. Arsenic <1.11>—Prepare the test solution with 0.40 g of Sodium Iodide according to Method 1, and per-form the test (not more than 5 ppm). Loss on drying <2.41> Not more than 5.0z (2 g, 1209C, 2 hours).
Weigh accurately about 0.4 g of Sodium Iodide, pre-viously dried, in an iodine flask, dissolve in 10 mL of water, add 35 mL of hydrochloric acid and 5 mL of chloroform, and titrate <2.50> with 0.05 mol/L potassium iodate VS while shaking vigorously until the red-purple color of the chloroform layer disappears. The end point is attained when the red-purple color does not reappear in the chloroform ‌layer within 5 minutes after the layer has been decolorized.‌‌‌‌ Each mL of 0.05 mol/L potassium iodate VS ＝ 14.99 mg of NaI
Sodium Lauryl Sulfate is a mixture of sodium alkyl sulfate consisting chiefly of sodium lauryl sulfate. It contains not less than 85.
Sodium Lauryl Sulfate occurs as white to light yellow, crystals or powder. It has a slightly characteris-tic odor. It is sparingly soluble in ethanol (95). A solution of 1 g of Sodium Lauryl Sulfate in 10 mL of water is a clear or an opalescent solution.◆
(1) Determine the infrared absorption spec-trum of Sodium Lauryl Sulfate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. Put 2.5 g of Sodium Lauryl Sulfate in a platinum or quartz crucible, and add 2 mL of 5 mol/L sulfuric acid TS. Heat on a water bath, cautiously raise the temperature gradually with a burner, and ignite. Ignite, preferably in an electric furnace, at 600 ± 259C and incinerate the residue completely. After cooling, add a few drops of 1 mol/L sulfu-ric acid TS, and heat and ignite as above. After cooling, add a few drops of ammonium carbonate TS, evaporate to dry-ness, and further ignite as above. After cooling, dissolve the residue in 50 mL of water, and stir. To 2 mL of this solution add 4 mL of potassium hexahydroxoantimonate (V) TS. If necessary, rub the inside wall of the vessel with a glass rod: a white crystalline precipitate is formed. Acidify a solution of Sodium Lauryl Sulfate (1 in 10) with hydrochloric acid, and boil for 20 minutes: no precipi-tate is formed. To this solution add barium chloride TS: a white precipitate is formed.
(1) Alkalinity—Dissolve 1.0 g of Sodium Lauryl Sulfate in 100 mL of water, add 0.1 mL of phenol red TS, and titrate <2.50> with 0.1 mol/L hydrochloric acid VS: the consumed volume is not more than 0.5 mL. Sodium chloride—Dissolve about 5 g of Sodium Lauryl Sulfate, accurately weighed, in 50 mL of water, neu-tralize the solution with dilute nitric acid, if necessary, add exactly 5 mL of 0.1 mol/L sodium chloride TS, and titrate <2.50> with 0.1 mol/L silver nitrate VS until the color of the solution changes from yellow-green through yellow to ‌orange (indicator: 2 drops of fluorescein sodium TS). Per-form a blank determination in the same manner, and make any necessary correction.‌ Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl The combined content of sodium chloride (NaCl: 58.44) and sodium sulfate (Na2SO4: 142.04) obtained in (3) is not more than 8.0z. Sodium sulfate—Dissolve about 1 g of Sodium Lauryl Sulfate, accurately weighed, in 10 mL of water, add 100 mL of ethanol (95), and heat at a temperature just below the boiling point for 2 hours. Filter through a glass filter (G4) while hot, and wash with 100 mL of boiling ethanol (95). Dissolve the residue on the glass filter by washing with 150 mL of water, collecting the washings in a beaker. Add 10 mL of dilute hydrochloric acid, heat to boiling, add 25 mL of barium chloride TS, and allow to stand overnight. Collect the precipitate, and wash with water until the last washing produces no opalescence with silver nitrate TS. Dry the pre-cipitate together with the filter paper, ignite to a constant mass between 5009C and 6009C by raising the temperature gradually, and weigh as barium sulfate (BaSO4: 233.39). Amount (mg) of sodium sulfate (Na2SO4) ＝ amount (mg) of barium sulfate (BaSO4) × 0.6086 Unsulfated alcohols—Dissolve about 10 g of Sodium Lauryl Sulfate, accurately weighed, in 100 mL of water, add 100 mL of ethanol (95), and transfer to a separator. Extract the solution with three 50-mL portions of pentane. If an emulsion forms, sodium chloride may be added to promote separation of the two layers. Combine the pentane extracts, wash with three 50-mL portions of water, dehydrate with an-hydrous sodium sulfate, and filter. Put the filtrate to a tared beaker, and evaporate the pentane on a water bath. Dry the residue at 1059C for 30 minutes, cool, and weigh: the mass of the residue is not more than 4.0z. Water <2.48> Not more than 5.0z (0.5 g, volumetric titration, direct titration).◇ Total alcohol content Dissolve about 5 g of Sodium Lauryl Sulfate, accurately weighed, in 150 mL of water and 50 mL of hydrochloric acid, and boil under a reflux con-denser for 4 hours. Cool, extract with two 75-mL portions of diethyl ether, and evaporate the combined diethyl ether ex-tracts on a water bath. Dry the residue at 1059C for 30 minutes, and weigh: the mass of the residue is not less than 59.0z.◇
Weigh accurately about 1.15 g of Sodium Lauryl Sulfate, and dissolve in water to make exactly 1000 mL, by warming if necessary. Transfer exactly 20 mL of this solu-tion to a 100-mL stoppered graduated cylinder, add 15 mL of dichloromethane and 10 mL of dimidium bromide-patent blue TS, and shake. Titrate <2.50> with 0.004 mol/L ben-zethonium chloride VS until the color of the dichlorometh-ane layer changes from light red to grayish blue, while shak-ing vigorously. Allow the layers to separate before each titra-tion. Each mL of 0.004 mol/L benzethonium chloride VS ＝ 1.154 mg of C12H25NaO4S
Sodium Pyrosulfite occurs as white, crystals or crystalline powder. It has the odor of sulfur dioxide. It is freely soluble in water, very slightly soluble in ethanol (95), and practically insoluble in diethyl ether. A solution of Sodium Pyrosulfite (1 in 20) is acid. It is hygroscopic. It decomposes slowly on exposure to air.
A solution of Sodium Pyrosulfite (1 in 20) responds to Qualitative Tests <1.09> for sodium salt and for bisulfite.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Pyrosulfite in 10 mL of water: the solution is clear and colorless. Thiosulfate—Dissolve 1.0 g of Sodium Pyrosulfite in 15 mL of water, add slowly 5 mL of dilute hydrochloric acid, shake, and allow to stand for 5 minutes: no turbidity is produced. Heavy metals <1.07>—Dissolve 1.0 g of Sodium Pyrosulfite in 10 mL of water, and evaporate with 5 mL of hydrochloric acid on a water bath to dryness. Dissolve the residue in 10 mL of water, add 1 drop of phenolphthalein TS, and add ammonia TS until the solution becomes slightly red. Add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: evaporate 5 mL of hydrochloric acid on a water bath to dryness, and to the residue add 2 mL of dilute acetic acid, 2.0 mL of Standard Lead Solution and water to make 50 mL (not more than 20 ppm). Iron <1.10>—Prepare the test solution with 1.0 g of Sodium Pyrosulfite according to Method 1, and perform the test according to Method A. Prepare the control solution with 2.0 mL of Standard Iron Solution (not more than 20 ppm). Arsenic <1.11>—Dissolve 0.5 g of Sodium Pyrosulfite in 10 mL of water, heat with 1 mL of sulfuric acid on a sand bath until white fumes are evolved, and add water to make 5 mL. Perform the test with this solution as the test solution (not more than 4 ppm).
Weigh accurately about 0.15 g of Sodium Pyrosul-fite, and transfer to an iodine flask containing an exactly measured 50 mL of 0.05 mol/L iodine VS. Stopper tightly, shake well, and allow to stand for 5 minutes in a dark place. Add 1 mL of hydrochloric acid, and titrate <2.50> the excess of iodine with 0.1 mol/L sodium thiosulfate VS (indicator: 1 mL of starch TS). Perform a blank determination in the same manner. Each mL of 0.05 mol/L iodine VS ＝ 4.753 mg of Na2S2O5
Sodium Picosulfate Hydrate contains not less than 98.
Sodium Picosulfate Hydrate occurs as a white crystalline powder. It is odorless and tasteless. It is very soluble in water, soluble in methanol, slightly soluble in ethanol (99.5), and practically insoluble in diethyl ether. It is gradually colored by light. The pH of a solution of 1.0 g of Sodium Picosulfate Hy-drate in 20 mL of water is between 7.4 and 9.4.
(1) Mix 5 mg of Sodium Picosulfate Hy-drate with 0.01 g of 1-chloro-2,4-dinitrobenzene, and melt Heavy metals <1.07>—Proceed with 2.0 g of Sodium Picosulfate Hydrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Sodium Picosulfate Hydrate according to Method 3, and perform the test (not more than 1 ppm). Related substances—Dissolve 0.25 g of Sodium Picosulfate Hydrate in 5 mL of methanol, and use this solu-tion as the sample solution. Pipet 1 mL of the sample solu-tion, add methanol to make exactly 500 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent indicator for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water and acetic acid (100) (74:20:19) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu-tion. Water <2.48> 3.0 – 4.5z (0.5 g, volumetric titration, direct titration).
Weigh accurately about 0.4 g of Sodium Picosulfate Hydrate, dissolve in 50 mL of methanol, add 7 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Perform a blank determina-tion in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 48.14 mg of C H NNa O S by gentle heating for 5 to 6 seconds. After cooling, add 4 mL 18 13 8 2 of potassium hydroxide-ethanol TS: an orange-red color de-velops. To 0.2 g of Sodium Picosulfate Hydrate add 5 mL of dilute hydrochloric acid, boil for 5 minutes, cool, and add 1 mL of barium chloride TS: a white precipitate is formed. Determine the absorption spectrum of a solution of Sodium Picosulfate Hydrate (1 in 25,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of So-dium Picosulfate Hydrate, previously dried at 105°C in
ポリスチレンスルホン酸ナトリウム
Sodium Polystyrene Sulfonate occurs as a yel-low-brown powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95), in ace-tone and in diethyl ether.
(1) Determine the infrared absorption spec-trum of Sodium Polystyrene Sulfonate as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum: both spectra exhibit similar intensities of ab-sorption at the same wave numbers. (2) To 1 g of Sodium Polystyrene Sulfonate add 10 mL of dilute hydrochloric acid, stir, and filter. Add ammonia TS to the filtrate to neutralize: the solution responds to Qualita-tive Tests <1.09> for sodium salt.
(1) Ammonium—Place 1.0 g of Sodium Polysty-rene Sulfonate in a flask, add 5 mL of sodium hydroxide TS, cover the flask with a watch glass having a moistened strip of red litmus paper on the underside, and boil for 15 minutes: the gas evolved does not change the red litmus paper to blue. Heavy metals <1.07>—Proceed with 2.0 g of Sodium Polystyrene Sulfonate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Stand-ard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Prepare the test solution with 2.0 g of Sodium Polystyrene Sulfonate according to Method 3, and perform the test (not more than 1 ppm). Styrene—To 10.0 g of Sodium Polystyrene Sulfonate add 10 mL of acetone, shake for 30 minutes, centrifuge, and use the supernatant liquid as the sample solution. Separately, dissolve 10 mg of styrene in acetone to make exactly 100 mL. Pipet 1 mL of this solution, add acetone to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine peak areas, AT and AS, of styrene in each solution: AT is not larger than AS. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4 mm in inside diameter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of water and acetonitrile (1:1). Flow rate: Adjust so that the retention time of styrene is about 8 minutes. System suitability— System performance: Dissolve 20 mg each of styrene and butyl parahydroxybenzoate in 100 mL of acetone. To 5 mL of this solution add acetone to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, butyl parahydroxybenzoate and sty-rene are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of styrene is not more than 2.0z. Water <2.48> Not more than 10.0z (0.2 g, volumetric titration, direct titration).
(1) Sodium—Weigh accurately about 1 g of So-dium Polystyrene Sulfonate, calculated on the anhydrous basis, in a glass-stoppered flask, add exactly 50 mL of 3 mol/L hydrochloric acid TS, shake for 60 minutes, and filter. Discard the first 20 mL of the filtrate, pipet 5 mL of the subsequent filtrate, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add water to make exactly 1000 mL, and use this solution as the sample solution. Sepa-rately, pipet a suitable quantity of Standard Sodium Stock Solution, dilute exactly with water so that each ml of the so-lution contains 1 to 3 mg of sodium (Na: 22.99), and use these solutions as the standard solutions. Perform the test with the sample solution and the standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and calculate the amount of sodium in the sample solution using the calibration curve obtained from the standard solutions. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A sodium hollow-cathode lamp. Wavelength: 589.0 nm. (2) Potassium exchange capacity—Weigh accurately about 1.5 g of Sodium Polystyrene Sulfonate, calculated on the anhydrous basis, in a glass-stoppered flask, add exactly 100 mL of Standard Potassium Stock Solution, shake for 15 minutes, and filter. Discard the first 20 mL of the filtrate, pipet 10 mL of the subsequent filtrate, and add water to make exactly 100 mL. Pipet 10 mL of this solution, add water to make exactly 1000 mL, and use this solution as the sample solution. Separately, pipet a suitable quantity of Standard Potassium Stock Solution, dilute exactly with water so that each mL of the solution contains 1 to 5 mg of potassium (K: 39.10), and use these solutions as the standard solutions. Perform the test with these solutions as directed under Atomic Absorption Spectrophotometry <2.23> accord-ing to the following conditions, and calculate the amount Y (mg) of potassium in 1000 mL of the sample solution using the calibration curve obtained from the standard solution. The quantity of potassium absorbed on each g of Sodium Polystyrene Sulfonate, calculated on the anhydrous basis, is calculated from the following equation: it is between 0.110 g and 0.135 g. Quantity (mg) of potassium (K) absorbed on 1 g of Sodium Polystyrene Sulfonate, calculated on the anhydrous basis ＝ (X － 100Y )/M X: Amount (mg) of potassium in 100 mL of the Standard Potassium Stock Solution before exchange M: Mass (g) of Sodium Polystyrene Sulfonate taken, cal-culated on the anhydrous basis Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: A potassium hollow-cathode lamp. ‌Wavelength: 766.5 nm.‌
Sodium Risedronate Hydrate contains not less than 98.
Sodium Risedronate Hydrate occurs as a white crystalline powder. It is soluble in water, and practically insoluble in ethanol (99.5). It dissolves in diluted dilute sodium hydroxide TS (1 in 20).
(1) Determine the absorption spectrum of a solution of Sodium Risedronate Hydrate in diluted dilute so-dium hydroxide TS (1 in 20) (1 in 20,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra ex-hibit similar intensities of absorption at the same wave-lengths. Determine the infrared absorption spectrum of So-dium Risedronate Hydrate as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Sodium Risedronate Hydrate responds to Qualitative Tests <1.09> (1) for sodium salt.
(1) Heavy metals—To 0.50 g of Sodium Rise-dronate Hydrate in a quartz crucible add 0.50 g of magne-sium oxide, mix, heat until the content becomes a light gray while mixing occasionally with a glass rod, then incinerate at 8009C. After cooling, dissolve the residue with 3 mL of hy-drochloric acid, and add 3 mL of water. Adjust this solution to pH 8.5 with ammonia TS, then adjust to pH 4 with acetic acid (100), and adjust the pH to 3.4 with dilute hydrochloric acid. Filter the solution into a Nessler tube using a filter paper, rinse the crucible and filter with water, add the rins-ings to the Nessler tube, then add water to make 50 mL, and use this solution as the test solution. Separately, to 1.0 mL of Standard Lead Solution add 0.50 g of magnesium oxide, dryness at 1109C, and proceed with the residue in the same manner as for the test solution, and use the solution so ob-tained as the control solution. To the test and control solu-tions add 1 drop each of sodium sulfide TS, mix, and allow to stand for 5 minutes, and compare the colors of both solu-tions against a white background: the color of the test solu-tion is not more intense than that of the control solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution by dissolv- ing 1.0 g of Sodium Risedronate Hydrate in 5 mL of a solu-tion of sodium hydroxide (1 in 5), and perform the test (not more than 2 ppm). Related substance 1—Dissolve 50 mg of Sodium Risedronate Hydrate in 1.5 mL of 0.2 mol/L sodium hy-droxide TS, add the mobile phase to make 25 mL, and use this solution as the sample solution. Pipet 2.5 mL of the sample solution, and add the mobile phase to make exactly 50 mL. Pipet 2 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine each peak area by the automatic in-tegration method: the area of the peak other than risedronic acid obtained from the sample solution is not larger than the peak area of risedronic acid from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of risedronic acid, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risedronic acid are not less than 4500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risedronic acid is not more than 5.0z. Related substance 2—Dissolve 0.10 g of Sodium Risedronate Hydrate in 3 mL of 0.2 mol/L sodium hydrox-ide TS, add the diluting solution below to make 50 mL, and use this solution as the sample solution. Pipet 2.5 mL of the sample solution, and add the diluting solution to make ex- acid is about 5 minutes. Time span of measurement: About 10 times as long as the retention time of risedronic acid, beginning after the solvent peak. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of risedronic acid are not less than 5000 and not more than 1.4, respectively. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of risedronic acid is not more than 2.0z. Water <2.48> 11.9 – 13.9z (40 mg, volumetric titration, direct titration. Use a mixture of formamide for water deter-mination and methanol for water determination (1:1) instead of methanol for water determination).
Weigh accurately about 50 mg of Sodium Rise-dronate Hydrate, dissolve in 1.5 mL of 0.2 mol/L sodium hydroxide TS, and add the mobile phase to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Risedronic Acid RS (separately determine the water <2.48> using 80 mg, in the same manner as Sodium Risedronate Hydrate), dissolve in 3 mL of 0.2 mol/L sodium hydroxide TS, and add the mobile phase to make exactly 25 mL. Pipet 10 mL of this solution, add exactly 5 mL of the internal standard solution, then add the mobile phase to make 25 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and calculate the ratios, QT and QS, of the peak area of risedronic acid to that of the internal standard. actly 50 mL. Pipet 2 mL of this solution, add the diluting so- Amount (mg) of sodium risedronate (C H NNaO P ) lution to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 50 mL each ＝ MS × QT/QS × 1.078 7 10 7 2 of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the area of the peak other than risedronic acid obtained from the sample solution is not larger than the peak area of risedronic acid from the stand-ard solution. Diluting solution: Dissolve 0.11 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate and 2.47 g of tetra-decyl trimethylammonium bromide in 1000 mL of water, and adjust to pH 6.5 with 0.2 mol/L sodium hydroxide TS. To 700 mL of this solution add 300 mL of acetonitrile. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 0.14 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate, 3.16 g of tetradecyl trimethylammonium bromide, 4.81 g of ammonium dihy-drogen phosphate and 2.93 g of diammonium hydrogen phosphate in 1280 mL of water, and add 720 mL of aceto-nitrile. Flow rate: Adjust so that the retention time of risedronic MS: Amount (mg) of Risedronic Acid RS taken, calcu-lated on the anhydrous basis Internal standard solution—A solution of sodium benzoate in the mobile phase (1 in 125). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 263 nm). Column: A polyether ether ketone column 4 mm in inside diameter and 25 cm in length, packed with quaternary alkylaminated styrene-divinylbenzene copolymer for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.8 g of disodium dihydrogen ethylenediamine tetraacetate dihydrate in 1000 mL of water, and adjust to pH 9.5 with 0.2 mol/L sodium hydroxide TS. Flow rate: Adjust so that the retention time of risedronic acid is about 14 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the internal standard and risedronic acid are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat- ‌ing conditions, the relative standard deviation of the peak area of risedronic acid is not more than 1.0z.
Sodium Salicylate, when dried, contains not less than 99.
Sodium Salicylate occurs as white, crystals or crystalline powder. It is very soluble in water, freely soluble in acetic acid (100), and soluble in ethanol (95). It is gradually colored by light.
(1) Determine the infrared absorption spec-trum of Sodium Salicylate, previously dried, as directed in the potassium bromide disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Sodium Salicylate (1 in 20) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> The pH of a solution of 2.0 g of Sodium Salicy-late in 20 mL of water is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Salicylate in 10 mL of water: the solution is clear, and its absorbance at 420 nm determined as directed under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.02. Chloride <1.03>—Dissolve 0.5 g of Sodium Salicylate in 15 mL of water, add 6 mL of dilute nitric acid and ethanol (95) to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.30 mL of 0.01 mol/L hydrochloric acid VS, 28 mL of ethanol (95), 6 mL of dilute nitric acid and water to make 50 mL (not more than 0.021z). Sulfate—Dissolve 0.25 g of Sodium Salicylate in 5 mL of water, and add 0.5 mL of barium chloride TS: the solution shows no change. Sulfite and thiosulfate—Dissolve 1.0 g of Sodium Salicylate in 20 mL of water, add 1 mL of hydrochloric acid, and filter. Add 0.15 mL of 0.05 mol/L iodine VS to the fil-trate: a yellow color develops. Heavy metals <1.07>—Proceed with 1.0 g of Sodium Salicylate according to Method 2, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—To 1.0 g of Sodium Salicylate in a decomposition flask add 5 mL of nitric acid and 2 mL of sul-furic acid, and heat carefully until white fumes are evolved. After cooling, add 2 mL of nitric acid, and heat. After cool-ing, add several 2-mL portions of hydrogen peroxide (30), and heat until the solution is colorless to pale yellow. Repeat the procedure of adding nitric acid and hydrogen peroxide (30) and heating, if necessary. After cooling, add 2 mL of a saturated solution of ammonium oxalate monohydrate, and heat until white fumes are evolved. After cooling, add water to make 5 mL, and perform the test with this solution (not more than 2 ppm). Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours).
Weigh accurately about 0.3 g of Sodium Salicylate, previously dried, dissolve in 50 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.01 mg of C7H5NaO3
デンプングリコール酸ナトリウム
Sodium Starch Glycolate is the sodium salt of a car-boxymethyl ether of starch or of a cross-linked car-boxymethyl ether of starch. There are two neutralization types of Sodium Starch Glycolate, Type A and Type B, and their insoluble matter in a mixture of ethanol (99.5) and water (8:2), when dried, contains not less than 2.
Sodium Starch Glycolate occurs as a white powder, and has a characteristic salty taste. It practically insoluble in ethanol (99.5). It swells with water, and becomes viscous, pasty liquid. It is hygroscopic.◆
(1) Acidify 5 mL of a solution of Sodium Starch Glycolate (1 in 500) with dilute hydrochloric acid, then add one drop of iodine TS, and stir: a blue to violet color is produced. (2) Determine the infrared absorption spectrum of So- dium Starch Glycolate as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers.◆ The sample solution obtained in the Purity (2) re-sponds to Qualitative Tests <1.09> (2) for sodium salt. Per-form the test using 2 mL of the sample solution and 4 mL of potassium hexahydroxoantimonate (V) TS. pH <2.54> To 1 g of Sodium Starch Glycolate add 30 mL of water and stir: the pH of the resulting suspension of Type A is 5.5 – 7.5, and that of Type B is 3.0 – 5.0.
◆(1) Heavy metals <1.07>—Proceed with 1.0 g of Sodium Starch Glycolate according to Method 2, and per-form the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).◆ Iron Sample solution Take 2.5 g of Sodium Starch Glycolate in a silica or platinum crucible, add 2 mL of 5 mol/L sulfuric acid TS. Heat on a water bath, then ignite cautiously with a gas burner or preferably in an electric fur-nace at 600 ± 259C, and incinerate the residue completely. Allow to cool, add a few drops of 1 mol/L sulfuric acid TS, and heat and ignite as above. Allow to cool, add a few drops of ammonium carbonate TS, evaporate to dryness on a water bath, and heat and ignite as above. After cooling, dis-solve the residue by adding 50 mL of water. Standard solution Weigh accurately 863.4 mg of ammonium iron (III) sulfate dodecahydrate, dissolve in water, add 25 mL of 1 mol/L sulfuric acid TS, and add water to make exactly 500 mL. Pipet 10 mL of this solution, and add water to make exactly 100 mL. Pipet 5 mL of this solution, and add water to make exactly 100 mL. Each mL of this solution contains 1.0 mg of iron (Fe). Procedure Pipet 10 mL each of the sample solution and standard solution, and to each solution add 2 mL of citric acid solution (1 in 5) and 0.1 mL of thioglycolic acid. Then add ammonia solution (28) dropwise to render the so-lution alkaline, using litmus paper as an indicator. Add water to make 20 mL, and use these solutions as the test so-lution and the control solution, respectively. Allow these so-lutions to stand for 5 minutes, and compare the color of the solutions using white background: the color of the test solu-tion is not deeper than that of the control solution (not more than 20 ppm). Sodium glycolate—Conduct this procedure without exposure to light, using light-resistant vessels. Sample solution Weigh accurately 0.200 g of So-dium Starch Glycolate in a beaker, add 4 mL of 6 mol/L acetic acid TS and 5 mL of water, and stir to dissolve. Add 50 mL of acetone and 1 g of sodium chloride, stir, and filter through a filter paper previously soaked with acetone. Rinse the beaker and the filter paper with acetone, combine the fil-trate and washings, and add acetone to make exactly 100 mL. Allow to stand for 24 hours, and use the supernatant liquid as the sample solution. Standard solution To exactly 0.310 g of glycolic acid, previously dried in a desiccator (silica gel) for 18 hours, add water to dissolve to make exactly 500 mL. Pipet 5 mL of this solution, add 4 mL of 6 mol/L acetic acid TS, and allow to stand for 30 minutes. Add 50 mL of acetone and 1 g of so-dium chloride, proceed as (i) above, and use the supernatant liquid as the standard solution. Procedure Pipet 2.0 mL each of the sample solu-tion and standard solution into 25-mL stoppered test tubes, and heat on a water bath for 20 minutes to remove acetone. After cooling, add 20.0 mL of 2,7-dihydroxynaphthalene TS to the residue, stopper the test tube, and heat on a water bath for 20 minutes. Cool under running water, and transfer whole quantity of the content to a 25-mL volumetric flask. Maintain the flask under running water, and add sulfuric acid to make 25 mL. Within 10 minutes, determine the ab-sorbance of these solutions at 540 nm using water as the blank as directed under Ultraviolet-visible Spectrophotome-try <2.24>; the absorbance of the sample solution is not larger than that of the standard solution (not more than 2.0z). Sodium chloride—Weigh accurately about 0.5 g of Sodium Starch Glycolate in a beaker, disperse in 100 mL of water, and add 1 ml of nitric acid. Titrate <2.50> with 0.1 mol/L silver nitrate VS (potentiometric titration): the amount of sodium chloride (NaCl: 58.44) is not more than 7.0z. Each mL of 0.1 mol/L silver nitrate VS ＝ 5.844 mg of NaCl Loss on drying <2.41> Not more than 10.0z (1 g, 1309C, 90 minutes). Microbial limits <4.05> Salmonella and Escherichia coli are not observed.
To about 1 g of Sodium Starch Glycolate add 20 mL of a mixture of ethanol (99.5) and water (8:2), stir for 10 minutes, and filter. Repeat this procedure until no more tur-bidity is produced by adding silver nitrate TS, and dry the residue on the filter paper at 1059C to constant mass. Weigh accurately 0.7 g of the mass, add 80 mL of acetic acid (100), and heat the mixture under a reflux condenser on a water bath for 2 hours. After cooling, titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Content (z) of sodium (Na) ＝ V × 2.299 × 100/M V: Consumed amount (mL) of 0.1 mol/L perchloric acid VS M: Mass (mg) of the dried residue ◆Containers and storage Containers—Tight containers.◆ ‌Dried Sodium Sulfite‌‌
Sodium Thiosulfate Hydrate, when dried, contains not less than 99.
Sodium Thiosulfate Hydrate occurs as color-less, crystals or crystalline powder. It is odorless. It is very soluble in water, and practically insoluble in ethanol (99.5). It effloresces in dry air, and is deliquescent in moist air.
(1) A solution of Sodium Thiosulfate Hy-drate (1 in 10) responds to Qualitative Tests <1.09> for thio-sulfate. (2) A solution of Sodium Thiosulfate Hydrate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> Dissolve 1.0 g of Sodium Thiosulfate Hydrate in 10 mL of water: the pH of the solution is between 6.0 and 8.0.
(1) Clarity and color of solution—Dissolve 1.0 g of Sodium Thiosulfate Hydrate in 10 mL of water: the solu-tion is clear and colorless. Heavy metals <1.07>—Dissolve 1.0 g of Sodium Thiosulfate Hydrate in 10 mL of water, add slowly 5 mL of dilute hydrochloric acid, and evaporate on a water bath to dryness. Add 15 mL of water to the residue, boil gently for 2 minutes, and filter. Heat the filtrate to boil, and add bro-mine TS to the hot filtrate to produce a clear solution and provide a slight excess of bromine. Boil the solution to expel the bromine. Cool, add 1 drop of phenolphthalein TS, and add dropwise sodium hydroxide TS until a slight red color is produced. Add 2 mL of dilute acetic acid and water to make 50 mL. Perform the test using this solution as the test solu-tion. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). Calcium—Dissolve 1.0 g of Sodium Thiosulfate in 10 mL of water, add 2 mL of ammonium oxalate TS, and allow to stand for 4 minutes: no turbidity is produced. Arsenic <1.11>—To 0.40 g of Sodium Thiosulfate add mL of nitric acid and 5 mL of water, evaporate on a water bath to dryness, and perform the test with the residue. Pre-pare the test solution according to Method 2, and perform the test (not more than 5 ppm). Loss on drying <2.41> 32.0 – 37.0z (1 g, in vacuum, 40 – 459C, 16 hours).
Weigh accurately about 0.4 g of Sodium Thiosul-fate, previously dried, dissolve in 30 mL of water, and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 15.81 mg of Na2S2O3
Sodium Valproate, when dried, contains not less than 98.
Sodium Valproate occurs as a white crystalline powder. It is very soluble in water, freely soluble in ethanol (99.5) and in acetic acid (100). It is hygroscopic.
(1) To 5 mL of a solution of Sodium Val-proate (1 in 20) add 1 mL of a solution of cobalt (II) nitrate hexahydrate (1 in 20) and warm on a water bath: a purple precipitate is formed. Dissolve 0.5 g of Sodium Valproate in 5 mL of water, add 5 mL of diethyl ether and 1 mL of 2 mol/L hydrochloric acid TS, and shake vigorously for 1 minute. Separate the diethyl ether layer, dehydrate with anhydrous sodium sul-fate, and filter. Evaporate the solvent of the filtrate, deter-mine the infrared spectrum of the residue as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Sodium Valproate (1 in 10) responds to Qualitative Tests <1.09> for sodium salt. pH <2.54> Dissolve 1.0 g of Sodium Valproate in 20 mL of water: the pH of this solution is between 7.0 and 8.5.
(1) Heavy metals <1.07>—Dissolve 2.0 g of So-dium Valproate in 44 mL of water, shake with 6 mL of dilute hydrochloric acid, allow to stand for 5 minutes, and filter. Discard the first 5 mL of the filtrate, neutralize 25 mL of the subsequent filtrate with ammonia TS, and add 2 mL of di-lute acetic acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid and water to make 50 mL (not more than 20 ppm). (2) Related substances—Dissolve 0.10 g of Sodium Val-proate in 10 mL of a mixture of formic acid and methyl ace-tate (1:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add a mixture of formic acid and methyl acetate (1:1) to make exactly 200 mL, and use this solution as the standard solution. Perform the test with exactly 2 mL each of the sample solution and standard solu-tion as directed under Gas Chromatography <2.02> accord-ing to the following conditions. Determine each peak area of both solutions by automatic integration method: the total area of the peaks other than valproic acid obtained from the sample solution is not larger than the peak area of valproic acid from the standard solution. Operating conditions— Detector: A hydrogen flame-ionization detector. Column: A glass column 3 mm in inside diameter and 2 m in length, packed with siliceous earth for gas chromatogra-phy (150 to 180 mm in particle diameter) coated with diethy-lene glycol adipate ester for gas chromatography and phos-phoric acid at the ratios of 5z and 1z, respectively. Column temperature: A constant temperature of about 1459C. Carrier gas: Nitrogen. Flow rate: Adjust so that the retention time of valproic acid is about 7 minutes. Time span of measurement: About 2 times as long as the retention time of valproic acid, beginning after the solvent peak. System suitability— System performance: To 2 mL of the sample solution and 8 mL of n-valerianic acid, add a mixture of formic acid and methyl acetate (1:1) to make 10 mL. When the procedure is run with 2 mL of this solution under the above operating conditions, n-valerianic acid and valproic acid are eluted in this order with the resolution between these peaks being not less than 5. System repeatability: Pipet 2 mL of the standard solution and add a mixture of formic acid and methyl acetate (1:1) to make exactly 10 mL. When the test is repeated 6 times with 2 mL of this solution under the above operating conditions, the relative standard deviation of the peak area of valproic acid is not more than 5.0z. Loss on drying <2.41> Not more than 1.0z (1 g, 1059C, 3 hours).
Weigh accurately about 0.2 g of Sodium Valproate, previously dried, dissolve in 80 mL of acetic acid (100), and titrate <2.50> with 0.1 mol/L perchloric acid VS (potenti-ometric titration). Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 16.62 mg of C8H15NaO2
Spironolactone, when dried, contains not less than respectively, then evaporate methanol to dryness, and repeat the test on the residues.
Spironolactone occurs as a white to light yel-low-brown fine powder. It is freely soluble in chloroform, soluble in ethanol (95), slightly soluble in methanol, and practically insoluble in water. Melting point: 198 – 2079C (Insert the capillary tube into a bath at about 1259C, and continue the heating so that the temperature rises at a rate of about 109C per minute in the range between 1409C and 1859C, and when the temperature is near the expected melting range, reduce the heating so that the temperature rises at a rate of about 39C per minute.) MS: amount (mg) of Spironolactone RS taken
(1) Mercapto compounds—Shake 2.0 g of Spironolactone with 20 mL of water, and filter. To 10 mL of the filtrate add 1 mL of starch TS and 0.05 mL of 0.01 mol/L iodine VS, and mix: a blue color develops. (2) Related substances—Dissolve 0.20 g of Spironolac-tone in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add ethanol (95) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with n-butyl acetate to a distance of about 15 cm, and air-dry the plate. Spray evenly a solution of sulfuric acid in methanol (1 in 10) on the plate, and heat the plate at 1059C for 10 minutes: the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (1 g, 1059C, 2 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 50 mg each of Spironolac-tone and Spironolactone RS, previously dried at 1059C for 2 hours, dissolve in methanol to make exactly 250 mL. Pipet 5 mL each of these solutions, add methanol to make exactly 100 mL, and use these solutions as the sample solution and the standard solution, respectively. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, and determine the absorbances, AT and AS, of the sample solution and standard solution at 238 nm. 97.0z and not more than 103.0z of spironolactone (C24H32O4S). Amount (mg) of spironolactone (C ＝ MS × AT/AS 24H 32O4S) Description Spironolactone occurs as a white to light yel-low-brown fine powder. It is freely soluble in chloroform, soluble in ethanol (95), slightly soluble in methanol, and practically insoluble in water. Melting point: 198 – 2079C (Insert the capillary tube into a bath at about 1259C, and continue the heating so that the temperature rises at a rate of about 109C per minute in the range between 1409C and 1859C, and when the temperature is near the expected melting range, reduce the heating so that the temperature rises at a rate of about 39C per minute.) MS: amount (mg) of Spironolactone RS taken
Sulbactam Sodium contains not less than 875 mg (potency) and not more than 941 mg (potency) per mg, calculated on the anhydrous basis.
Sulbactam Sodium occurs as a white to yellow-ish white crystalline powder. It is freely soluble in water, sparingly soluble in methanol, very slightly soluble in ethanol (99.5), and practically insolu-ble in acetonitrile.
(1) Determine the infrared absorption spec-trum of Sulbactam Sodium as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. (2) Sulbactam Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋219 – ＋2339(1 g, water, 100 mL, 100 mm). ‌pH <2.54> Dissolve 1.0 g of Sulbactam Sodium in 20 mL of water: the pH of the solution is between 5.2 and 7.2.
(1) Clarity and color of solution—Dissolve 1.0 g of Sulbactam Sodium in 20 mL of water: the solution is clear, and its absorbance at 430 nm determined as directed under Ultraviolet-visible Spectrophotometry <2.24> is not more than 0.10. Heavy metals <1.07>—Proceed with 1.0 g of Sulbac- tam Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Sulbactam penicillamine—Weigh accurately about 0.2 g of Sulbactam Sodium, dissolve in the mobile phase to make exactly 50 mL, and use this solution as the sample so-lution. Separately, weigh accurately about 40 mg of sulbac-tam sodium for sulbactam penicillamine, dissolve in 2 mL of water, and add 0.5 mL of sodium hydroxide TS. Allow to stand at room temperature for 10 minutes, add 0.5 mL of 1 mol/L hydrochloric acid TS, and then add the mobile phase to make exactly 100 mL. Pipet 5 mL of this solution, add the mobile phase to make exactly 50 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of sulbactam penicillamine by the automatic integration method: the amount of sulbactam penicillamine is not more than 1.0z. Amount (z) of sulbactam penicillamine ＝ MS/MT × AT/AS × 5 MS: Amount (mg) of sulbactam sodium for sulbactam penicillamine taken MT: Amount (mg) of Sulbactam Sodium taken Operating conditions— Column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Detector: An ultraviolet absorption photometer (wave-length: 230 nm). System suitability— System performance: Proceed as directed in the system suitability in the Assay. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sulbactam penicillamine is not more than 2.0z. Water <2.48> Not more than 1.0z (0.5 g, volumetric titra-tion, direct titration).
Weigh accurately amounts of Sulbactam Sodium and Sulbactam RS, equivalent to about 50 mg (potency), dis-solve each in a suitable amount of the mobile phase, add ex-actly 5 mL of the internal standard solution, then add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respectively. Per-form the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of sulbactam to that of the internal standard. Amount [mg (potency)] of sulbactam (C8H11NO5S) ＝ MS × QT/QS × 1000 MS: Amount [mg (potency)] of Sulbactam RS taken Internal standard solution—A solution of ethyl parahy-droxybenzoate in the mobile phase (7 in 1000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 220 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: To 750 mL of 0.005 mol/L tetrabutylam-monium hydroxide TS add 250 mL of acetonitrile for liquid chromatography. Flow rate: Adjust so that the retention time of sulbactam is about 6 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, sulbactam and the internal standard are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of sulbactam is not more than 1.0z.
Sulbenicillin Sodium contains not less than 900 mg (potency) and not more than 970 mg (potency) per mg, calculated on the anhydrous basis.
Sulbenicillin Sodium occurs as white to light yellow-white powder. It is very soluble in water, freely soluble in methanol, and slightly soluble in ethanol (99.5). It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Sulbenicillin Sodium as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- trum or the spectrum of Sulbenicillin Sodium RS: both spec-tra exhibit similar intensities of absorption at the same wave numbers. (2) Sulbenicillin Sodium responds to Qualitative Tests <1.09> (1) for sodium salt. D Optical rotation <2.49> [a]20: ＋167 – ＋1829(1 g calculated on the anhydrous basis, water, 20 mL, 100 mm). ‌pH <2.54> The pH of a solution obtained by dissolving 0.20 g of Sulbenicillin Sodium in 10 mL of water is between 4.5 and 7.0.
(1) Clarity and color of solution—Dissolve 2.5 g of Sulbenicillin Sodium in 5 mL of water: the solution is clear and colorless to pale yellow. Heavy metals <1.07>—Proceed with 1.0 g of Sul- benicillin Sodium according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Sulbenicillin Sodium according to Method 1, and perform the test (not more than 2 ppm). Related substances—Dissolve 0.10 g of Sulbenicillin Sodium in 15 mL of the mobile phase, and use this solution as the sample solution. Perform the test with 10 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, determine each peak area by the automatic integration method, and calcu-late the amount of these peaks by the area percentage method: the amount of the each peak other than the two peaks of sulbenicillin is not more than 2.0z, and the total amount of the peaks other than the two peaks of sulbenicil-lin is not more than 5.0z. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 3.9 mm in inside diame-ter and 30 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 10 g of potassium dihydrogen phosphate in 750 mL of water, adjust the pH to 6.0 ± 0.1 with sodium hydroxide TS, and add water to make 1000 mL. To 940 mL of this solution add 60 mL of acetonitrile. Flow rate: Adjust so that the retention time of the lately eluted peak of sulbenicillin is about 18 minutes. Time span of measurement: About 1.5 times as long as the retention time of the lately eluted peak of sulbenicillin, be-ginning after the solvent peak. System suitability— Test for required detectability: Measure exactly 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suita-bility test, and add the mobile phase to make exactly 10 mL. Confirm that the total area of the two peaks of sulbenicillin obtained with 10 mL of this solution is equivalent to 7 to 13z of that with 10 mL of the solution for system suitability test. System performance: When the procedure is run with 10 mL of the sample solution under the above operating condi-tions, the resolution between the two peaks of sulbenicillin is not less than 2.0. System repeatability: When the test is repeated 6 times with 10 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the total areas of the two peaks of sulbenicillin is not more than 5.0z. Water <2.48> Not more than 6.0z (0.5 g, volumetric titra-tion, direct titration).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism—Bacillus subtilis ATCC 6633 Culture medium—Use the medium i in 1) under (1) Agar media for seed and base layer. Adjust the pH of the medium so that it will be 6.4 to 6.6 after sterilization. Standard solutions—Weigh accurately an amount of Sulbenicillin Sodium RS, equivalent to about 50 mg (po-tency), dissolve in phosphate buffer solution (pH 6.0) to make exactly 50 mL, and use this solution as the standard stock solution. Keep the standard stock solution in a freezer, and use within 4 days. Take exactly a suitable amount of the standard stock solution before use, add phosphate buffer so-lution (pH 6.0) to make solutions so that each mL contains 40 mg (potency) and 10 mg (potency), and use these solutions as the high concentration standard solution and the low con-centration standard solution, respectively. Sample solutions—Weigh accurately an amount of Sulbenicillin Sodium, equivalent to about 50 mg (potency), and dissolve in phosphate buffer solution (pH 6.0) to make exactly 50 mL. Take exactly a suitable amount of this solu-tion, add phosphate buffer solution (pH 6.0) to make solu-tions so that each mL contains 40 mg (potency) and 10 mg (potency), and use these solutions as the high concentration sample solution and the low concentration sample solution, respectively.
Sulfobromophthalein Sodium, when dried, contains not less than 96.
Sulfobromophthalein Sodium occurs as a white crystalline powder. It is odorless. It is soluble in water, and practically insoluble in ethanol (95) and in diethyl ether. It is hygroscopic.
(1) Dissolve 0.02 g of Sulfobromophthalein Sodium in 10 mL of water, and add 1 mL of sodium carbon-ate TS: a blue-purple color is produced. Add 1 mL of dilute hydrochloric acid to the solution: the color of the solution disappears. Transfer 0.2 g of Sulfobromophthalein Sodium to a porcelain crucible, mix well with 0.5 g of anhydrous sodium carbonate, and ignite until the mixture is charred. After cooling, add 15 mL of hot water to the residue, heat for 5 minutes on a water bath, filter, and render the filtrate slightly acid with hydrochloric acid: the solution responds to Qualitative Tests <1.09> for bromide, and (1) and (2) for sul-fate. Sulfobromophthalein Sodium responds to Qualita-tive Tests <1.09> (1) for sodium salt. pH <2.54> The pH of a solution of 1.0 g of Sulfobromo-phthalein Sodium in 20 mL of water is between 4.0 and 5.5.
(1) Clarity and color of solution—Dissolve 0.5 g of Sulfobromophthalein Sodium in 10 mL of water: the so-lution is clear and colorless to pale yellow. Loss on drying <2.41> Not more than 5.0z (0.5 g, 1059C, hours). Residue on ignition <2.44> 14 – 19z (after drying, 0.5 g, 700 – 7509C).
Dissolve about 0.1 g of Sulfobromophthalein So-dium, previously dried and accurately weighed, in water to make exactly 500 mL. Pipet 5 mL of this solution, and add a solution of anhydrous sodium carbonate (1 in 100) to make exactly 200 mL. Perform the test with this solution as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Determine the absorbance A of this solution at the wave-length of maximum absorption at about 580 nm, using water as the blank. Amount (mg) of sulfobromophthalein sodium (C20H8Br4Na2O10S2) ＝ A/881 × 200,000
Synthetic Aluminum Silicate occurs as a white powder. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95) and in diethyl ether. Synthetic Aluminum Silicate (1 g) dissolves when heated in 20 mL of a solution of sodium hydroxide (1 in 5), leaving a small amount of insoluble substance .
(1) To 0.5 g of Synthetic Aluminum Sili-cate add 3 mL of diluted sulfuric acid (1 in 3), heat until white fumes evolve, cool, add 20 mL of water, and filter. Render the filtrate slightly acid with ammonia TS: the solu-tion responds to Qualitative Tests <1.09> for aluminum salt. (2) Prepare a bead by fusing ammonium sodium hydrogenphosphate tetrahydrate on a platinum loop. Place the bead in contact with Synthetic Aluminum Silicate, and fuse again: an infusible material appears in the bead, producing, upon cooling, an opaque bead with a web-like structure.
(1) Acidity or alkalinity—Shake 1.0 g of Synthetic Aluminum Silicate with 20 mL of water, and centrifuge: the supernatant liquid so obtained is neutral. Chloride <1.03>—To 5.0 g of Synthetic Aluminum Silicate add 100 mL of water, boil gently for 15 minutes while shaking, then cool, add water to restore the original volume, and centrifuge. To 10 mL of the supernatant liquid add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solution. Pre-pare the control solution with 0.30 mL of 0.01 mol/L hydro-chloric acid VS (not more than 0.021z). Sulfate <1.14>—To 2.0 mL of the supernatant liquid obtained in (2) add 1 mL of dilute hydrochloric acid and water to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 1.0 mL of 0.005 mol/L sulfuric acid VS (not more than 0.480z). Heavy metals <1.07>—To 3.0 g of Synthetic Alumi-num Silicate add 50 mL of water and 5 mL of hydrochloric acid, boil gently for 20 minutes while shaking, then after cooling, centrifuge, remove the supernatant liquid, wash the precipitate with two 10-mL portions of water, centrifuging ‌444 each time, combine these washings with the filtrate, and add ammonia solution (28) dropwise until a precipitate just ap-pears. Add dropwise dilute hydrochloric acid with vigorous shaking to redissolve the precipitate. Heat the solution with 0.45 g of hydroxylammonium chloride, and after cooling, add 0.45 g of sodium acetate trihydrate, 6 mL of dilute ace-tic acid and water to make 150 mL. Perform the test with 50 mL of this solution as the test solution. Prepare the control solution with 3.0 mL of Standard Lead Solution, 0.15 g of hydroxylammonium chloride, 0.15 g of sodium acetate trihy-drate, 2 mL of dilute acetic acid and water to make 50 mL (not more than 30 ppm). Arsenic <1.11>—To 1.0 g of Synthetic Aluminum Sili- cate add 10 mL of dilute hydrochloric acid, heat gently to boiling while shaking well, cool rapidly, and centrifuge. Mix the residue with 5 mL of dilute hydrochloric acid with shak-ing, centrifuge, then add 10 mL of water to the residue, and repeat the extraction in the same manner. Concentrate the combined extracts on a water bath to 5 mL. Use this solution as the test solution, and perform the test (not more than 2 ppm). Loss on drying <2.41> Not more than 20.0z (1 g, 1059C, 3 hours). Acid-consuming capacity <6.04> Weigh accurately about 1 g of Synthetic Aluminum Silicate, transfer to a glass-stoppered flask, add 200 mL of 0.1 mol/L hydrochloric acid VS, exactly measured, stopper the flask, and shake at 37 ± 29C for 1 hour. Filter, pipet 50 mL of the filtrate, and titrate <2.50> by stirring well the excess hydrochloric acid with 0.1 mol/L sodium hydroxide VS until the pH of the so-lution changes to 3.5. The volume of 0.1 mol/L hydrochloric acid VS consumed is not less than 50.0 mL per g of Synthetic Aluminum Silicate.
Talc is a powdered, selected, natural, hydrated mag-nesium silicate. Pure talc is Mg3Si4O10(OH)2: 379.27. It may contain related mineral substances consisting chiefly of chlorite (hydrous magnesium aluminum silicate), magnesite (magnesium carbonate), calcite (calcium carbonate) and dolomite (calcium magnesium carbonate). It contains no asbestos.
Talc occurs as a white to grayish white, fine, crystalline powder. It is unctuous, and adheres readily to the skin. It is practically insoluble in water and in ethanol (99.5).◆
Determine the infrared absorption spectrum of Talc as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>: it exhibits absorp-tion at the wave numbers of about 3680 cm－1, 1018 cm－1 and 669 cm－1.
(1) Acidity or alkalinity—To 2.5 g of Talc add 50 mL of freshly boiled and cooled water, and heat under a reflux condenser. Filter the liquid by suction, add 0.1 mL of bromothymol blue-sodium hydroxide-ethanol TS to 10 mL of the filtrate, and add 0.01 mol/L hydrochloric acid VS until the color of the solution changes: the necessary volume of the VS is not more than 0.4 mL. Separately, to 10 mL of the filtrate add 0.1 mL of phenolphthalein TS, and add 0.01 mol/L sodium hydroxide VS until the color of the solution changes to light red: the necessary volume of the VS is not more than 0.3 mL. (2) Acid-soluble substances—Weigh accurately about g of Talc, heat with 20 mL of dilute hydrochloric acid at 509C for 15 minutes with stirring. Cool, add water to make exactly 50 mL, and filter. Centrifuge, if necessary, until the filtrate becomes clear. To 25 mL of the filtrate add 1 mL of dilute sulfuric acid, evaporate to dryness, and ignite to con-stant mass at 800 ± 259C: the amount of the residue is not more than 2.0z.◆ (3) Water-soluble substances—To 10.0 g of Talc add 50 mL of water, weigh the mass, and boil for 30 minutes, sup-plying water lost by evaporation. Cool, add water to restore and heat under a reflux condenser on a water bath for 30 minutes. After cooling, transfer the content to a beaker, and allow to settle the insoluble matter. Filter the supernatant liquid through a filter paper for quantitative analysis (No. 5B), leaving the precipitate in the beaker as much as possi-ble, wash the remaining precipitate in the beaker with three 10-mL portions of hot water, and also wash the filter paper with 15 mL of hot water, and combine the washings and the filtrate. After cooling, add water to make exactly 100 mL, and use this solution as the sample stock solution. Pipet 2.5 mL of the stock solution, add 50 mL of 0.5 mol/L hydro-chloric acid TS, then add water to make exactly 100 mL, and use this solution as the sample solution. Separately, to 50 mL each of 0.5 mol/L hydrochloric acid TS add exactly 2 mL, 2.5 mL, 3 mL and 4 mL of Standard Iron Solution for Atomic Absorption Spectrophotometry, add water to make them exactly 100 mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions, and calculate the amount of iron from the calibration curve prepared from the absorbances of the standard solu-tions: not more than 0.25z. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Iron hollow-cathode lamp. Wavelength: 248.3 nm. Aluminum—Pipet 5 mL of the sample stock solution obtained in the Assay, add 10 mL of cesium chloride TS and 10 mL of hydrochloric acid, then add water to make exactly 100 mL, and use this solution as the sample solution. Sepa-rately, to 10 mL of hydrochloric acid and 10 mL of cesium chloride TS add exactly 5 mL, 10 mL, 15 mL and 20 mL of Standard Aluminum Solution for Atomic Absorption Spec-trophotometry, add water to make them exactly 100 mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following conditions, and calculate the amount of aluminum from the calibration curve pre-pared from the absorbances of the standard solutions: not more than 2.0z. Gas: Combustible gas—Acetylene. Supporting gas—Nitrous oxide. Lamp: Aluminum hollow-cathode lamp. Wavelength: 309.3 nm. Lead—Use the sample stock solution obtained in (4) as the sample solution. Separately, to 50 mL of 0.5 mol/L hydrochloric acid TS add exactly 5 mL, 7.5 mL, 10 mL and 12.5 mL of Standard Lead Solution, add water to make them exactly 100 mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions, and calculate the amount of lead from the calibration curve prepared from the absorbances of the standard solu-tions: not more than 10 ppm. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Lead hollow-cathode lamp. Wavelength: 217.0 nm. ‌Calcium—Pipet 5 mL of the sample stock solution obtained in the Assay, add 10 mL of hydrochloric acid and 10 mL of lanthanum chloride TS, then add water to make exactly 100 mL, and use this solution as the sample solution. Separately, to 10 mL of hydrochloric acid and 10 mL of lan-thanum chloride TS add exactly 1 mL, 2 mL, 3 mL and 5 mL of Standard Calcium Solution, add water to make them exactly 100 mL, and use these solutions as the standard solu-tions. Perform the test with the sample solution and stand-ard solutions as directed under Atomic Absorption Spectro-photometry <2.23> according to the following conditions, and calculate the amount of calcium from the calibration curve prepared from the absorbances of the standard solu-tions: not more than 0.9z. Gas: Combustible gas—Acetylene. Supporting gas—Nitrous oxide. Lamp: Calcium hollow-cathode lamp. Wavelength: 422.7 nm. (8) Arsenic <1.11>—To 0.5 g of Talc add 5 mL of dilute sulfuric acid, and heat gently to boiling with shaking. Cool immediately, filter, and wash the residue with 5 mL of dilute sulfuric acid, then with 10 mL of water. Combine the filtrate and the washings, evaporate to 5 mL on a water bath, and perform the test with this solution as the test solution (not more than 4 ppm).◆ Loss on ignition <2.43> Not more than 7.0z (1 g, 1050 – 11009C, constant mass).
Weigh accurately about 0.5 g of Talc in a poly-tetrafluoroethylene dish, add 5 mL of hydrochloric acid, 5 mL of nitric acid and 5 mL of perchloric acid, then add 35 mL of hydrofluoric acid while mixing gently, and evaporate to dryness on a hot plate by heating gradually. Add 5 mL of hydrochloric acid to the residue, cover the dish with a watch glass, and heat to boil. After cooling, transfer the content to a volumetric flask while washing the watch glass and dish with water, further wash the dish with water, transfer the washings to the flask, then add water to make exactly 50 mL, and use this solution as the sample stock solution. Pipet 0.5 mL of the sample stock solution, and add water to make exactly 100 mL. Pipet 4 mL of this solution, add 10 mL of hydrochloric acid and 10 mL of lanthanum chloride TS, then add water to make exactly 100 mL, and use this solution as the sample solution. Separately, to 10 mL of hydrochloric acid and 10 mL of lanthanum chloride TS add exactly 2.5 mL, 3 mL, 4 mL and 5 mL of Standard Magnesium Solution for Atomic Absorption Spectrophotometry, add water to make them exactly 100 mL, and use these solutions as the standard solutions. Perform the test with the sample solution and standard solutions as directed under Atomic Absorption Spectrophotometry <2.23> according to the following condi-tions, and calculate the amount of magnesium from the calibration curve prepared from the absorbances of the standard solutions. Gas: Combustible gas—Acetylene. Supporting gas—Air. Lamp: Magnesium hollow-cathode lamp. Wavelength: 285.2 nm.
Thiamylal Sodium contains not less than 97.
Thiamylal Sodium occurs as light yellow, crys-tals or powder. It is very soluble in water, and freely soluble in ethanol (95). The pH of a solution of 1.0 g of Thiamylal Sodium in 10 mL of water is between 10.0 and 11.0. It is hygroscopic. It is gradually decomposed by light. Its solution in ethanol (95) (1 in 10) shows no optical rota-tion.
(1) Determine the absorption spectrum of Amount (mg) of thiamine nitrate (C ＝ MS × QT/QS × 0.971 12H17N5O4S) a solution of Thiamylal Sodium in ethanol (95) (7 in 1,000,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare spectrum with the Reference MS: Amount (mg) of Thiamine Chloride Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution of methyl benzoate in methanol (1 in 50). Operating conditions— Detector: An ultraviolet spectrophotometer (wavelength: 254 nm). Column: A stainless steel column 4.6 mm in inside diame- Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wavelengths. Determine the infrared absorption spectrum of Thiamylal Sodium, previously dried, as directed in the potas-sium bromide disk method under Infrared Spectrophotome-try <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorp-tion at the same wave numbers. ‌A solution of Thiamylal Sodium (1 in 10) responds to Qualitative Tests <1.09> for sodium salt.
(1) Clarity and color of solution—To 1.0 g of Thiamylal Sodium in a 11- to 13-mL glass-stoppered test tube add 10 mL of freshly boiled and cooled water, stopper tightly, allow to stand, and dissolve by occasional gentle shaking: the solution is clear and light yellow. Heavy metals <1.07>—Proceed with 1.0 g of Thiamylal Sodium according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Thiamylal So-dium in 10 mL of ethanol (95), and use this solution as the sample solution. Pipet 1 mL and 3 mL of the sample solu-tion, add ethanol (95) to make exactly 200 mL, and use these solutions as the standard solution (1) and the standard solu-tion (2). Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography, develop with a mixture of toluene, methanol and ethyl acetate (40:7:3) to a distance of about 12 cm, and air-dry the plate. Allow the plate to stand in iodine vapor for a night: the spot at an Rf value of about 0.1 obtained from the sample solu-tion is not more intense than the spot from the standard so-lution (2), and the spot other than the principal spot, the spot at origin and the spot mentioned above from the sample solution is not more intense than the spot from the standard solution (1). Loss on drying <2.41> Not more than 2.0z (1 g, 1059C, 1 hour).
Weigh accurately about 0.25 g of Thiamylal So-dium, dissolve in 50 mL of methanol and 5 mL of dilute hy-drochloric acid, and add methanol to make exactly 100 mL. Pipet 10 mL of this solution, and add methanol to make ex-actly 100 mL. Pipet 5 mL of this solution, add exactly 10 mL of the internal standard solution and the mobile phase to make 200 mL, and use this solution as the sample solution. Separately, weigh accurately about 23 mg of Thiamylal RS, previously dried at 1059C for 1 hour, dissolve in 50 mL of methanol and 0.5 mL of dilute hydrochloric acid, and add methanol to make exactly 100 mL. Pipet 5 mL of this solu-tion, add exactly 10 mL of the internal standard solution and the mobile phase to make 200 mL, and use this solution as the standard solution. Perform the test with 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and calculate the ratios, QT and QS, of the peak area of thiamylal to that of the internal standard. Amount (mg) of thiamylal sodium (C12H17N2NaO2S) ＝ MS × QT/QS × 10 × 1.086 MS: Amount (mg) of Thiamylal RS taken Internal standard solution—A solution of phenyl benzoate in methanol (3 in 500). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 289 nm). Column: A stainless steel column about 4 mm in inside diameter and about 15 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: A mixture of methanol and 0.05 mol/L ace- tic acid-sodium acetate buffer solution (pH 4.6) (13:7). Flow rate: Adjust so that the retention time of thiamylal is about 6 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, thiamylal and the internal standard are eluted in this order with the resolution between these peaks being not less than 12. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of thiamylal to that of the internal standard is not more than 1.0z.
Thiopental Sodium, when dried, contains not less than 97.
Thiopental Sodium occurs as a light yellow powder. It has a faint, characteristic odor. It is very soluble in water, freely soluble in ethanol (95), ＝ MSa × QTa/QSa × 1/5 7 6 3 and practically insoluble in diethyl ether. A solution of Thiopental Sodium (1 in 10) is alkaline. Amount (mg) of phenol (C6H6O) ＝ MSb × QTb/QSb × 1/5 MSa: Amount (mg) of salicylic acid for assay taken MSb: Amount (mg) of phenol for assay taken Internal standard solution—A solution of theophylline in methanol (1 in 10,000). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 270 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 25 to 30 cm in length, packed with octadecyl-silanized silica gel for liquid chromatography (5 mm in parti-cle diameter). Column temperature: Room temperature. Mobile phase: A mixture of 0.1 mol/L phosphate buffer solution (pH 7.0) and methanol (3:1). Flow rate: Adjust so that the retention time of salicylic acid is about 6 minutes. Selection of column: Dissolve 0.2 g of benzoic acid, 0.2 g of salicylic acid and 0.05 g of theophylline in 100 mL of di-luted methanol (1 in 2). To 10 mL of this solution add 90 mL of diluted methanol (1 in 2). Proceed with 10 mL of this solu-tion under the above operating conditions. Use a column giving elution of benzoic acid, salicylic acid and theophylline in this order, and clearly dividing each peak.
(1) Dissolve 0.2 g of Thiopental Sodium in 5 mL of sodium hydroxide TS, and add 2 mL of lead (II) acetate TS: a white precipitate, which dissolves upon heat-ing, is produced. Boil the solution thus obtained: a black precipitate forms gradually, and the precipitate responds to Qualitative Tests <1.09> for sulfide. Dissolve 0.5 g of Thiopental Sodium in 15 mL of water, add 10 mL of dilute hydrochloric acid to produce white precipitate, and extract with four 25-mL portions of chloroform. Combine the chloroform extracts, evaporate on a water bath, and dry at 1059C for 2 hours: the residue melts <2.60> between 1579C and 1629C. A solution of Thiopental Sodium (1 in 10) responds to Qualitative Tests <1.09> (1) and (2) for sodium salt.
(1) Clarity and color of solution—Dissolve 1.0 g of Thiopental Sodium in 10 mL of freshly boiled and cooled water: the solution is clear and light yellow. Heavy metals <1.07>—Dissolve 2.0 g of Thiopental Sodium in 76 mL of water, add 4 mL of dilute hydrochloric acid, shake, and filter through a glass filter (G4). To 40 mL of the filtrate add 2 mL of ammonium acetate TS, dilute with water to 50 mL, and perform the test using this solution as the test solution. Prepare a control solution as follows: to 2.0 mL of Standard Lead Solution add 2 mL of dilute acetic acid, 2 mL of ammonium acetate TS and water to make 50 mL (not more than 20 ppm). Neutral and basic substances—Weigh accurately about 1 g of Thiopental Sodium, dissolve in 10 mL of water and 5 mL of sodium hydroxide TS, and shake vigorously with 40 mL of chloroform. Separate the chloroform layer, wash with two 5-mL portions of water, filter, and evaporate the filtrate on a water bath to dryness. Dry the residue at 1059C for 1 hour: the amount of the residue is not more than 0.50z. Related substances—Dissolve 50 mg of Thiopental Sodium in 50 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 200 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and the standard so-lution as directed under Liquid Chromatography <2.01> ac- ‌cording to the following conditions. Determine each peak area by the automatic integration method: the total area of peaks other than thiopental obtained from the sample solu-tion is not larger than the peak area of thiopental from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel (5 mm in particle diameter). Column temperature: A constant temperature of about 409C. Mobile phase: Dissolve 1 g of potassium dihydrogen phos-phate in 1000 mL of water, and adjust the pH to 3.0 with phosphoric acid. To 700 mL of this solution add 300 mL of acetonitrile. Flow rate: Adjust so that the retention time of thiopental is about 15 minutes. Time span of measurement: About 1.5 times as long as the retention time of thiopental. System suitability— Test for required detectability: To exactly 2 mL of the standard solution add the mobile phase to make exactly 10 mL. Confirm that the peak area of thiopental obtained with 20 mL of this solution is equivalent to 15 to 25z of that with 20 mL of the standard solution. System performance: Dissolve 5 mg each of isopropyl par-ahydroxybenzoate and propyl parahydroxybenzoate in 50 mL of acetonitrile, and add water to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating conditions, isopropyl parahydroxybenzoate and propyl parahydroxybenzoate are eluted in this order with the resolution between these peaks being not less than 1.9. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of thiopental is not more than 2.0z. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, 809C, 4 hours).
Weigh accurately about 0.5 g of Thiopental Sodium, previously dried, transfer to a separator, dissolve in 20 mL of water, add 5 mL of ethanol (95) and 10 mL of dilute hy-drochloric acid, and extract with 50 mL of chloroform, then with three 25-mL portions of chloroform. Combine the chlo-roform extracts, wash with two 5-mL portions of water, and extract the washings with two 10-mL portions of chlo-roform. Filter the combined chloroform extracts into a coni-cal flask, and wash the filter paper with three 5-mL portions of chloroform. Combine the filtrate and the washings, and add 10 mL of ethanol (95). Titrate <2.50> with 0.1 mol/L po-tassium hydroxide-ethanol VS until the color of the solution changes from yellow through light blue to purple (indicator: 2 mL of alizarin yellow GG-thymolphthalein TS). Perform a blank determination in the same manner with a mixture of 160 mL of chloroform and 30 mL of ethanol (95), and make any necessary correction. Each mL of 0.1 mol/L potassium hydroxide-ethanol VS ＝ 26.43 mg of C11H17N2NaO2S
Containers—Tight containers. Storage—Light-resistant, and not exceeding 259C.
Triclofos Sodium, when dried, contains not less than 97.
Triclofos Sodium is a white crystalline powder. It is freely soluble in water, slightly soluble in ethanol (95), and practically insoluble in diethyl ether. It is hygroscopic.
(1) Determine the infrared absorption spec-trum of Triclofos Sodium as directed in the potassium bro-mide disk method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. To 0.5 g of Triclofos Sodium add 10 mL of nitric acid, evaporate on a water bath to dryness, and ignite fur-ther over a flame. Dissolve the residue in 5 mL of water, and filter it necessary: the filtrate responds to Qualitative Tests <1.09> for sodium salt. To 0.1 g of Triclofos Sodium add 1 g of anhydrous sodium carbonate, and heat for 10 minutes. After cooling, dissolve the residue in 40 mL of water, filter if necessary, and render the filtrate acidic with dilute nitric acid: the solu-tion responds to Qualitative Tests <1.09> (2) for chloride. The remainder of the filtrate responds to Qualitative Tests <1.09> (1) for chloride and to Qualitative Tests <1.09> for phosphate. pH <2.54> Dissolve 1.0 g of Triclofos Sodium in 50 mL of water: the pH of this solution is between 3.0 and 4.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Triclofos Sodium in 50 mL of water: the solution is clear and colorless. Chloride <1.03>—Perform the test with 0.20 g of Triclofos Sodium. Prepare the control solution with 1.0 mL of 0.01 mol/L hydrochloric acid VS (not more than 0.178z). Heavy metals <1.07>—Proceed with 1.0 g of Triclofos Sodium according to Method 1, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Triclofos Sodium according to Method 1, and perform the test (not more than 2 ppm). Free phosphoric acid—Weigh accurately about 0.3 g of Triclofos Sodium, previously dried, dissolve in water to make exactly 100 mL, and use this solution as the sample so-lution. Pipet 5 mL each of the sample solution and Standard Phosphoric Acid Solution, add 2.5 mL of hexaammonium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS, shake, add water to make ex-actly 25 mL, and allow to stand at 209C for 30 minutes. Per-form the test with these solutions, using a solution obtained in the same manner with 5 mL of water as the blank, as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Determine the absorbances, AT and AS, of each solution from the sample solution and Standard Phosphoric Acid So-lution at 740 nm: the content of the free phosphoric acid is not more than 1.0z. Content (z) of the free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Triclofos Sodium taken Loss on drying <2.41> Not more than 5.0z (1 g, in vacu-um, 1009C, 3 hours).
(1) Triclofos sodium—Weigh accurately about 0.2 g of Triclofos Sodium, previously dried, place in a Kjel-dahl flask, add 2 mL of sulfuric acid and 2.5 mL of nitric acid, and heat until brown gas are not evolved. After cool-ing, add 1 mL of nitric acid, heat until white fumes are pro-duced, and cool. Repeat this procedure until the solution becomes colorless. Transfer this solution to a flask using 150 mL of water, add 50 mL of molybdenum (VI) oxide-citric acid TS, heat gently to boil, add gradually 25 mL of quino-line TS with stirring, and heat on a water bath for 5 minutes. After cooling, filter the precipitate, and wash repeatedly with water until the washing does not indicate acidity. Trans-fer the precipitate to a flask using 100 mL of water, add exactly 50 mL of 0.5 mol/L sodium hydroxide VS, dissolve, and titrate <2.50> with 0.5 mol/L hydrochloric acid VS until the color of the solution changes from purple to yellow (indicator: 3 drops of phenolphthalein-thymol blue TS). Perform a blank determination in the same manner. Each mL of 0.5 mol/L sodium hydroxide VS ＝ 4.834 mg of C2H3Cl3NaO4P (2) Chlorine—Weigh accurately about 10 mg of Triclo-fos Sodium, previously dried, perform the test according to the procedure of determination for chlorine as directed under Oxygen Flask Combustion Method <1.06>, using 1 mL of sodium hydroxide TS and 20 mL of water as the absorb-ing liquid.
Warfarin Potassium, when dried, contains not less than 98.
Warfarin Potassium occurs as a white crystal-line powder. It is very soluble in water, and freely soluble in ethanol (95). It dissolves in sodium hydroxide TS. The pH of a solution prepared by dissolving 1.0 g of Warfarin Potassium in 100 mL of water is between 7.2 and 8.3. It is colored to light yellow by light. A solution of Warfarin Potassium (1 in 10) shows no opti-cal rotation.
(1) Determine the absorption spectrum of a solution of Warfarin Potassium in 0.02 mol/L potassium hydroxide TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Warfarin Potassium RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Warfarin Potassium, previously dried, as directed in the potassium bromide disk method under Infrared Spectropho-tometry <2.25>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of Warfarin Potassium RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Warfarin Potassium (1 in 250) responds to Qualitative Tests <1.09> (1) for potassium salt.
(1) Alkaline colored substances—Dissolve 1.0 g of Heavy metals <1.07>—Dissolve 2.0 g of Warfarin Po-tassium in 30 mL of ethanol (95), add 2 mL of dilute acetic acid and ethanol (95) to make 50 mL. Perform the test using this solution as the test solution. Prepare the control solution with 2.0 mL of Standard Lead Solution, 2 mL of dilute ace-tic acid and ethanol (95) to make 50 mL (not more than 10 ppm). Related substances—Dissolve 0.10 g of Warfarin Po-tassium in 100 mL of a mixture of water and methanol (3:1), and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mixture of water and methanol (3:1) to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the automatic integration method: each peak area other than warfarin obtained from the sample solution is not larger than 1/10 times the peak area of warfarin from the standard solution, and the total area of the peaks other than warfarin from the sample solution is not larger than 1/2 times the peak area of warfarin from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 2 times as long as the retention time of warfarin, beginning after the solvent peak. System suitability— Test for required detectability: To exactly 1 mL of the standard solution add the mixture of water and methanol (3:1) to make exactly 20 mL. Confirm that the peak area of warfarin obtained with 20 mL of this solution is equivalent to 3.5 to 6.5z of that with 20 mL of the standard solution. System performance: Dissolve 20 mg of propyl parahy- droxybenzoate in 50 mL of methanol, and add water to make 200 mL. To 5 mL of this solution add 4 mL of a solu-tion of Warfarin Potassium in the mixture of water and methanol (3:1) (1 in 2000), and add the mixture of water and methanol (3:1) to make 100 mL. When the procedure is run with 20 mL of this solution under the above operating condi-tions, propyl parahydroxybenzoate and warfarin are eluted in this order with the resolution between these peaks being not less than 7 and the symmetry factor is not more than 1.5. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of warfarin is not more than 2.0z. Loss on drying <2.41> Not more than 4.5z (1 g, 1059C, 3 hours).
Weigh accurately about 25 mg each of Warfarin Po-tassium and Warfarin Potassium RS, previously dried, and separately dissolve in the mixture of water and methanol (3:1) to make exactly 50 mL. Pipet 10 mL each of these solu-tions, add the mixture of water and methanol (3:1) to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine the peak areas, AT and AS, of warfarin in each solution. Warfarin Potassium in a solution of sodium hydroxide (1 in 20) to make exactly 10 mL, and determine the absorbance at 385 nm within 15 minutes as directed under Ultraviolet- Amount (mg) of warfarin potassium (C ＝ MS × AT/AS 19H15KO4) visible Spectrophotometry <2.24>, using a solution of sodium hydroxide (1 in 20) as a blank: it does not exceed 0.20. MS: Amount (mg) of Warfarin Potassium RS taken ‌Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 260 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with cyanopropylsilanized silica gel for liquid chromatography (5 mm in particle diame-ter). Column temperature: A constant temperature of about 409C. Mobile phase: A mixture of water, acetonitrile and acetic acid (100) (68:32:1). Flow rate: Adjust so that the retention time of warfarin is about 10 minutes. System suitability— System performance: When the procedure is run with 20 RS, previously dried at 1059C for 3 hours, and dissolve in water to make exactly 100 mL. Pipet 5 mL of this solution, add water to make exactly 100 mL, and use this solution as the standard solution. Pipet 20 mL each of the sample solu-tion and standard solution, add 0.05 mol/L hydrochloric acid TS to make exactly 25 mL, and use these solutions as the solution T1 and the solution S1, respectively. Separately, pipet 20 mL each of the sample solution and standard solu-tion, add 0.05 mol/L potassium hydroxide TS to make ex-actly 25 mL, and use these solutions as the solution T2 and the solution S2, respectively. Determine the absorbances, AT and AS, of the solution T1 and the solution S1 at 272 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>, using the solution T2 and the solution S2 as the blank, respec-tively. mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of warfarin are not less than 8000 and not Amount (mg) of warfarin potassium (C ＝ MS × AT/AS × V/2000 19H15KO4) more than 1.5, respectively. System repeatability: When the test is repeated 6 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak area of warfarin is not more than 1.0z.
Zinc Chloride contains not less than 97.
Zinc Chloride occurs as white, crystalline pow-der, rods, or masses. It is odorless. It is very soluble in water, and freely soluble in ethanol (95), and its solution may sometimes be slightly turbid. The solution becomes clear on addition of a small amount of hy-drochloric acid. The pH of a solution of 1.0 g of Zinc Chloride in 2 mL of water is between 3.3 and 5.3. It is deliquescent.
A solution of Zinc Chloride (1 in 30) re-sponds to Qualitative Tests <1.09> for zinc salt and chloride. allow to stand for 5 minutes, and immediately observe from the top downward against a white background: the solution has no more color than the following control solution. Control solution: To 2.5 mL of Standard Lead Solution add 3 mL of water and 15 mL of potassium cyanide TS, shake thoroughly, and add 1 drop of sodium sulfide TS (not more than 50 ppm). Alkali earth metals and alkali metals—Dissolve 2.0 g of Zinc Chloride in 120 mL of water, add ammonium sulfide TS to complete precipitation, add water to make 200 mL, shake thoroughly, and filter through dry filter paper. Dis-card the first 20 mL of the filtrate, take the following 100 mL of the filtrate, evaporate with 3 drops of sulfuric acid to dryness, and heat the residue strongly at 6009C to constant mass: the mass is not more than 10.0 mg. Arsenic <1.11>—Prepare the test solution with 0.40 g of Zinc Chloride according to Method 1, and perform the test (not more than 5 ppm). Oxychloride—Shake gently 0.25 g of Zinc Chloride with 5 mL of water and 5 mL of ethanol (95), and add 0.3 mL of 1 mol/L hydrochloric acid VS: the solution is clear.
Weigh accurately about 0.3 g of Zinc Chloride, add 0.4 mL of dilute hydrochloric acid and water to make exactly 200 mL. Measure exactly 20 mL of the solution, add 80 mL of water, 2 mL of ammonia-ammonium chloride buffer so-lution (pH 10.7) and titrate <2.50> with 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.01 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 1.363 mg of ZnCl2
Zinc Oxide Oil contains not less than 45.
Zinc Oxide Oil is a white to whitish, slimy sub-stance, separating a part of its ingredients when stored for a prolonged period.
Mix thoroughly, and place 0.5 g of Zinc Oxide Oil in a crucible, heat gradually raising the tempera-ture until the mass is thoroughly charred, and then ignite it strongly: a yellow color is produced, and disappears on cool-ing. To the residue add 10 mL of water and 5 mL of dilute hydrochloric acid, shake well, and filter. To the filtrate add 2 to 3 drops of potassium hexacyanoferrate (II) TS: a white precipitate is formed (zinc oxide).
Weigh accurately about 0.8 g of Zinc Oxide Oil, mixed well, place in a crucible, heat gradually raising the temperature until the mass is thoroughly charred, and then ignite until the residue becomes yellow, and cool. Dissolve the residue in 1 mL of water and 1.5 mL of hydrochloric acid, and add water to make exactly 100 mL. Pipet 20 mL of this solution, add 80 mL of water, and add a solution of so-dium hydroxide (1 in 50) until a small amount of precipitates begins to form in the solution. Add 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylene- diamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.069 mg of ZnO
Zinc Oxide, when ignited, contains not less than 99.
Zinc Oxide occurs as a white, amorphous pow-der. It is odorless and tasteless. It is practically insoluble in water, in ethanol (95), in acetic acid (100) and in diethyl ether. It dissolves in dilute hydrochloric acid and in sodium hy-droxide TS. It gradually absorbs carbon dioxide from air.
(1) Heat Zinc Oxide strongly: a yellow color develops on strong heating, and disappears on cooling. ‌(2) A solution of Zinc Oxide in dilute hydrochloric acid (1 in 10) responds to Qualitative Tests <1.09> for zinc salt.‌
(1) Carbonate, and clarity and color of solu-tion—Mix 2.0 g of Zinc Oxide with 10 mL of water, add 30 mL of dilute sulfuric acid, and heat on a water bath with stirring: no effervescence occurs, and the solution obtained is clear and colorless. Alkalinity—To 1.0 g of Zinc Oxide add 10 mL of water, and boil for 2 minutes. Cool, filter through a glass filter (G3), and to the filtrate add 2 drops of phenolphthalein TS and 0.20 mL of 0.1 mol/L hydrochloric acid VS: no color develops. Sulfate <1.14>—Shake 0.5 g of Zinc Oxide with 40 mL of water, and filter. Take 20 mL of the filtrate, add 1 mL of dilute hydrochloric acid and water to make 50 mL, and perform the test using this solution as the test solution. Prepare the control solution with 0.50 mL of 0.005 mol/L sulfuric acid VS (not more than 0.096z). Iron—Dissolve 1.0 g of Zinc Oxide in 50 mL of di-luted hydrochloric acid (1 in 2), dissolve 0.1 g of ammonium peroxodisulfate in this solution, and extract with 20 mL of 4-methyl-2-pentanone. Add 30 mL of acetic acid-sodium ace-tate buffer solution for Iron Limit Test (pH 4.5) to the 4-methyl-2-pentanone layer, extract again, and use the layer of the buffer solution as the test solution. Separately, perform the test in the same manner with 1.0 mL of Standard Iron Solution, and use the layer so obtained as the control solu-tion. Add 2 mL each of L-ascorbic acid solution for Iron Limit Test (1 in 100) to the test solution and the control solu-tion, respectively, mix, allow to stand for 30 minutes, add 5 mL of a solution of 2,2?-bipyridyl in ethanol (95) (1 in 200) and water to make 50 mL. After allowing to stand for 30 minutes, compare the color of the both liquids against a white back: the color of the liquid from the test solution is not stronger than that from the control solution (not more than 10 ppm). Lead—To 2.0 g of Zinc Oxide add 20 mL of water, then add 5 mL of acetic acid (100) with stirring, and heat on a water bath until solution is complete. Cool, and add 5 drops of potassium chromate TS: no turbidity is produced. Arsenic <1.11>—Dissolve 0.5 g of Zinc Oxide in 5 mL of dilute hydrochloric acid, use this solution as the test solu-tion, and perform the test (not more than 4 ppm). Loss on ignition <2.43> Not more than 1.0z (1 g, 8509C, 1 hour).
Weigh accurately about 0.8 g of Zinc Oxide, previ-ously ignited at 8509C for 1 hour, dissolve in 2 mL of water and 3 mL of hydrochloric acid, and add water to make exactly 100 mL. Pipet 10 mL of this solution, add 80 mL of water, then add a solution of sodium hydroxide (1 in 50) until a slight precipitate is produced. Add 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodium dihydrogen ethylene- diamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.069 mg of ZnO
Zinc Sulfate Hydrate contains not less than 99.
Weigh accurately about 2 g of Zinc Oxide Ointment, place in a crucible, melt by warming, heat gradually raising the temperature until the mass is thoroughly charred, and then ignite until the residue becomes uniformly yellow, and cool. Dissolve the residue in 1 mL of water and 1.5 mL of hydrochloric acid, and add water to make exactly 100 mL. Add 80 mL of water to exactly 20 mL of this solution, and add a solution of sodium hydroxide (1 in 50) until a small amount of precipitates begins to form in the solution. Add 5 mL of ammonia-ammonium chloride buffer solution (pH 10.7), and titrate <2.50> with 0.05 mol/L disodium dihy-drogen ethylenediamine tetraacetate VS (indicator: 0.04 g of eriochrome black T-sodium chloride indicator). Each mL of 0.05 mol/L disodium dihydrogen ethylenediamine tetraacetate VS ＝ 4.069 mg of ZnO
Akebia quinata Decaisne or Akebia trifoliata Koidzumi
Angelica dahurica Bentham et Hooker filius ex Franchet et Savatier
Artemisia princeps Pampanini or Artemisia montana Pampanini
Asiasarum heterotropoides F. Maekawa var. mandshuricum F. Maekawa or Asiasarum sieboldii F. Maekawa
Astragalus membranaceus Bunge or Astragalus mongholicus Bunge
Catalpa ovata G. Don or Catalpa bungei C. A. Meyer
Prunus jamasakura Siebold ex Koidzumi or Prunus verecunda Koehne
Citrus unshiu Marcowicz or Citrus reticulata Blanco
Codonopsis pilosula Nannfeldt or Codonopsis tangshen Oliver
Corydalis turtschaninovii Basser forma yanhusuo Y. H. Chou et C. C. Hsu
Dioscorea japonica Thunberg or Dioscorea batatas Decaisne
Fritillaria verticillata Willdenow var. thunbergii Baker
Geranium thunbergii Siebold et Zuccarini
Glehnia littoralis Fr. Schmidt ex Miquel
Cephaelis ipecacuanha A. Richard or Cephaelis acuminata Karsten
Angelica acutiloba Kitagawa or Angelica acutiloba Kitagawa var. sugiyamae Hikino
Ziziphus jujuba Miller var. inermis Rehder
Ziziphus jujuba Miller var. spinosa Hu ex H. F. Chow
Lithospermum erythrorhizon Siebold et Zuccarini
Notopterygium incisum Ting ex H. T. Chang or Notopterygium forbesii Boissieu
Prunus persica Batsch or Prunus persica Batsch var. davidiana Maximowicz
Perilla frutescens Britton var. crispa W. Deane
Phellodendron amurense Ruprecht or Phellodendron chinense Schneider
Aconitum carmichaeli Debeaux or Aconitum japonicum Thunberg
Rehmannia glutinosa Liboschitz var. purpurea Makino or Rehmannia glutinosa Liboschitz
Cassia angustifolia Vahl or Cassia acutifolia Delile
Hydrangea macrophylla Seringe var. thunbergii Makino
Bufo gargarizans Cantor or Bufo melanostictus Schneider
solution as directed in the Assay under Powdered Opium
a clear, yellowish liquid, having a bitter taste
Vitamin C
L-Ascorbic Acid
ビタミンC
Ascorbic Acid, when dried, contains not less than 99.0% of C6H8O6.
White crystals or crystalline powder, odorless, having an acid taste
Freely soluble in water, sparingly soluble in ethanol (95), slightly soluble in glycerin, and practically insoluble in diethyl ether.
To 5 mL of a solution of Ascorbic Acid (1 in 50) add 1 drop of potassium permanganate TS: the color of the TS disappears immediately.
To 2 mL of a solution of Ascorbic Acid (1 in 100) add 2 mL of sodium carbonate TS and 0.5 mL of sodium nitroferricyanide TS, then add 0.5 mL of sodium hydroxide TS: a blue color is produced.
Weigh accurately about 2.5 g of Ascorbic Acid, dissolve in water to make exactly 25 mL, and determine the optical rotation at 20°C in a 100-mm cell: [α]D20 = +20.5 to +21.5°
Dissolve 1.0 g of Ascorbic Acid in 10 mL of water: the solution is clear and colorless.
Proceed according to Method 2 using 2.0 g of Ascorbic Acid. Prepare the control solution with 2.0 mL of Standard Lead Solution.
Prepare the test solution with 1.0 g of Ascorbic Acid according to Method 1, and perform the test.
Dissolve 0.25 g of Ascorbic Acid in 5 mL of water, add 0.2 mL of dilute acetic acid and 0.5 mL of calcium chloride TS, and allow to stand for 1 minute: the solution is clear.
Dry in a desiccator (silica gel) for 3 hours
Weigh accurately about 0.2 g of Ascorbic Acid, previously dried, dissolve in 50 mL of freshly boiled and cooled water, add 2 mL of dilute sulfuric acid and 2 mL of starch TS, and titrate with 0.05 mol/L iodine VS until the color of the solution becomes pale blue and persists for 30 seconds. Perform a blank determination in the same manner, and make any necessary correction. Each mL of 0.05 mol/L iodine VS = 4.403 mg of C6H8O6.
Store in a cool, dark place, protected from light and moisture
Biotin, when dried, contains not less than 98.
Biotin occurs as white crystals or a white crys-talline powder. It is very slightly soluble in water and in ethanol (99.5). It dissolves in dilute sodium hydroxide TS. Melting point: about 2319C (with decomposition).
Determine the infrared absorption spectrum of Biotin as directed in the potassium bromide disc method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. D Optical rotation <2.49> [a]20: ＋89 – ＋939(after drying, 0.4 g, dilute sodium hydroxide TS, 20 mL, 100 mm).
(1) Clarity and color of solution—Dissolve 1.0 g of Biotin in 10 mL of 0.5 mol/L sodium hydroxide TS: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 2.0 g of Biotin JP XVIII Official Monographs / Biperiden Hydrochloride 553 ‌according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Arsenic <1.11>—Place 0.7 g of Biotin in a Kjeldahl flask, add 5 mL of nitric acid and 2 mL of sulfuric acid, place a small funnel on the mouth of the flask, and carefully heat until white fumes are evolved. After cooling, add 2 mL of nitric acid twice, heat, add 2 mL of hydrogen peroxide (30) several times, and heat until the color of the solution becomes colorless or pale yellow. After cooling, add 2 mL of saturated ammonium oxalate solution, and heat to concen-trate until white fumes are evolved again. After cooling, add water to make 5 mL, and perform the test using this solution as the test solution (not more than 2.8 ppm). Related substances—Dissolve 0.10 g of Biotin in 10 mL of diluted ammonia solution (28) (7 in 100), and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add diluted ammonia solution (28) (7 in 100) to make exactly 100 mL. Pipet 10 mL of this solution, add diluted ammonia solution (28) (7 in 100) to make exactly 50 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solu-tion and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of 1-butanol, water, and acetic acid (100) (5:2:1) to a distance of about 10 cm, air-dry the plate, and then dry for 30 minutes at 1059C. Spray the plate evenly with a mixture of a solution of 4-dimethylaminocinnamaldehyde in ethanol (99.5) (1 in 500) and a solution of sulfuric acid in ethanol (99.5) (1 in 50) (1:1): the spots other than the principal spot obtained from the sample solution are not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.5z (0.5 g, 1059C, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.25 g of Biotin, previously dried, dissolve by adding exactly 20 mL of 0.1 mol/L sodium hydroxide VS, and titrate <2.50> the excess sodium hydrox-ide with 0.1 mol/L hydrochloric acid VS (indicator: 2 drops of phenolphthalein TS). Perform a blank determination in the same manner. Each mL of 0.1 mol/L sodium hydroxide VS ＝ 24.43 mg of C10H16N2O3S
Folic Acid occurs as a yellow to orange-yellow crystalline powder. It is odorless. It is practically insoluble in water, in methanol, in ethanol (95), in pyridine and in diethyl ether. It dissolves in hydrochloric acid, in sulfuric acid, in dilute sodium hydroxide TS and in a solution of sodium carbonate decahydrate (1 in 100), and these solutions are yellow in color. It is slowly affected by light.
(1) Dissolve 1.5 mg of Folic Acid in dilute sodium hydroxide TS to make 100 mL. Determine the ab-sorption spectrum of the solution as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Folic Acid RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) To 10 mL of the solution obtained in (1) add 1 drop of potassium permanganate TS, and mix well until the color changes to blue, and immediately observe under ultraviolet light (main wavelength: 365 nm): a blue fluorescence is pro-duced.
(1) Clarity and color of solution—Dissolve 0.10 g of Folic Acid in 10 mL of dilute sodium hydroxide TS : the solution is clear and yellow in color. (2) Free amines—Pipet 30 mL of the sample solution ob-tained in the Assay, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of p-Aminobenzoyl Glutamic Acid for Purity RS, previ-ously dried in a desiccator (in vacuum, silica gel) for 4 hours, dissolve in diluted ethanol (2 in 5) to make exactly 100 mL. Pipet 3 mL of this solution, add water to make exactly 1000 mL, and use this solution as the standard solution. Pipet 4 mL each of the sample solution and standard solution, proceed as directed in the Assay, and perform the test as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Determine the absorbances, AT and AS, of subsequent solu-tions of the sample solution and standard solution at 550 nm: the content of free amines is not more than 1.0z. solution of sodium nitrite (1 in 1000), mix well, and allow to stand for 2 minutes. To each solution add 1 mL of a solution of ammonium amidosulfate (1 in 200), mix thoroughly, and allow to stand for 2 minutes. To each of these solutions, add 1 mL of a solution of N-(1-naphthyl)-N?-diethylethylenedia-mine oxalate (1 in 1000), shake, allow to stand for 10 minutes, and add water to make exactly 20 mL. Separately, to 30 mL of the sample solution, accurately measured, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solution, add 18 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 4 mL of this solution, and prepare the blank solution in the same manner as the sample solution. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared with 4 mL of water in the same manner as a blank. Determine the absor-bances, AT, AS and AC, of the subsequent solution of the sample solution, the standard solution and the blank solu-tion at 550 nm. Amount (mg) of folic acid (C19H19N7O6) ＝ MS × (AT － AC)/AS MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis
Weigh accurately about 50 mg each of Folic Acid and Folic Acid RS (separately, determine the water <2.48> in the same manner as Folic Acid). To each add 50 mL of dilute sodium hydroxide TS, mix well to dissolve, add dilute so-dium hydroxide TS to make exactly 100 mL, and use these solutions as the sample solution and standard solution. To 30 mL each of these solutions, accurately measured, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 60 mL each of these solutions add 0.5 g of zinc powder, and allow to stand with frequent shaking for 20 minutes. Filter each mixture through a dry filter paper, and discard the first 10 mL of the filtrate. Pipet 10 mL each of the subsequent filtrate, and add water to make exactly 100 lated on the anhydrous basis. 19 19 7 6 mL. To 4 mL each of solutions, accurately measured, add 1 mL of water, 1 mL of dilute hydrochloric acid and 1 mL of a Description Folic Acid occurs as a yellow to orange-yellow crystalline powder. It is odorless. It is practically insoluble in water, in methanol, in ethanol (95), in pyridine and in diethyl ether. It dissolves in hydrochloric acid, in sulfuric acid, in dilute sodium hydroxide TS and in a solution of sodium carbonate decahydrate (1 in 100), and these solutions are yellow in color. It is slowly affected by light. Identification (1) Dissolve 1.5 mg of Folic Acid in dilute sodium hydroxide TS to make 100 mL. Determine the ab-sorption spectrum of the solution as directed under Ultravio-let-visible Spectrophotometry <2.24>, and compare the spec-trum with the Reference Spectrum or the spectrum of a solu-tion of Folic Acid RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) To 10 mL of the solution obtained in (1) add 1 drop of potassium permanganate TS, and mix well until the color changes to blue, and immediately observe under ultraviolet light (main wavelength: 365 nm): a blue fluorescence is pro-duced. Purity (1) Clarity and color of solution—Dissolve 0.10 g of Folic Acid in 10 mL of dilute sodium hydroxide TS : the solution is clear and yellow in color. (2) Free amines—Pipet 30 mL of the sample solution ob-tained in the Assay, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of p-Aminobenzoyl Glutamic Acid for Purity RS, previ-ously dried in a desiccator (in vacuum, silica gel) for 4 hours, dissolve in diluted ethanol (2 in 5) to make exactly 100 mL. Pipet 3 mL of this solution, add water to make exactly 1000 mL, and use this solution as the standard solution. Pipet 4 mL each of the sample solution and standard solution, proceed as directed in the Assay, and perform the test as di-rected under Ultraviolet-visible Spectrophotometry <2.24>. Determine the absorbances, AT and AS, of subsequent solu-tions of the sample solution and standard solution at 550 nm: the content of free amines is not more than 1.0z. solution of sodium nitrite (1 in 1000), mix well, and allow to stand for 2 minutes. To each solution add 1 mL of a solution of ammonium amidosulfate (1 in 200), mix thoroughly, and allow to stand for 2 minutes. To each of these solutions, add 1 mL of a solution of N-(1-naphthyl)-N?-diethylethylenedia-mine oxalate (1 in 1000), shake, allow to stand for 10 minutes, and add water to make exactly 20 mL. Separately, to 30 mL of the sample solution, accurately measured, add 20 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 10 mL of this solution, add 18 mL of dilute hydrochloric acid and water to make exactly 100 mL. Pipet 4 mL of this solution, and prepare the blank solution in the same manner as the sample solution. Perform the test with these solutions as directed under Ultraviolet-visible Spectro-photometry <2.24>, using a solution prepared with 4 mL of water in the same manner as a blank. Determine the absor-bances, AT, AS and AC, of the subsequent solution of the sample solution, the standard solution and the blank solu-tion at 550 nm. Amount (mg) of folic acid (C19H19N7O6) ＝ MS × (AT － AC)/AS MS: Amount (mg) of Folic Acid RS taken, calculated on the anhydrous basis
Retinol Acetate occurs as pale yellow to yel-low-red, crystals or an ointment-like substance, and has a faint, characteristic odor, but has no rancid odor. It is freely soluble in petroleum ether, soluble in ethanol (95), and practically insoluble in water. It is decomposed by air and by light.
Dissolve Retinol Acetate and Retinol Ace-tate RS, equivalent to 15,000 Units each, in 5 mL of petrole-um ether, and use these solutions as the sample solution and standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop with a mixture of cyclohexane and diethyl ether (12:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly antimony (III) chloride TS: the principal spot obtained from the sample solution is the same in color tone and Rf value with the blue spot from the standard solution.
(1) Acid value <1.13>—Take exactly 5.0 g of Retinol Acetate, and perform the test: not more than 2.0. (2) Peroxide—Weigh accurately about 5 g of Retinol Acetate, transfer in a 250-mL glass-stoppered conical flask, add 50 mL of a mixture of acetic acid (100) and isooctane (3:2), and gently mix to dissolve completely. Replace the air of the inside gradually with about 600 mL of Nitrogen, then add 0.1 mL of saturated potassium iodide TS under a cur-rent of Nitrogen. Immediately stopper tightly, and mix with a swirling motion for 1 minute. Add 30 mL of water, stopper tightly, and shake vigorously for 5 to 10 seconds. Titrate <2.50> this solution with 0.01 mol/L sodium thiosulfate VS until the blue color of the solution disappears after addition of 0.5 mL of starch TS near the end point where the solution is a pale yellow color. Calculate the amount of peroxide by the following formula: not more than 10 mEq/kg. Amount (mEq/kg) of peroxide ＝ V/M × 10 V: Volume (mL) of 0.01 mol/L sodium thiosulfate VS consumed M: Amount (g) of Retinol Acetate taken
Proceed as directed in Method 1-1 under Vitamin A Assay <2.55>. ‌Containers and storage Containers—Tight containers. Storage—Light-resistant, and almost well-filled, or under‌ Nitrogen atmosphere, and in a cold place. Retinol Palmitate Vitamin A Palmitate
Vitamin A Oil is synthetic vitamin A esters diluted with fixed oils. It contains not less than 30,000 vitamin A Units per g.
Vitamin A Oil is a yellow to yellow-brown, clear or slightly turbid oil. It is odorless or has a faint, char-acteristic odor. It is decomposed upon exposure to air or light.
Dissolve Vitamin A Oil, Retinol Acetate RS and Retinol Palmitate RS, equivalent to 15,000 Units, in 5 mL of petroleum ether, and use these solutions as the sample solution, the standard solution (1) and the standard solution (2), respectively. Perform the test with these solutions as di-rected under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solutions (1) and (2) on a plate of silica gel for thin-layer chromatography. Develop with a mixture of cyclohexane and diethyl ether (12:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly antimony (III) chloride TS: the principal spot ‌obtained from the sample solution has the same color tone and the same Rf value with the blue spot from the standard solution (1) or the standard solution (2).
(1) Acidity—Dissolve 1.2 g of Vitamin A Oil in 30 mL of a mixture of neutralized ethanol and diethyl ether (1:1), boil gently for 10 minutes under a reflux condenser, cool, and add 5 drops of phenolphthalein TS and 0.60 mL of 0.1 mol/L sodium hydroxide VS: a red color develops. (2) Rancidity—No unpleasant odor of rancid oil is per-ceptible by warming Vitamin A Oil.
Proceed as directed in Method 1-1 under Vitamin A Assay <2.55>.
Containers—Tight containers. Storage—Light-resistant, and almost well-filled, or under
Retinol Palmitate occurs as a light yellow to yellow-red, ointment-like or an oily substance. It has a faint, characteristic odor, but has no rancid odor. It is very soluble in petroleum ether, slightly soluble in ethanol (95), and practically insoluble in water. It is decomposed by air and by light.
Dissolve Retinol Palmitate and Retinol Palmitate RS, equivalent to 15,000 Units each, in 5 mL of petroleum ether, and use these solutions as the sample solu-tion and standard solution. Perform the test with these solu-tions as directed under Thin-layer Chromatography <2.03>. Spot 5 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Devel-op with a mixture of cyclohexane and diethyl ether (12:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly antimony (III) chloride TS: the principal spot ob-tained from the sample solution is the same in color tone and Rf value with the blue spot from the standard solution.
(1) Acid value <1.13>—Take exactly 5.0 g of Retinol Palmitate, and perform the test: not more than 2.0. (2) Peroxide—Weigh accurately about 5 g of Retinol Palmitate, transfer in a 250-mL glass-stoppered conical flask, add 50 mL of a mixture of acetic acid (100) and isooc-tane (3:2), and gently mix to dissolve completely. Replace the air of the inside gradually with about 600 mL of Nitro-gen, then add 0.1 mL of saturated potassium iodide TS under a current of Nitrogen. Immediately stopper tightly, and mix with a swirling motion for 1 minute. Add 30 mL of water, stopper tightly, and shake vigorously for 5 to 10 se-conds. Titrate <2.50> this solution with 0.01 mol/L sodium thiosulfate VS until the blue color of the solution disappears after addition of 0.5 mL of starch TS near the end point where the solution is a pale yellow color. Calculate the amount of peroxide by the following formula: not more than 10 mEq/kg. Amount (mEq/kg) of peroxide ＝ V/M × 10 V: Volume (mL) of 0.01 mol/L sodium thiosulfate VS M: Amount (g) of Retinol Palmitate taken
Proceed as directed in Method 1-1 under the Vita-min A Assay <2.55>.
Thiamine Chloride Hydrochloride occurs as white, crystals or crystalline powder. It is odorless or has a slight, characteristic odor. It is freely soluble in water, sparingly soluble in methanol, and slightly soluble in ethanol (95). Melting point: about 2459C (with decomposition). It shows crystal polymorphism.
(1) To 5 mL of a solution of Thiamine Chloride Hydrochloride (1 in 500) add 2.5 mL of sodium hy-droxide TS and 0.5 mL of potassium hexacyanoferrate (III) TS. Then add 5 mL of 2-methyl-1-propanol, shake the mix-ture vigorously for 2 minutes, allow to stand, and examine under ultraviolet light (main wavelength: 365 nm): the 2-methyl-1-propanol layer shows a blue-purple fluorescence. This fluorescence disappears when the mixture is acidified, but reappears when it is again made alkaline. Determine the absorption spectrum of a solution of Thiamine Chloride Hydrochloride (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Thiamine Chloride Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Thia-mine Chloride Hydrochloride, previously dried at 1059C for 2 hours, as directed in the potassium chloride disk method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum, or the spectrum of Thiamine Chloride Hydrochloride RS previously dried at 1059C for 2 hours: both spectra exhibit similar intensities of absorption at the same wave numbers. In case when some differences are found between the spectra, repeat the test with residues obtained by dissolving these substances in ‌water, evaporating to dryness, and drying at 1059C for 2 hours. A solution of Thiamine Chloride Hydrochloride (1 in 500) responds to Qualitative Tests <1.09> for chloride. pH <2.54> Dissolve 1.0 g of Thiamine Chloride Hydrochlo-ride in 100 mL of water: the pH of this solution is between 2.7 and 3.4.
(1) Clarity and color of solution—Dissolve 1.0 g of Thiamine Chloride Hydrochloride in 10 mL of water: the solution is clear, and has no more color than the following control solution. Control solution: To 1.5 mL of 1/60 mol/L potassium dichromate VS add water to make 1000 mL. Sulfate <1.14>—Weigh 1.5 g of Thiamine Chloride Hydrochloride, and perform the test. Prepare the control so-lution with 0.35 mL of 0.005 mol/L sulfuric acid VS (not more than 0.011z). Nitrate—Dissolve 0.5 g of Thiamine Chloride Hydro-chloride in 25 mL of water. Add 2 mL of sulfuric acid to 2 mL of this solution, shake, cool, and superimpose iron (II) sulfate TS: no dark brown ring is produced at the junction of the two layers. Heavy metals <1.07>—Proceed with 1.0 g of Thia- mine Chloride Hydrochloride according to Method 1, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Related substances—Dissolve 0.10 g of Thiamine Chloride Hydrochloride in 100 mL of the mobile phase, and use this solution as the sample solution. Pipet 1 mL of the sample solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the area of each peak by the automatic integration method: the total area of the peaks other than thiamine obtained from the sample solution is not larger than the peak area of thiamine from the standard solution. Operating conditions— Detector, column, column temperature, mobile phase, and flow rate: Proceed as directed in the operating conditions in the Assay. Time span of measurement: About 3 times as long as the retention time of thiamine. System suitability— System performance: Proceed as directed in the system suitability in the Assay. Test for required detectability: To exactly 5 mL of the each of the solutions, accurately measured, add exactly 5 mL each of the internal standard solution, add the mobile phase to make 50 mL, and use these solutions as the sample solu-tion and standard solution. Perform the test with 10 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the peak area of thiamine to that of the internal standard. Amount (mg) of thiamine chloride hydrochloride (C12H17ClN4OS.HCl) ＝ MS × QT/QS MS: Amount (mg) of Thiamine Chloride Hydrochloride RS taken, calculated on the anhydrous basis Internal standard solution—A solution methyl benzoate in methanol (1 in 50). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 259C. Mobile phase: Dissolve 1.1 g of sodium 1-octanesulfonate in 1000 mL of diluted acetic acid (100) (1 in 100). To 600 mL of this solution add 400 mL of a mixture of methanol and acetonitrile (3:2). Flow rate: Adjust so that the retention time of thiamine is about 12 minutes. System suitability— System performance: When the procedure is run with 10 mL of the standard solution under the above operating con-ditions, thiamine and the internal standard are eluted in this order with the resolution between these peaks being not less than 6. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the peak area of thiamine to that of the internal standard is not more than 1.0z.
Weigh accurately about 0.1 g each of Thiamine Chloride Hydrochloride and Thiamine Chloride Hydrochlo-ride RS (separately determine the water <2.48> in the same manner as Thiamine Chloride Hydrochloride), and dissolve them in the mobile phase to make exactly 50 mL. To 10 mL
Cyanocobalamin occurs as dark red, crystals or powder. It is sparingly soluble in water, and slightly soluble in ethanol (99.5). It is hygroscopic.
(1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cyanocobalamin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Mix 1 mg of Cyanocobalamin with 50 mg of potas-sium hydrogen sulfate, and fuse by igniting. Cool, break up the mass with a glass rod, add 3 mL of water, and dissolve by boiling. Add 1 drop of phenolphthalein TS, then add dropwise sodium hydroxide TS until a light red color just develops. Add 0.5 g of sodium acetate trihydrate, 0.5 mL of dilute acetic acid and 0.5 mL of a solution of disodium 1-nitroso-2-naphthol-3,6-disulfonate (1 in 500): a red to orange-red color is immediately produced. Then add 0.5 mL of hydrochloric acid, and boil for 1 minute: the red color does not disappear. Transfer 5 mg of Cyanocobalamin to a 50-mL distil-ling flask, dissolve in 5 mL of water, and add 2.5 mL of hypophosphorous acid. Connect the flask with a short con-denser, and dips its tip into a test tube containing 1 mL of a solution of sodium hydroxide (1 in 50). Heat gently for 10 minutes, then distil 1 mL into a test tube. To the test tube add 4 drops of a saturated solution of ammonium iron (II) sulfate hexahydrate, shake gently, then add about 30 mg of sodium fluoride, and heat the contents to boil. Immediately Flow rate: Adjust so that the retention time of cyanocobalamin is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of cyanocobalamin, beginning after the sol-vent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cyanocobalamin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: Perform this procedure quickly after the solution is prepared. To 25 mg of cyanocobalamin add 10 mL of water, and warm, if necessary, to dissolve. After cooling, add 0.5 mL of sodium toluenesulfonchloramide TS, 0.5 mL of 0.05 mol/L hydrochloric acid TS and water to make 25 mL, mix, and allow the solution to stand for 5 minutes. To 1 mL of the solution add the mobile phase to make 10 mL. When the procedure is run with 20 mL of the solution under the above operating conditions, two principal peaks appear with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cyanocobalamin is not more than 3.0z. Loss on drying <2.41> Not more than 12z (50 mg, in vacu-um at a pressure not exceeding 0.67 kPa, phosphorus (V) ‌JP XVIII Official Monographs / Cyclopentolate Hydrochloride 821‌ oxide, 1009C, 4 hours).
(1) Clarity and color of solution—Dissolve 20 mg of Cyanocobalamin in 10 mL of water: the solution is clear and red in color. (2) Related substances—Conduct this procedure using light-resistant vessels. Dissolve 10 mg of Cyanocobalamin in 10 mL of the mobile phase, and use this solution as the sam-ple solution. Pipet 3 mL of the sample solution, add the mo-bile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with exactly 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and determine each peak area by the au-tomatic integration method: the total area of the peak other than cyanocobalamin obtained from the sample solution is not larger than the peak area of cyanocobalamin from the standard solution. Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 361 nm). C H CoN O P: 1355.37 Column: A stainless steel column 4.6 mm in inside diame- 63 88 14 14 ter and 25 cm in length, packed with octylsilanized silica gel Coa-[a-(5,6-Dimethyl-1H-benzimidazol-1-yl)]-Cob- cyanocobamide [68-19-9] Cyanocobalamin contains not less than 96.0z and not more than 102.0z of cyanocobalamin for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: Dissolve 10 g of anhydrous disodium hydrogen phosphate in 1000 mL of water, and adjust to pH (C63H 88CoN14 O14 P), calculated on the dried basis. 3.5 with phosphoric acid. To 147 mL of this solution add 53 mL of methanol. Description Cyanocobalamin occurs as dark red, crystals or powder. It is sparingly soluble in water, and slightly soluble in ethanol (99.5). It is hygroscopic. Identification (1) Determine the absorption spectrum of the sample solution obtained in the Assay as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Cyanocobalamin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. Mix 1 mg of Cyanocobalamin with 50 mg of potas-sium hydrogen sulfate, and fuse by igniting. Cool, break up the mass with a glass rod, add 3 mL of water, and dissolve by boiling. Add 1 drop of phenolphthalein TS, then add dropwise sodium hydroxide TS until a light red color just develops. Add 0.5 g of sodium acetate trihydrate, 0.5 mL of dilute acetic acid and 0.5 mL of a solution of disodium 1-nitroso-2-naphthol-3,6-disulfonate (1 in 500): a red to orange-red color is immediately produced. Then add 0.5 mL of hydrochloric acid, and boil for 1 minute: the red color does not disappear. Transfer 5 mg of Cyanocobalamin to a 50-mL distil-ling flask, dissolve in 5 mL of water, and add 2.5 mL of hypophosphorous acid. Connect the flask with a short con-denser, and dips its tip into a test tube containing 1 mL of a solution of sodium hydroxide (1 in 50). Heat gently for 10 minutes, then distil 1 mL into a test tube. To the test tube add 4 drops of a saturated solution of ammonium iron (II) sulfate hexahydrate, shake gently, then add about 30 mg of sodium fluoride, and heat the contents to boil. Immediately Flow rate: Adjust so that the retention time of cyanocobalamin is about 7 minutes. Time span of measurement: About 4 times as long as the retention time of cyanocobalamin, beginning after the sol-vent peak. System suitability— Test for required detectability: To 1 mL of the sample so-lution, add the mobile phase to make 100 mL, and use this solution as the solution for system suitability test. Pipet 1 mL of the solution for system suitability test, and add the mobile phase to make exactly 10 mL. Confirm that the peak area of cyanocobalamin obtained with 20 mL of this solution is equivalent to 7 to 13z of that with 20 mL of the solution for system suitability test. System performance: Perform this procedure quickly after the solution is prepared. To 25 mg of cyanocobalamin add 10 mL of water, and warm, if necessary, to dissolve. After cooling, add 0.5 mL of sodium toluenesulfonchloramide TS, 0.5 mL of 0.05 mol/L hydrochloric acid TS and water to make 25 mL, mix, and allow the solution to stand for 5 minutes. To 1 mL of the solution add the mobile phase to make 10 mL. When the procedure is run with 20 mL of the solution under the above operating conditions, two principal peaks appear with the resolution between these peaks being not less than 2.5. System repeatability: When the test is repeated 6 times with 20 mL of the solution for system suitability test under the above operating conditions, the relative standard devia-tion of the peak area of cyanocobalamin is not more than 3.0z. Loss on drying <2.41> Not more than 12z (50 mg, in vacu-um at a pressure not exceeding 0.67 kPa, phosphorus (V) ‌JP XVIII Official Monographs / Cyclopentolate Hydrochloride 821‌ oxide, 1009C, 4 hours).
Weigh accurately about 20 mg each of Cyanocobala-min and Cyanocobalamin RS (previously determine the loss on drying <2.41> under the same conditions as Cyanocobala-min), dissolve in water to make exactly 1000 mL, respec-tively, and use these solutions as the sample solution and the standard solution. Determine the absorbances, AT and AS, of the sample solution and standard solution, at 361 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of cyanocobalamin (C63H88CoN14O14P) ＝ MS × AT/AS MS: Amount (mg) of Cyanocobalamin RS taken, calcu-lated on the dried basis
Riboflavin Butyrate occurs as orange-yellow, crystals or crystalline powder. It has a slight, characteristic odor and a slightly bitter taste. It is freely soluble in methanol, in ethanol (95) and in chlo-roform, slightly soluble in diethyl ether, and practically in-soluble in water. It is decomposed by light.
(1) A solution of Riboflavin Butyrate in ethanol (95) (1 in 100,000) shows a light yellow-green color with a strong yellowish green fluorescence. To the solution add dilute hydrochloric acid or sodium hydroxide TS: the fluorescence disappears. (2) Dissolve 0.01 g of Riboflavin Butyrate in 5 mL of ethanol (95), add 2 mL of a mixture of a solution of sodium hydroxide (3 in 20) and a solution of hydroxylammonium tion. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Chloride—Dissolve 2.0 g of Riboflavin Butyrate in 10 mL of methanol, and add 24 mL of dilute nitric acid and water to make 100 mL. After shaking well, allow to stand for 10 minutes, filter, discard the first 10 mL of the filtrate, and use the subsequent filtrate as the sample solution. To 25 mL of the sample solution add water to make 50 mL, then add 1 mL of silver nitrate TS, and allow to stand for 5 minutes: the turbidity of the solution is no more than that of the following control solution. Control solution: To 25 mL of the sample solution add 1 mL of silver nitrate TS, allow to stand for 10 minutes, and filter. Wash the precipitate with four 5-mL portions of water, and combine the washings with the filtrate. To this solution add 0.30 mL of 0.01 mol/L hydrochloric acid VS and water to make 50 mL, add 1 mL of water, and mix (not more than 0.021z). Heavy metals <1.07>—Proceed with 2.0 g of Riboflavin Butyrate according to Method 2, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 10 ppm). Free acid—To 1.0 g of Riboflavin Butyrate add 50 mL of freshly boiled and cooled water, shake, and filter. To 25 mL of the filtrate add 0.50 mL of 0.01 mol/L sodium hy-droxide VS and 2 drops of phenolphthalein TS: the solution shows a red color. Related substances—Dissolve 0.10 g of Riboflavin Butyrate in 10 mL of chloroform, and use this solution as the sample solution. Pipet 1 mL of the sample solution, and add chloroform to make exactly 50 mL. Pipet 5 mL of this solution, add chloroform to make exactly 20 mL, and use this solution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatogra-phy <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel with fluorescent in-dicator for thin-layer chromatography. Develop the plate with a mixture of chloroform and 2-propanol (9:1) to a dis-tance of about 10 cm, and air-dry the plate. Examine under ultraviolet light (main wavelength: 254 nm): the spots other than 98.5z of riboflavin butyrate (C33 H44 N4O10). than the principal spot obtained from the sample solution are not more intense than the spot from the standard solu- Description Riboflavin Butyrate occurs as orange-yellow, crystals or crystalline powder. It has a slight, characteristic odor and a slightly bitter taste. It is freely soluble in methanol, in ethanol (95) and in chlo-roform, slightly soluble in diethyl ether, and practically in-soluble in water. It is decomposed by light. Identification (1) A solution of Riboflavin Butyrate in ethanol (95) (1 in 100,000) shows a light yellow-green color with a strong yellowish green fluorescence. To the solution add dilute hydrochloric acid or sodium hydroxide TS: the fluorescence disappears. (2) Dissolve 0.01 g of Riboflavin Butyrate in 5 mL of ethanol (95), add 2 mL of a mixture of a solution of sodium hydroxide (3 in 20) and a solution of hydroxylammonium tion. Loss on drying <2.41> Not more than 0.5z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 40 mg of Riboflavin Butyrate, previously dried, dissolve in ethanol (95) to make exactly 500 mL, and pipet 10 mL of this solu-tion, add ethanol (95) to make exactly 50 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of Riboflavin RS, previously dried at 1059C for 2 hours, dissolve in 150 mL of diluted acetic acid (100) (2 in 75) by warming, and after cooling, add water to make ex-actly 500 mL. Pipet 5 mL of this solution, add ethanol (95) ‌to make exactly 50 mL, and use this solution as the standard solution. Determine the absorbances, AT and AS, of the sam-ple solution and standard solution at 445 nm as directed under Ultraviolet-visible Spectrophotometry <2.24>. Amount (mg) of riboflavin butyrate (C33H44N4O10) ＝ MS × AT/AS × 1/2 × 1.745 MS: Amount (mg) of Riboflavin RS taken
Riboflavin Sodium Phosphate is a yellow to orange-yellow crystalline powder. It is odorless, and has a slightly bitter taste. It is soluble in water, and practically insoluble in ethanol (95), in chloroform and in diethyl ether. It is decomposed on exposure to light. It is very hygroscopic.
(1) A solution of Riboflavin Sodium Phos-phate (1 in 100,000) is light yellow-green in color and has an intense yellow-green fluorescence. The color and fluores-cence of the solution disappear upon the addition of 0.02 g of sodium hydrosulfite to 5 mL of the solution, and reappear on shaking the mixture in air. This fluorescence disappears upon the addition of dilute hydrochloric acid or sodium hy-droxide TS. To 10 mL of a solution of Riboflavin Sodium Phos-phate (1 in 100,000) placed in a glass-stoppered test tube add 1 mL of sodium hydroxide TS, and after illumination with a fluorescence lamp of 10 to 30 watts at 20-cm distance for 30 minutes between 209C and 409C, acidify with 0.5 mL of ace-tic acid (31), and shake with 5 mL of chloroform: the chlo-roform layer shows a yellow-green fluorescence. Determine the absorption spectrum of a solution of Riboflavin Sodium Phosphate in phosphate buffer solution (pH 7.0) (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar inten-sities of absorption at the same wavelengths. To 0.05 g of Riboflavin Sodium Phosphate add 10 mL of nitric acid, evaporate on a water bath to dryness, and ignite. Boil the residue with 10 mL of diluted nitric acid (1 in 50) for 5 minutes, after cooling, neutralize this solution with ammonia TS, and filter, if necessary: the solution responds to Qualitative Tests <1.09> for sodium salt and phosphate. D Optical rotation <2.49> [a]20: ＋38 – ＋439(0.3 g calculated on the anhydrous basis, 5 mol/L hydrochloric acid TS, 20 mL, 100 mm). pH <2.54> Dissolve 0.20 g of Riboflavin Sodium Phosphate in 20 mL of water: the pH of the solution is between 5.0 and 6.5.
(1) Clarity and color of solution—Dissolve 0.20 g of Riboflavin Sodium Phosphate in 10 mL of water: the so-lution is clear and yellow to orange-yellow in color. Lumiflavin—To 35 mg of Riboflavin Sodium Phos-phate add 10 mL of ethanol-free chloroform, and shake for 5 minutes, then filter: the filtrate has no more color than the control solution. Control solution: To 3.0 mL of 1/60 mol/L potassium dichromate VS add water to make 1000 mL. Free phosphoric acid—Weigh accurately about 0.4 g of Riboflavin Sodium Phosphate, dissolve in water to make exactly 100 mL, and use this solution as the sample solution. Measure exactly 5 mL each of the sample solution and Standard Phosphoric Acid Solution, transfer to separate 25-mL volumetric flasks, add 2.5 mL of hexaammonium heptamolybdate-sulfuric acid TS and 1 mL of 1-amino-2-naphthol-4-sulfonic acid TS to each of these flasks, mix, and add water to make 25 mL. Allow to stand for 30 minutes at 20 ± 19C, and perform the test with these solutions as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, using a solution prepared with 5 mL of water in the same manner as a blank. Determine the absorbances, AT and AS, of the subsequent solutions of the sample solution and Standard Phosphoric Acid Solution at 740 nm: the free phosphoric acid content is not more than 1.5z. Content (z) of free phosphoric acid (H3PO4) ＝ 1/M × AT/AS × 258.0 M: Amount (mg) of Riboflavin Sodium Phosphate taken, calculated on the anhydrous basis Water <2.48> Place 25 mL of a mixture of methanol for water determination and ethylene glycol for water determi-nation (1:1) in a dry flask for titration, and titrate with Karl Fischer TS for water determination to the end point. Weigh accurately about 0.1 g of Riboflavin Sodium Phosphate, place quickly into the flask, add a known excess volume of Karl Fischer TS for water determination, mix for 10 minutes, and perform the test: the water content is not more than 10.0z.
Conduct this procedure without exposure to light, using light-resistant vessels. To about 0.1 g of Riboflavin So-dium Phosphate, accurately weighed, dissolve in diluted ace-tic acid (100) (1 in 500) to make exactly 1000 mL, then pipet 10 mL of this solution, and add diluted acetic acid (100) (1 in 500) to make exactly 50 mL. Use this solution as the sample solution. Separately, dry Riboflavin RS at 1059C for 2 hours, weigh accurately about 15 mg, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) by warming, cool, add water to make exactly 1000 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spectrophotometry <2.24>, using water as the blank, and de-termine the absorbances, AT and AS, at 445 nm. Add 0.02 g ‌of sodium hydrosulfite to 5 mL of each solution, shake until decolorized, and immediately measure the absorbances, AT? and AS?, of the solutions.‌ Amount (mg) of riboflavin sodium phosphate (C17H20N4NaO9P) ＝ MS × (AT － AT?)/(AS － AS?) × 5 × 1.271 MS: Amount (mg) of Riboflavin RS taken
Riboflavin occurs as yellow to orange-yellow crystals. It has a slight odor. It is very slightly soluble in water, practically insoluble in ethanol (95), in acetic acid (100), and in diethyl ether. It dissolves in sodium hydroxide TS. A saturated solution of Riboflavin is neutral. It is decomposed by light. Melting point: about 2909C (with decomposition).
(1) A solution of Riboflavin (1 in 100,000) is light yellow-green in color and has an intense yellow-green fluorescence. The color and fluorescence of the solution dis-appear upon the addition of 0.02 g of sodium hydrosulfite to 5 mL of the solution, and reappear on shaking the mixture in air. This fluorescence disappears upon the addition of dilute hydrochloric acid or sodium hydroxide TS. To 10 mL of a solution of Riboflavin (1 in 100,000) placed in a glass-stoppered test tube add 1 mL of sodium hy-droxide TS, and after illumination with a fluorescence lamp of 10 to 30 watts at 20-cm distance for 30 minutes between 209C and 409C, acidify with 0.5 mL of acetic acid (31), and shake with 5 mL of chloroform: the chloroform layer shows a yellow-green fluorescence. Determine the absorption spectrum of a solution of Riboflavin in phosphate buffer solution (pH 7.0) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Rivoflavin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]20: －128 – －1429 Weigh accu-rately about 0.1 g of dried Riboflavin, dissolve in exactly 4 mL of dilute sodium hydroxide TS, add 10 mL of freshly boiled and cooled water, add exactly 4 mL of aldehyde-free ethanol while shaking, add freshly boiled and cooled water to make exactly 20 mL, and determine the rotation in a 100-mm cell within 30 minutes after preparing the solution.
Lumiflavin—Shake 25 mg of Riboflavin with 10 mL of ethanol-free chloroform for 5 minutes, and filter: the fil-trate has no more color than the following control solution. Control solution: To 2.0 mL of 1/60 mol/L potassium dichromate VS add water to make 1000 mL. MS: Amount (mg) of Riboflavin RS taken
Conduct this procedure without exposure to light, using light-resistant vessels. Weigh accurately about 15 mg of Riboflavin, previously dried, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) by warming, cool, add water to make exactly 1000 mL, and use this solution as the sample solution. Dry Riboflavin RS at 1059C for 2 hours, weigh accurately about 15 mg, dissolve in 800 mL of diluted acetic acid (100) (1 in 400) by warming, cool, add water to make exactly 1000 mL, and use this solution as the standard solution. Perform the test with the sample solution and standard solution as directed under Ultraviolet-visible Spec-trophotometry <2.24>, using water as the blank, and deter-mine the absorbances, AT and AS, at 445 nm. Add 0.02 g of sodium hydrosulfite to 5 mL of each solution, shake until decolorized, and immediately measure the absorbances, AT? and AS?, of the solutions. Amount (mg) of riboflavin (C17H20N4O6) ＝ MS × (AT － AT?)/(AS － AS?) 17 20 4 6 Description Riboflavin occurs as yellow to orange-yellow crystals. It has a slight odor. It is very slightly soluble in water, practically insoluble in ethanol (95), in acetic acid (100), and in diethyl ether. It dissolves in sodium hydroxide TS. A saturated solution of Riboflavin is neutral. It is decomposed by light. Melting point: about 2909C (with decomposition). Identification (1) A solution of Riboflavin (1 in 100,000) is light yellow-green in color and has an intense yellow-green fluorescence. The color and fluorescence of the solution dis-appear upon the addition of 0.02 g of sodium hydrosulfite to 5 mL of the solution, and reappear on shaking the mixture in air. This fluorescence disappears upon the addition of dilute hydrochloric acid or sodium hydroxide TS. To 10 mL of a solution of Riboflavin (1 in 100,000) placed in a glass-stoppered test tube add 1 mL of sodium hy-droxide TS, and after illumination with a fluorescence lamp of 10 to 30 watts at 20-cm distance for 30 minutes between 209C and 409C, acidify with 0.5 mL of acetic acid (31), and shake with 5 mL of chloroform: the chloroform layer shows a yellow-green fluorescence. Determine the absorption spectrum of a solution of Riboflavin in phosphate buffer solution (pH 7.0) (1 in 100,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Rivoflavin RS prepared in the same manner as the sample solution: both spectra exhibit similar intensities of absorption at the same wavelengths. D Optical rotation <2.49> [a]20: －128 – －1429 Weigh accu-rately about 0.1 g of dried Riboflavin, dissolve in exactly 4 mL of dilute sodium hydroxide TS, add 10 mL of freshly boiled and cooled water, add exactly 4 mL of aldehyde-free ethanol while shaking, add freshly boiled and cooled water to make exactly 20 mL, and determine the rotation in a 100-mm cell within 30 minutes after preparing the solution. Purity Lumiflavin—Shake 25 mg of Riboflavin with 10 mL of ethanol-free chloroform for 5 minutes, and filter: the fil-trate has no more color than the following control solution. Control solution: To 2.0 mL of 1/60 mol/L potassium dichromate VS add water to make 1000 mL. MS: Amount (mg) of Riboflavin RS taken
Pyridoxine Hydrochloride occurs as a white to pale yellow crystalline powder. It is freely soluble in water, slightly soluble in ethanol (99.5), and practically insoluble in acetic anhydride and in acetic acid (100). It is gradually affected by light. Melting point: about 2069C (with decomposition). (65:13:13:9) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of sodium carbonate in diluted ethanol (3 in 10) (1 in 20) on the plate. After air-dry ing, spray evenly a solution of 2,6-dibromo-N-chloro-1,4-benzoquinone monoimine in ethanol (99.5) (1 in 1000) on the plate, and air-dry: the spot other than the principal spot ob-tained from the sample solution is not more intense than the spot from the standard solution. Loss on drying <2.41> Not more than 0.30z (1 g, in vacu-um, silica gel, 4 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
(1) Determine the absorption spectrum of a solution of Pyridoxine Hydrochloride in 0.1 mol/L hydro- Vitamin B6 Injection chloric acid TS (1 in 100,000) as directed under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum or the spectrum of a solution of Pyridoxine Hydrochloride RS prepared in the same manner as the sample solution: both spectra exhibit similar inten-sities of absorption at the same wavelengths. Determine the infrared absorption spectrum of Pyridoxine Hydrochloride, previously dried, as directed in the potassium chloride disk method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum or the spectrum of Pyridoxine Hydrochlo-ride RS: both spectra exhibit similar intensities of absorption at the same wave numbers. A solution of Pyridoxine Hydrochloride (1 in 10) responds to Qualitative Tests <1.09> for chloride. pH <2.54> The pH of a solution prepared by dissolving 1.0 g of Pyridoxine Hydrochloride in 50 mL of water is be-tween 2.5 and 3.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Pyridoxine hydrochloride in 20 mL of water: the solution is clear and colorless. Heavy metals <1.07>—Proceed with 1.0 g of Pyridoxine Hydrochloride according to Method 1, and per-form the test. Prepare the control solution with 3.0 mL of Standard Lead Solution (not more than 30 ppm). Related substances—Dissolve 1.0 g of Pyridoxine Hydrochloride in 10 mL of water, and use this solution as the sample solution. Pipet 2.5 mL of the sample solution, and add water to make exactly 100 mL. Pipet 1 mL of this solution, add water to make exactly 10 mL, and use this so-lution as the standard solution. Perform the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 2 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatogra-phy, and air-dry the plate. Develop the plate with a mixture of acetone, tetrahydrofuran, hexane and ammonia solution
Weigh accurately about 0.2 g of Pyridoxine Hydro-chloride, previously dried, add 5 mL of acetic acid (100) and 5 mL of acetic anhydride, dissolve by gentle boiling, cool, add 30 mL of acetic anhydride, and titrate <2.50> with 0.1 mol/L perchloric acid VS (potentiometric titration). Per-form a blank determination in the same manner, and make any necessary correction. Each mL of 0.1 mol/L perchloric acid VS ＝ 20.56 mg of C8H11NO3.HCl
Ascorbic Acid occurs as white crystals or a white crystalline powder. It is odorless, and has an acid taste. It is freely soluble in water, sparingly soluble in ethanol (95), and practically insoluble in diethyl ether. Melting point: about 1909C (with decomposition).
(1) To 5 mL each of a solution of Ascorbic Acid (1 in 50) add 1 drop of potassium permanganate TS or 1 to 2 drops of 2,6-dichloroindophenol sodium TS: the color of the solution is discharged immediately in each case. (2) Dissolve 0.1 g of Ascorbic Acid in 100 mL of a solu-tion of metaphosphoric acid (1 in 50). To 5 mL of the solu-tion add iodine TS until the color of the solution becomes light yellow. Then add 1 drop of a solution of copper (II) sulfate pentahydrate (1 in 1000) and 1 drop of pyrrole, and warm the mixture at 509C for 5 minutes: a blue color de-velops. D Optical rotation <2.49> [a]20: ＋20.5 – ＋21.59 (2.5 g, water, 25 mL, 100 mm). pH <2.54> Dissolve 1.0 g of Ascorbic Acid in 20 mL of water: the pH of this solution is between 2.2 and 2.5.
(1) Clarity and color of solution—Dissolve 1.0 g of Ascorbic Acid in 20 mL of water: the solution is clear and colorless. (2) Heavy metals <1.07>—Perform the test with 1.0 g of Ascorbic Acid according to Method 1. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Loss on drying <2.41> Not more than 0.20z (1 g, silica gel, 24 hours). Residue on ignition <2.44> Not more than 0.1z (1 g).
Weigh accurately about 0.2 g of Ascorbic Acid, previously dried, dissolve in 50 mL of a solution of meta-phosphoric acid (1 in 50), and titrate <2.50> with 0.05 mol/L iodine VS (indicator: 1 mL of starch TS). Each mL of 0.05 mol/L iodine VS ＝ 8.806 mg of C6H8O6
Ergocalciferol occurs as white crystals. It is odorless, or has a faint, characteristic odor. It is freely soluble in ethanol (95), in diethyl ether and in chloroform, sparingly soluble in isooctane, and practically insoluble in water. It is affected by air and by light. Melting point: 115 – 1189C Transfer Ergocalciferol to a capillary tube, and dry for 3 hours in a desiccator (in vacu-um at a pressure not exceeding 2.67 kPa). Immediately fire-seal the capillary tube, put it in a bath fluid, previously heated to a temperature about 109C below the expected melting point, and heat at a rate of rise of about 39C per minute, and read the melting point.
(1) Dissolve 0.5 mg of Ergocalciferol in 5 mL of chloroform, add 0.3 mL of acetic anhydride and 0.1 mL of sulfuric acid, and shake: a red color is produced, and rapidly changes through purple and blue to green. (2) Determine the infrared absorption spectrum of Ergocalciferol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Ergocalciferol RS: both spectra exhibit similar inten-sities of absorption at the same wave numbers. has been prepared.
Ergosterol—Dissolve 10 mg of Ergocalciferol in 2.0 mL of diluted ethanol (9 in 10), add a solution of 20 mg of digitonin in 2.0 mL of diluted ethanol (9 in 10), and allow the mixture to stand for 18 hours: no precipitate is formed.
Weigh accurately about 30 mg each of Ergocal-ciferol and Ergocalciferol RS, and dissolve each in isooctane to make exactly 50 mL. Pipet 10 mL each of these solutions, add exactly 3 mL each of the internal standard solution, then add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution, respec-tively. Perform the test with 10 to 20 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following condi-tions, and calculate the ratios, QT and QS, of the peak area of ergocalciferol to that of the internal standard. Perform the procedure rapidly avoiding contact with air or other oxidizing agents and using light-resistant containers. Amount (mg) of ergocalciferol (C28H44O) ＝ MS × QT/QS MS: Amount (mg) of Ergocalciferol RS taken Internal standard solution—A solution of dimethyl phtha-late in isooctane (1 in 100). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with a silica gel for liquid chromatography (10 mm particle diameter). Column temperature: A constant temperature of about 209C. Mobile phase: A mixture of hexane and n-amylalcohol (997:3). Flow rate: Adjust so that the retention time of ergocal-ciferol is about 25 minutes. System suitability— System performance: Dissolve 15 mg of Ergocalciferol RS in 25 mL of isooctane. Transfer this solution to a flask, heat in an oil bath under a reflux condenser for 2 hours, and cool immediately to room temperature. Transfer the solution to a quartz test tube, and irradiate with a short-wave lamp (main wavelength: 254 nm) and a long-wave lamp (main wave-length: 365 nm) for 3 hours. To 10 mL of this solution add the mobile phase to make 50 mL. When the procedure is run with 10 mL of this solution under the above operating condi-tions, the ratios of the retention time of previtamin D2, trans-vitamin D2 and tachysterol2 to that of ergocalciferol are about 0.5, about 0.6 and about 1.1, respectively, and the resolution between previtamin D2 and trans-vitamin D2 is not less than 0.7, and that between ergocalciferol and tachysterol2 is not less than 1.0. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratios of the peak area of ergocalciferol to that of the internal standard is not more than 1.0z.
Containers—Hermetic containers. Storage—Light-resistant, under Nitrogen atmosphere, and
Cholecalciferol occurs as white crystals. It is odorless. It is freely soluble in ethanol (95), in chloroform, in diethyl ether and in isooctane, and practically insoluble in water. It is affected by air and by light. Melting point: 84 – 889C Transfer Cholecalciferol to a capillary tube, and dry for 3 hours in a desiccator (in vacu-um at a pressure not exceeding 2.67 kPa). Immediately fireseal the capillary tube, put it in a bath fluid, previously heated to a temperature about 109C below the expected melting point, and heat at a rate of rise of about 39C per minute, and read the melting point. Amount (mg) of chlorpropamide (C ＝ MS × AT/AS 10H13ClN2O3S)
(1) Dissolve 0.5 mg of Cholecalciferol in 5 mL of chloroform, add 0.3 mL of acetic anhydride and 0.1 mL of sulfuric acid, and shake: a red color is produced, and MS: Amount (mg) of chlorpropamide for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). rapidly changes through purple and blue to green. Determine the infrared absorption spectrum of Cholecalciferol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cholecalciferol RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Column temperature: A constant temperature of about Absorbance <2.24> E 1z (265 nm): 450 – 490 (10 mg, 1 cm 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of chlor-propamide is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of chlorpropamide are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times ethanol (95), 1000 mL). D Optical rotation <2.49> [a]20 ＋103 – ＋1129 (50 mg, ethanol (95), 10 mL, 100 mm). Prepare the solution without delay, using Cholecalciferol from a container opened not longer than 30 minutes, previously, and determine the rota-tion within 30 minutes after the solution has been prepared.
7-Dehydrocholesterol—Dissolve 10 mg of Cholecal-ciferol in 2.0 mL of diluted ethanol (95) (9 in 10), add a solu-tion prepared by dissolving 20 mg of digitonin in 2.0 mL of diluted ethanol (95) (9 in 10), and allow the mixture to stand for 18 hours: no precipitate is formed. ‌JP XVIII Official Monographs / Cholesterol 749
Weigh accurately and powder not less than 20 Chlorpropamide Tablets. Weigh accurately a quantity of the powder, equivalent to about 50 mg of chlorpropamide (C10H13ClN2O3S), add 75 mL of the mobile phase, shake for 10 minutes, and add the mobile phase to make exactly 100 mL. Centrifuge this solution, pipet 10 mL of the supernatant liquid, add the mobile phase to make exactly 100 mL, and use this solution as the sample solution. Separately, weigh accurately about 50 mg of chlorpropamide for assay, previ-ously dried at 1059C for 3 hours, dissolve in the mobile phase to make exactly 100 mL. Pipet 10 mL of this solution, add the mobile phase to make exactly 100 mL, and use this solution as the standard solution. Perform the test with ex-actly 20 mL each of the sample solution and standard solu-tion as directed under Liquid Chromatography <2.01> ac-cording to the following conditions. Determine the peak areas, AT and AS, of chlorpropamide in each solution. (3S,5Z,7E )-9,10-Secocholesta-5,7,10(19)-trien-3-ol [67-97-0] Cholecalciferol contains not less than 97.0z and not more than 103.0z of cholecalciferol (C27H44O). Description Cholecalciferol occurs as white crystals. It is odorless. It is freely soluble in ethanol (95), in chloroform, in diethyl ether and in isooctane, and practically insoluble in water. It is affected by air and by light. Melting point: 84 – 889C Transfer Cholecalciferol to a capillary tube, and dry for 3 hours in a desiccator (in vacu-um at a pressure not exceeding 2.67 kPa). Immediately fireseal the capillary tube, put it in a bath fluid, previously heated to a temperature about 109C below the expected melting point, and heat at a rate of rise of about 39C per minute, and read the melting point. Amount (mg) of chlorpropamide (C ＝ MS × AT/AS 10H13ClN2O3S) Identification (1) Dissolve 0.5 mg of Cholecalciferol in 5 mL of chloroform, add 0.3 mL of acetic anhydride and 0.1 mL of sulfuric acid, and shake: a red color is produced, and MS: Amount (mg) of chlorpropamide for assay taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 240 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (10 mm in particle diameter). rapidly changes through purple and blue to green. Determine the infrared absorption spectrum of Cholecalciferol as directed in the potassium bromide disk method under Infrared Spectrophotometry <2.25>, and com-pare the spectrum with the Reference Spectrum or the spec-trum of Cholecalciferol RS: both spectra exhibit similar in-tensities of absorption at the same wave numbers. Column temperature: A constant temperature of about Absorbance <2.24> E 1z (265 nm): 450 – 490 (10 mg, 1 cm 259C. Mobile phase: A mixture of diluted acetic acid (100) (1 in 100) and acetonitrile (1:1). Flow rate: Adjust so that the retention time of chlor-propamide is about 5 minutes. System suitability— System performance: When the procedure is run with 20 mL of the standard solution under the above operating con-ditions, the number of theoretical plates and the symmetry factor of the peak of chlorpropamide are not less than 1500 and not more than 1.5, respectively. System repeatability: When the test is repeated 6 times ethanol (95), 1000 mL). D Optical rotation <2.49> [a]20 ＋103 – ＋1129 (50 mg, ethanol (95), 10 mL, 100 mm). Prepare the solution without delay, using Cholecalciferol from a container opened not longer than 30 minutes, previously, and determine the rota-tion within 30 minutes after the solution has been prepared. Purity 7-Dehydrocholesterol—Dissolve 10 mg of Cholecal-ciferol in 2.0 mL of diluted ethanol (95) (9 in 10), add a solu-tion prepared by dissolving 20 mg of digitonin in 2.0 mL of diluted ethanol (95) (9 in 10), and allow the mixture to stand for 18 hours: no precipitate is formed. ‌JP XVIII Official Monographs / Cholesterol 749 Assay Proceed with the operation avoiding contact with air or other oxidizing agents and using light-resistant containers. Dissolve separately about 30 mg each of Cholecalciferol and Cholecalciferol RS, accurately weighed, in isooctane to make exactly 50 mL. Pipet 10 mL each of these solutions, add 3 mL each of the internal standard solution, then add the mobile phase to make 50 mL, and use these solutions as the sample solution and the standard solution. Perform the test with 10 mL each of the sample solution and standard so-lution as directed under Liquid Chromatography <2.01> ac-cording to the following conditions, and calculate the ratios, QT and QS, of the peak area of cholecalciferol to that of the internal standard. Cholesterol
Tocopherol Acetate is a clear, colorless or yel-low, viscous and odorless liquid. It is miscible with ethanol (99.5), with acetone, with chlo-roform, with diethyl ether, with hexane and with vegetable oils. It is freely soluble in ethanol (95), and practically insoluble in water. It is optically inactive. It is affected by air and light.
(1) Dissolve 0.05 g of Tocopherol Acetate in 10 mL of ethanol (99.5), add 2 mL of nitric acid, and heat at 759C for 15 minutes: a red to orange color is produced. (2) Determine the infrared absorption spectrum of Tocopherol Acetate as directed in the liquid film method under Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Tocopherol Acetate RS: both spectra exhibit similar inten- Refractive index <2.45> n20: 1.494 – 1.499 D Specific gravity <2.56> d 20: 0.952 – 0.966
(1) Clarity and color of solution—Dissolve 0.10 g of Tocopherol Acetate in 10 mL of ethanol (99.5): the solu-tion is clear, and has no more color than the following con-trol solution. Control solution: To 0.5 mL of Iron (III) Chloride CS add 0.5 mol/L hydrochloric acid TS to make 100 mL. Heavy metals <1.07>—Carbonize 1.0 g of Tocopherol Acetate by gentle heating. Cool, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and ignite the ethanol to burn. Cool, add 1 mL of sulfuric acid, proceed according to Method 4, and perform the test. Pre-pare the control solution with 2.0 mL of Standard Lead So-lution (20 ppm). a-Tocopherol—Dissolve 0.10 g of Tocopherol Ace-tate in exactly 10 mL of hexane, and use this solution as the sample solution. Separately, dissolve 50 mg of Tocopherol RS in hexane to make exactly 100 mL. Pipet 1 mL of this so-lution, add hexane to make exactly 10 mL, and use this solu-tion as the standard solution. Perform the test with these so-lutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solu-tion on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of toluene and acetic acid (100) (19:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of iron (III) chloride hexahy-drate in ethanol (99.5) (1 in 500) on the plate, then spray evenly a solution of 2,2?-bipyridyl in ethanol (99.5) (1 in 200) on the same plate, and allow to stand for 2 to 3 minutes: the spot obtained from the sample solution corresponding to that from the standard solution is not larger and not more intense than the spot from the standard solution.
Dissolve 50 mg each of Tocopherol Acetate and Tocopherol Acetate RS, accurately weighed, in ethanol (99.5) to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of these solutions as directed under Liq-uid Chromatography <2.01> according to the following con-ditions, and determine the peak heights, HT and HS, of tocopherol acetate in each solution. Amount (mg) of tocopherol acetate (C31H52O3) ＝ MS × HT/HS MS: Amount (mg) of Tocopherol Acetate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 284 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and water (49:1). Flow rate: Adjust so that the retention time of tocopherol acetate is about 12 minutes. System suitability— System performance: Dissolve 0.05 g each of Tocopherol Acetate and tocopherol in 50 mL of ethanol (99.5). When the procedure is run with 20 mL of this solution under the above operating conditions, tocopherol and tocopherol ace- ‌tate are eluted in this order with the resolution between these peaks being not less than 2.6. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak heights of tocopherol acetate is not more than 0.8z.
Tocopherol Calcium Succinate occurs as a white to yellowish white powder. It is odorless. It is freely soluble in chloroform and in carbon tetrachlo-ride, and practically insoluble in water, in ethanol (95) and in acetone. Shake 1 g of Tocopherol Calcium Succinate with 7 mL of acetic acid (100): it dissolves, and produces a turbidity after being allowed to stand for a while. It dissolves in acetic acid (100). It is optically inactive.
(1) Dissolve 0.05 g of Tocopherol Calcium Succinate in 1 mL of acetic acid (100), add 9 mL of ethanol (99.5), and mix. To this solution add 2 mL of fuming nitric acid, and heat at 759C for 15 minutes: a red to orange color develops. Dissolve 0.08 g of Tocopherol Calcium Succinate, previously dried, in 0.2 mL of carbon tetrachloride. Deter-mine the infrared absorption spectrum of the solution as directed in the liquid film method under Infrared Spectro-photometry <2.25>, and compare the spectrum with the Ref-erence Spectrum: both spectra exhibit similar intensities of absorption at the same wave numbers. Dissolve 5 g of Tocopherol Calcium Succinate in 30 0.5 mol/L hydrochloric acid TS to make 100 mL. Alkalinity—To 0.20 g of Tocopherol Calcium Suc-cinate add 10 mL of diethyl ether, 2 mL of water, 1 drop of phenolphthalein TS and 0.10 mL of 0.1 mol/L hydrochloric acid VS, and shake: no red color develops in the water layer. Chloride <1.03>—Dissolve 0.10 g of Tocopherol Cal- cium Succinate in 4 mL of acetic acid (100), add 20 mL of water and 50 mL of diethyl ether, shake thoroughly, and col-lect the water layer. To the diethyl ether layer add 10 mL of water, shake, and collect the water layer. Combine the water layers, add 6 mL of dilute nitric acid and water to make 50 mL, and perform the test using this solution as the test solu-tion. Prepare the control solution in the same manner using 0.60 mL of 0.01 mol/L hydrochloric acid VS in place of Tocopherol Calcium Succinate (not more than 0.212z). Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol Calcium Succinate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Arsenic <1.11>—Prepare the test solution with 1.0 g of Tocopherol Calcium Succinate according to Method 3, and perform the test (not more than 2 ppm). a-Tocopherol—Dissolve 0.10 g of Tocopherol Cal-cium Succinate in exactly 10 mL of chloroform, and use this solution as the sample solution. Separately, dissolve 50 mg of Tocopherol RS in chloroform to make exactly 100 mL. Pipet 1 mL of this solution, add chloroform to make exactly 10 mL, and use this solution as the standard solution. Per-form the test with these solutions as directed under Thin-layer Chromatography <2.03>. Spot 10 mL each of the sample solution and standard solution on a plate of silica gel for thin-layer chromatography. Develop the plate with a mixture of toluene and acetic acid (100) (19:1) to a distance of about 10 cm, and air-dry the plate. Spray evenly a solution of iron (III) chloride hexahydrate in ethanol (99.5) (1 in 500) on the plate, then spray evenly a solution of 2,2?-bipyridyl in ethanol (99.5) (1 in 200) on the same plate, and allow to stand for 2 to 3 minutes: the spots obtained from the sample solution corresponding to the spots from the standard solu-tion is not larger and not more intense than the spots from the standard solution. Loss on drying <2.41> Not more than 2.0z (1 g, in vacu-um, phosphorus (V) oxide, 24 hours).
(1) Clarity and color of solution—Dissolve 0.10 g of Tocopherol Calcium Succinate in 10 mL of chloroform: the solution is clear, and has no more color than the follow-ing control solution. Control solution: To 0.5 mL of Iron (III) Chloride CS add length: 284 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 15 to 30 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: Room temperature. ‌Mobile phase: A mixture of methanol, water and acetic acid (100) (97:2:1). Flow rate: Adjust so that the retention time of tocopherol succinate is about 8 minutes. Selection of column: Dissolve 0.05 g each of tocopherol succinate and tocopherol in 50 mL of a mixture of ethanol (99.5) and diluted acetic acid (100) (1 in 5) (9:1). Proceed with 20 mL of this solution under the above operating condi-tions, and calculate the resolution. Use a column giving elu-tion of tocopherol succinate and tocopherol in this order with the resolution between these peaks being not less than 2.0. System repeatability: Repeat the test 5 times with 20 mL of the standard solution under the above operating conditions: the relative standard deviation of the peak height of tocopherol succinate is not more than 0.8z.
Weigh accurately about 50 mg each of Tocopherol Calcium Succinate and Tocopherol Succinate RS, previously dried, dissolve in a mixture of ethanol (99.5) and diluted ace-tic acid (100) (1 in 5) (9:1) to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Pipet exactly 20 mL each of the sample solution and standard solution, and perform the test as directed under Liquid Chromatography <2.01> according to the following operat-ing conditions. Determine the peak heights, HT and HS, of tocopherol succinate in each solution. Amount (mg) of tocopherol calcium succinate mL of chloroform, add 10 mL of hydrochloric acid, shake for 10 minutes, then draw off the water layer, and neutralize (C66 H106 CaO 10) with ammonia TS: the solution responds to Qualitative Tests <1.09> for calcium salt. ＝ MS × HT/HS × 1.036 MS: Amount (mg) of Tocopherol Succinate RS taken 1 cm Absorbance <2.24> E1z chloroform, 100 mL). (286 nm): 36.0 – 40.0 (10 mg, Operating conditions— Detector: An ultraviolet absorption photometer (wave- Purity (1) Clarity and color of solution—Dissolve 0.10 g of Tocopherol Calcium Succinate in 10 mL of chloroform: the solution is clear, and has no more color than the follow-ing control solution. Control solution: To 0.5 mL of Iron (III) Chloride CS add length: 284 nm). Column: A stainless steel column about 4 mm in inside di-ameter and 15 to 30 cm in length, packed with octadecyl-silanized silica gel (5 to 10 mm in particle diameter). Column temperature: Room temperature. ‌Mobile phase: A mixture of methanol, water and acetic acid (100) (97:2:1). Flow rate: Adjust so that the retention time of tocopherol succinate is about 8 minutes. Selection of column: Dissolve 0.05 g each of tocopherol succinate and tocopherol in 50 mL of a mixture of ethanol (99.5) and diluted acetic acid (100) (1 in 5) (9:1). Proceed with 20 mL of this solution under the above operating condi-tions, and calculate the resolution. Use a column giving elu-tion of tocopherol succinate and tocopherol in this order with the resolution between these peaks being not less than 2.0. System repeatability: Repeat the test 5 times with 20 mL of the standard solution under the above operating conditions: the relative standard deviation of the peak height of tocopherol succinate is not more than 0.8z.
Tocopherol Nicotinate occurs as a yellow to orange-yellow, liquid or solid. It is freely soluble in ethanol (99.5), and practically insolu-ble in water. A solution of Tocopherol Nicotinate in ethanol (99.5) (1 in 10) shows no optical rotation. It is affected by light.
(1) Determine the absorption spectrum of a solution of Tocopherol Nicotinate in ethanol (99.5) (1 in 20,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum or the spectrum of a solution of Tocopherol Nicotinate RS prepared in the same manner as the sample so-lution: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared spectrum of Tocopherol under the above operating conditions, tocopherol and tocopherol nicotinate are eluted in this order with the resolu-tion between these peaks being not less than 8. System repeatability: When the test is repeated 6 times with 10 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak areas of tocopherol nicotinate is not more than 2.0z.
(1) Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol Nicotinate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand- MS: Amount (mg) of Tocopherol Nicotinate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 264 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ‌gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Methanol. Flow rate: Adjust so that the retention time of tocopherol nicotinate is about 10 minutes. System suitability— System performance: Dissolve 0.05 g of Tocopherol Nicotinate and 0.25 g of tocopherol in 100 mL of ethanol (99.5). When the procedure is run with 5 mL of this solution under the above operating conditions, tocopherol and tocopherol nicotinate are eluted in this order with the resolu-tion between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions: the relative standard deviation of the peak areas of tocopherol nicotinate is not more than 0.8z
Weigh accurately about 50 mg each of Tocopherol Nicotinate and Tocopherol Nicotinate RS, dissolve each in ethanol (99.5) to make exactly 50 mL, and use these solu-tions as the sample solution and the standard solution, respectively. Perform the test with exactly 5 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas, AT and AS, of tocopherol nicotinate in each solution. Nicotinate, if necessary melt by warming, as directed in the liquid film method under the Infrared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spec- Amount (mg) of tocopherol nicotinate (C ＝ MS × AT/AS 35H53NO3) trum or the spectrum of Tocopherol Nicotinate RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Purity (1) Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol Nicotinate according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Stand- MS: Amount (mg) of Tocopherol Nicotinate RS taken Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 264 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica ‌gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: Methanol. Flow rate: Adjust so that the retention time of tocopherol nicotinate is about 10 minutes. System suitability— System performance: Dissolve 0.05 g of Tocopherol Nicotinate and 0.25 g of tocopherol in 100 mL of ethanol (99.5). When the procedure is run with 5 mL of this solution under the above operating conditions, tocopherol and tocopherol nicotinate are eluted in this order with the resolu-tion between these peaks being not less than 3. System repeatability: When the test is repeated 6 times with 5 mL of the standard solution under the above operating conditions: the relative standard deviation of the peak areas of tocopherol nicotinate is not more than 0.8z
Tocopherol is a clear, yellow to red-brown, viscous liquid. It is odorless. It is miscible with ethanol (99.5), with acetone, with chlo-roform, with diethyl ether and with vegetable oils. It is freely soluble in ethanol (95), and practically insoluble in water. It is optically inactive. It is oxidized by air and light, and acquires a dark red color.
(1) Dissolve 0.01 g of Tocopherol in 10 mL of ethanol (99.5), add 2 mL of nitric acid, and heat at 759C for 15 minutes: a red to orange color develops. Determine the infrared absorption spectrum of Tocopherol as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum or the spectrum of Tocopherol RS: both spectra exhibit similar intensities of absorption at the same wave numbers. Bacterial endotoxins <4.01> Less than 0.50 EU/mg (po-tency). Absorbance <2.24> E1z 1 cm ethanol (99.5), 200 mL). (292 nm): 71.0 – 76.0 (10 mg, Extractable volume <6.05> It meets the requirement. Foreign insoluble matter <6.06> Perform the test according to Method 1: it meets the requirement. Insoluble particulate matter <6.07> It meets the require-ment. Sterility <4.06> Perform the test according to the Mem-brane filtration method: it meets the requirement.
(1) Clarity and color of solution—Dissolve 0.10 g of Tocopherol in 10 mL of ethanol (99.5): the solution is clear and has no more color than Matching Fluid C. (2) Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm).
Perform the test according to the Cylinder-plate method as directed under Microbial Assay for Antibiotics <4.02> according to the following conditions. Test organism, culture medium, and standard solu-tions—Proceed as directed in the Assay under Tobramycin. Sample solutions—To exactly 5 mL of Tobramycin Injection add 0.1 mol/L phosphate buffer solution (pH 8.0) so that each mL contains 1 mg (potency) of Tobramycin. Take exactly a suitable amount of this solution, add 0.1 mol/L phosphate buffer solution (pH 8.0) to make solutions Refractive index <2.45> n20: 1.503 – 1.507 D Specific gravity <2.56> d 20: 0.947 – 0.955 Purity (1) Clarity and color of solution—Dissolve 0.10 g of Tocopherol in 10 mL of ethanol (99.5): the solution is clear and has no more color than Matching Fluid C. (2) Heavy metals <1.07>—Proceed with 1.0 g of Tocopherol according to Method 4, and perform the test. Prepare the control solution with 2.0 mL of Standard Lead Solution (not more than 20 ppm). Assay Dissolve about 50 mg each of Tocopherol and Tocopherol RS, accurately weighed, in ethanol (99.5) to make exactly 50 mL, and use these solutions as the sample solution and standard solution. Perform the test with exactly 20 mL each of these solutions as directed under Liquid Chro-matography <2.01> according to the following conditions, and determine the peak heights, HT and HS, of tocopherol in each solution. so that each mL contains 8 mg (potency) and 2 mg (potency), and use these solutions as the concentration sample solution Amount (mg) of tocopherol (C 29H50O2) ＝ M S × HT/HS ‌MS: Amount (mg) of Tocopherol RS taken sities of absorption at the same wave numbers. Operating conditions— Detector: An ultraviolet absorption photometer (wave- Absorbance <2.24> E1z 1 cm ethanol (99.5), 100 mL). (284 nm): 41.0 – 45.0 (10 mg, length: 292 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 15 cm in length, packed with octadecylsilanized silica gel for liquid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 359C. Mobile phase: A mixture of methanol and water (49:1). Flow rate: Adjust so that the retention time of tocopherol is about 10 minutes. System suitability— System performance: Dissolve 0.05 g each of Tocopherol and tocopherol acetate in 50 mL of ethanol (99.5). When the procedure is run with 20 mL of this solution under the above operating conditions, tocopherol and tocopherol acetate are eluted in this order with the resolution between these peaks being not less than 2.6. System repeatability: When the test is repeated 5 times with 20 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the peak heights of tocopherol is not more than 0.8z.
Containers—Tight containers. Storage—Light-resistant, and well-filled, or under nitro-
Phytonadione is a clear yellow to orange-yel-low viscous liquid. It is miscible with isooctane. It is soluble in ethanol (99.5), and practically insoluble in water. It decomposes gradually and changes to a red-brown by light. Specific gravity d 20: about 0.967
(1) Determine the absorption spectrum of a solution of Phytonadione in isooctane (1 in 100,000) as di-rected under Ultraviolet-visible Spectrophotometry <2.24>, and compare the spectrum with the Reference Spectrum 1: both spectra exhibit similar intensities of absorption at the same wavelengths. Separately, determine the absorption spectrum of a solution of Phytonadione in isooctane (1 in 10,000) as directed under Ultraviolet-visible Spectropho-tometry <2.24>, and compare the spectrum with the Refer-ence Spectrum 2: both spectra exhibit similar intensities of absorption at the same wavelengths. (2) Determine the infrared absorption spectrum of Phytonadione as directed in the liquid film method under In-frared Spectrophotometry <2.25>, and compare the spectrum with the Reference Spectrum: both spectra exhibit similar in-tensities of absorption at the same wave numbers. D Refractive index <2.45> n20: 1.525 – 1.529
(1) Ratio of absorbances—Determine the absor-bances, A1, A2 and A3, of a solution of Phytonadione in isooctane (1 in 100,000) at 248.5 nm, 253.5 nm and 269.5 nm, respectively, as directed under Ultraviolet-visible Spec-trophotometry <2.24>: the ratio A2/A1 is between 0.69 and 0.73, and the ratio A2/A3 is between 0.74 and 0.78. Deter-mine the absorbances, A4 and A5, of a solution of Phytonadione in isooctane (1 in 10,000) at 284.5 nm and 326 nm, respectively: the ratio A4/A5 is between 0.28 and 0.34. Heavy metals <1.07>—Carbonize 1.0 g of Phytona- dione by gentle heating. Cool, add 10 mL of a solution of magnesium nitrate hexahydrate in ethanol (95) (1 in 10), and ignite the ethanol to burn. Cool, add 1 mL of sulfuric acid, proceed according to Method 4, and perform the test. Pre- lution (not more than 20 ppm). Menadione—Dissolve 20 mg of Phytonadione in 0.5 mL of a mixture of water and ethanol (95) (1:1), add 1 drop of a solution of 3-methyl-1-phenyl-5-pyrazolone in ethanol (95) (1 in 20) and 1 drop of ammonia solution (28), and al-low to stand for 2 hours: no blue-purple color develops. Isomer ratio Conduct this procedure rapidly and without exposure to light. Dissolve 30 mg of Phytonadione in 50 mL of the mobile phase. To 4 mL of this solution add the mobile phase to make 25 mL. To 10 mL of this solution add the mobile phase to make 25 mL, and use this solution as the sample solution. Perform the test with 50 mL of the sample solution as directed under Liquid Chromatography <2.01> according to the following conditions, and determine the peak areas of Z-isomer and E-isomer, ATZ and ATE: ATZ/ (ATZ ＋ ATE) is between 0.05 and 0.18. Operating conditions— Proceed as directed in the operating conditions in the Assay. System suitability— System performance: When the procedure is run with 50 mL of the sample solution under the above operating condi-tions, Z-isomer and E-isomer are eluted in this order with the resolution between these peaks being not less than 1.5. System repeatability: When the test is repeated 6 times with 50 mL of the sample solution under the above operating conditions, the relative standard deviation of the total area of the peaks of Z-isomer and E-isomer is not more than 2.0z.
Conduct this procedure rapidly and without expo-sure to light. Weigh accurately about 30 mg each of Phytonadione and Phytonadione RS, and dissolve each in the mobile phase to make exactly 50 mL. Pipet 4 mL each of these solutions, and add the mobile phase to make exactly 25 mL. To exactly 10 mL each of these solutions add exactly 7 mL of the internal standard solution and the mobile phase to make 25 mL, and use these as the sample solution and the standard solution, respectively. Perform the test with 50 mL each of the sample solution and standard solution as directed under Liquid Chromatography <2.01> according to the fol-lowing conditions, and calculate the ratios, QT and QS, of the total area of the peaks of Z-isomer and E-isomer to the peak area of the internal standard. Amount (mg) of phytonadione (C31H46O2) ＝ MS × QT/QS MS: Amount (mg) of Phytonadione RS taken Internal standard solution—A solution of cholesterol benzo-ate in the mobile phase (1 in 400). Operating conditions— Detector: An ultraviolet absorption photometer (wave-length: 254 nm). Column: A stainless steel column 4.6 mm in inside diame-ter and 25 cm in length, packed with porous silica gel for liq-uid chromatography (5 mm in particle diameter). Column temperature: A constant temperature of about 309C. Mobile phase: A mixture of hexane and n-amyl alcohol (4000 : 3). Flow rate: Adjust so that the retention time of the peak of E-isomer of phytonadione is about 25 minutes. System suitability— System performance: When the procedure is run with 50 mL of the standard solution under the above operating con-ditions, the internal standard, Z-isomer and E-isomer are ‌eluted in this order with the resolution between the peaks of‌ Z-isomer and E-isomer being not less than 1.5. System repeatability: When the test is repeated 6 times with 50 mL of the standard solution under the above operat-ing conditions, the relative standard deviation of the ratio of the total area of the peaks of Z-isomer and E-isomer to the peak area of the internal standard is not more than 1.0z.